南京林业大学学报（自然科学版）2019，Vol.43Issue(1)：39-44,6.DOI:10.3969/j.issn.1000-2006.201808007
灌木柳SlSIP基因的克隆和功能验证
Cloning and functional characterization of SlSIP from shrub Salix
摘要
Abstract
[Objective]The objectives of this study were to elucidate the structural characteristics and biological functions of the Sl SIP gene from shrub Salix and to provide a theoretical basis for the new function of alkaline alpha-galactosidase gene.[Method]An alkalineα-galactosidase gene encoding Sl SIP was cloned from the c DNA template of Salix jiangsuensis'2345'after salt stress for 48 h.A bioinformatics method was used to analyze the protein structure and relationships.The expression level and function of Sl SIP were conducted through agrobacterium-mediated transformation into Arabidopsis thaliana and real-time PCR.[Result]This gene encoded 776 amino acids with a molecular weight of 84.88ku, a theoretical isoelectric point of 5.85, an unstable coefficient of 35.74, and the average hydrophilic coefficient of-0.154.It was located on the endoplasmic reticulum.The conserved domains of Sl SIP were WFGWCTW and IDDGWQ with the catalytic active site V.In total, 11 positive T3 generation transgenic homozygotes were obtained.qRT-PCR showed that the expression level of Sl SIP increased by 9, 28, 29 times in 3 Arabidopsis transgenic lines.The seed germination rate of the transgenic lines was higher than that of the wild type on 85 mmol/L Na Cl medium, and the growth state was well, but the chlorophyll content of the wild-type plants was higher than that of the transgenic lines.[Conclusion]Sl SIP belongs to the GH27 gene family.This indicated that the Sl SIP gene not only increased the salt tolerance of seed germination, but also participated in leaf growth and senescence.This is the first study identifying the novel function of an alkalineα-galactosidase gene.关键词
苏柳'2345'/碱性α-半乳糖苷酶/耐盐/转基因/拟南芥/功能验证Key words
Salix jiangsusuensis &apos/2345&apos/alkaline α-galactosidase/salt tolerance/transgenic/Arabidopsis thaliana/functional annotation分类
生物科学引用本文复制引用
周洁,黄婧,邢玮,王保松,何旭东,教忠意..灌木柳SlSIP基因的克隆和功能验证[J].南京林业大学学报（自然科学版）,2019,43(1):39-44,6.基金项目
国家自然科学基金项目(31400572) （31400572）
江苏省自然科学基金项目(BK20141039) （BK20141039）
