木薯SCARECROW-LIKE(SCL)基因克隆、生物信息学及非生物胁迫下表达分析  

Cloning,bioinformatics analysis and expression under abiotic stress of SCARECROW-LIKE(SCL)gene in cassava

作　　者：樊铸硼 罗兴录[1,2] 单忠英 黄堂伟[1] 程隆祥 吴美艳 FAN Zhu-peng;LUO Xing-lu;SHAN Zhong-ying;HUANG Tang-wei;CHENG Long-xiang;WU Mei-yan(College of Agriculture,Guangxi University,Nanning 530004,China;State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,Nanning 530004,China)

机构地区：[1]广西大学农学院,南宁530004 [2]亚热带农业生物资源保护与利用国家重点实验室,南宁530004

出　　处：《南方农业学报》2020年第8期1888-1895,共8页Journal of Southern Agriculture

基　　金：广西科技计划项目(桂科AB18221127);亚热带农业生物资源保护与利用国家重点实验室开放课题(SKLCUSAb201609,SKLCUSA-b201704);亚热带农业生物资源保护与利用国家重点实验室自主研究课题(SKLCuSA-a201802)。

摘　　要：【目的】克隆木薯SCARECROW-LIKE(MeSCL)基因,对其进行生物信息学分析,并检测其在非生物胁迫下的表达情况,为深入研究木薯MeSCL基因响应非生物胁迫的调控机制提供理论参考。【方法】以木薯品种D346为材料,采用RT-PCR克隆MeSCL基因编码区(CDS)序列,并利用生物信息学分析软件进行序列特征分析,采用实时荧光定量PCR检测其在干旱、盐、氧化和低温胁迫下木薯叶片中的表达情况。【结果】克隆获得的MeSCL基因编码区(CDS)序列全长1655 bp,与参考序列(GenBank登录号LOC110627921)仅存在2个碱基的差异,开放阅读框(ORF)的长度为1560 bp,编码519个氨基酸,编码蛋白分子量为57.84 kD,等电点(pI)为6.08,脂肪系数为80.46%,总平均亲水性指数为-0.195,为亲水性蛋白,含有1个信号肽、6个从内部到外部的跨膜螺旋区和5个从外部到内部的跨膜螺旋区,定位于细胞核和内质网中,属于GRAS蛋白家族成员,具有该家族的保守结构域。MeSCL蛋白三级结构模型显示,该蛋白含有14个典型的α螺旋、10个β-折叠和36个β-转角。MeSCL蛋白与橡树HbSCL蛋白的相似性最高,为90.80%,与蓖麻RcSCR、胡杨PeSCL、毛果杨PtSCR、可可树Tc SCR和哥伦比亚锦葵HuSCL蛋白的相似性在80.00%左右。MeSCL基因受干旱、盐、氧化和低温胁迫诱导表达量整体呈升高趋势,但在不同处理时间的表达量存在明显差异,其中,干旱和氧化胁迫下,MeSCL基因均在处理24 h时表达量最高,分别是对照的6.05和11.17倍,而盐和低温胁迫下,MeSCL基因均在处理6 h时表达量最高,分别是对照的11.76和3.80倍。【结论】MeSCL基因参与木薯植株的非生物胁迫响应,正向调控其抗非生物胁迫能力。【Objective】In this study,the SCARECROW-LIKE(MeSCL)gene of cassava was cloned,then its bioinformatics analysis was carried out and its expression under abiotic stress was detected to provide theoretical reference for further study on the regulatory mechanism of MeSCL gene in response to abiotic stress.【Method】Taking cassava variety D346 as material,the coding region(CDS)sequence of MeSCL gene was cloned by RT-PCR,and its structural characteristics were analyzed by bioinformatics analysis software,then observed the expression of MeSCL under drought,salt,oxidation and low temperature stresses by real-time fluorescence quantitative PCR(qPCR).【Result】The full-length CDS sequence of MeSCL gene in cassava was 1655 bp.According to alignment with existing target gene sequences on NCBI(GenBank accession No.:LOC110627921),there were two different bases.The length of open reading frame(ORF)was 1560 bp and encoded 519 amino acids,the encoded protein molecular weight was 57.84 kD and the isoelectric point(pI)was 6.08.The fat coefficient of MeSCL protein was 80.46%,total average hydrophilic index was-0.195,which belonged to hydrophilic protein.The MeSCL protein had a signal peptide with six possible internal to external transmembrane helix regions and five possible external to internal transmembrane helix regions.The subcellular localization predicted that the protein was in the nucleus and endoplasmic reticulum and was a member of the GRAS family with the conserved structural domain of the GRAS family.The three-dimensional structure model of MeSCL protein showed that the protein had 14 typical alpha helices,10 beta folds and 36 beta turns.MeSCL amino acid sequence shared the highest similarity of 90.80%with Hevea brasiliensis(HbSCL)protein sequence,about 80.00%similarity with Ricinus communis(RcSCR),Populus euthoides(PeSCL),P.euthoides(PtSCR),Theobroma cacao(TcSCR)and Colombian mallow(HuSCL).The qPCR data showed that MeSCL gene expression level were increased under drought,salt,oxidation and low temperature stress.

关 键 词：木薯 SCARECROW-LIKE(SCL)基因 基因克隆 生物信息学分析 非生物胁迫 表达分析 

分 类 号：S533.035.3[农业科学—作物学]