Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (4): 184-192.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1198

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Molecular Cloning and Characterization of an Acetolactate Synthase Gene(CeALS)from Cyperus esculentus L.

XIAO Yan-hua1,2(), ZOU Zhi2(), ZHAO Yong-guo3, GUO An-ping2, ZHANG Li1()   

  1. 1. 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
    2. 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
    3. School of Biological and Food Engineering,Guangdong University of Petrochemical Technology,Maoming 525000
  • Received:2021-09-16 Online:2022-04-26 Published:2022-05-06
  • Contact: ZOU Zhi, ZHANG Li E-mail:1740329672@qq.com;zouzhi2008@126.com;zhangli0624@mail.scuec.edu.cn

Abstract:

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,RT-PCR 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.

Key words: Cyperus esculentus L., oil crop, acetolactate synthase, herbicide resistance