Cloning Of Three NAC Transcription Factors From Lilium Pumilum And Genetic Transformation To Tobacco

L.pumilum,which originates in the cold areas of Northern China,is striking due to its graceful flowers and bright red color,and it has a ornamental value.L.Pumilum was tolerant to abiotic stresses,such as cold,drought,salt and alkalinity,and has an edible and medicinal value.In this study,21 NAC genes were screened from the transcriptome of L.pumilum,we analyzed the bioinformatics,then their domains,protein structure prediction and predicted the function of LpNAC protein.Analyzed the 21 NAC genes expression in various stress.The results are as follows:1.There were 21 NAC genes filtered from the transcriptome sequencing of L.pumilum cryogenic dormancy and named LpAC1-LpNAC21.Bioinformatic analysis of 21 NAC genes showed that most NAC genes contained the complete NAM domain.The amino acid number of NAC proteins were between 51-890 and the molecular weight were 5966.5-100080.4 Da.The protein structure of the LpNAC protein,which was mainly composed of an irregular curl,?-helix,?-angle,was scattered throughout the protein.The transmembrane domain prediction shows that LpNAC19 and LpNAC20 have a single transmembrane domain.All LpNAC proteins were divided into seven different groups(named NACa to NACg)along with Arabidopsis NAC proteins.The PlantTFDB further predicted the function of LpNAC genes showed that NAC genes had the function of transcriptional regulation,as well as being involved in flower development,fruit maturation and abscisic acid signal transduction in response to abiotic stresses.2.The expression patterns of 21 NAC transcription factors were analyzed in roots,bulbs and leaves of L.pumilum under salt,PEG,low temperature(2 ?)and abiotic stress conditions of ABA.Under salt stress conditions LpNAC5,LpNAC10 and LpNAC20 were dominant expression genes;LpNAC8,LpNAC10,LpNAC17 and LpNAC21 were the dominant expression genes under low temperature stress conditions;LpNAC5,LpNAC13,LpNAC1 7 and LpNAC20 were dominantly expressed genes under drought stress conditions;LpNAC5,LpNAC14 and LpNACl 7 were dominant expression genes under ABA stress conditions.3.Three NAC genes with complete open reading frames were cloned by PCR.The LpNAC5,LpNAC13 and LpNAC21 were respectively ligated to the pBI121-GFP vector,and the expression vector was transformed into tobacco by the leaf disc method.The resistant plants were obtained through the screening of kanamycin and PCR identification,and finally there were 12 tobacco plants transformed with LpNAC5 gene,10 tobacco plants transformed with LpNAC13 gene,and 10 tobacco plants transformed with LpNAC21 gene.