Functional Analysis Of GmWRKY16 Enhances Drought And Salt Tolerance In Arabidopsis

Soybean（Glycine max L.Merr）is an important oil crop and economic crop in the world.Drought,salt stress is the main abiotic stress that limits the growth and yield of soybean.In this study,the GmWRKY16 gene was cloned from soybean Huachun2.The gene was subjected to bioinformatics analysis,transcription verification,subcellular localization,tissue expression and other tests,and to its sodium chloride,sodium bicarbonate,The expression level of PEG-6000 under ABA stress was analyzed.The main findings are as follows:1 The GmWRKY16 gene has a CDS sequence of 1014 bp and encodes 337 amino acids with a WRKY domain.Phylogenetic tree construction analysis showed that Phylogenetic tree construction analysis showed that GmWRKY16,GmWRKY129,AtWRKY8,AtWRKY28,AtWRKY48 and AtWRKY71 were in the same branch and all belonged to WRKY IIc subgroup.The similarity of GmWRKY16 and GmWRKY129 protein was as high as 93%.The GmWRKY16 gene CDS sequence was inserted into the vector Pgbkt-7 and the yeast was transformed.The experimental results show that GmWRKY16 has transcriptional autoactivation activity.The GmWRKY16 gene CDS sequence was inserted into the vector PCAMBIA1302 and the recombinant plasmid was transformed into tobacco.The results showed that GmWRKY16 was expressed on the nucleus.2 Real-time RT-PCR was used to analyze the tissue expression pattern of GmWRKY16 gene.The results showed that the relative expression of GmWRKY16 gene was higher in old leaves,flowers,roots and seeds,and the expression level was highest in old leaves.The stress response and expression patterns of the GmWRKY16 gene were analyzed with NaCl（200 mM）,NaHCO₃（80 mM）,PEG（20%）and ABA（100μM）,and the gene expression was significant under various stresses.3 GmWRKY16 gene was heterologously transformed into Arabidopsis to obtain transgene positive seedlings.The phenotypes of Arabidopsis thaliana seedlings,seedlings and seedlings were analyzed.After WT treatment with 10uM ABA,the WT significantly increased the germination rate and root length ratio of GmWRKY16-overexpressing Arabidopsis lines.Under the same drought and high salt stress,Arabidopsis plants over-expressing GmWRKY16 were more resistant to drought than transgenic WT Arabidopsis plants.Compared with the GmWRKY16 transgenic line,the stomatal density is larger,and the GmWRKY16 transgenic line has a smaller pore size than WT,and the length and width are significantly different.The rate of water loss in the leaf was related to the regulation of air and air.The results showed that the slower water loss in GmWRKY16transgenic plants was due to the closure of stomata.4 GmWRKY16 induced Arabidopsis stress-related downstream gene expression.These related genes were not treated with drought stress in WT Arabidopsis,and there was no significant difference between the relative gene expression and the expression level under drought stress treatment.The relative expression level of GmWRKY16 transgenic Arabidopsis plants after drought stress treatment was significantly higher than that of non-stress treatment.This indicates that the expression of GmWRKY16 induced Arabidopsis stress-related downstream genes enhances the stress resistance of Arabidopsis thaliana.