Whereas William Kent may have ‘leaped the fence, and saw that all nature was a garden’, to jump or destroy the wall of a horticultural expo is to typically find the periphery of a city, complete with its own implied social delineations. It is in this context that the dissolution of the physical and psychological frame of the institutionalized expo itself— rather than the frames of the individual gardens within—that is the more potent force in contemporary landscape and urbanism.During the past decade, grocery stores have expanded into underserved urban communities with growing awareness of ‘food deserts’ and their associations with urban health disparities and economic disinvestment.Even though scholars contest whether physical access to grocery stores truly improves food accessibility and health , policy advocates continue to promote grocery stores as the ultimate solution to urban food deserts. Federal, state, and local governments have followed suit with the creation of incentives and tax breaks for food retailers that locate in underserved urban communities.Despite numerous studies on food deserts and their potential impacts, there are few explorations of the prescribed “solution” of grocery stores. Moreover, recent studies on the impacts of new and recently closed urban grocery stores tend to focus on resident behavioral changes . Thus, scholars and policymakers alike have limited understandings of the broader neighborhood implications of grocery stores newly introduced into underserved urban communities. This dissertation addresses this gap by analyzing how local organizations and agencies pursue grocery development in response to neighborhood issues. By exploring local processes, I aim to understand whether governments should continue to incentivize urban grocery stores and if so, under what conditions. As such, the ultimate goal of this dissertation is to understand the conditions for successful grocery development across contexts. Using a comparative case study approach, I analyze the historical drivers, planning processes, and outcomes of recently completed ground-floor grocery stores in two distressed San Francisco Bay Area communities: a Fresh and Easy Neighborhood Market in San Francisco’s Bayview Hunters Point and the Mandela Foods Cooperative in Oakland’s West Oakland. Despite their marked differences, both stores were cited in national advocacy efforts around the federal Healthy Food Financing Initiative . In 2005, British supermarket magnate, Tesco created the Fresh and Easy chain primarily for untapped urban markets and health and cost-conscious consumers in California, Arizona, and Nevada.

In 2006, amidst massive grocery closings citywide, former San Francisco Mayor Gavin Newsom,flower pot wholesale the Office of Economic and Workforce Development, and the Department of Public Health coordinated Fresh and Easy’s entry into San Francisco’s distressed Bayview Hunters Point neighborhood. As the first new grocery store in the neighborhood in decades, Fresh and Easy represented a convergence of previously disparate efforts around public health, economic and workforce development. As such, the City’s vision of grocery development was consistent with the rhetoric of federal food access programs. Yet ultimately, national and global capitalist forces led to the store’s demise. In September 2013, Tesco sold the Fresh and Easy chain to a private equity firm, following two years of massive profit losses. The Bayview Hunters Point location was one of numerous stores that were closed as a result. Despite similar underlying socio-economic conditions, West Oakland’s first coordinated grocery store efforts took a markedly different form. Beginning in the late 1990s, West Oakland residents and activists created farmers markets, mobile grocery stores, and community gardens—all with the goal of establishing a full service cooperative grocery store. For the next decade, environmental justice activist Dana Harvey led efforts to realize this vision. Following financial hurdles and disputes with developers over retail space, Harvey and her team of consultants negotiated a 2300 square foot ground floor retail space. In June 2009, the worker-owned Mandela Foods Cooperative and café opened for business. As of 2013, Harvey is exploring possibilities for expansion with funding support from the California Fresh Works program. Through an institutional analysis of the planning processes, I find that both Fresh and Easy and Mandela Foods reflect distinctive neighborhood revitalization legacies, critical moments of institutional capacity building, localized versions of national policy narratives, and the role of charismatic leaders in grocery store implementation. While national narratives shape the rhetoric of urban grocery development, ultimately local context dictates how food access issues are defined, who addresses them, and how. In Bayview, a city-led grocery agenda entailed top-down planning processes that aligned with the corporate model of the Fresh and Easy store. Ultimately, collaborations across government agencies and a systematic role of the Department of Public Health failed to sustain the community’s first new grocery store. In West Oakland, an activist-defined grocery agenda produced grassroots planning processes consistent with the community-based model of the Mandela Foods Cooperative. Conflict and community protests eventually won out—albeit for a scaled-down version of West Oakland activists’ original vision. These findings suggest that federal grocery incentive programs should: 1) maintain a broad framework that enables local communities to define food access problems and their solutions on a case-by-case basis, 2) encourage diverse solutions not limited to grocery stores and supermarkets, and 3) emphasize community reinvestment goals.As mentioned in the beginning of this chapter, urban grocery stores are now on federal, state, and local policy agendas. Much of this current momentum is the result of an expansive body of research on ‘food deserts,’ which has pointed to urban grocery stores as a comprehensive solution to food access, health disparities, and economic disinvestment.

As early as the 1970s, government agencies and researchers profiled the phenomenon of “supermarket flight” due to deindustrialization and the consolidation of the grocery industry . The combined impacts of these trends contributed to the continued “suburbanization” of grocery stores with that of housing and employment centers. Already since the 1960s, Community Development Corporations helped reintroduce grocery stores and other urban retail into underserved areas with expanding federal programs for neighborhood revitalization . Urban grocery stores entered more squarely into federal policy agendas with the push for “inner city revitalization” beginning in the 1990s and new evidence of the “urban grocery gap” . By the late 1990s and early 2000s, researchers conducted the first detailed accounts of the socio-economic and health impacts of lacking urban grocery stores in the UK. This research had a resounding impact on domestic research, recasting the “urban grocery gap” as the phenomenon of food deserts. In the U.S., the visceral metaphor of the ‘food desert’ effectively shifted discourses on the function of urban grocery stores from economic reinvestment to health promotion. The greater extent of food desert research that followed suggested a positive correlation between the lack of grocery stores and poor health outcomes in poor urban communities . A separate but related body of literature linked food environments more generally to growing rates of childhood obesity in the U.S. . These studies directly informed the first comprehensive federal efforts focused dually on health promotion and retail revitalization, namely First Lady Michelle Obama’s Let’s Move childhood obesity initiative and the Healthy Food Financing Initiative discussed at the beginning of this chapter. Already since the 1990s, community-based organizations nationwide addressed urban food access issues through “community food security” initiatives—alternative modes of food production and consumption ranging from farmers markets, produce stands, mobile grocery stores, and community gardens. Overall, policy and practice were built on similar assumptions about the contributions of new food retail outlets on the health and livelihoods of disenfranchised urban residents. However, researchers, policymakers, and journalists—with varied technical backgrounds and political leanings—continued to contest the claims of research and policies that purported the promise of urban grocery stores. For example,berry pots some researchers found that areas defined as food deserts are not devoid of food stores as the term would suggest. Others found that areas otherwise defined as food deserts can in fact be replete with corner markets and ethnic food stores that meet culturally preferred food needs .

Even in spite of this evidence, scholars have agreed that poor communities tend to lack stores that provide greater fresh food options . Furthermore, disinvested communities become targets for fast food establishments, convenience stores, and liquor stores, which may contribute to poor diets and health disparities . As such, some prefer to characterize underserved communities as ‘food swamps’ as opposed to food deserts to emphasize the ubiquity of “low quality” food options . These debates came to a head in late 2011 and early 2012, when The Economist and The New York Times highlighted several longitudinal studies claiming that the introduction of grocery stores into poor communities had no effect on health outcomes and that cost was more of a barrier to healthy food access than physical access . While these suffer from methodological shortcomings, even proponents of urban grocery stores urged policymakers to fully consider non-physical means of improving urban food access. This includes greater attention to nutrition education, food affordability and the role of food assistance programs, and efforts to expand healthy food offerings at corner markets and liquor stores in underserved areas . This raises the question—should grocery stores be developed in poor communities at all? Even the critics say yes. It is an undisputed fact that poor communities tend to lack grocery stores, by their very nature of being under-resourced, disinvested areas. The issue is the definition of the problem and how it points to a rather simple solution that is ultimately inconsequential to the broader issues faced by poor communities. For example, community activists and researchers argue that perhaps the root of the problem is the use and operationalization of the term ‘food desert,’ a term that may not fully capture food access issues as experienced by disenfranchised residents. Along these lines, researchers argue that a purely spatial definition of food deserts fails to capture the broader political-economic factors that contribute to inequitable access to resources in poor communities. For example, Donald argues that singularly “built environment solutions” to food access issues fail to consider the problem of food cost—healthier food items cost more due to agricultural subsidies that favor the production and distribution of low-cost, highly processed, “unhealthy” food. argues that the mere designation of food deserts constitutes ‘neoliberal paternalism’—the assumption that the plight of urban communities are ultimately only solved through top-down market solutions .Scholars cast urban grocery stores as either a normative solution or a faulty “build-it-and-theywill-come” approach that requires redefinition and reassessment. Still, the practical reality is that grocery incentive programs continue to expand at local and state levels. Leading corporate retailers such as Walmart, Target, Kroger, and SuperValu continue to reenter urban markets by creating “urban format” versions of their largely suburban supermarket chains. Community based organizations are pursuing their own alternatives to chain supermarkets through corner store conversions, food cooperatives, mobile grocery stores, and farmers markets. In sum, urban grocery stores will continue to be developed in under served urban areas if not for current policies but due to industry and urban trends. What does this mean for future research? Shannon argues for research that “questions the naturalizing language of market relations that positions low-income communities as just another emerging market” . On a practical level, Donald calls for studies that explore: 1) before/after assessments of communities where new grocery stores are introduced, 2) how the changing grocery retail environment shapes local implementation, and 3) the institutional context and “localized geographies” of food access issues. In line with planning scholars and critical geographers, this dissertation reinforces the idea that future research must account for the complexity of food access problems and its solutions. First, this dissertation provides a detailed account of grocery planning processes to complement spatial and quantitative studies of food deserts. The greater extent of literature on urban grocery stores has involved assessments of the phenomenon of food deserts—namely evaluations of the “problem” of lacking food deserts and its possible impacts. In light of recent critiques, scholars have made strides in redefining food deserts and how food accessibility more generally might be measured. While there is ample evidence to suggest the “problem” of lacking grocery stores exists, there are few detailed accounts16 of how local communities and municipalities themselves define the problem and choose to address it through grocery development. Second, this dissertation explores grocery store development as a set of processes instead of as a finite solution.

Since all three variants showed similar binding kinetics in BLI experiments, we hypothesized that the differences in EC50 may result from rCMG2-Fc stability differences between variants during the 8-h cell incubation period at 37°C. With a less stable variant, the fraction of functional rCMG2-Fc decays over time, together with the fact that CMG2 and PA binding is reversible , resulting in a higher EC50 compared to more stable variants. The observed binding to PA was the same before and after incubation at 37°C for APO and Agly variants . For the ER variant, the fraction of functional rCMG2-Fc decayed over time, and a significant drop was observed for the overnight incubation sample , which explains the higher EC50 for the ER variant in the TNA . GnGnXF, MAN8, and Agly rCMG2-Fc glycoforms were each independently simulated for 100 ns. Figure 8 shows the initial and final conformations of all glycoforms from the simulations. The images at t = 0 are all aligned by the Fc CH2 domain; the images at t = 100 ns are each independently aligned to best illustrate macro-structural orientation. For all simulations, we see the core structure of the protein remains intact. However, there is significant variability in the macro-structural orientation of the final structures’ CMG2 and Fc domains. The final GnGnXF and Agly structures exhibit significantly contracted linkers with respect to the final MAN8 structure, where the linker is fully extended. All glycoforms retain accessibility of the PA binding site after simulation. This is in agreement with the BLI , where all three variants have similar ka and kd, and the functional ELISA , where all three variants have control curves with similar absorbance level. To quantitatively characterize the macro-structural differences between the GnGnXF, MAN8, and Agly glycoforms, we report the center of mass distance between the CMG2 and Fc domains in Figure 9. Among the glycoforms, MAN8 has the highest COM distance with the narrowest spread at around 7.2 nm, GnGnXF and Agly have lower COM distances with wider spreads, roughly centered around 6 nm,vertical plant rack with the Agly COM spread being the widest.

The significantly higher COM of MAN8 than GnGnXF and Agly is visually consistent with the final conformations in Figure 8. The large spread in Agly rCMG2-Fc COM distance is largely due to refolding of the flexible residues in and near the LNK region. The COM distance temporal profile in Supplementary Figure S10 displays a gradual increase in Agly COM distance, which tapers off around 80 ns. Thus, the larger spread in Agly COM is due to conformational changes during the simulation, not a single conformation with increased flexibility. The root mean square fluctuation data in Supplementary Figure S11 show increased Agly RMSF in and around the LNK region during the first 50 ns and a reduction in Agly RMSF during the latter 50 ns, which is consistent with the trending exhibited in the Agly COM distance.The backbone root mean square deviation of ordered domains referenced from the initial and final conformations of the CMG2 and Fc regions for all three simulated glycoforms is shown in Figure 10. A low RMSD indicates protein folding transition is minor and the protein structure is stable. For each monomer, the ordered domain of the CMG2 region were defined as residues 10–181, while the ordered domains of the Fc region were defined as the CH2 and CH3 regions: residues 210–308 and 315–414, respectively. Each domain’s RMSD was fit and referenced independently, and subsequently averaged to produce the plots in Figure 10. Two RMSD profiles were averaged to generate the CMG2 RMSD, and four RMSDs were averaged to generate the Fc RMSD for each glycoform. All RMSDs are below 1.6 Å, indicating a generally conserved fold for all the ordered domains of each glycoform. This is further confirmed by the secondary structure data, provided in Supplementary Figure S12. In general, the RMSD from the final structure is lower than that of the initial, indicating conformational convergence is progressing throughout the simulations. The low RMSD and conserved secondary structure in all three simulated glycoforms indicate that there were no major refolding events; thus, the reduced activity in the TNA and the functional ELISA of the ER variant is likely not due to reduced fold stability. The Fc RMSD is noticeably lower than the CMG2 RMSD for the Agly and GnGnXF glycoforms, indicating minor fold transitions in the CMG2 domains of these glycoforms.The hydrophobic solvent accessible surface area distributions are reported in Figure 11.

The MAN8 glycoform has the highest amount of hydrophobic SASA in the simulation, while the aglycosylated has the lowest. Increased hydrophobic SASA likely yields a greater aggregation propensity . To elucidate which residues contribute most to the hydrophobic SASA difference between MAN8 and Agly, the average per-residue hydrophobic SASA was calculated, and the five residues with the greatest positive difference of MAN8 minus Agly hydrophobic SASA were obtained. These residues were Pro199, Pro201, Leu205, and Pro300 of one monomer, and Leu206 of the other monomer. All of these residues are located in close proximity to the N-terminal region of the Fc domain, shown in Figure 12. We see that these residues are highly spread out in the MAN8 structure, moderately spread out in the GnGnXF structure, and closely associating in the Agly structure. The increased SASA in the MAN8 glycoform is consistent with the existence of the high molecular weight band in the ER variant at ~250 kDa in the SDS-PAGE results .In this paper, experimental and computational techniques were employed to study the effects of N-glycosylation on the expression, structure, function, and stability for anthrax decoy protein rCMG2-Fc.N-glycosylation was found to strongly stabilize rCMG2-Fc in planta as both APO and ER variants have over two-fold higher expression level than the Agly variant, shown in Figure 1. The increase in expression level of glycosylated variants could be attributed to a decreased susceptibility of the APO and ER variants to proteases, where the steric hindrance of oligosaccharides inhibits proteolytic degradation of glycosylated rCMG2-Fc. From a manufacturing standpoint, producing glycosylated rCMG2-Fc would require less than half the production capacity of the aglycosylated form. Thus, when glycosylation is not detrimental, preserving natural N-glycosylation sites can enhance protein production. Alternatively, retaining the aglycosylated protein in the ER can also help improve yield, since the ER has fewer types of proteases than the apoplast . Furthermore, unlike the apoplast, the ER has chaperone proteins to provide folding support . It is also possible to enhance a glycoprotein’s function via modification of the N-glycosylation profile during expression. This can be achieved using subcellular targeting and/or the co-expression of glycan-processing enzymes or addition of enzyme inhibitors in the agroinfiltration buffer. For example, high mannose Fc N-glycosylation has been shown to enhance antibody-dependent cell-mediated cytotoxicity , which can be achieved by targeting the protein to the ER or addition of mannosidase I inhibitor to the agroinfiltration buffer or cell culture medium .

It is worth noting that N-glycosylation of Fc is not strictly required when Fc is fused to the target protein for the purpose of increasing circulation half-life . SDS-PAGE and Western blot confirmed that intact rCMG2-Fc was produced, with bands near 50 kDa . There is also a band around 250 kDa in the non-reducing SDS-PAGE for the ER variant , which may correspond to a high molecular weight protein aggregate. The hypothesis of increased aggregation propensity of the ER variant is supported by the hydrophobic SASA predicted from our MD simulations. MAN8 was found to have significantly higher hydrophobic SASA,growing strawberries vertical system with the five strongest contributing residues in the N-terminal region of the Fc domain. Protein aggregation might reduce PA binding capacity, as it is evident in the functional ELISA , which is later discussed in the rCMG2-Fc function section.The ability for rCMG2-Fc variants to sequester anthrax PA and prevent cell death was assessed using a cell-based TNA. Results are shown in Figure 5, where the ER variant has a statistically higher EC50 value than APO and Agly variants. The possibility of FcγR dependent toxin neutralization was ruled out as the EC50 values did not change upon FcγR blocking. This is likely because previous studies used antibodies or serum against PA that bind to PA but not necessarily blocks the its binding site to the anthrax cellular receptor CMG2. Thus, when incubated with cells, the antibody-bound PA can still bind to CMG2 and form prepore, resulting in LF and EF endocytosis. In this situation, Fc and FcγR could form an immune complex that is then sorted and degraded in the lysosome . In our experiments, rCMG2-Fc competitively inhibits binding between cellular receptor CMG2 and PA. This completely eliminates LF and EF internalization. The binding kinetics between rCMG2-Fc variants and PA were determined by BLI, results shown in Figure 6. The 37°C KD values were slightly lower than the 25°C KD values, but no appreciable difference in binding kinetics as a function of glycosylation was observed at 25 or 37°C. Considering the CMG2 domain is linked to Fc through a flexible linker, it is not surprising that the glycosylation of the Fc domain has minimal impact on the binding kinetics of the CMG2 domain with PA. Moreover, the sub-nanomolar affinity reported in this work is consistent with previous work on rCMG2 and PA binding kinetics , which is direct evidence that neither the fused Fc domain nor its glycosylation interferes with CMG2/PA binding kinetics. Even though the kinetics of all three variants were unaffected by glycosylation, it is possible that the fraction of functional protein changes over time.

The BLI experiments only characterized the interaction kinetics, which are independent of the fraction of functional rCMG2-Fc on the sensor tip. The hypothesis that the fraction of functional protein at 37°C is glycosylation-dependent was confirmed with the functional rCMG2-Fc ELISA, where the ER variant lost the most activity overnight, consistent with the ER variant having the highest EC50. However, the MD simulation data exhibit high fold stability in all glycoforms, according to the RMSD of ordered domains as well as the secondary structure . Thus, we hypothesize that the reduction in activity is not due reduced fold stability. Moreover, the MAN8 had the highest hydrophobic SASA among the three simulated glycoforms, indicating a higher aggregation propensity. The residues that contributed the most to the decreased hydrophobic SASA in the simulated Agly from the MAN8 were located in the N-terminal region of the Fc domain, just after the C-terminal region of the linker . This region is also more extended in the MAN8 glycoform, as shown in the final conformation and the COM distance distribution . This exposure of hydrophobic residues in the thinly extended N-terminal region of the Fc domain could facilitate the aggregation with other rCMG2-Fc or protein fragments. This could explain the reduced activity for ER variant in the functional ELISA and the 250 kDa band observed in the non-reducing SDS-PAGE gel for the ER variant . Fusion protein aggregation has been observed in the literature for another Fc fusion protein ALK1-Fc, where a high abundance of MAN5 glycoform was found as high molecular weight aggregates . It is worth noting that a recent study on high-mannose type IgG showed a decrease in protection factors of backbone amide nitrogen in the CH2 domain , which could also contribute to the reduced activity of the ER variant. Meanwhile, Lu et al., found that highmannose glycans have no detrimental effect on antibody stability and aggregate rate . The antibodies used in their study were IgG1 and IgG2, which has a molecular weight ~150 kDa. Since both the protein size and structure can affect protein stability, it is not surprising to see diverse protein stability results performed on different molecules. Within Fc-fusion proteins, structure can still vary depending on the fusion partner size and structure. However, we do expect Fc-fusion proteins with similar structure, molecular weight and conserved glycosylation site as rCMG2-Fc to likely exhibit similar behaviors. The APO and Agly variants had no significant difference in fraction of functional protein after being incubated overnight at 37°C. This result is in agreement with a previous study where human IgG1s stored at 37°C for 21 days had no difference in aggregation or fragmentation , suggesting the absence of glycans had no major impact on stability under physiological temperature.

For plants inoculated by the drop method, the number of culturable cells declined by 4 to 5 log CFU/leaf by day 2 to an average of 10 CFU/leaf . After inoculation by spray, the average number of culturable E. coli O157:H7 ATCC 700728 cells remained higher and ranged from 100 to 1000 CFU/plant, even on day 7 of the study .Although there were significantly higher numbers of culturable EC4045 cells than strain ATCC 700728 cells at 24 h after inoculation, the culturable amounts of both strains declined to equivalently low levels within 48 h on lettuce . Real-time PCR was used to quantify total E. coli O157:H7 cells on the lettuce. The number of E. coli O157:H7 cells remained equivalent to the inoculum cell density for 7 days, regardless of the strain or the inoculation method used . The addition of PMA prior to real-time PCR amplification revealed that viable cell quantities were equal to the numbers of total and culturable cells at the time of inoculation but then declined by an average of 100- fold within the first day. For both ATCC 700278 and EC4045 strains, the number of viable cells detected using PMA real-time PCR remained above the detection limit and was significantly higher than estimated by the standard plate count method .Lettuce plants in the field were spray inoculated with E. coli O157:H7 ATCC 700278 at a density of 106 CFU/plant. E. coli O157:H7 was not detected on uninoculated lettuce plants either by culture or by real-time PCR with or without PMA. Within 2 h after application onto the lettuce, culturable cell amounts declined by 2 and 4 log for plants inoculated in the spring and summer trials, respectively . Two days after inoculation, the amounts of culturable E. coli ATCC 700278 were reduced to the lower limit of detection by plating for most plants . At that time, E. coli O157:H7 was not retrieved by plating or enrichment for 3% and 13% of the spring and summer plants, respectively. In contrast to colony enumerations of E. coli O157:H7, total cell numbers determined by real-time PCR quantification were equivalent to the inoculum levels 2 h after application to lettuce plants.

Two days after application the numbers of E. coli O157:H7 estimated by real-time PCR decreased,vertical farming supplies on average, by 2 log CFU/plant and were below the limit of detection for six out of 24 plants tested for both field trials.Accurate measurements of pathogen amounts and viability are important for risk assessments and control mechanisms aimed at preventing outbreaks of food borne illness. This study was the first to apply a culture-independent method to quantify the number of E. coli O157:H7 cells on plants inoculated with the pathogen in the field. We also compared the utility of the method for quantifying virulent and avirulent pathogen survival on laboratory-grown lettuce. The first step in the development of culture-independent methods for bacterial enumeration is to ensure that the technique is optimized for the specific organism and environmental conditions of interest. While numerous assessment methods are available, we selected PMA combined with real-time PCR because this technique offered the possibility of rapid enumeration of viable cells in a variety of environments. For E. coli O157:H7 ATCC 700728, we first identified appropriate real-time PCR primers, PMA concentrations, and PMA incubation times with a goal of selective and sensitive detection of viable E. coli O157:H7 cells in the presence of high levels of dead E. coli O157:H7 cells. Among the results was the confirmation that PMA-mediated PCR-inhibition is incomplete when the product size is less than 200 bp. Although our tests on PMA real-time PCR revealed some potential short-comings with the method, most notably the interference of high quantities of inactivated cells and the relatively high numbers of cells required for detection, these parameters were within the range found in other PCR-based studies. E. coli O157:H7 strains ATCC 700728 and EC4045 were inoculated onto leaves of potted lettuce plants and incubated in a growth chamber under temperature and low humidity conditions that favored the decline of cell numbers in trends similar to those recorded in field trials performed in the Salinas Valley. This approach differs from most other laboratory studies that examined plants exposed to high moisture and temperatures supportive of pathogen growth. Within 2 days after inoculation on lettuce plants, the number of E. coli O157:H7 ATCC 700728 cells able to form a colony on TSA declined 1000-fold when the pathogen was initially applied at a level of 6 log CFU per plant or per leaf using an aerosolized spray and declined .The numbers of culturable cells remained similar for the subsequent 5 days, suggesting that the initial events after contact with the plant are crucial for determining the survival of the organism on lettuce.

In contrast with plate counts, real-time PCR targeting of the E. coli O157:H7 ATCC 700278 and EC4045 inoculants showed that the genomic DNA of the organisms remained on the lettuce in quantities nearly equivalent to the inoculum concentrations over the duration of the 7-day experiment. This finding confirmed that both strains adhered to the leaf surfaces and the recovery method was sufficient to remove the majority of the cell inoculants from the plant. Moreover, the results indicated the presence of high numbers of intact E coli O157:H7 cells on lettuce even when very few were able to form colonies on TSA. Exposure of E. coli O157:H7 suspensions to PMA prior to realtime PCR quantification facilitated the detection of cells with an intact cell membrane, and these cells were likely viable. Application of this method to E. coli ATCC 700728 and EC4045 recovered from growth chamber lettuce indicated that a significant fraction of the pathogen inoculants were viable, even though colony-based assessments indicated otherwise. This finding was particularly evident for E. coli O157:H7 inoculated onto leaves by direct drop inoculation. While an average of 5 log CFU per leaf were viable as measured by PMA real-time PCR, less than 1 log CFU per leaf were enumerated by culturing. These results are supported by previous studies reporting that E. coli O157:H7 enters into a viable but not culturable state on lettuce leaves incubated at cold temperatures. Similar outcomes were reported for other E. coli including E. coli O104:H4 in water or after exposure to toxic concentrations of copper. Once the stress condition was removed, a fraction of the cells recovered the capacity to grow in laboratory media, thereby indicating a potential of the E. coli O1O4:H4 cells to cause human disease. The formation of VBNC bacterial cells on plants also was previously described for Listeria monocytogenes on parsley. The number of viable L. monocytogenes cells was 1 to 2 log higher than the culturable cells recovered from parsley grown in greenhouses at 20uC under low relative humidity ; growth of the VBNC cells was not restored on the plants when the RH was increased to 100%. Although we did not examine for E. coli O157:H7 recovery from the VBNC-like state,vertical lettuce tower future efforts might investigate whether those cells can recover and resume growth either under growth-conducive conditions on lettuce or after removal of the cells from the plants and prior to or after plating for viable cell enumeration.

Overall, the toxigenic strain EC4045 survived in similar quantities as ATCC 700728 on lettuce. A recent study showed that certain lineages of E. coli are more commonly associated with plants and presumably have evolved the capacity to tolerate plant associated environments better than E. coli isolated from other sites. Because we only compared two strains, subsequent investigations should examine multiple attenuated and virulent O157:H7 strains isolated from different sites for their capacity to colonize and persist on lettuce under field relevant conditions. Survival of E. coli O157:H7 on lettuce also was measured in two field studies. Culturable amounts of strain ATCC 700728 declined shortly after inoculation onto plants in the field, as we reported previously. Rates of cell decline were similar to E coli O157:H7 on lettuce in the growth chamber directly inoculated in drops with a pipette. Real-time PCR estimates of E. coli O157:H7 ATCC 700728 in lettuce leaf washes showed that this strain was present on the plants immediately after inoculation and 2 h later in quantities equivalent to the inoculum levels. Importantly, the rapid decline in culturable E. coli during the first hours after application onto plants in the field was not due to an inability to remove the organism from the lettuce or from dispersal and lack of strain attachment. Rather, it appears that the majority of the E. coli cells in the inoculum either died shortly after application or entered a VBNC state. In contrast to the growth chamber experiment results, the numbers of E. coli O157:H7 cells were below detection by real time PCR within 2 days after inoculation onto field lettuce. These findings suggest that the E. coli O157:H7 cells and genomic DNA were degraded rapidly. Environmental parameters such as solar radiation, heat, and water stress could be responsible for the differences in the stability of E. coli O157:H7 DNA in the field compared with the laboratory. Alternatively, cell maintenance might depend on other microorganisms on the leaves. Also, it is notable that different lettuce cultivars were used in the field and growth chamber studies, which may have impacted survival. The potential for cultivar-dependent effects was shown for E. coli O157:H7 on lettuce cultivars grown under axenic conditions in the laboratory.

Because the E. coli O157:H7 ATCC 700728 DNA was degraded on field-grown plants within 2 days after inoculation, it is unlikely that the organism developed a VBNC state, particularly over longer time scales . However, this possibility could not be directly addressed using the PMA realtime PCR assay in the field trials. Viable cell amounts measured by culturing and PMA real-time PCR were in agreement immediately after application of ATCC 700728 onto laboratory grown lettuce, but PMA-mediated detection was impaired on plants from the field. For those plants, the viable cell number estimates for strain ATCC 700278 were 10-fold lower as measured by PMA real-time PCR than by colony enumeration and total cell numbers estimated by real-time PCR. These differences might have been due to the higher turbidity or opacity of the lettuce plant washes from the field samples, thereby preventing light from penetrating the suspension during the PMA photo inactivation step. This interference would prevent the inactivation of free PMA, resulting in sufficient quantities of the compound to bind genomic DNA released from viable cells during the subsequent DNA extraction and amplification steps. In addition, the PMA real-time PCR assay was unable to detect low numbers of cells. Attempts to detect the ATCC 700278 strain after concentrating the leaf washes were unsuccessful . Similarly, PMA real-time PCR was found to be more reliable for viable cell detection in diluted wastewater than in pooled and concentrated wastewater samples. Such factors strongly limit the overall usefulness of this approach for field-grown plants. However, this method is informative for examination of E. coli O157:H7 on ‘‘cleaner’’ plants grown in the growth chamber and not exposed to the biotic conditions that are common outdoors. In conclusion, this study illustrates the similarities and differences between controlled studies of human pathogens on plants in a growth chamber and experiments examining the population dynamics of pathogens on plants under production-like conditions in the field. By applying relevant environmental conditions and droplet inoculation in the growth chamber, we were able to more closely mimic the rapid decline in E. coli O157:H7 culturability that was observed after inoculation of this organism onto lettuce plants in the Salinas Valley. Culture-independent assessments confirmed that the pathogen remains on the plant long after application. However, field studies showed that at least for the majority of E. coli O157:H7 cell inoculants, the loss in culturability was most likely due to cell death rather than an inability to form colonies on standard laboratory media. Hence, this work confirmed our observations that low numbers of E. coli O157:H7 persist on lettuce grown in the Salinas Valley and variations in pathogen survival among individual plants are dependent on other unknown factors .Ethnically diverse populations are disproportionately exposed to hazardous environmental materials by virtue of living in close proximity to toxic waste materials.

Finally, the environmental cost to Colombia of chemical eradication has not been sufficiently studied and quantified. However, it is estimated that “for every hectare of poppies sown, an average of 2.5 hectares of woodlands are destroyed; in the case of coca plantations the ratio is 1 to 4, and for marijuana it is 1 to 1.5.”However the negative effects of herbicide fumigation have not been assessed in this process of forest destruction. What we do know is that the mere fact of fumigation forces the growers to move elsewhere in order to plant their illegal crops, and that entails necessarily a further environmental disaster.Despite the fact that organizations such as Greenpeace, the Worldwide Fund for Nature and Dow Agrosciences are opposed to the use of this herbicide, the United States authorities have insisted that it is quite harmless. They have gone even further; during the Pastrana administration especially, they have been putting pressure on Bogotá to apply a dangerous fungus, fusarium oxysporum, in the process of obligatory eradication. Since its launch in 2009, Evergreen Co-operative Corporation, a network of worker-owned co-operatives in Cleveland Ohio, has magnetized media, political elite, and academic attention. Evergreen has garnered supportive coverage in the Economist, Harper’s, The Nation, The New York Times, Fast Company, Time, and Business Week. Sarah Raskin lauded the initiative in 2013 while she was serving on the Board of Governors for the Federal Reserve System .Ron Sims, then Deputy Secretary for the U.S. Department of Housing and Urban Development, referred to theEvergreen network as “brilliant” during a 2011 interview . Intellectuals on the Left have also been attracted to the initiative: Noam Chomsky has celebrated Evergreen in interviews and public talks, and the initiative has been cited by numerous academics as a hopeful alternative to the capitalist firm and its social and environmental externalities . Evergreen is currently comprised of three worker-owned co-operative enterprises: Evergreen Laundry , Evergreen Energy Solutions , and Green City Growers . Evergreen was designed to capture procurement flows from area “anchor” institutions: large hospitals and universities that are unlikely to leave the community, have a general commitment to improving it, and can do so by leveraging their purchasing power in support of local economic development . While Evergreen currently employs approximately 120 people, the vision is that it will become a large network of worker co-operatives that can rejuvenate the depressed regional economy in Cuyahoga County and inspire replications in other regions across the United States. Evergreen’s key features are ensuring worker ownership, harnessing the local wealth of anchor institutions,vertical hydroponic garden and prioritizing sustainable service delivery.

Supporters refer to Evergreen as the “Cleveland Model,” an approach that can be pursued in communities across the country . According to leaders with Evergreen, “What’s especially promising about the Cleveland model is that it could be applied in hard-hit industries and working-class communities around the nation” . Despite all of the attention, the Evergreen case has not yet been studied in a sustained way. Furthermore, there is a dearth of literature on co-operative development in general . With this article we aim to contribute to the collective learning that can happen from successful and failed co-op development experiments. Building this knowledge is especially important at a time when heightened contestation over neoliberal capitalism has intensified interest in the co-operative model . Our primary finding is that Evergreen’s development depended on contextual factors that might not be replicable: a supportive and wealthy community foundation and champions within local government. The post-2008 period of contested neoliberalism in which Evergreen emerged created opportunities for new alliances, as diverse actors were willing to consider alternative economic models. These alliances were critical to Evergreen’s emergence, but similar connections might not be available elsewhere. The fact that Evergreen’s start-up relied so heavily on context-specific private and ad hoc arrangements suggests that more systematic, government-supported programs of financing and technical support are needed if worker co-operatives are to thrive in North America. We conclude that bottomup, movement-driven action often precedes – and creates a climate for – policy change. Our analysis therefore falls within the social movement approach to co-operative development, which argues that robust popular movements are integral to successful development of co-operatives and often predicate policy breakthroughs . While Evergreen’s replicability may be limited, its social movement orientation and ambition to scale up the co-operative alternative to neoliberal capitalism position it as a contributor to the important project of movement building that can facilitate the policy change needed to grow the co-operative economy. To contextualize our case study we conducted an extensive literature review on cooperative policy, focused specifically on co-op dense regions . Our research team then visited Cleveland in May 2013. We did site visits to Green City Growers and Evergreen Laundry, interviewing management and speaking with employees at each location. We also conducted semi-structured interviews with key actors involved in the conception and implementation of the project. We sought from the outset to make our findings relevant not only to academics, but also to practitioners in the co-operative movement.We thus undertook this project as a form of “movement-relevant” research . According to Bevington and Dixon, movement-relevant research “emerges out of a dynamic and reciprocal engagement with the movements themselves. This engagement not only informs the scholarship but also provides … accountability” . Our research team would like to see Evergreen and the co-operative movement in general thrive, and this article is an effort to understand the conditions that might enable this success. We believe, following Bevington and Dixon, that this commitment to the co-operative movement does not lead to bias, but instead adds incentive to provide the “best possible information” to movement participants and supporters. Interest in the “Cleveland Model” has cut across the political spectrum, coming not only from progressive media and academics on the Left, but also from conservative venues like the Economist and the Federal Reserve Board.

Evergreen emerged one year after the 2008 financial crisis, during heightened contestation over the philosophy and policy mix that has guided political economic affairs for the past forty years: neoliberalism. Neoliberalism involves a significant reduction in the state’s social and environmental welfare role coupled with an expansion in the state’s facilitation of private capital accumulation . While neoliberalism has been consistently challenged since it arose in the late 1970s, contestation became mainstream after 2008, as the financial crisis raised questions about the viability of under-regulated financial markets and the growing inequality that helped fuel increased consumer reliance on credit . During this period, too, climate change moved into the mainstream of political debate: in 2007 Al Gore’s film An Inconvenient Truth won an Academy Award, and Gore shared the Nobel Peace Prize with the Intergovernmental Panel on Climate Change. As Gore himself articulates, neoliberal philosophy and policy has been a significant impediment to strong government action on climate change . This more mainstream contestation of neoliberalism – fuelled by economic crisis, rising inequality, and climate change – has not facilitated the emergence of broadly accepted alternatives, leading some critics to worry about a “zombie neoliberalism” that will not die . While the post-2008 period of contestation has not enabled a consensus solution to neoliberal capitalism’s contradictions, it has powered the search for alternatives . Ideological perspective necessarily conditions the kinds of solutions different actors seek and support. American political elites like Sarah Raskin and Ron Sims, for example, are interested in innovative ways of addressing inequality and ecological strain that leave in place the fundamentals of capitalism . Radical critics like Chomsky are interested in systemic alternatives to not only neoliberalism but also capitalism itself . Co-operatives, Evergreen founders note, provide alternative economic models that “cut across ideological lines – especially at the local level,vertical home farming where practicality, not rhetoric, is what counts in distressed communities” . Evergreen, then, is an ideologically flexible initiative: an innovative market-based poverty alleviation strategy or the germ of capitalism’s successor, depending on one’s point of view. While Evergreen has benefitted from the surge of interest in economic alternatives post-2008, the whole co-operative movement is experiencing resurgence. The General Assembly of the United Nations declared 2012 the International Year of Co-operatives. The International Co-operative Alliance , an organization representing the global co-operative movement, recently reflected that “rarely has the argument in favor of cooperatives looked stronger” . Co-ops can be read as either an ethical supplement to neoliberal capitalism, one that evens out its contradictions in distressed communities, or they can be read as the basis for a systemic alternative. Leaders of the Cleveland Model explicitly subscribe to the latter, more radical view, even as they strategically benefit from the former. Evergreen is modeled after the Mondragon Cooperative Corporation in Spain, which has long been a model for large-scale co-operative development worldwide. Founded in 1956, MC is now a conglomerate including 110 worker co-operatives, and employing more than 80,000 workers . Mondragon does business in manufacturing, retail, finance, and knowledge . As a worker owned co-operative system, Mondragon has several features that distinguish it from traditional capitalist firms: for example, a pay cap specifies that top earners with MC can only earn six times the pay of those in the bottom bracket . By comparison, CEOs for US corporations regularly make 400 times an average worker’s salary – a rate that has increased twenty fold since 1965 . Mondragon is one of the largest employers in the Basque region of Spain where it is centered . The Mondragon model is not without its challenges, including the recent bankruptcy of Fagor, one of its larger companies , but it remains an example of how co-operatives can operate on a large scale, produce considerable wealth, share it equitably, and promote relative worker satisfaction. As such, Mondragon is a longstanding inspiration for movements and intellectuals interested in alternatives to the capitalist firm and economy . Replicating Mondragon’s successes, however, is no easy task. The region’s political culture is an enabling factor: Basque country is home to a robust nationalist and separatist movement, and considerable associational energy is generated from feelings of marginalization at the hands of a dominant majority .3 Political culture supportive of co-operative development is not easily replicated, a fact that limits the ability of the co-operative movement to transplant successes from one region to another . Evergreen has generated popular and movement excitement partly because it appears to have successfully adapted the Mondragon model for North America. Northeast Ohio is not home to a political culture distinctively supportive of co-operatives. As Ted Howard, one of Evergreen’s leaders, told us, “Some people think there must have been something about the Cleveland community that would welcome this co-operative development, but it was a foreign concept” . As we learned, however, key supports were available in Cleveland – mainly a wealthy community foundation and champions in local government – that might not exist in other North American communities.Cleveland is still struggling to recover from the economic decline that began in the late 1960s. Once the fifth-largest city in the US and a center for manufacturing, Cleveland was hit hard by forces of economic globalization and the deindustrialization they brought. Plant closures, unemployment, and out-migration contributed to a depressed urban economy. Between 1970 and 1980, the city lost 24 percent of its population, one of the steepest drops in US urban history . Those who left generally had the means to do so, “with the poor, elderly, structurally unemployed, or marginally unemployed remaining behind” . White flight, the large-scale migration of whites from racially mixed urban neighborhoods to more suburban regions, was also a factor in the hollowing out of Cleveland . In the 1990s Cleveland began to slowly recover economically, and the city is currently a hub for health care services, biotech, and polymer manufacturing . The two largest employers in the region are the Cleveland Clinic and University Hospitals, together employing 46,000 people . Both institutions are located in the Greater University Circle , an area four miles east of downtown that is also home to Case Western University, the Cleveland Museum of Art, and the Museum of Contemporary Art. While the GUC is an economic and cultural engine, it is surrounded by seven low income and racially segregated neighborhoods.

Crew-based hiring explains why recruitment and retention are often part of the same labor market function in agriculture. Indeed, an analogy to obtaining irrigation water may be helpful to understand the recruitment and retention options. There are two major ways to supply irrigation water to crops: a field can be “flooded” with water so that some trickles to each tree or vine, or fields can be irrigated with a drip system that involves laying plastic pipes down or under the rows and dripping water and nutrients to each tree or vine. If water is cheap, farmers flood fields with water; if water is expensive, farmers may invest in drip irrigation systems. The analogy to recruitment and retention is clear: farmers more often work collectively to flood the labor market with workers, usually by getting border gates opened or left ajar, instead of recruiting and retaining the best farm workers for their operation, the drip irrigation model. The best way to ensure plenty of irrigation water is to invest in more dams and canals; the best way to flood the labor market is to invest in politicians willing to ease access to foreign workers.Farm workers were not granted federal collective bargaining rights in the 1935 National Labor Relations Act, and remain excluded from the NLRA. In 1975, California enacted the Agricultural Labor Relations Act to provide state-level organizing and bargaining rights: the purpose of the ALRA was to end a decade of strife in the fields, to “ensure peace in the agricultural fields by guaranteeing justice for all agricultural workers and stability in labor relations.” The ALRA includes three major elements: organizing and bargaining rights for farm workers, unfair labor practices that employers and unions can commit when they interfere with these worker rights, and a state agency,plant benches the Agricultural Labor Relations Board , to supervise elections in which farm workers decide if they want to be represented by unions and to remedy ULPs.

Between 1975 and 1984, there were over 1000 elections on California farms, and unions were certified by the ALRB to represent workers on 70 percent of these farms. Since then, there have been fewer than 250 elections, and unions were certified on less than 50 percent of the farms on which they requested elections .Farm worker unions were often unable to negotiate first agreements with most of the farms on which they were certified to represent workers, and in many cases, were unable to re-negotiate first agreements. The number of collective bargaining agreements in California agriculture has never exceeded 300 at any time, and in 2002 was about 225—80 percent of the current contracts cover 3-4 workers under Christian Labor Association contracts with dairy and poultry farms. The United Farm Workers , Teamsters, and other unions representing field workers have fewer than 30 contracts covering less than 25,000 workers. Unions such as the UFW charge that farm employers are able to avoid reaching first or subsequent contracts by refusing to bargain toward agreement. In 2002, the UFW led an effort to amend the ALRA to provide for state intervention to ensure contracts on farms on which workers voted for union representation. The UFW’s original goal was binding arbitration, under which a union and employer that cannot negotiate a contract typically go through a three-step procedure. First is mediation, when a neutral third party listens to each party separately and makes suggestions to narrow differences and allow them to reach a voluntary settlement. Second is fact finding, when a neutral party listens to both sides and proposes a non-binding settlement. Third is binding arbitration, when a neutral party proposes either any settlement deemed best or when the arbitrator is required to recommend one of the party’s final offers at the bargaining table. Binding arbitration is normally restricted to public employees such as police and firefighters who cannot strike lawfully.

The California Legislature approved binding arbitration in agriculture, but Governor Gray Davis threatened to veto the bill, so a last-minute compromise, “mandatory mediation,” was approved. Mandatory mediation, which went into effect January 1, 2003, requires unions and farm employers to bargain for at least 180 days for a first contract. If they cannot reach agreement, a mediator tries to help the parties to resolve their differences for another 30 days but, if mediation fails to produce an agreement, the mediator must, within 21 days, recommend the terms of a collective bargaining agreement that the ALRB can then impose on the parties. Although mandatory mediation might result in a greater number of collective bargaining agreements, other factors suggest that the new law will not affect a large number of agricultural employers or employees while it is in effect through at least 2007.The hired farm workers of tomorrow are growing up today outside the U.S., usually in rural Mexico and Central America. A major federal policy issue is what conditions, including what housing provisions, U.S. farm employers should satisfy to get access to these foreign workers. The U.S. has a guest worker program for farm workers, known as the H-2A program. It requires DOL to certify a farmer’s need for H-2A guest workers. In order to obtain certification, a farmer must satisfy certain recruitment, wage, and housing regulations, including applying for certification and trying to recruit U.S. workers at least 45 days before they are needed, offering to pay the higher of the minimum, prevailing, or Adverse Effect Wage Rate, and offering to provide free and approved housing to out-of-area U.S. and H-2A workers.Except for sheep farmers, California farm employers have traditionally not obtained workers through the H-2A program; most admissions have been in eastern states such as North Carolina. But the number of H-2A admissions in these eastern states has been rising, and H-2A workers for non-shepherding jobs were approved in California in March 2002, when a Ventura county custom harvester/FLC brought 38 H-2A workers from Mexico to California to harvest lemons, possibly a precursor to more H-2A farm workers.

If the H-2A program expands, there would likely be an increased demand for barracks or dorm style housing, and inspectors to check it. Instead of expanding the H-2A program, three other concepts are being debated to regulate the access of farmers to foreign farm workers: temporary guest workers, legalization, and earned legalization. Temporary guest workers are non-immigrants, persons in the U.S. to work generally for one employer, who must leave when the work ends—guest workers, under U.S. law, do not generally obtain any preference for admission as immigrants. During the 1990s, the SAWs—unauthorized farm workers legalized in 1987- 88—and their replacement with newly arrived unauthorized workers increased the risk to farmers that they may be fined or lose their workers at critical harvest times. Farmers could avoid such risks by having DOL certify their need for H-2A workers, but certification required offering at least a DOL-set wage and free housing. Many California farmers want an alternative guest worker program that does not require certification, and they do not want to offer free housing to legal guest workers. In July 1998, the U.S. Senate approved one grower proposal, the Agricultural Job Opportunity Benefits and Security Act ,rolling bench which avoided the need for farmers to be certified by creating a registry in each state to enroll legally authorized farm workers. Under AgJOBS, farmers would apply to the registry, for example, requesting 100 workers. If only 60 registry workers were available, the farmer would be automatically “certified” to recruit and have admitted to the U.S. 40 foreign workers. AgJOBS would also end the housing requirement by allowing the governor to certify that there is “sufficient” farm worker housing in the area, and then the farmer could offer a housing allowance equivalent to “the statewide average fair market rental for existing housing for nonmetropolitan counties for the State…based on a two bedroom dwelling unit and an assumption of two persons per bedroom,” about $500 a month in the northern Sacramento Valley and $800 a month in San Benito in 2000. However, most California agriculture is in metro counties, where 40th percentile fair market rents in 2000 are about $525 to $1,100 for two-bedroom units. Under AgJOBS, typical housing payments for guest workers would have been $125 to $150 per worker per month in California. President Clinton opposed AgJOBS, and issued a statement: “When these programs were tried in the past, many temporary guest workers stayed permanently and illegally in this country. Hundreds of thousands of immigrants now residing in the U.S. first came as temporary workers, and their presence became a magnet for other illegal immigration.” In 1999, after consultations with worker advocates, a new concept was added to AgJOBS: earned legalization. Legalizing unauthorized farm workers might encourage many of them to leave for  non-farm jobs, as SAWs did in the 1990s, so farmers who wanted guest workers and worker advocates who wanted legalization agreed to a program that would grant unauthorized workers a temporary legal status.

Under their compromise, unauthorized workers who could prove that they did 100 or 150 days of farm work in the preceding year would get a temporary legal status that permitted them to live and work in the U.S.In order to maintain this temporary legal status, and eventually apply to become a regular U.S. immigrant, the temporary worker would have to do a certain amount of farm work each year for several years, e.g., 80 or 100 days of farm work for three to five years. Thus, after several years and 240 or 500 days of farm work, the temporary legal worker could earn an immigrant status. Farmers and worker advocates argued over the details of a revised AgJOBS program that included earned legalization throughout 2000, with farmers wanting more days of farm work to qualify for eventual immigrant status, and worker advocates fewer days. After the November 2000 elections, some worker advocates, noting that both U.S. President Bush and Mexican President Fox favored a new guest worker program, agreed to a compromise that won the endorsement of the United Farm Workers and the National Council of Agricultural Employers. Under this December 2000 compromise, unauthorized workers who did at least 100 days of farm work in the preceding 18 months could qualify for temporary legal status, and they could convert this temporary legal status into an immigrant status if they did at least 360 days of farm work in the next six years. The compromise included freezing the minimum wage that had to be paid to foreign workers for several years and giving farmers the option of providing a housing allowance rather than housing to workers. The AgJOBS compromise came close to Congressional approval in December 2000, but was blocked by those opposed to any type of amnesty for unauthorized foreigners. The atmosphere changed in 2001, especially after U.S. President Bush and Mexican President Fox met in Mexico in February 2001 and agreed to establish a migration working group that was charged with creating “an orderly framework for [Mexico-U.S.] migration that ensures humane treatment [and] legal security, and dignifies labor conditions.” Senator Phil Gramm became the leading proponent of the guest worker-only approach, favoring a program that would permit unauthorized Mexicans already in the U.S. to obtain seasonal or year-round work permits: seasonal workers could return to the U.S. indefinitely, and year-round workers could remain in the U.S. three years, and then they would have to stay in Mexico at least one year before returning legally. U.S. employers and guest workers would pay social security taxes to a trust fund that would reimburse U.S. hospitals that provided emergency medical care for injured guest workers; the balance of the social security taxes paid would be placed in individual IRA-type accounts that workers could receive when they surrendered their work permits to U.S. consulates in Mexico. Gramm’s proposal covers Mexicans employed in all U.S. industries, but does not include a path to immigrant status. The other extreme is legalization. Under a plan embraced by the AFL-CIO and many church and ethnic groups, unauthorized foreigners in the U.S. from any country, and employed in any industry, could become immigrants, and then sponsor their families for admission. Rep. Luis V. Gutierrez introduced a bill that would grant immigrant status to all persons who were in the U.S. at least five years, and temporary legal status to those in the U.S. less than five years.

COOL regulations do not affect restaurants, but have implications for nearly everyone else within the unprocessed food chain. The law states that “the Secretary may require that any person that…distributes a covered commodity for retail sale maintain a verifiable record keeping audit trail…to verify compliance” for a period of up to two years . This includes foreign and domestic farmers and ranchers, distributors and processors, and retailers. We discuss the ramifications of this audit trail requirement for the cost of compliance below.The cost of COOL implementation can only be estimated at this time. The major direct costs of the program include the costs of segregating and tracking product origins, the physical cost of labels, and enforcement costs. AMS itself projects that domestic producers, food-handlers, and retailers will spend $2 billion and 60 million labor hours on COOL in the first year, though these figures were questioned by the GAO in a 2003 report. The GAO reports that the Food and Drug Administration has estimated that the cost of monitoring COOL for producers will be about $56 million annually. The costs of implementation for produce will likely be lower than the costs of implementation for meats as some fruits and vegetables are already labeled by country of origin. From a policy perspective, whether these uncertain costs outweigh the benefits to society of the program, and the extent to which retailers, producers and consumers will share these costs, are of equal importance. The extent to which COOL may benefit domestic producers depends on two considerations, whether country-of-origin information will induce and/or allow consumers to demand more domestic products relative to their foreign counterparts , and whether the costs of COOL implementation will be differentially higher for foreign suppliers than domestic suppliers. If COOL costs foreign suppliers more to comply than domestic suppliers,aeroponic tower garden system the transaction costs imposed by COOL will be lower for domestic suppliers than for foreign suppliers.

Even if the price elasticity of demand for foreign and domestic goods is the same, demand for foreign products will fall more than demand for domestic products, and some consumers who previously bought foreign goods will switch to buying domestic ones. This effect will be exacerbated to the extent that labels themselves affect consumers’ preferences or allow them to act upon preferences that were unsatisfied before mandatory labeling. If consumers truly prefer domestic products relative to foreign ones, all other characteristics being equal, COOL will be accompanied by increased demand for domestic goods. If this effect and the differentially higher compliance costs for foreign goods are large enough, this could theoretically offset the reduced demand for labeled goods occasioned by the transactions costs imposed by COOL. Gains to domestic producers are limited by the size of the market share claimed by foreign producers prior to the introduction of COOL, but in this case domestic producers would benefit from the regulation. Consumers could be net beneficiaries as well if mandatory labeling satisfied a preference that the market previously failed to serve. Economic theory and empirical evidence both suggest that the benefits of COOL are unlikely to outweigh the costs of compliance. Both consumers and suppliers are likely to be worse off as a result of this regulation. In the absence of market failures, the fact that producers have not found it profitable to provide COOL to customers voluntarily is strong evidence that willingness to pay for this information does not outweigh the cost of providing it. If the benefits outweighed the costs, profit maximizing firms would have already exploited this opportunity. Of course, this argument depends on whether the market for agricultural products functions well and would be responsive to consumer demands for COOL if it existed. In this section, we argue that this is indeed the case, and provide empirical support for the theoretical argument that the costs of COOL exceed its benefits. These findings are consistent with the conclusion of the U.S. Food Safety and Inspection Service , that there is no evidence that “a price premium engendered by country of origin labeling will occur, and, if it does, [that it] will be large or persist over the long term.” There is little evidence that imperfections in the food market prevent producers from providing country-of-origin-labeling. Asymmetric information, where one party in a potential transaction has better information than the other, can indeed lead to inefficient outcomes. However, in standard economic theory this result arises either because a seller would like to signal that his product is of high quality but is unable to do so convincingly, or because a seller that has a low-quality product can pretend that it is high quality.But this situation does not plausibly apply in the case of COOL in agriculture.

There is nothing now that inhibits producers from “signaling” the national origin of their products. Whatever their revealed preference, do consumers have a stated preference for country-of-origin labeling? The GAO summarizes survey evidence as indicating that American consumers claim they would prefer to buy U.S. food products if all other factors were equal, and that consumers believe American food products are safer than foreign ones. However, surveys also suggest that labeling information about freshness, nutrition, storage, and preparation tips is more important to consumers than country of origin . Revealed preference arguments in their simplest form suggest that if consumers truly preferred domestic food products, it would only take one grocer to limit store items to domestic-only products before other stores saw this grocer’s success and followed suit . Many producers have voluntarily provided labeling information for a variety of reasons. Producers of organic products have voluntarily labeled their products to attempt to capture a premium, as have producers of “dolphin-safe tuna.” If demand for information exists, agricultural producers have generally been adept at seizing this opportunity. Similarly, many lamb imports from Australia and New Zealand already bear obvious country-of-origin labels going beyond legal requirements because consumers prefer this product to domestic lamb and lamb from the rest of the world . Thus, Australian and New Zealand suppliers have an incentive to label their lamb products because they infer a positive net benefit to doing so, while producers and retailers who abstain from the practice must know that sales will not increase enough from offset labeling costs. There are other non-economic arguments used to support mandatory COOL that relate to food safety. It is possible that COOL would make tracing disease outbreaks easier, thus reducing the health costs of food-related diseases. This is less likely than might initially seem to be the case, because of the long delay between disease outbreaks and the shipment of contaminated products . If domestic products are systematically safer than foreign products, substitution towards domestic goods could also increase the average safety level of food consumed. However,hydroponic net pots there is little evidence that foreign food products are systematically less safe than domestic products.

Existing inspection rules ensure that foreign and domestic meats meet the same standards.Foreign fruits and vegetables do not systemically carry more pesticide residue than their domestic counterparts . There is insufficient evidence to determine if bacteria levels differ between foreign and domestic produce .Not surprisingly, in light of revealed preference arguments, many retailers have argued that the cost of COOL implementation will be excessive and burdensome. As noted above, AMS has forecast an annual cost of $2 billion to implement the regulation. These costs will be borne by the private sector as the Farm Bill provides no funds to alleviate industry costs for developing and maintaining the necessary record-keeping systems . In addition, the statute prohibits the development of a mandatory identification system for certification purposes. Instead, USDA must “use as a model certification programs in existence on the date of this Act” . As discussed earlier, USDA is also allowed to require a verifiable record keeping audit trail from retailers to verify compliance.” These seemingly contradictory directions to the USDA—no mandatory identification system is allowed, but an audit trail from retailers may be required—could limit the AMS’s ability to implement the COOL legislation, but is likely intended to act as a prohibition against any efforts to mandate full-scale “trace back” requirements that would track products from the farm gate to the grocery store . Such a formal trace back requirement would impose costs with legal incidence on producers in the field unlike a certification program, where the legal incidence of the costs of regulation falls mostly on retailers and processors. Of course, the economic incidence of the costs of this regulation will be determined by the price elasticity of demand for products, as explained in the discussion that conceptualized COOL as a transaction cost. While retailers’ organizations, like the Food Marketing Institute, have generally been against mandatory COOL, perhaps the loudest complaints about the cost of COOL have come from the meat packing and processing industry. In particular, the costs of tracking and labeling the origin of ground meat products are expected to be relatively high. For example, the president of the American Meat Institute, a trade group representing meat packers and processors has claimed that COOL regulation will be costly and complicated and that it will “force companies to source their meat not based on quality or price, but based on what will simplify their labeling requirements” . The National Pork Producer’s Council also opposed COOL legislation , and has since funded a study that estimates that the cost of COOL implementation will translate into a $0.08 per pound increase in the average retail cost of pork .

A key element of this study is an argument that, whatever the intention of the authors of the COOL legislation, implementation will in practice require complete “trace back” capability from the farm to the retail level. With the 2003 discovery of BSE in the U.S., a comprehensive trace back system for livestock may receive greater political support. Agricultural ranchers and growers have largely welcomed the COOL legislation. The California Farm Bureau , the Rocky Mountain Farmers Union , and the Western Growers Association , among other such organizations, have endorsed this regulation. These organizations generally argue that consumers “want” labeling, , consumers have a “right” to country-of-origin information , and that the legislation is a valuable “marketing tool” . The first of these arguments is weakened by the logic of revealed preference. In the case of meat products, the comments of the president of the American Meat Institute above explain the logic of the third justification; packers may demand more domestic inputs if this lowers the cost of COOL compliance. There is also some suggestion that the alleged market power exercised by the relatively concentrated meat-packing industry has created rents that COOL will dissipate . That is, the bargaining position of producers relative to packers will be improved as a result of these rules. This is at least in part because legal liability for failure to comply with COOL will rest with retailers, not with suppliers closer to the farm gate.COOL has been justified as an attempt to favor domestic products in the U.S. market, and early indications suggest that foreign suppliers believe it will do so. Canadian cattle groups have suggested that beef be given a “North American” label if it comes from any country in NAFTA . Meat producers in New Zealand have stated their disappointment with the regulation . International trade considerations may have made COOL more politically palatable in 2002 than it had been in the past. In 2002, the EU required member states to label all beef at the retail level, including ground beef, with information about the country of birth, fattening, and slaughter. This tightens regulations that have been in place since 2000 . Canada, Mexico, and Japan all have some version of COOL regulation. Other labeling initiatives have also been introduced in the EU, particularly for foods containing genetically modified organisms , regulations which are generally thought to be detrimental to U.S. products . One of the main arguments in favor of COOL, discussed above, has also been used to justify mandatory GMO labeling in Europe. That is, the consumer has a “right to know” what they are eating. Ironically, the U.S. government has strongly opposed mandatory GMO labeling, and for good reason.

Thus, processors have an incentive to procure production near their processing facilities. Timing of production is also critical. Tomatoes must be harvested immediately upon ripening and then processed quickly to avoid spoilage. The efficient operation of processing facilities and the effective processing of the harvest require that a processor’s deliveries be spread uniformly over an extended harvest period of 20 or more weeks. Similarly, processors specialize in producing different types of tomato products. Some plants produce only bulk tomato paste, which is then remanufactured at other locations into various processed tomato products, while others produce whole tomato products. The preferred type of tomato to grow depends upon the intended finished product. Delivery dates and product characteristics cannot be communicated effectively through spot markets. Nor will a central market work when processors are interested in procuring product only in the vicinity of their plants.Thus, the California processing tomato industry transacts essentially its entire production through grower-processor contracts. These contracts specify the specific acreage the product is to be grown on, variety of tomato to be grown, delivery dates, and premiums and discounts for various quality characteristics. Price terms in these contracts are set with the intervention of a producer bargaining cooperative.Cooperatives are firms that are owned by the producers who patronize them,blueberry grow pot although many cooperatives also do business with nonmembers. California is home to many large and important food-marketing cooperatives.

Producers who are members of a marketing cooperative essentially have vertically integrated their operation downstream into the processing and marketing of their production. A number of incentives can account for producer cooperative integration, including avoidance of processor market power, margin reduction, and risk reduction . Cooperatives are the leading marketing firm in several California agricultural industries including almonds , walnuts , prunes , citrus and raisins . However, the recent years have represented difficult times for some California marketing cooperatives. Tri Valley Growers , a fruit and tomato processing cooperative, formerly the second largest cooperative in California, declared bankruptcy in the summer of 2000. Around this same time the Rice Growers Association, a large and long-lived rice milling operation closed its doors, as did Blue Anchor, a diversified fresh fruit marketer. Reverberations from the failure of these prominent California cooperatives were felt nationally and caused some to wonder whether the model of cooperative marketing was well suited for 21st century agriculture. Indeed cooperatives do face some important challenges competing in the market environment we have described here. As noted, retailers prefer suppliers who can both provide products across an entire category and provide them year around. Cooperatives are traditionally organized around a single or limited number of commodities and member production is likely to be seasonal. Cooperatives can attempt to surmount these difficulties by undertaking marketing joint ventures with, for example, other cooperatives, and sourcing product from nonmembers, including internationally. However, cooperatives may face impediments relative to investor-owned competitors in pursuing such strategies. For example, various laws affecting cooperatives specify that at least 50 percent of business volume must be conducted with members.

Joint ventures with firms that are not cooperatives are not afforded legal protection under the Capper-Volstead Act.2 Doing business with nonmembers may also adversely affect a cooperative’s membership, if it is perceived that most of the benefits of the cooperative can be obtained without incurring the financial commitment associated with membership. This issue was important for TVG when it appeared that tomato producers selling to TVG under nonmember contracts received a better deal than member growers. Cooperatives may also face challenges in procuring the consistent high-quality production that the market place now demands. Cooperatives usually employ some form of pooling mechanism to determine payments to members. In essence, revenues from product sales and costs of processing and marketing flow into one or more “pools.” A producer’s payment is then determined by his/her share of the total production marketed through each pool. The problem with some pools is that high quality and low-quality products are commingled and producers receive a payment based upon the average quality of the pool. Such an arrangement represents a classic adverse selection problem, and its consequence is to drive producers of high-quality products out of a cooperative to the cooperative’s ultimate detriment. Cooperatives can obviate this pooling problem through operating multiple pools and/or by designing a system of premiums and discounts based upon quality, but the key point is that investor-owned competitors face no similar hurdles in paying directly for the qualities of products they desire. Offsetting these limitations on cooperative marketing in the 21st Century at least to an extent is the recognition that the marketing clout producers can attain from joint action may be as important now as ever. As we have documented in this chapter, the food retailing and processing sectors have consolidated. Although producers, too, have generally gotten larger in absolute scale, the typical producer’s power in the market place pales in comparison to that of the firms with whom he/she conducts business. Also worthy of mention is that some cooperatives have evolved the structure of their organization to “keep up with the times.”

Such cooperatives are usually known as“new generation cooperatives” or NGCs. Typical features of an NGC include grower contracts that include both delivery rights and delivery obligations. Delivery rights, however, are transferable and can function somewhat analogously to capital stock, i.e., if a cooperative is successful, its delivery rights will increase in value. These rules are intended to give the cooperative assurance of a stable supply but also to regulate the amount of product it must process and sell in line with market conditions. To date, the NGC structure is most common among cooperatives in the mid western U.S. and has made few inroads in California. Interestingly, TVG underwent a re-organization to an NGC structure in 1995-96. Although it is doubtful that this re-organization had much impact on TVG’s ultimate demise, its failure may have made Californians skeptical of the NGC structure. Two types of cooperative organizations are relatively unique to California. They are information-sharing cooperatives and bargaining cooperatives. Information sharing cooperatives perform no handling or other traditional marketing activities for their members. Rather, they serve as devices for their members to communicate, share information on production plans and market conditions, and formulate pricing strategies. Industries where these cooperatives have emerged include iceberg lettuce, melons, kiwifruit, table grapes, fresh stone fruits, mushrooms,hydroponic bucket and fresh tomatoes. The activities undertaken by these cooperatives would ordinarily be illegal under the U.S. antitrust laws but are rendered lawful due to the Capper-Volstead Act, which grants an exemption from the antitrust laws to farmers acting collectively through a cooperative. The major examples of this form of cooperative are industries where the product is highly perishable and production is concentrated in the hands of relatively few grower-shippers. Successful coordination of production and marketing in these industries can be a major advantage in terms of managing the flow of product to the market to avoid the periods of over supply and low prices that have been common in these industries. Membership in these organizations tends to fluctuate, however, and there is little evidence to date that they have been successful in either raising or stabilizing grower prices.3 Bargaining cooperatives also engage in little or no actual handling of product. Rather, they function to enable growers to bargain collectively the terms of trade with processors. Iskow and Sexton identified 10 active bargaining cooperatives in California and 29 nationwide. Prominent California bargaining cooperatives are the California Tomato Growers, California Canning Peach Association, California Pear Growers, Prune Bargaining Association, and Raisin Bargaining Association. These cooperatives are a response to the asymmetry in power that might otherwise characterize dealings between farmers and processors. Bargaining associations are especially common in processed fruit and vegetable industries, where products are generally grown on a contract basis and there is no active spot market. In addition to increasing growers’ relative bargaining power, these associations play a valuable role in facilitating exchange and minimizing transaction costs. Rather than having to negotiate terms of trade with each individual grower, a processor need strike only a single agreement with the bargaining association. Generally the bargaining association will negotiate first with a single leading processor, with similar contract terms then applying to other processors. In no case is a cooperative the sole marketer or bargainer in California. Farmers always retain the option not to participate in a cooperative. In fact, many of the benefits that a cooperative provides are available to a grower whether or not he/she is a member of the cooperative. For example, Blue Diamond was a leader in opening new export markets for almonds. However, once these markets were established, other handlers were easily able to sell into them.In industries with cooperative bargaining, a farmer who is not a member of the bargaining association generally receives the same terms of trade as growers who are members. Thus, farmers have an incentive to free ride on the activities of the cooperative.In addition to undertaking joint action in marketing through cooperatives, U.S. legislation at both the national and state levels allows producers and marketers of many agricultural products to act collectively through a legal structure to control various aspects of the marketing of their products.

Enabling legislation for federal marketing orders is provided by the Agricultural Marketing Agreement Act of 1937 , while state orders and agreements are governed by the California Marketing Act of 1937, with amendments. California also has more than 20 individual laws for the formation of commodity commissions and councils. There are differences between the use of federal and state marketing programs. Federal marketing orders can cover a production region in more than one state, while state orders are effective only within the state boundaries. Federal marketing orders tend to focus on quality regulations and sometimes volume controls, while California state marketing programs tend to focus on research programs and promotion. Several California commodities utilize different programs for different activities. For example, California-grown kiwifruit has a federal marketing order program that administers grades and standards and a state commission that conducts advertising and promotion; California walnuts have a federal marketing order with provisions for grades and standards and quantity control and a state commission used only for export advertising and promotion. California agricultural producers were at the forefront in adopting both federal and state marketing order programs when they first became available in the 1930s. The mandatory nature of the programs overcame the free-rider problems that had earlier led to a breakdown of cooperative-organized quality and supply control marketing efforts. The popularity of government-mandated commodity programs is clearly reflected by their continued use by a large number of commodity producers. California had 17 federal marketing orders and 48 state marketing programs effective in 1993 . Those programs covered commodities that accounted for 54 percent of California’s 1990 agricultural output, based on value. There are 12 federal orders and 51 California commodity marketing programs effective in 2003. As shown in Table 4, these commodities accounted for 55 percent of California’s total agricultural output in 2002. The largest proportional drop in marketing program coverage between 1990 and 2002 was for vegetables .Among the 17 federal marketing orders operating in California in 1993, four were eliminated and coverage of California production was dropped by two. The terminated federal programs included the marketing order for Tokay grapes and the long-standing marketing orders for California-Arizona Navel oranges, Valencia oranges, and lemons. The marketing orders for Northwest winter pears and spearmint oil, while still in effect, no longer apply to California production. There is one new federal order, the Hass Avocado Promotion, Research and Information Order, and another has been proposed for pistachios. Thus, there was a net loss of 5 federal marketing orders for California commodities between 1993 and 2003. There have also been changes in the coverage of California marketing programs. The marketing orders for apricots and fresh tomatoes have been dropped and there is a new state order for garlic and onion dehydrators and a “Buy California” marketing agreement. The California Egg Commission is no longer operating but there are new California Commissions for dates, rice, tomatoes, and sheep. The total number of California marketing programs went from 48 in 1993 to 51 in 2003 for a net gain of three.

Eighty-two percent of Sacramento Valley cropland is irrigated. Irrigation water sources include private and cooperatively developed surface water supplies along the western slope of the Sierras, riparian sources along the major rivers, e.g., the Sacramento, Feather, Yuba, Bear and others, and more recent additions of federally developed water supplying the western valley via the Tehama-Colusa Canal. The Sacramento River and its tributaries are the initial components of the conveyance system for federal and state water systems which, from the Delta southwards, delivers surface water via pumping plants and canals to the San Joaquin Valley and Southern California for agricultural, municipal, and industrial uses. Groundwater sources are also significant. Cooler winters, higher rainfall, and less productive soils than the San Joaquin underlie the continued importance of field crops in the Sacramento Valley. Rice is grown in areas with more impervious basin soils; both wheat and corn are included in irrigated crop rotations; and alfalfa, dry beans, sunflowers, safflower, and vine seeds are among other important field and seed crops. Field corn is grown extensively in the Delta. A variety of fruit and nut crops—mainly almonds, peaches, pears, prunes and walnuts—are grown on the deeper, better-drained and more fertile soils of the region. Fruits and nuts amount to 33 percent of the region’s value of production in 1995. Vegetable crops, mostly processing tomatoes, contributed 16 percent, and livestock and livestock products, an additional 11 percent, of the regional production total.About a third of California’s farmland and 55 percent of its irrigated lands lie in the San Joaquin Valley. Nearly 90 percent of valley cropland is irrigated. The eight counties of the San Joaquin Valley accounted for $12.75 billion of the $22.1 billion total value of California agricultural production reported for 1995 . Unlike the Sacramento Valley,vertical hydroponic nft system the San Joaquin does not have a single river system that runs through the entire valley.

The southern portion of the valley is two lake basins, historically fed by seasonal runoff from the Sierra Nevada Mountains to the east. Early farming depended on private and cooperative development of water supplies from Sierra rivers to irrigate alluvial lands on the east side of the valley, and on the reclamation of the Tulare and Buena Vista Lake Basins in the south valley bringing more acreage into agricultural production. In the post-World War II period, federal and state surface water development brought additional water supplies to the most southern area and to the entire western San Joaquin Valley, which had formerly depended on limited and often poor quality groundwater. Because much of the valley is either of a desert or steppe climatic type, irrigation is the major factor that has made the San Joaquin the most extensive and productive of the agricultural regions of California. The west side of the San Joaquin Valley was the region most affected by the 1987-93 drought and by reduced allocations from CVPIA and CALFED decisions. Consequently, this area is among the most innovative in implementing market transfer initiatives and adopting water-conserving irrigation technologies. Clearly the economic fate of this region, and the others, is closely tied to long run supplies of irrigation water and to current initiatives that seek to reallocate surface water supplies among competing agricultural, municipal and industrial, and non consumptive environmental uses.With the majority of the state’s agricultural production located in “The Valley,” most kinds of production can be found somewhere within its confines. What is surprising is the diversity in types of farming enterprises, ranging from older, smaller, more intensively cultivated farms on the east side to the larger, more extensive farms on the west. Fruit and nut crops, including grapes and citrus, are important to the region, contributing 39 percent of the total value of production in 1995. While the majority of permanent plantings lies on the east side of the valley, recent plantings of nuts and some deciduous fruits have been made on the west side.

Livestock and livestock products are located throughout the valley and contribute an additional 28 percent of the region’s agricultural production.Cotton is the most important field crop. Recent introductions of pima varieties have augmented traditional upland cotton production but total cotton acreage has fallen due to poor profitability. The region is an important producer of most field crops . Irrigation and a long growing season have also led over time to increased vegetable production . Summer melon production is important, as is seasonal production for many of the major vegetables . Some seasonal production is timed to fill marketing niches as the fresh produce industry moves in the spring from desert to coastal areas and in the fall back toward the desert. Of the major categories, nursery products and cut flowers appear relatively insignificant in comparison with the total value of agricultural commodities .This region of the state is very similar to that of the North, being largely dominated by livestock and livestock-related economic activity an private and leased public lands. The Mountain region covers about 15 percent of the state’s land area, and land is mostly in public ownership; less than 10 percent of the total land area is in farms. Together, livestock , livestock products , and field crops—mainly rangeland and pastureland production —, amount to about three quarters of the value of the region’s agricultural activity in 1995. In truth, the dominance of these commodities in the region’s agricultural economy is larger because the geographic location of fruit and nut production , and nursery products recorded for the region, actually occur on the west slope, foothill “valley” portions of several mountain counties.Including the eastern areas of the Los Angeles area , this region also extends across the more remote desert valleys—the Coachella, Palo Verde, and Imperial Valleys—irrigated by early diversion rights to Colorado River water.

Only 28 percent of the land area is in private ownership, and only 10 percent of the land area is in farms. Because of the severe climatic conditions, a high proportion of cropland is irrigated . The western San Bernardino and Riverside areas include remnants of the once-dominant citrus and drylot dairying industries, which are gradually being displaced by urban expansion. Livestock and livestock product activities contribute the greatest proportion of the value of production in the South Coast region by capitalizing on the region’s proximity to markets and a long tradition of cattle feeding in the Imperial Valley and other desert valley areas. Vegetable production , predominantly in the irrigated desert valleys, includes important winter and early season production of asparagus, carrots, lettuce, melons, and sweet corn. Highly productive desert lands with irrigation benefit from temperate winters and nearly frost-free growing seasons to produce a variety of high-valued fruit and vegetable crops that are in supply during the off- and early seasons of the major production regions. Fruit production is mainly in the western areas and in the Coachella Valley . Field crop production includes alfalfa hay production for the region’s livestock activities, cotton, sugar beets, and wheat, including durum.Risk is substantially greater in the production and marketing of perishable fruits and vegetables than in more stable commodities.Investments in permanent plantings are large and must be paid back over the period of economic production. Figure 2 shows the pronounced change in the distribution of field crop,nft hydroponic system tree fruit and nut, and vegetable acreages and value of production over the decade of the 1980s. In 1980, production of fruit, nuts, and vegetables contributed over half of the value of production , but only used 27.9 percent of the acreage in production. In 1990, these more intensive, higher-valued, higher-risk crops amounted to 73 percent of the value of production, while using 38.7 percent of acreage. The residual nature of field crops is evident as farmers and ranchers seek more intensive production enterprises. Shifts toward increased acreages of vegetables and permanent plantings continued through the decade of the 1990s, most noticeably with substantial increased acreages of nut crops , deciduous tree fruits , and wine grapes. The composition of California agricultural production is compared for the years 1955, 1975 and 1995 in Figure 3. Total value of agricultural production grew three-fold from 1955 to 1975, from $2.68 to $7.43 billion.

Change in composition between 1955 and 1975 was not as dramatically different as that which has occurred over the last period, 1975-95, partly due to an overall increase in irrigated acreage through most of the first period.16 By 1995, high-valued fruit and nut, vegetable, and nursery and greenhouse products contributed 60 percent of the aggregate value of production for the state, and total value of agriculture production amounted to almost $22 billion. Field crop and livestock/livestock product categories were reduced by about one-half and one-third, respectively, in terms of their relative contribution to the value of California agricultural production. In 2001, the value of nursery, greenhouse and floriculture exceeded the value of field crops, and the dairy sector alone accounted for 17 percent of the state’s value of agricultural production . As a consequence, the share produced by livestock, poultry, and products actually rose from 25 percent in 1995 to 28 percent in 2001.The shifting composition of agricultural production is also reflected in changes in the state’s “Top Twenty” agricultural commodities over time. Table 2 shows the “Top Twenty” commodities ranked by gross farm income for the 2001 crop year, with comparisons for 1981 and 1961. Comparison of the 1961 and 2001 lists shows that whereas there were a total of 12 livestock/livestock products, and field crops identified in 1961, only 5 were on the 2001 list. In sharp contrast, there are now 13 fruit, nut, and vegetable crops on the 2001 list, compared to only 8 on the 1961 list. Nursery products and foliage and cut flowers have been added since 1961, appearing on both the 1981 and 2001 lists.California is now the number one milk producer in the United States. California’s dairies and the dairy processing sector are part of a dynamic system that has progressively become more efficient, larger, and more specialized over its history. Herd sizes are, on the average, ten times larger than the national average, and cows are, on the average, significantly more productive. Dairy processing capacity has more than doubled during the 1990s. The state’s dairy industry evolved from “local” dairies that originally provided fluid milk to nearby growing population centers in the San Francisco and the Los Angeles area milk sheds. The San Joaquin Valley milk shed was first a center for lower-valued manufacturing milk used mainly for butter and cheese production. With improved transportation systems and reduced land available for dairies in or near the main population centers, the San Joaquin Valley is now the major source of fluid milk serving both the Bay Area and the Los Angeles Basin. Processing continues to be concentrated there as well. Continuing urbanization and waste disposal challenges have caused more dairies to move into Central Valley and South desert areas, principally into the San Joaquin Valley. California’s dairies are highly specialized. As the number of dairies decreased, their size has become significantly larger, requiring more capital-intensive specialized production systems based on genetics, herd health, nutrition, and high levels of management. Urban expansion in the Los Angeles area led to the development of the drylot, feedlot style dairy using concentrates and feed stuffs often grown in other areas. Modern dairies often milk 3,000 or more cows daily and use waste effluents and solids on silage and forage crops on adjacent cropland.Wine grape production occurs throughout the state. California’s premium wines come from grapes grown predominantly in cooler, coastal valleys, most notably in the Napa Valley, but also in other North Coast areas as well as in some Central Coast areas. Higher yielding vineyards in the San Joaquin Valley produce standard and mid-quality table wines often marketed in larger-sized bottles and containers. The California wine-grape vineyard and wine-production industries have grown sporadically over the last half century. Following World War II, about 80 percent of the wine produced was in the fortified appetizer or dessert wine category with production chiefly in the San Joaquin Valley. Americans did not then know much about quality wines, but gradually, as tastes changed, the industries also changed toward the production of both standard table and world-class premium quality wines.

Studies show that between 1980 and 2000, growth in non-farm jobs reduced poverty, but growth in farm jobs did the opposite. In the past, increased demand for farm labor induced new immigration from Mexico, while increases in the supply of farm labor through immigration stimulated growth in the agricultural sector, thereby increasing the demand for farm labor. The key to this circular relationship between farm labor demand and immigration was that the supply of immigrant farm labor was elastic; that is, immigration was responsive to changes in U.S. farm wages. Our findings suggest that Mexico’s farm labor supply is not as elastic as it once was. Raising worker productivity is a prerequisite for increasing farm wages and enabling farm worker families to rise above the poverty line. Rising farm wages, in turn, create an incentive for farmers to make investments that will make farm workers more productive. In California, grapes rank as the highest-valued agricultural crop and the second-highest valued agricultural product after milk and cream. Wine grapes alone contributed roughly $2.1 billion, or 5.9%, to the total value of California farm production in 2010, with a further $0.9 billion contributed by table grapes, raisin grapes, and grapes crushed for other uses. California produced 86% of both the volume and value of U.S. wine grapes in 2010.Measures of demand response to economic factors, including price and income, are often used in economic analysis of markets and policies. The elasticity of demand for wine grapes is useful for estimating the price, quantity, and economic welfare effects of anything that causes a change in the production or consumption of wine grapes—new policy, disease, or pests, for example. Despite the economic importance of this industry,hydroponic grow table and the usefulness of elasticities, estimates of demand response for California wine grapes are scarce. In our recent article in the Journal of Wine Economics we report estimates of demand response for California wine grapes.

We also discuss the pitfalls and challenges of the estimation of demand response for commodities that are highly differentiated, with huge variation in price by agronomic variety, geographic location of production, and other characteristics that affect “quality” and end-use of wine grapes. Here, we summarize the main findings of that work, leaving aside the technical details, which can be found in the longer article in the Journal of Wine Economics. We focus on price elasticities of demand for wine grapes, which measure the percentage change in quantity demanded in response to a one-percent increase in price.Several aspects of the demand for California wine grapes are pertinent when estimating elasticities that will be useful for policy and market analysis, and in interpreting the results from estimation. First, it is appropriate to estimate an “inverse” demand model, in which the market price varies in response to variations in market quantity, rather than vice versa. Wine grapes are a perennial crop, for which current production is determined to a great extent by decisions made years, or even decades, earlier. Thus, variations in the current market price have comparatively little influence over the quantity supplied in the current season. Consequently, we can treat year-to-year quantity variation as determined by factors other than the current price, including past vineyard investment decisions, as well as current pest and disease incidence and weather, and treat the market-clearing price as responding to these quantity variations. Second, as for most farm commodities, the demand for California wine grapes does not reflect final consumer demand, but rather demand from processors who use grapes to produce a consumer product. This is important for how we approach the estimation problem and how we interpret the resulting estimates. Third, California wine is sold in the rest of the United States and exported, and competes in these markets—even in California—with wine produced in other states and other countries. Thus, global supply and demand conditions influence the demand for California wine and hence the demand for California wine grapes from which California wine is derived. With close substitutes in the market , we expect the quantity of California wine grapes demanded to be more sensitive to price than it would be otherwise. Fourth, wine is highly differentiated, made from highly differentiated wine grapes of many varieties produced across a diverse range of agroecologies.

Reflecting this differentiation and diversity, the California Department of Food and Agriculture collects detailed data for each of the 17 geographically based California “crush districts.” Broadly speaking, Napa and Sonoma vineyards produce comparatively few tons per acre at comparatively high cost per ton. In the Central Valley, especially in the southern San Joaquin Valley, yields are up to 10 times higher and grape prices per ton are in the range of one tenth of prices in the Napa and Sonoma crush districts. The rest of the state has a range of yields, costs and prices that fall between these extremes. For the purposes of our demand analysis, we aggregated the 17 crush districts into three regions that we defined as “High,” “Medium,” and “Low” based on their average wine grape prices, while noting that every region produces a range of wine grape varieties and characteristics. The regions are depicted in Figure 1. Table 1 presents regional statistics on value of production, average price per ton, total crush, and average vineyard yield in 2010.As noted, in this work we focus on price elasticities of demand for wine grapes, which measure the percentage change in quantity demanded in response to a one-percent increase in price. James Fogarty reviewed the worldwide literature on demand for alcohol. He reported estimates of the own-price elasticities of demand for beer, wine, and spirits from 141 studies. He reported 177 estimates of the elasticity of demand for wine with respect to its own price ranging from –1.86 to –0.18. These are measures of price responsiveness of demand for wine, a finished product, which is different from the demand for winegrapes, an input. In what follows we use an average value of –0.80 for the elasticity of demand for wine together with other information to derive estimates of elasticities or price responsiveness of the demand for California wine grapes. The demand for California wine grapes as an aggregate category is derived from the demand for California wine in conjunction with technology of wine making and the supply of wine making inputs.We evaluated this equation using a range of values for the parameters related to ROW wine grape supply response, supply response of wine making inputs, and international price transmission, combined with a value of –0.80 for the elasticity of demand for all wine.

The resulting estimates of the own price elasticity of demand for California wine grapes range from –0.4 to –4.5. The range reflects alternative assumptions about the elasticity of supply of wine grapes from the rest of the world, price transmission, and the elasticity of supply of other wine making inputs. Using intermediate values for these key parameters and available data, we estimated the overall elasticity of demand for California wine grapes as –2.2. The demand for California wine grapes can be further decomposed into interdependent demands for wine grapes by quality category. The corresponding elasticities of demand for wine grapes from different quality regions can be measured as a function of the overall elasticity of demand for California wine grapes,flood tray market shares, and the extent to which the different quality categories can substitute for one another in wine making. We derived the equations for these disaggregated elasticities and evaluated them using data on market shares, the intermediate value for the overall elasticity of demand for California wine grapes , and a range of substitutability between the different qualities of wine grapes. Allowing for quality differentiation and imperfect substitution among wine grapes from the three different regions—as defined Figure 1—gives a full set of own- and cross-price elasticities as shown in Table 2.The own price elasticities are in boldface.In addition to the “derived” estimates just discussed, we estimated elasticities using an econometric model of demand. We estimated inverse demand system models for the three quality cum-regional categories of wine grapes defined in Figure 1 and with differences in average prices and yields as illustrated by the summary statistics in Table 3. The models were estimated using annual data on prices and quantities of California wine grapes taken from the annual NASS/CDFA Crush Reports for the years 1985–2010. Table 2 shows the elasticities estimated using this method in Column . The own-price elasticity of demand for high-priced wine grapes is fairly large in magnitude , suggesting that a one percent increase in price for wine grapes from Napa and Sonoma counties, holding all other prices constant, would induce a 9.5% decrease in quantity demanded. The other own-price elasticities are substantially smaller in absolute value ; a one percent increase in price for mediumor low-priced wine grapes, holding all other prices constant, would result in roughly a 5.2% or 2.6% decrease in quantity demanded, respectively. Thus, demands for all three categories are fairly elastic. The econometric estimates indicate that demand for high-priced wine grapes is the most elastic and the demand for low-priced wine grapes, mostly from areas in the southern San Joaquin Valley, is the least elastic. We might have anticipated the converse, given the very strong international competition in the bulk wine market, and we have some reservations about putting too much credence in any particular disaggregated elasticities for particular quality categories estimated in this fashion.

Several points are clear from the comparison of the econometric estimates in Column and the derived estimates in Columns,and —the latter computed using a range of assumptions about substitutability among different qualities of wine grapes and an elasticity of aggregate demand for California wine grapes of –2.2. First, reflecting our assumptions, the derived estimates of cross-price elasticities are all positive numbers whereas some of the econometric estimates are negative numbers, indicating complementary relationships—though small values relative to the negative own-price and positive cross-price effects. While cross-price elasticities are of some interest, analysts are typically more concerned with own-price elasticities, and for this comparison we would place greater weight on ownprice elasticities while giving some weight to cross-price elasticities. Second, while the econometric estimates are broadly comparable to the derived estimates they are not completely consistent with any particular assumption about the degree of substitutability, denoted s, among wine grape qualities. The econometric estimates for the “Low”-price region are closest to the derived estimates assuming low substitutability ; those for the “Medium”-price region are closest to the derived estimates assuming moderate or high substitutability ; and those for the “High”-price region are closest to the derived estimates assuming high substitutability . Olive oil is hot. From the New York Timesto the New England Journal of Medicineto the U.S. International Trade Commission , olive oil is making news and raising important economic and policy issues. The California olive oil industry is also hot. Planting, production, and the reputation for quality are all increasing. Leaders in the California industry have plans to improve its competitive position in the market and become a major crop in the state. To act on their plans, however, they need more information and analysis. To understand how the market for olive oil in the United States is evolving and the importance of economic trends and policy initiatives, we must examine the market in some detail, including how factors such as prices, income, and new information affect quantities of olive oil demanded. This article begins this detailed examination. Global production of olive oil increased from less than 1.5 million tons in 1990 to over 3 million tons in 2012. Major producing regions, which surround the Mediterranean Sea, produce about 95% and consume about two-thirds of the world’s olive oil . About 300,000 tons of olive oil were sold in the United States in 2012, tripling the quantity sold in 1990. Thus, U.S. consumption now accounts for about 10% of world production. Spain is the world’s largest olive oil producer, with Italy a distant second.Italy is a major exporter to destinations outside of the EU and some of what is exported from Italy is oil produced elsewhere. North Africa is a major producer and an even more important exporter of olive oil. The U.S. industry produces only 4,000 tons of olive oil annually, or only about 1.3% of U.S. consumption.

DNA endoreduplication does not appear to affect this overall growth rate but may be required to sustain it beyond a critical cell size, giving rise to the robust continued growth of optoBem1 cells. It has been shown in other organisms, for example, that DNA endoreduplication enables large increases in cell size. One possibility by which our findings can be reconciled with prior observations of exponential growth in wild type budding yeast is that cells become surface area-limited at sizes just above that of wild type cells, thereby inducing a shift from volume proportional growth to surface area-proportional growth.Cell size control pathways exist to correct for deviations from a set-point size, yet most previously-identified size control pathways specifically operate on cells that are born too small, delaying cell cycle progression to enable further growth to occur. Because the light and temperature-shift stimuli with which we prepared ‘giant’ yeast are fully reversible, we reasoned that we could monitor the return to a steady-state size distribution after releasing giant cells from their block. We prepared giant optoBem1 cells by incubating them in red light for 8 h and monitored them by live-cell microscopy after releasing them into infrared light. Strikingly, we found that cell populations rapidly returned to their unperturbed state , with individual daughter cells reaching the set-point volume in as few as three rounds of division .Return to the set-point size is not driven by cell shrinking, as giant mothers maintained their maximum volume over multiple rounds of budding . Instead, the giant mothers are eventually diluted out as successive generations are born, an effect that is especially prominent in cell populations at least 10 h post-Bem1 release .

In these populations,hydroponic bucket size distributions have a single mode near the set-point volume but exhibit long tails towards larger volumes . Our observation that cell size recovers after only a few generations strongly supports the existence of size control acting on large cells and demonstrates that size homeostasis across a cell population is robust even to extreme increases in cell volume.Quantitatively monitoring cell growth in yeast—as well bacterial, archaeal, and mammalian cells—has shown that the behavior of many organisms is consistent with an adder that monitors size across an entire cell cycle to correct for deviations in cell size and maintain size homeostasis in the population. However, a recent study argued that in budding yeast, the adder behavior could arise from independent regulation of pre- and post-Start events, without a cell needing to keep track of its added volume across all cell cycle phases, and may fail under various perturbations. To test whether adder-based mechanisms could account for size control in giant yeast, we quantified inter division volume change in successive cell division cycles after releasing optoBem1 cells into infrared light. For this experiment we prepared optoBem1 cells that also expressed fluorescently-labeled septin rings, which enabled us to time both bud emergence and cytokinesis and thus separate pre-Start and post Start size regulation . The ‘adder’ model predicts that the cell volume at division should be proportional to cell volume at birth with a slope of 1. Indeed, for unperturbed cells, we found that cell volume at division was linearly related to volume at birth with a slope of 1.19 . However, we found that the adder model poorly explained the cell size relationships in our giant cells, where the volume at division was related to volume at birth with a slope of 1.73 . This relationship was also evident when individual cells were tracked over time: the interdivision volume change, Δ, was positively correlated with the volume at birth . This size-dependent volume change occurred entirely during S/G2/M phase, as cells added a minimal volume during G1 that did not vary with cell size . We also performed analogous experiments in cdk1-ts giant cells that were shifted back to the permissive temperature. These experiments revealed a similar relationship: large cells grew more than small cells, exhibiting a linear relationship between volume at division and volume at birth with a slope of 1.70 .

These results are broadly consistent with recent work showing that although size control in unperturbed cells resembles an adder-based mechanism, no mechanistic adder regulates volume addition across the entire cell cycle. Our data also suggest that any size regulation limiting the growth of large cells is likely a consequence of regulation in S/G2/M, as growth during G1 is negligible.If an adder is unable to explain size homeostasis in giant yeast, what regulatory mechanisms or growth laws might operate on the daughters of giant cells during S/G2/M? Two possibilities include a bud ‘sizer’, where bud growth would be restricted after reaching a critical size; and a bud ‘timer’ in which cytokinesis would occur at a fixed duration following the beginning of S/G2/M . Such ‘sizers’ and ‘timers’ have been proposed to operate in a variety of biological systems. To distinguish between these possibilities, we tracked the timing of bud emergence and cytokinesis by septin ring appearance and disappearance, respectively, following reactivation of Bem1 in giant optoBem1 cells . Daughter volume strongly correlated with mother volume , inconsistent with a bud sizer mechanism. Our prior observation that the inter division volume change scales positively with cell birth size further argues against a bud sizer for cell volume control. In contrast, our data were consistent with a timer specifying the duration of S/G2/M: the time from bud emergence to cytokinesis did not vary as a function of mother cell volume and took average 95 min across cells of all volumes .Similar experiments performed using cdk1-ts cells were consistent with our observations in optoBem1 cells, revealing a size-independent duration of budding. However, we observed one notable difference: the duration of the size-invariant bud timer in giant cdk1-ts cells was substantially longer than that of giant optoBem1 cells . As Cdk1 is a key driver of mitosis in eukaryotes, the increased duration of the bud timer in cdk1-ts cells may arise from the need to refold or synthesize new Cdk1 molecules to complete S/G2/M following a shift from the restrictive to permissive temperature. Furthermore, even when grown at the permissive temperature, the doubling time of cdk1-ts cells is longer than an isogenic wild type strain , suggesting that cdk1-ts may not be able to fully complement CDK1. In summary, we find that a timer specifying a constant budding duration describes how a cell population founded by ‘giant’ cells returns to their set-point volume.

Although mother and daughter sizes are correlated across a broad size range, daughters are always born smaller than mother cells. After cytokinesis, daughter cells remaining larger than the set-point volume exhibit a G1 phase with virtually no growth and bud rapidly, leading to a geometric shrinking in successive generations . Indeed, a back-of-the-envelope calculation demonstrates that if newly-budded daughters are each 50% smaller than their mothers, a 32-fold decrease in cell volume can be achieved in 5 generations . Assuming a 100 min doubling time , a return to the set-point size would take ~8 h. A fixed budding time, even in the absence of active molecular size sensors in S/G2/M, is sufficient to buffer against persistence of abnormally large cell sizes in the population. We also note that the bud duration timer we describe is quite complementary to G1-phase size sensors such as Whi5, which compensate for a small size at birth by elongating G1 phase.Our conclusions are derived from cells prepared using two independent perturbations: optogenetic inactivation of the Bem1 polarity factor and a temperature-sensitive cdk1 allele. Importantly,stackable planters each of these perturbations targets distinct cellular processes and thus produces distinct physiological defects. Cells lacking Bem1 activity exhibit weakened cells walls and undergo successive rounds of DNA endoreduplication following their initial arrest in G1 . In contrast, loss of Cdk1 does not produce such defects but its disruption requires incubating cells at 37˚C, which may broadly activate environmental stress response pathways. Furthermore, cdk1-ts may not fully complement CDK1, even at the permissive temperature . That each of these perturbations reveals similar mother-daughter size correlations as well as a size-invariant bud timer strongly supports the generality of our conclusions. The bud timer we describe here is consistent with prior work suggesting that the duration of budding tends to be invariant to changes in growth rates. However, such a timer need not be a dedicated biochemical circuit to sense budding duration, compare it to a set-point, and dictate the transition to cytokinesis. Its existence could simply arise due to the time required by independent cellular processes that coincide with bud growth, such as the combined duration of S-phase or mitosis. Nevertheless, one observation suggests more complex regulation: the duration of the size-invariant bud timer is markedly longer in enlarged cdk1-ts vs. optoBem1 cells , yet mother-daughter sizes are nearly identical in these two backgrounds . These data suggest that the duration of the bud timer may be inter-related to Cdk1 activity and cells’ growth rate during S/G2/M. Recent work has found that mitosis and bud growth rate are closely coordinated and that cells may extend the duration of mitosis to compensate for slow growth that occurs under poor nutrient conditions. Dissecting the dependencies between growth rate, Cdk1 activity and the duration of post-Start events presents a promising direction for future study.All yeast strains used are isogenic to an ‘optoBem1’ strain which was created in the w303 genetic background and contained exogenous PhyB-mCherry-Tom7 with endogenous Bem1 C-terminally tagged with mCherry-PIF, as previously described. The cdc28-13strain was a kind gift from David Morgan. A pACT1-CDC10-eGFP expression vector was created by Gibson assembly, with the CDC10 expression cassette inserted between the NotI and XmaI sites of the pRS316 vector.

For the experiments described in Figs 3 and 4; Fig D, E, F, and G in S1 Fig; and S2 Fig; the indicated vector was transformed into our optoBem1 or cdk1-ts strain and selection was maintained by growing yeast in synthetic complete media lacking uracil . For all other experiments, yeast were cultured in synthetic complete media .Preparation of yeast prior to optogenetic experiments was performed, in general, as previously described. Yeast undergoing exponential growth in synthetic media were treated with 31.25 μM phycocyanobilin and incubated in foil-wrapped tubes at 30˚C for a minimum of 2 h. For all microscopy experiments, yeast were spun onto glass-bottom 96-well plates coated with Concanavalin A and washed once with fresh PCB-containing media to remove floating cells. Cells remained approximately spherical following this procedure, as assessed by Concanavalin A staining .Imaging was performed atroom temperature. For experiments where isotropic growth was measured , yeast were plated and imaged immediately following PCB treatment. For experiments where growth following Bem1 reactivation was examined , PCB-treated yeast were first placed in clear culture tubes and incubated at room temperature for >6 h while undergoing constant illumination with a red LED panel . Cells were then plated and imaged. For experiments involving the cdk1-ts strain, cells were maintained in liquid cultures of synthetic complete media at 25˚C for at least 24 h and plated as described for the optoBem1 strain. Imaging was performed at 37˚C for experiments where isotropic growth during G1 was measured . For experiments where size control was assessed , cells were incubated at 37˚C for 8 hr, then shifted to 25˚C 30 min prior to imaging.For isotropic growth experiments, samples were imaged on a Nikon Eclipse Ti inverted microscope equipped with a motorized stage , a Lamba XL Broad Spectrum Light Source , a 60x 1.4 NA Plan Apo objective , and a Clara interline CCD camera . Samples were imaged by bright-field microscopy every 10 min for 12 h. Throughout experiments involving optoBem1 cells, a red LED panel was carefully balanced against the motorized stage and microscope body to provide oblique illumination to the cells and ensure that Bem1 remained deactivated. Generous amounts of lab tape were applied to the LED panel and scope to prevent slippage during image acquisition and stage movement. For the remaining experiments, samples were imaged on one of two spinning disk confocal microscopes, both of which were Nikon Eclipse Ti inverted microscopes with motorized stages . The first microscope was equipped with a Diskovery 50-μm pinhole spinning disk , a laser merge module with 405, 440, 488, 514, and 561-nm laser lines, a 60x 1.49 NA TIRF Apo objective , and a Zyla sCMOS camera . The second microscope was equipped with CSU-X1 spinning disk , a MLC400B monolithic laser combiner with 405, 488, 561, and 640-nm laser lines, a 60x 1.4 NA Plan Apo objective , and a Clara interline CCD camera.