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There are few published studies on the genetics of tolerance to chilling temperatures in tomato

Our results reveal that BrpHMA2 could be activated by Cd2+ , which is similar to the results found for HMA2 in Arabidopsis. Results suggest that BrpHMA2 is involved in the Cd response of plants. BrpHMA2 was also found to be expressed explicitly in the vascular tissues of roots, stems, leaves, flowers, siliques, and carpopodia, and its protein was localized in the plasma membrane . These results are consistent with previous findings for HMA2 in Arabidopsis, OsHMA2 in rice, and TaHMA2 in wheat. The protein plasma membrane localization and the vascular-specific expression pattern of the genes revealed that HMA2 might function as a membrane transporter in long-distance transport in plants. In recent years, some studies have investigated the function of HMA2. Most of these studies demonstrated that HMA2 is involved in Zn2+ and Cd2+ transmembrane transport and influences root-to-shoot Zn/Cd translocation. For example, HMA2 in Arabidopsis is thought to be involved in the outward transport of Zn2+ and Cd2+ from the cell cytoplasm, and HMA2 mutants are more sensitive to Cd stress and exhibit higher Zn or Cd accumulation than wild-type plants in the presence of high levels of Zn2+ or Cd2+ 14,15. The over expression of OsHMA2 in wheat, rice, and Arabidopsis improves root-to-shoot Zn/Cd translocation. In addition, the transformation of TaHMA2 in yeast enhances the resistance of cells to Zn/Cd. In rice, the suppression of OsHMA2 decreases the Zn and Cd concentrations in leaves, increases the retention of Zn in roots and reduces the translocation of Cd and Zn from roots to shoots compared with the results obtained with wild type plants. According to the literature, HMA2 is responsible for Zn2+/Cd2+ efflux from cells, plays roles in Zn and Cd loading to the xylem,procona buckets and participates in the root-to-shoot translocation of Zn/Cd. However, Yamaji et al. found that OsHMA2 is localized at the pericycle of the roots and in the phloem of enlarged and diffuse vascular bundles in the nodes. Their insertion lines of rice showed decreased concentrations of Zn and Cd in the upper nodes and reproductive organs.

The study revealed that the heterologous expression of OsHMA2 in yeast is associated with the influx transport of Zn and Cd. These researchers suggested that OsHMA2in the nodes plays an important role in the preferential distribution of Zn and Cd through the phloem to the developing tissues. Our results also revealed that, in the presence of Cd2+, transgenic Arabidopsis seedlings and yeast over expressing BrpHMA2 showed higher concentrations of Cd and enhanced Cd2+ sensitivity compared with the controls . Thus, we propose that BrpHMA2 functions in Cd2+ transport in the phloem tissue of vascular systems through influx into cells, and the efflux from phloem cells during long-distance transport may be performed by other transporters. The differential function of HMA2 from various plants might come from the tiny difference in amino acids in their function domains; this puzzle requires further investigation.In this study, we identified the NAC TF gene BrpNAC895, a homolog of Arabidopsis ANAC087 , which could be induced by Cd2+ stress . We confirmed that BrpNAC895 has a role in the response of B. parachinensis to Cd2+ stress by upregulating BrpHMA2 expression through direct binding to the BrpHMA2 promoter using EMSA, ChIP–qPCR, and the transient transformation method with B. parachinensis protoplasts . Previous studies have demonstrated that Arabidopsis ANAC087 is associated with plant programmed cell death . It functions along with the TF ANAC046 to show partial redundancy in coregulating the expression of some PCD genes in the root columella, including ZEN1, BFN1, and RNS3. Whether ANAC087 could participate in regulating Cd transporters in plants has not been reported. Our findings on BrpNAC895 show that this NAC TF has a novel role in upregulating BrpHMA2 expression in response to Cd2+ stress. We also identified the Cd-responsive AREB TF BrpABI 449 , which is a homolog of Arabidopsis ABF3 and can bind to the promoter of BrpHMA2 . ABF3 modulates the response to drought, salt, and other osmotic stresses as a master component in ABA signaling. This TF can also regulate the expression of multiple genes, such as the AGAMOUS like MADS-box TF family gene SOC1, which is a floralintegrator regulating flowering in response to drought, and the AREB TF ABI5, which is a core component in the ABA signaling pathway in the regulation of seed germination and early seedling growth during exposure to ABA and abiotic stresses .

In general, ABF3 can form protein complexes with other TFs. For example, ABF3 forms homodimers or heterodimers with AREB1/AREB2 and acts cooperatively to regulate ABRE dependent gene expression. ABF3 forms a complex with NF-YC3 to promote the expression of the SOC1 gene and thus accelerate flowering and drought-escape responses; ABF3 interacts with NAC072 to regulate RD29A and RD29B expression in response to ABA. Thus, complex formation might be the important functional mechanism by which ABF3 regulates gene transcription. Using EMSAs and ChIP–qPCR assays, we found that BrpABI449 could directly bind to regions of the BrpHMA2 promoter . The interaction of BrpABI449 and BrpNAC895 was further confirmed by pull-down and BiFC assays . The inhibition of BrpABI449 on the transcriptional regulatory role of BrpNAC895 was detected in the B. parachinensis protoplast transient system . The inhibition by BrpABI449 of the transcriptional regulatory role of BrpNAC895 complex, likely interferes with BrpNAC895’s activity in the transcriptional activation of BrpHMA2 in response to Cd stress. It has also been reported that Cd stress can induce a stress response via ABA signaling. Our results showing that BrpNAC895 and BrpABI449 are upregulated by Cd stress also support this point. The uptake or homeostatic regulation of heavy metals needs proper modulation to ensure plant health. Previous studies have shown that Cd stress induces the MYB TF gene MYB49 in Arabidopsis. This TF may further positively regulate the downstream TF gene bHLH38 and bHLH101 by directly binding to their promoters, and activate iron-regulated transporter 1 to enhance Cduptake. In contrast, Cd stress upregulates the expression of ABI5. ABI5 interacts with MYB49, prevents its binding to the promoters of downstream genes, and functions as a negative regulator to control Cd uptake and accumulation. Our present results also demonstrate a mechanism for controlling the expression of the heavy metal transporter gene BrpHMA2 under Cd stress. We propose that Cd2+ induces the expression of BrpNAC895 and BrpABI449, which might be mediated by ABA signaling. BrpNAC895 then promotes the transcription of BrpHMA2 by binding directly to its promoter . The activation of BrpHMA2 enhances Cd2+ uptake and may induce cell damage. Negative regulation of BrpHMA2 is then achieved by the upregulation of another AREB TF, BrpABI449, which interacts with BrpNAC895 and forms BrpNAC895-BrpABI449 protein complexes to inhibit the BrpHMA2 transcription activated by BrpNAC895 .

BrpABI449 could also bind to the promoter of BrpHMA2 directly to compete with BrpNAC895 in binding to the BrpHMA2 promoter. This negative regulation may play a supplementary role in the uptake and transport of Cd.Many plant species of Brassicaceae, including Arabidopsis, turnip, and oil seed rape, can be genetically modified, but the creation of transgenic B. parachinensis remains difficult. Therefore, we over expressed BrpHMA2 in Arabidopsis to investigate the function of BrpHMA2 and established a transient transformation system in B. parachinensis protoplasts to perform gene regulatory network analysis. Protoplasts have been widely used for sub-cellular protein localization and gene regulation analyses. In this study,procona florida container the transient transformation of B. parachinensis protoplasts was demonstrated to be a powerful system for ChIP–qPCR analysis. Previous studies have applied a similar approach to Populus trichocarpa and Brassica napus. Although the transient transformation system of B. parachinensis protoplasts was successfully used in this study of molecular mechanisms, the system cannot be easily used for phenotype and physiological analyses. The lack of BrpNAC895 and BrpABI449 transgenic B. parachinensis is a problem that severely limits research on this plant. New techniques, such as the transient reprogramming of plant traits via the transfection of RNA based viral vectors using Agrobacterium and gene editing combined with fast-treated Agrobacterium coculture, may be useful approaches for comprehending gene function concerning physiology and for the further application of modifications of gene function to effectively control the accumulation of Cd in B. parachinensis.Abiotic stresses, especially those which affect the water relations of the plant such as low temperatures, may decrease plant growth and yield. The majority of plants will suffer damage when exposed to freezing temperatures , but plants of tropical or sub-tropical origin also suffer damage when exposed to chilling temperatures . Exposure of roots to chilling temperatures decreases root hydraulic conductance , and can result in water stress and chilling injury within a few hours of exposure . The susceptibility to water stress induced by root chilling in species of tropical and sub-tropical origin is a concern for agricultural production in Mediterranean climates such as California, where exposure to cold soils in the spring can affect seedling establishment because soil temperatures under an open canopy may be colder than air temperatures . Cultivated tomato is a classic example of a chilling-sensitive crop . It was domesticated from the wild cherry tomato, which is native to mesic, tropical environments . A related wild tomato species, S. habrochaites, grows in the Peruvian Andes at altitudes up to 3300 m and thrives in xeric habitats and at chilling temperatures detrimental to S. lycopersicum . Upon exposure to root chilling conditions, the root hydraulic conductance of both tomato species decreases, but S. habrochaites closes its stomata rapidly in response to chilling stress, thereby maintaining water potential and shoot turgor, whereas the stomata of S. lycopersicum remains open and the shoots wilt . Other agronomically important crops of tropical or sub-tropical origin such as maize and rice respond to root chilling in a manner consistent with that of cultivated tomato . An improved understanding of the underlying mechanisms of root chilling tolerance in wild S. habrochaites would contribute to a better general understanding of chilling sensitivity in crops of tropical and sub-tropical origins.A review by Venema et al. focused on physiological effects of chilling and noted that wild tomato species were promising sources of genetic tolerance to chilling.

Oyanedel et al. evaluated a back cross inbred line population derived from S. habrochaites acc. LA1777 for growth traits under chilling temperatures and reported QTL for higher biomass accumulation on chromosomes 2, 3, and 9. Elizondo and Oyanedel evaluated tomato introgression lines containing S. habrochaites acc. LA1777 introgressions on chromosomes 2 and 3 in the field under low temperatures . The ILs had higher growth rates but lower fruit set than the parental lines in response to an increase in the number of hours of chilling temperatures. To investigate the genetic basis of shoot turgor maintenance under root chilling, Truco et al. used an interspecific BC1 population derived from chilling-susceptible S. lycopersicum cv. T5 and chilling-tolerant wild S. habrochaites acc. LA1778 to map QTL for this trait. Three QTL for shoot turgor maintenance under root chilling were identified on chromosomes 5, 6, and 9. The largest effect QTL located on chromosome 9 accounted for 33 % of the trait phenotypic variance . We designated this QTL stm9 for shoot turgor maintenance, chromosome 9. Subsequently, QTL stm9 was fine-mapped to a 2.7-cM region on the short arm of chromosome 9 between markers T1670 and T1673 . Easlon et al. determined that tomato ILs containing the short arm of chromosome 9 from chilling-tolerant S. lycopersicoides and S. habrochaites maintained shoot turgor under root chilling. Here we high-resolution mapped QTL stm9 using recombinant sub-near-isogenic lines and compared high resolution mapped QTL stm9 to the S. lycopersicum reference genome for initial identification of potential candidate genes and regulatory sequences . Our longer term goal is to identify and functionally test candidate genes and regulatory sequences from S. habrochaites and determine the causal gene or polymorphisms for QTL stm9.A population of near-isogenic lines containing the chromosome 9 region from S. habrochaites acc. LA1778 in an otherwise completely S. lycopersicum cv. T5 background was marker-selected and used for fine-mapping, as described in Goodstal et al. . For high-resolution mapping of stm9, we created and marker selected recombinant sub-near-isogenic lines as follows.

An understanding of the depth to the groundwater table is also needed

As is the case with any model, and with soil survey information in particular, ground-truthing at the field scale is necessary to verify results. We acknowledge limitations to our model. It does not consider proximity to a surface water source, which is an issue especially in areas that are irrigated solely from groundwater wells and are not connected to conveyance systems that supply surface water. The SAGBI also does not consider characteristics of the vadose zone or depth to groundwater. In arid regions, deep vadose zones may contain contaminants such as salts or agricultural pollutants that have accumulated over years of irrigation and incomplete leaching. These deep accumulations of contaminants could be flushed into the water table when excess water is applied during groundwater banking events. Furthermore, deep sediment likely contains hydraulically restrictive horizons that have not been documented, creating uncertainty as to where the water travels.Given these issues, SAGBI may be most useful when used in concert with water infrastructure models and hydrogeologic models — which generally do not incorporate soil survey information in a comprehensive way — to develop a fuller assessment of the processes and limitations involved in a potential groundwater banking effort.Selenium received recognition as an environmental contaminant in the 1980s,procona system as a result of the unprecedented events at the Kesterson Reservoir in California , a national wildlife refuge at the time . Large amounts of this trace element had been mobilized through irrigation of selenium-rich soils in the western San Joaquin Valley, transported along with agricultural runoff, and accumulated at the Reservoir.

Toxic selenium concentrations brought about death and deformities for as much as 64% of the wild aquatic birds hatched at the reservoir, including both local migratory species. Within a few years, the habitat of a variety of fish and waterfowl was classified as a toxic waste site . Today, the Reservoir’s ponds are drained and covered beneath a layer of soil fill , yet the mechanisms of selenium release now known as “the Kesterson effect” are still a threat in California and around the world . The environmental and management conditions creating irrigation-induced selenium contamination have been characterized in Theresa Presser’s seminal work . In brief, problems arise when seleniferous soils, such as those formed from Cretaceous marine sedimentary deposits along the Western side of the San Joaquin basin are subjected to irrigated agriculture. Salts, including selenium, naturally present in such soils are mobilized through irrigation, and high evaporation rates concentrate them in the root zone. In order to avoid negative effects on plant growth, subsurface drainage systems are used to export excess salts from the soil. This is particularly necessary in places where deep percolation is inhibited by a shallow impermeable layer. Such subsurface runoff routinely contains selenium in concentrations that exceed the US Environmental Protection Agency designation of toxic waste and thus poses an acute threat to aquatic ecosystems that receive it . The irrigation runoff feeding into the evaporation ponds of the Kesterson reservoir averaged 300 µg Se/L . The discovery of widespread deformities among waterfowl hatched near these ponds in 1983 led to a shift in the perception of selenium. While research had thus far been focused on farm-scale problems related to crop accumulation and toxicity to livestock, it became clear that excessive selenium concentrations in agricultural runoff was a watershed-scale resource protection issue that would greatly complicate irrigation management throughout the Western United States . As a result, California has been a hot spot for global research and management of environmental selenium contamination .

As selenium load management in the San Joaquin basin has made significant progress, new major sites of concern, such as the San Francisco Bay-Delta and the Salton Sea , have emerged in California. Current regulatory standards for selenium as aquatic contaminant are insufficient to be protective of sensitive ecosystems because they do not account for amplified exposure through bio-accumulation . There are many other pathways of anthropogenic selenium contamination – the San Francisco Bay-Delta for example receives half of its input from refineries . However, the diffuse agricultural sources are particularly hard to control , are the principal source of selenium in western US surface waters , and have shaped California’s history like no other selenium source. This paper analyzes what can be learned from the last three decades of seleniferous drainage management and regulatory approaches developed in California. In particular I seek to answer two key questions: 1) What were the greatest achievements and shortfalls of seleniferous drainage management in California? 2) To what extent may the current development of site-specific selenium water quality criteria for the San Francisco Bay and Delta serve as a model for future regulation?Selenium is a naturally occurring trace element heterogeneously distributed across terrestrial and marine environments . On land, seleniferous soils and those marked by selenium deficiency sometimes occur as close as 20 km from one another . Selenium contamination of natural ecosystems is linked to an array of human activities including irrigated agriculture, mining and smelting of metal ores, as well as refining and combusting of fossil fuels. The bio-spheric enrichment factor, which is computed as the ratio of anthropogenic to estimated “natural” emissions of a substance, was found to be 17 for selenium , highlighting the dominance of the anthropogenic component in the modern selenium cycle . Anthropogenic fluxes are expected to keep increasing in the foreseeable future as energy and resource demands increase . Selenium bio-accumulates, with tissue concentrations in animals and plants typically 1-3 orders of magnitude above those found in water.

Consequently, the predominant selenium uptake pathway for animals is through the consumption of food rather than water. Bio-accumulation and bio magnification are particularly intense in aquatic ecosystems and selenium contamination of such habitats is a global concern . In the Western United States alone, nearly 400,000 km2 of land are susceptible to irrigation induced contamination by the same mechanisms that led to the demise of the Kesterson Reservoir . Other nations where irrigation induced selenium contamination has been observed include Canada, Egypt, Israel, and Mexico . The environmental impacts of selenium depend on the element’s chemical speciation. The element’s primary dissolved forms, selenate and selenite , are mobile and bio-available . They can be sequestered in soils or sediments upon microbial reduction to solid elemental Se, metal selenides, or volatilized to the atmosphere upon reduction to gaseous methylated Se. Both selenate and selenite are toxic at elevated concentration,procona valencia buckets selenite however was found to be more toxic than selenate in direct exposure studies involving invertebrates and fish and also to bio-accumulate more readily at the base of aquatic food chains . Additionally, once any dissolved form of selenium is assimilated by an organism it is converted into highly bio-available organo-selenide species, Se . Exposure studies comparing organo-selenides to selenite in the diets of water birds established lower toxicity thresholds for the former . Organo-selenides are released from decaying organisms and organic matter during decomposition and can then persist in solution or be oxidized to selenite, while the conversion back to selenate does not occur at relevant rates in aquatic environments . Thus, recycling of selenium at the base of aquatic food webs through assimilation and decomposition usually leads to a buildup of the more bio-available and toxic forms over time . This buildup of bio-available selenium species may also explain why tissue concentrations in the upper trophic levels of stagnant or low-flowing ecosystems typically exceed those of fast flowing ecosystems with comparable selenium inputs, but shorter residence times . The complex environmental cycling of selenium has been a major obstacle in creating water quality regulations for this element . Regulatory concentration guidelines vary widely between jurisdictions and there are significant opportunities for new regulatory approaches . The Californian office of the EPA is currently working on site-specific water quality criteria for the protection of wildlife in the San Francisco Bay and Delta . These criteria are to be based on a modeling approach developed by USGS scientists, capable of translating tissue limits to dissolved concentration limits . There is hope among aquatic toxicologists that California’s new site-specific approach may become a model for national standards . For all contaminants regulated since 1985, aquatic life criteria under the Clean Water Act have been defined through separate dissolved concentration limits for longer term “continuous” and short term “maximum” limits . The selenium criteria that were established in 1987 defined continuous concentration limits of 5 µg/L as acid-soluble selenium with maximum concentrations not exceeding 20 µg/L more than once every three years for freshwater environments, but allowed up to 71 µg/L with up to one three-year exceedance of 300 µg/L for saltwater environments.These selenium limits became legally binding for 14 states including California after promulgation with the 1992 Water Quality Standards.

A central problem with the current criteria is that they were predominantly based on data drawn from direct exposure laboratory studies and thus failed to take into account the more ecologically relevant toxic effects due to bio-accumulation and trophic transfer. The freshwater criteria were based on field data from a contamination event , while the saltwater criteria were purely based on laboratory studies which did not account for bio-accumulation. The resulting difference of more than one order of magnitude between fresh- and saltwater criteria is not supported by field data . In fact, the saltwater criteria have widely been regarded as under protective of wildlife, including waterfowl . In addition, the freshwater criteria appear under productive of particularly sensitive ecosystems and species . To be protective of waterfowl in the wetlands of the Central Valley Region, a 2 µg Se/L monthly mean water quality criterion was deemed necessary by the Regional Water Quality Control Board and this objective was officially approved for the region by the EPA in 1990 . For the wetlands of the Central Valley Region, this criterion overrides the statewide criteria promulgated in 1992 and remains in effect today. However, given the wide range of bio-availability between different selenium species and the complex transfer processes between environmental compartments and trophic levels, regulation based solely on dissolved or acid-soluble concentrations has been characterized as inadequate . In response to such criticism the EPA proposed in 2004 a new tissue-based criterion for selenium with a 7.91 µg/g fish tissue limit to supersede the previous national water quality guidelines for selenium. This limit is based on the lowest level of effect in juvenile bluegill sunfish under simulated overwintering conditions . Whereas there is little doubt that tissue concentrations are more representative of exposure than dissolved concentrations for individual species, it is unclear if a single fish tissue limit will be protective across entire food webs including a diversity of fish and waterfowl . The proposed tissue based criteria have to date remained at draft stage due to objection by the US Fish and Wildlife Service. The historic developments that lead to the rise of selenium contamination in the San Joaquin Valley can be traced to the passage of the California Water Resources Development Act of 1960. The Act laid the financial foundation for the State Water Plan providing for the construction of the nation’s largest water distribution system and including also infrastructure measures for “the removal of drainage water” . The State Water Projects funded under this plan began delivering water to 4,000 km2 in the Southern San Joaquin Valley as of 1968 . To prevent salinization and manage agricultural runoff, the Bureau of Reclamation constructed collector drains, a main drainage canal , and a regulating reservoir, Kesterson . Originally, the San Luis Drain was planned to deliver drainage out of the San Joaquin Valley all the way to the San Francisco Bay Delta, however the northern part of the drain was never completed . Instead, from the time of the San Luis Drain’s completion in 1975 until its temporary closure in 1986, all runoff water channeled through the drain was delivered to the evaporation ponds of the Kesterson Reservoir, which had become part of a newly created national wildlife refuge in 1970 . There, in the early 1980s, high rates of embryo deformity and mortality, as well as large numbers of adult deaths among waterfowl were identified as caused by the elevated selenium concentrations in the evaporation ponds . This led to the closure of the Reservoir to all runoff inputs in 1986 .