Study Of StOSM And Function On Drought Tolerance In Potato

Drought stress has become a worldwide problem, harm to crops than the rest of adversity hazards, so the drought tolerance molecular mechanisms of plant genetic engineering has become the focus of research. Osmotin is an integral membrane protein of the plant.Studies have shown that OSM in plants response to low temperature and fungi infection, but it is little known whether its drought stress, especially the different isoforms of OSM is response to Water shortage and how to respond.In this study, from potato water stress subtracted cDNA library, access to the water shortage stress response genes OSM EST. By BLAST searches.its components 99% similarity with the membrane protein StOSM-3B, BLAST and bioinformatic analysis revealed that there are at least eleven StOSMs in the potato genome.. This study defined the evolutionary relationships of 11 StOSM encoding genes distribution, as well as conserved amino acid sequence regions on the chromosome 12.Qualitative determination of StOSM gene expression show eight StOSMs were upregulated and three were downregulated in response to water deficits. Quantitative expression determination showed among eleven StOSMs, eight StOSM mRNAs accumulated in leaf at the drought-lethal critical point (DLP) were at least 4-fold higher than in the control. The peak StOSM-8E mRNA accumulation was 49-fold higher than the control. This result of qRT-PCR quantification was identical with the FPKM values of all eleven StOSMs.Isolates were obtained from OSM-3b full-length cDNA by PCR, wherein ORF is 744bp, speculate its encoding 244 amino acid residues osmotin, with GenBank described-StOSM-3B deduced amino acid mutations exist 6,Using isolated OSM-3b cDNA constructed prokaryotic expression vector pET-28c-OSM,and to convert it into a host bacterium Escherichia coli BL21 (DE3), obtaining a recombinant strain BL21-OSM.PEG6000 concentration gradient in osmotic stress, recombinant strain BL21-OSM colony survival statistics show that at each concentration of the control strain BL21 colony survival was significantly lower than the recombinant strain BL21-OSM; peak survival under 2% PEG6000 recombinant bacteria is controlled 3 times; NaCl concentration gradient in osmotic stress, BL21-OSM colony survival was significantly higher than the control strain, under 1% NaCl recombinant bacteria survival is 2 times the peak control.Constructed StOSM inhibition and overexpression vector (pCAMBIA1305-OSM-, pCAMBIA1305-OSM). Expression in plants transformed StORM. Positive transformed lines StOSM mRNA expression was detected and phenotypes observed show water stress life of seedling of transformed StOSM inhibit lines was significantly shorter than the control. In water stress overexpressing transformed lines than in the control of small blades, but its growth was significantly higher than the control state of health.Vivo expression analysis showed that water-induced membrane protein-coding genes StOSM upregulated.Water stress response.enhanced StOSM expression can significantly enhance drought resistance.An full-length DNA of a high expression promoter KT was isolated, sequence alignment found isolated promoter sequences obtained from GenBank KT in Salt Solanum pennellii of HG975450 putative 91% similarity. Gene IAA5 placed downstream of the promoter KT and 35S, in vivo expression of transforming potato,screened positive for overexpression IAA5 transformed lines, phenotypic observation showed more significant than 35S KT Startup IAA5 control plant height and leaf lines. The results show that, KT startup gene expression stronger than 35S promoter.This study will provide the molecular breeding of new potato germplasm.