大豆茸毛突变体的鉴定及相关基因表达分析
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中国农业科学院作物科学研究所 /农作物基因资源与基因改良国家重大科学工程 / 农业部北京大豆生物学重点实验室

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转基因生物新品种培育重大专项(2016ZX08009003-003);中国农业科学院科技创新工程(作物分子标记技术及其应用)


Identification of Soybean Pubescence Mutants and Expression Analysis of Related Genes
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Institute of Crop Sciences, Chinese Academy of Agricultural Sciences / The National Key Facility for Crop Gene Resources and Genetic Improvement / Key Laboratory of Soybean Biology (Beijing), Ministry of Agriculture

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the National Transgenic Major Program of China;the the Agricultural Science and Technology Innovation Program (ASTIP) of Chinese Academy of Agricultural Sciences

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    摘要:

    茸毛是大豆的重要形态性状,对其自身的生长发育有着重要作用,也与百粒重等诸多农艺性状相关,因此对大豆茸毛的研究显得尤为重要。本研究从大豆 EMS 突变体库中筛选到 4 个茸毛发育异常的突变体,分别是短茸毛突变体 ddsp1,无茸毛突变体 ddsp2,茸毛部分空瘪突变体 ddsp3 和茸毛完全空瘪突变体 ddsp4。扫描电镜观察结果表明这些突变体中茸毛的长度或形态等都发生了明显的变化,同时突变体的籽粒大小、百粒重等也发生了明显的改变。选取了 10 个可能与大豆茸毛发育相关的同源基因,分别是参与正调控茸毛发育起始的基因 GL1、GL2、GL3 和 TCL1,参与核内复制负调控的基因 RBR1、SIM、KAK 和 SPY 以及参与核内复制正调控的基因 CPR5 和 RHL,通过 qRT-PCR 分析了这 10 个基因在 4 个茸毛突变体和野生型中的表达情况。结果表明,控制茸毛发育起始的基因除了 GL1 和 GL2 分别在突变体 ddsp2 和 ddsp4 中显著下调表达之外,其余控制茸毛发育起始的基因均显著上调或无显著变化。参与核内复制负调控的基因中,除了 RBR1 基因只在突变体 ddsp1中显著上调表达之外, SIM、KAK 和 SPY 在这 4 个突变体中的表达水平均显著升高。而参与核内复制正调控的基因 CPR5 和RHL 在 4 个突变体中的相对表达量均显著增加。本研究通过对茸毛相关基因的表达分析,为进一步挖掘控制大豆茸毛发育的基因以及研究基因的作用机理和调控模式奠定了基础。

    Abstract:

    Pubescence is an important morphological trait in soybean, which plays an important role in growth and development, and which is correlated to many agronomic traits such as 100-seed weight, and deserves investigation. In this study, four soybean mutants with developmental defect in pubescence were selected from EMS mutant library: the mutant ddsp1 with short pubescence, the glabrous mutant ddsp2, the mutant ddsp3 with partially shrunken pubescence, and the mutant ddsp4 with fully shrunken pubescence. Scanning electron microscope images showed that the length and morphology of pubescence were significantly altered in these mutants. In addition, the seed size and 100-seed weight were all significantly different from the wild type. Ten soybean orthologous genes, including GL1, GL2, GL3and TCL1 which act on the initiation of pubescence development, RBR1, SIM, KAK and SPY which participate in negative regulation of endoreduplication, and CPR5 and RHL which participate in positive regulation of endoreduplication, were selected for expression analysis in those four mutants and the wild type by qRT-PCR. The results showed that the genes controlling the initiation of pubescence development, except for GL1 and GL2 which significantly down-regulated the expression in mutants ddsp2 and ddsp4, up-regulated the expression or had no significant effect. The gene RBR1 significantly up-regulated the endoreduplication in mutant ddsp1 only, whereas the genes SIM, KAK and SPY significantly down-regulated the endoreduplication in all the four mutants, and both the genes CPR5 and RHL significantly up-regulated the endoreduplication in all the four mutants. Through the expression analysis of pubescence-related genes in soybean, this research has laid a foundation for further searching for the genes controlling the soybean pubescence development and understanding the regulation mechanisms of these genes.

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唐均勇,杨静,洪慧龙,等.大豆茸毛突变体的鉴定及相关基因表达分析[J].植物遗传资源学报,2020,21(1):121-129.

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  • 收稿日期:2019-05-07
  • 最后修改日期:2019-06-21
  • 录用日期:2019-07-24
  • 在线发布日期: 2020-01-17
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