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《Acta Botanica Boreali-Occidentalia Sinica》 2017-07
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Cloning and Functional Analysis of NtWRKYY2 Gene from Narcissus tazetta var. ‘Yunxiang'

WEN Xiuping;SUN Shenshen;YANG Feiying;CHEN Xiaojing;College of Horticulture,Fujian Agriculture and Forestry University;Institute of Genetics and Breeding in Horticultural Plants,FAFU;  
The WRKY transcription factors family gene NtWRKYY2(GenBank:KX056496)were cloned by PCR technology from Narcissus tazetta var.‘Yunxiang'.The Open Reading Frame(ORF)of NtWRKYY2 contained 867bp,encoding aprotein of 289 amino acid residues.Multiple alignments and phylogenetic analysis showed that NtWRKYY2 proteins containing one WRKY consecutive structural domain and C2H2 type zinc finger(Csx4Cx23HxH)belong toⅡd sub-group of WRKY transcription factor together.Quantitative real-time PCR(qRT-PCR)analysis demonstrated that the NtWRKYY2 gene was obviously induced by salt stress in‘Yunxiang'.We constructed overexpressing vector pMDC140-NtWRKYY2 using In-Fusion cloning technique,and transformed into tobacco by the method of Agrobacteriumthough leaf disc transformation.The carrier of PCR results indicated that the resistant plantlets were positive and the converting rate of 72%.Salt stress and chlorophyll fluorescence parameter analysis showed that after the salt stress treatment,overexpression of NtWRKYY2 transgenic tobacco showed less wilt and chlorosis than the wild type plants,and the decrease of Fv/Fmvalue was less than the wild type plants.Studies haveshown that overexpression of the NtWRKYY2 gene remarkably increased the salt tolerance in transgenic tobacco.This study offers an alternative gene for salt tolerant transgenic breeding of Narcissus.
【Key Words】: ‘Yunxiang' narcissus WRKY NtWRKYY2 gene expression salt stress
【Fund】: 福建省种业创新与产业化工程项目(K8114001B)
【CateGory Index】: Q943.2;S682.21
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