Cloning And Functional Preliminary Analysis Of Transcription Factor PwNAC11/PwNAC38 In Picea Wilsonii

Picea wilsonii,as an evergreen tree of Pinaceae spruce,has become an important tree spec ies for landscaping in recent years.With the changes in global environmental conditions and th e destruction of human factors,its distribution has been decreasing year by year.It has strong adaptability to the environment,is resistant to shade and cold,and has abundant anti-reverse ge ne resources.It is the dominant plant for abiotic stress research.NAC transcription factors are widely involved in the growth and development of plants,and play an important role in the re sponse of plants to various abiotic stresses such as drought and salt.However,there are few re ports on the study of NAC transcription factors in woody plants.Based on the laboratory-meas ured prion transcript data,this study cloned the coding region sequences of PwNAC11 and PwN AC38 genes from barley.The anti-reverse function of PwNAC11 and PwNAC38 was studied by t ransgenic,yeast one-hybrid and subcellular localization techniques,which provided a basis for i mproving the anti-reverse mechanism of Picea wilsonii The specific results are as follows:1.Bioinformatics analysis showed that both PwNAC11 and PwNAC38 proteins have typical NAM domains.Both proteins have threonine,serine and tyrosine phosphorylation sites,and the re are no signal peptides and transmembrane domains.Both are hydrophilic proteins.2.The results of transcriptional activation activity showed that the full-length and C-termin us of PwNAC11 protein have transcriptional activation activity,while the N-terminus has no tra nscriptional activation activity;the full length of PwNAC38 protein has transcriptional activation activity,while N-terminus and C-terminus none of them have transcriptional activation activity.Subcellular localization results showed that both PwNAC11 and PwNAC38 were expressed in t he nucleus.3.The overexpression vector of PwNAC11/PwNAC38 was successfully constructed,and Pw NAC11-T3 seeds and PwNAC38-T2 seeds of stable genetic expression were transformed into Ara bidopsis thaliana.At the DNA level,four PwNAC11 overexpressing plants and seven PwNAC38 overexpressing plants were obtained..Real time-quantitative PCR was used to detect the expres sion of PwNAC11 gene in overexpressing plants.The results showed that high expression of P wNAC11 was detected in overexpressing lines.4.A homozygous mutant plant of the homologous gene mutant atlg69490 of PwNAC38 in Arabidopsis thaliana was obtained by identification.The results of real-time PCR showed that the expression level of the gene in the atlg69490 homozygous mutant was down-regulated by 65.0 times compared with the wild type.5.Successful transformation of wild-type "Atlantic" potato,at the DNA level,identification of PwNAC11 overexpressing potato 3 strains,PwNAC38 overexpressing potato 5 strains.The r esults of phenotypic experiments showed that potato seedlings transformed from PwNAC11 and PwNAC38 were better than wild type potatoes under 10%PEG6000 conditions.6.Phenotypic experiments were performed using the PwNAC11-OE1 strain.The root length test of the seedlings treated with 0,100,200 and 300 mM mannitol showed that the root len gth of PwNAC11-OE1 was significantly longer than that of the wild type at 100 mM mannitol treatment,while the difference in other concentrations was not significantly different.