摘要
乙酰乳酸合酶(ALS)在植物的除草剂抗性中发挥重要作用。为揭示油莎豆ALS编码基因的序列特征、进化关系及表达特性,研究基于转录组数据,应用RT-PCR技术对其进行了克隆。序列分析显示,CeALS预测编码645个氨基酸,理论分子量为69.94 kD、等电点为6.10,属于叶绿体定位的亲水性蛋白。保守结构域和同源分析显示蛋白含有TPP_enzyme_N、TPP_enzyme_M和TPP_enzyme_C三个完整的乙酰乳酸合酶功能域,与其他莎草科植物中同源蛋白的一致性都在90%以上。进化分析显示,莎草科与禾本科属禾本目的姊妹科。qRT-PCR分析显示,CeALS主要在叶片和块茎中表达;在叶片发育过程中呈现逐步递增的趋势,其在成熟和衰老叶片中的表达水平显著高于幼嫩叶片。序列比对和SNP分析显示,团队收集的56份油莎豆种质均不存在抗药性变异。这些结果为除草剂抗性分子育种及油莎豆的开发与利用奠定了坚实的基础。
Acetolactate synthase(ALS)plays an important role in the herbicide resistance of plants.To uncover the sequence characteristics,evolutionary relationship,and expression pattern of the ALS gene in tigernut(Cyperus esculentus L.),in this study,RTPCR was adopted to isolate the corresponding gene CeALS on the basis of transcriptome data.Sequence analysis revealed that CeALS putatively encoded 646 amino acids with a theoretical molecular weight of 69.94 kD and an isolectric point of 6.10,which was predicted to be a hydrophilic protein with chloroplast localization.The protein was shown to harbor three conserved domains,i.e.,TPP_enzyme_N,TPP_enzyme_M,and TPP_enzyme_C,which were specific to acetolactate synthase.Homologous analysis suggested that CeALS shared an identity of>90%with homologs from other Cyperaceae plants.Phylogenetic analysis supported that Cyperaceae was a sister family to Poaceae within Poales.qRT-PCR analysis revealed that CeALS was mainly expressed in mature leaves and tubers.During leaf development,an increasing trend was observed and the expression level in mature and senescent leaves was significantly higher than that in young leaves.Sequence alignment and SNP analysis supported that no target site resistance mutations to acetolactate-synthase-inhibiting herbicides were found in 56 germplasms collected by our group.Results obtained in this study lay a solid foundation for molecular breeding for herbicide resistance as well as development and utilization of tigernut.
作者
肖艳华
邹智
赵永国
郭安平
张丽
XIAO Yan-hua;ZOU Zhi;ZHAO Yong-guo;GUO An-ping;ZHANG Li(College of Life Science,South-Central University for Nationalities,Hubei Provincial Key Laboratory for Protection and Application of Special Plants in Wuling Area of China,Wuhan 430074;Hainan Key Laboratory for Biosafety Monitoring and Molecular Breeding in Off-Season Reproduction Regions,Institute of Tropical Bioscience and Biotechnology,Sanya Research Institute of Chinese Academy of Tropical Agricultural Sciences,Haikou 571101;School of Biological and Food Engineering,Guangdong University of Petrochemical Technology,Maoming 525000)
出处
《生物技术通报》
CAS
CSCD
北大核心
2022年第4期184-192,共9页
Biotechnology Bulletin
基金
海南省自然科学基金(320RC705,319MS093)
国家自然科学基金项目(31971688)
2018年度广东省普通高校特色创新项目(2018KTSCX148)。
关键词
油莎豆
油料作物
乙酰乳酸合酶
除草剂抗性
Cyperus esculentus L.
oil crop
acetolactate synthase
herbicide resistance
