Bioinformatics Analysis Of C2H2-ZFP Of Larix Kaempferi And Function Analysis Of LkZFP36 Gene

Larix kaempferi is an important economic species,which occupies a dominant dominant position in short-cycle industrial timber forests.C2H2-ZFP gene family can regulate and participate in a variety of signal transduction pathways,which is essential for plant growth and stress response.The study of this gene family can provide an important reference for future breeding and improvement of larch.This study focuses on the subject of the C2H2-ZFP genes of L.kaempferi(LkZFPs)bioinformatics and expression pattern analysis.Using Agrobacterium-mediated method,LkZFP36 gene was transformed into hybrid larch callus to obtain transgenic callus,and its drought resistance was preliminarily analyzed.(1)Through bioinformatics analysis,C2H2-ZFP genes of L.kaempferi were identified to47 members and named LkZFP1-47.Based on phylogenetic analysis and conserved domain analysis,these genes were divided into four subfamilies.It was found that 8 LkZFPs were Qtype C2H2-ZFP,and 6 LkZFPs had EAR motif,that is,transcriptional inhibition domains.The results of promoter cis-element indicated that most LkZFP genes may respond to drought,ABA,Me JA,SA stresses.The GO annotations predict that LkZFPs are involved in a variety of biological processes,such as metabolic processes and biological regulation.(2)The results of real-time quantitative PCR(RT-q PCR)showed that LkZFP genes were significantly up-regulated under salt stress,drought stress and ABA treatment most of the time.LkZFP genes significantly upregulated at least at one time point when treated with Me JA and SA.The sensitivity of LkZFP genes under salt stress,drought stress and ABA treatment was relatively higher than that of Me JA and SA treatment.In particular,LkZFP36 gene was significantly up-regulated in roots under drought stress,which may play an important role in drought resistance mechanism of larix.(3)Overexpression LkZFP36 gene in embryonic callus of hybrid larch(Larix kaempferi 5×L.gmelinii 9)by Agrobacteriin-mediated method resulted in 10 resistant callus.DNA level detection and RT-q PCR detection were both positive,confirming that the transgenic callus was obtained.(4)Three overexpression transgenic and wild-type embryonic callus were subjected to 7%PEG6000 stress,and the unstressed callus were the control.Through phenotype observation,it was found that the wild-type callus was more damaged than the transgenic callus.By measuring relative water content,proline and soluble sugar content,SOD and POD activity,it was found that wild-type callus was more damaged than transgenic lines under drought stress.These results indicated that the overexpression of LkZFP36 gene in hybrid larch callus could significantly enhance drought resistance.