摘要
【目的】解析木薯脱落酸(ABA)受体PYR/PYL/RCARs家族基因(MePYL13)在木薯块根采后生理性变质(PPD)过程中的功能作用,为后续探索ABA信号通路在木薯PPD中的功能及作物抗逆育种打下基础。【方法】从木薯品种SC8中克隆MePYL13基因,应用生物信息学分析方法对其编码蛋白的理化性质、亲疏水性、保守结构域及二、三级结构等进行预测,对基因上游启动子进行元件分析,通过亚细胞定位观察MePYL13蛋白在植物细胞中的定位情况,并利用实时荧光定量PCR检测MePYL13基因在木薯PPD过程中的相对表达量。【结果】克隆获得的MePYL13基因编码区(CDS)长度663 bp,编码220个氨基酸,蛋白分子量23.957 kD,理论等电点(Ip)为6.74。MePYL13蛋白与巴西橡胶树(XP021654464.1)PYL家族蛋白的氨基酸序列同源性最高,为91.55%,表明MePYL13蛋白氨基酸序列具有高度保守性。从MePYL13基因启动子鉴定获得与非生物胁迫逆境相关的响应元件,包括厌氧诱导元件(ARE,AAACCA)、光响应元件(ATCT-motif,AATCTAATCC)、干旱MYB元件(MBS,TAACTG)及ABA应答元件(ABRE,AAACAGA)和分生组织表达相关元件(CAT-box,GCCACT)等;MePYL13蛋白在木薯细胞的细胞核和细胞质上均有表达。随着PPD进程的推移,MePYL13基因相对表达量整体上呈上升趋势,至PPD后期(48 h)其相对表达量是0 h时(PPD前)的16倍,即MePYL13基因受木薯块根PPD的显著诱导。【结论】MePYL13基因是PYR/PYL/RCARs家族成员,可能在木薯块根PPD过程中发挥正向调控作用,为培育木薯PPD改良品种提供了潜在基因资源。
【Objective】In this paper,abscisic acid(ABA)receptor PYR/PYL/RCARs family member(MePYL13)was cloned from cassava genome and its biological information and the expression characteristics in the process of post-harvest physiological deterioration(PPD)in cassava were analyzed,which provided a theoretical basis for further exploring the regulatory mechanism of ABA signaling pathway in cassava PPD and crop breeding for stress tolerance.【Method】MePYL13 was cloned from cassava variety SC8. The physicochemical properties,hydrophilic/hydrophobic,conserved domain,secondary and tertiary structures of the protein encoded by the gene were predicted,the elements in MePYL13 gene promoter were analyzed through bioinformatics analysis. Subcellular localization of MePYL13 protein was also identified.Besides,the expression pattern of MePYL13 in cassava PPD process was investigated by real-time fluorescence quantitative PCR.【Result】The coding regions(CDS)of this gene was 663 bp in length and encoded 220 amino acids,the protein molecular weight was 23.957 kD,and the theoretical isoelectric point(Ip)was 6.74. Multiple sequence alignment indicated that the similarity of MePYL13 protein with Hevea brasiliensis(XP021654464.1)was the highest(91.55%).Amino acid sequence of MePYL13 protein was highly conservative. Several elements related to abiotic stress response were identified from the promoter of MePYL13,including anaerobic induction elements(ARE,AAACCA),light response elements(ATCT-motif,AATCTAATCC),drought MYB element(MBS,TAACTG),ABA response elements(ABRE,AAACAGA)and meristem expression-related elements(CAT-box,GCCACT),and so on. Subcellular location study indicated that MePYL13 protein was distributed in the nucleus and cytoplasm. Totally,MePYL13 was up-regulated at all the three time points in storage roots after harvested,and its relative expression level at 48 h was 16 times of that at0 h(before PPD). Thus,MePYL13 was significantly increased in PPD process.【Conclusion】It is inferred that MePYL13 is a member of the PYR/PYL/RCARs family and may play a positive role in regulating PPD of cassava storage roots. This study is beneficial in providing potential genetic resources for cultivating improved cassava PPD varieties.
作者
赵思涵
颜彦
陈志晟
商桑
田丽波
胡伟
ZHAO Si-han;YAN Yan;CHEN Zhi-sheng;SHANG Sang;TIAN Li-bo;HU Wei(College of Horticulture,Hainan University,Haikou 570228,China;Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Science,Haikou 571101,China)
出处
《南方农业学报》
CAS
CSCD
北大核心
2019年第12期2629-2637,共9页
Journal of Southern Agriculture
基金
国家自然科学基金项目(31801419)
海南省自然科学基金面上项目(317036)
海南省研究生创新科研项目(Hys2018-34)
园艺“3+2”专业分段培养试点学生创新项目(YY17-02)
作者简介
通讯作者:田丽波(1976-),教授,主要从事园艺植物分子育种研究工作,E-mail:faiy7play@163.com;胡伟(1982-),副研究员,主要从事植物分子生物学研究工作,E-mail:huwei2010916@126.com;赵思涵(1995-),研究方向为木薯分子生物学,E-mail:1304368354@qq.com。
