Annotation, Subfunctionalization Of Cation/Oroton Antiporter 1(CPA1) Gene Families In Grapevine (Vitis Vinifera L.), And A Meta-analysis To Cpa1s Overexpression

Salt stress,as one of the common ecological disasters,limited the production of agriculture and threatened the environments of human beings.Salt stress not only impacts the plant growth,but destroys the soil source.Thus,making good use of salt tolerance/resistance varieties is a suitable method to solve this problem radically.Cation/proton antiporter 1?CPA1?is a key part of transport system.CPA 1 genes function in regulating ions and pH homeostasis in organisms,enhancing salt resistance/tolerance of plants through the vacuolar compartmentalization of Na+,Na+ efflux from the cell,and affecting K+ concentrations.Grapevine?Vitis spp.?is one of the most popular commercial fruit crops in the world.Grapevines are adapted to semiarid environments,where salinity is a common problem and became progressively worse.Salinity is an important factor which affetced the grapevine growth and limited the development of grapevine industry.Hence,it's very critical to study grapevine salt tolerance,however,fewer salt tolerance mechanism of grapevine on genes have been conducted.Recently,in the process of gene function studies,transgenic technology always be as a good way to determine the actual-function of genes.Transgenic breeding work has been carried out in some species.However,it's difficult to determine the function and effects of genes,due to many moderators during the process of transgenic experiment.Therefore,understand the interference effects of moderators clearly being a key step to understand the gene functions.In this research,the comprehensive bioinformatics and annotation analysis were applied,and the previous studies on the characterization of the CPA proteins were re-evaluated.We resolved the discordance of CPA family genes in grapevine,and revealed theduplications contributing to the expansion of CPA family genes in grapevine.Totally,29 CPA genes were identified in the grapevine reference genome.Furthermore,we investigated the structures of gene among the CPA1 subfamily genes in six species.We identified motifs between grapevine and Arabidopsis,and found some motifs were subgroup-specific motifs.The results would be helpful to decide the function of the CPA1 family on the physiological responses to salinity stress and breed the salt-tolerant grapevine cultivars.TheCPAl family genes of grapevine?Vitis vinifera?in response to short-term osmotic and NaC1 stresses were studied.We identified two genes that had sub-functionalized to be differentially expressed in response to NaC1 stress in the early stage in different organs.Specifically,VIT₁9s0090g01480 couldcontrol the Na+ compartmentation in grapevine roots and VIT₀5s0020g01960 could influence the Na+ transfer in stems.Based on the dynamics of ion concentration,the rate of electrolyte leakage,CPA1 gene expression in roots,stem and leaf tissues under osmotic and NaC1 stresses,we postulated a model that how grapevine responds physiologically and molecularly to the osmotic and ion toxicity under NaC1 stress in a short term.This work laid a foundation for future research on the CPA1 gene family regarding their evolutionary history and biological functions for modulating salt responses.Diversified roles of the multi-genic CPA1 family and the various parameters used in evaluating the transgenic plants making it challenging to assess the complex functions of CPA1 genes and their physiological mechanisms on salt tolerance.Using meta-analysis,we conducted a quantitative study of the literature on over-expression of CPA1 genes in model and crop plants.We determined how these genes have influenced several plant characteristics involved in response and resilience to NaC1 stress and evaluated which experimental conditions tended to favor or reduce CPA1s gene effects on transgenic plants.Viewed across all studies,over-expression of CPA1 genes have increased the size or amount of 14 of the 19 examined plant characteristics by 24%or more.Root K+/Na+ ratios were increased by 67%and 131%,respectively.Seed germination and chlorophyll concentrations were also increased markedly in transgenic plants,by 147%and 100%,respectively.Among 11 moderating variables tested,several had substantial impacts on the extent of CPA1s influence:tissue culture media?solid vs.liquid?,donor and recipient type?monocotyledonous vs dicotyledonous?,donor and recipient genus,and gene member.SOS1 and SOS1+ stimulated root and shoot K+ and Na+ dynamics much less than NHX1 and NHX1+.Longer exposure to the NaCl stress led to larger CPA1s-induced increases in most characteristics,with only shoot Na+,root length and seed germination showing a negative response to the period of exposure.These findings should assist in future investigations of the function of CPA1 gene family during plant salt tolerance and the use of genetic engineering for salt breeding.