三明野生蕉β-1,3-葡聚糖酶Mugsp7基因克隆及其在低温处理下的表达分析  

Cloning and Characterization of Mugsp7,α β-1,3-glucanasc Gene of Wild Banana Germplasm from Sanming,Fujian,China,Under Cold Treatment

作　　者：徐小萍[1] 谢燕萍[1] 陈芳兰[1] 陈晓慧[1] 陈裕坤[1] 张梓浩[1] 程春振[1] 林玉玲[1] 赖钟雄[1] XU Xiaoping;XIE Yanping;CHEN Fanglan;CHEN Xiaohui;CHEN Yukun;ZHANG Zihao;CHEN Chunzhen;LIN Yuling;LAI Zhongxiong(Institute of Horticultural Biotcchnology/Institutc of Subtropical Fruits,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China)

机构地区：[1]福建农林大学园艺植物生物工程研究所/亚热带果树研究所,福建福州350002

出　　处：《热带作物学报》2020年第2期292-299,共8页Chinese Journal of Tropical Crops

基　　金：国家重点研发计划(No.2019YFD1000901);国家现代农业产业技术体系专项资金(No.CARS-31-15);国家青年基金(No.31601713)。

摘　　要：以三明野生蕉组培苗为材料,通过RT-PCR技术克隆获得β-1,3-葡聚糖酶基因Mugsp7(β-1,3-glucanase)的cDNA和gDNA序列,并对该基因进行生物信息学分析和不同低温下的实时荧光定量PCR表达分析,并进一步测定8℃处理1、2、3、4和5d后三明野生蕉叶片β-1,3-葡聚糖酶活性。结果表明:MugsP7的gDNA长1132 bp,开放阅读框(ORF)长984 bp,具有一个148 bp的内含子,共编码327个氨基酸。cDNA、gDNA的登录号分别为KU363808和KU363809。生物信息学分析表明,Mugsp7属于酸性、亲水、稳定性蛋白,不具有信号肽,与小果野蕉、大叶藻、玉米、大麦、水稻等位于同一分支,与小果野蕉亲缘关系较近分为一类。实时荧光定量PCR表明,不同低温处理和8℃低温不同时间处理下,Mugsp7呈现不同表达模式,且三明野生蕉叶片β-1,3-葡聚糖酶活性的测定分析也进一步表明,Mugsp7能进一步响应低温胁迫。因此,推测在三明野生蕉抗寒响应中发挥重要作用。The tissue culture seedlings of a wild banana germplasm from Sanming were used to clone the cDNA and gDNA sequences of β-1,3-glucanase gene Mugsp7 by RT-PCR.Bioinformatics and qRT-PCR analysis of the expression at differ­ent low temperature were also carried out,and the enzyme activity of mpcraturcurccnt lowthc leaves was further deter­mined by treating at 8℃ for 1,2,3,4 and 5 days.The gDNA sequence of Mugsp7 was 1132 bp with an open reading frame(ORF)of 984 bp and one intron of 148 bp,which encoding 327 amino acids.Bioinformatics analysis showed that Mugsp7 was an acidic hydrophobic stable protein,and there was no signal peptide.The amino acid sequence of Mugsp7 shared significant similarity with that in Musa acuminata,Zostera marina.Maize,Hordeum vulgare L,Oryza saliva on the same branch,and it was closely related to Musa acuminata.The expression pattern of Mugsp7 under different low tem­perature treatments was different,suggesting that Mugsp7 was a low temperature stress related gene.

关 键 词：三明野生蕉 Β-1 3-葡聚糖酶 Mugsp7 基因克隆 低温胁迫 qPCR 

分 类 号：Q785[生物学—分子生物学] Q789