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作物学报 ›› 2022, Vol. 48 ›› Issue (8): 1926-1937.doi: 10.3724/SP.J.1006.2022.11067

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

小麦籽粒大小相关基因TaGS2克隆及功能分析

王沙沙1(), 黄超1, 汪庆昌1, 晁岳恩1,*(), 陈锋2,*(), 孙建国3, 宋晓4   

  1. 1河南省农业科学院小麦研究所 / 河南省小麦生物学重点实验室, 河南郑州 450002
    2河南农业大学农学院 / 省部共建小麦玉米作物学国家重点实验室 / 河南省粮食作物协同创新中心, 河南郑州 450002
    3濮阳市农业农村局, 河南濮阳 457000
    4河南省农业科学院植物营养与资源环境研究所, 河南郑州 450002
  • 收稿日期:2021-07-28 接受日期:2021-11-29 出版日期:2022-08-12 网络出版日期:2021-12-22
  • 通讯作者: 晁岳恩,陈锋
  • 作者简介:E-mail: shasha0391@126.com
  • 基金资助:
    河南省青年科学基金项目(202300410527);省部共建小麦玉米作物学国家重点实验室开放课题(30500772);国家青年科学基金项目(31801261)

Cloning and functional identification of TaGS2 gene related to kernel size in bread wheat

WANG Sha-Sha1(), HUANG Chao1, WANG Qing-Chang1, CHAO Yue-En1,*(), CHEN Feng2,*(), SUN Jian-Guo3, SONG Xiao4   

  1. 1Wheat Research Institute, Henan Academy of Agricultural Sciences / Henan Province Key Laboratory of Wheat Biology, Zhengzhou 450002, Henan, China
    2Agronomy College, National Key Laboratory of Wheat and Corn Crop Sciences / Collaborative Innovation Center of Henan Grain Crops / Agronomy College, Henan Agricultural University, Zhengzhou 450002, Henan, China
    3Department of Agricultural and Rural Affairs of Puyang, Puyang 457000, Henan, China
    4Institute of Plant Nutrient and Environmental Resources, Henan Academy of Agricultural Sciences, Zhengzhou 450002, Henan, China
  • Received:2021-07-28 Accepted:2021-11-29 Published:2022-08-12 Published online:2021-12-22
  • Contact: CHAO Yue-En,CHEN Feng
  • Supported by:
    Henan Province Science Foundation for Youths(202300410527);National Key Laboratory of Wheat and Maize Crop Science in Henan Agricultural University(30500772);National Science Foundation for Young Scientists of China(31801261)

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摘要：

籽粒大小影响小麦粒重, 进而影响产量。目前, 对小麦籽粒大小相关基因的研究已有报道, 但其潜在的分子机制尚不清楚。本研究通过同源克隆的方法从普通小麦中克隆了小麦籽粒大小相关基因TaGS2, 并对其序列进行生物信息学分析。烟草亚细胞定位结果表明, TaGS2定位在细胞核和细胞质内。不同组织材料qRT-PCR分析表明, TaGS2基因在小麦籽粒不同发育时期的表达量最高。构建TaGS2-PLGY-02-RNAi表达载体, 转化小麦。研究结果表明, TaGS2基因在RNAi转基因小麦中表达量显著降低。RNAi转基因小麦的籽粒长度变短, 籽粒宽度变窄, 千粒重也降低。因此推测TaGS2基因可能参与小麦籽粒大小或者千粒重的调控。本研究初步揭示了TaGS2基因功能, 并为小麦高产育种中千粒重的提高提供重要的基因资源。

关键词: 小麦, 籽粒大小, TaGS2基因, 亚细胞定位, 表达分析, RNAi干扰

Abstract:

The kernel size affects kernel weight in wheat, and then affects yield. Up to date, the genes related to kernel size have been reported in bread wheat. However, the underlying molecular mechanisms that regulates the size of wheat kernels remains unclear. In this study, the TaGS2 gene related to kernel size was successfully cloned from bread wheat based on in silico cloning and its sequence was analyzed by bioinformatics. Subcellular localization analysis of tobacco indicated that TaGS2 was localized in the nucleus and cytoplasm. Relative expression levels of different tissues showed that the TaGS2 gene was highly expressed at different developmental stages of the kernels. RNA interference vector TaGS2-PLGY-02-RNAi was constructed and transferred into wheat. The results indicated that the relative expression levels of TaGS2 gene was significantly reduced in RNAi transgenic wheat. In addition, the kernel length and width of RNAi transgenic wheat was shortened, and the thousand-kernel weight was reduced as well. Therefore, it was speculated that TaGS2 gene was probably involved in the regulation of wheat kernel size or thousand-kernel weight. This study preliminarily reveals the TaGS2 gene function and provides important genetic resources for the improvement of thousand-kernel weight in wheat breeding program.

Key words: Triticum aestivum L., kernel size, TaGS2 gene, subcellular localization, expression analysis, RNAi interference

附图1

小麦TaGS2-2A、TaGS2-2B和TaGS5-2D基因组DNA序列对比分析"

图1

小麦TaGS2基因结构图"

图2

小麦TaGS2基因生物信息学分析 A: 结构域预测; B: 蛋白二级结构预测; C: 跨膜结构预测。"

图3

小麦TaGS2蛋白的亚细胞定位"

图4

小麦TaGS2基因同源子不同组织材料的表达模式"

图5

转基因小麦的PCR检测 M: DL2000 marker; 1~14: T1代转基因植株; 15: 野生型小麦。"

图6

TaGS2转基因小麦相对表达量的检测(RNAi) 差异显著性分析(t-test方法), * P < 0.05、** P < 0.01、*** P < 0.001。"

图7

转基因小麦籽粒大小的比较(RNAi) 差异显著性分析(t-test方法), * P < 0.05、** P < 0.01、*** P < 0.001。"

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