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作物学报

作物学报 ›› 2018, Vol. 44 ›› Issue (05): 650-656.doi: 10.3724/SP.J.1006.2018.00650

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

水稻黄绿叶基因Yellow-Green Leaf 6 (YGL6)的表达模式与蛋白定位

施军琼1,2(), 王亚琴1, 张天泉1, 马玲1, 桑贤春1, 何光华1,*()   

  1. 1西南大学水稻研究所 / 转基因植物与安全控制重庆市重点实验室, 重庆 400716
    2西南大学生命科学学院, 重庆 400716
  • 收稿日期:2017-09-23 接受日期:2018-01-08 出版日期:2018-05-20 网络出版日期:2018-01-29
  • 通讯作者: 何光华
  • 作者简介:

    第一作者联系方式: E-mail: shijunqiong@163.com

  • 基金资助:
    本研究由科技部重点研发计划项目(2017YFD0100201), 重庆市科委项目(CSTCCXLJRC201713, cstc2016shms-ztzx0017)和中央高校基本业务费专项(XDJK2016C111)资助

Expression Pattern and Protein Localization of a Yellow-Green Leaf 6 (YGL6) Gene in Rice (Oryza sativa)

Jun-Qiong SHI1,2(), Ya-Qin WANG1, Tian-Quan ZHANG1, Ling MA1, Guang-Hua HE1,*()   

  1. 1 Rice Research Institute, Chongqing Key Laboratory of Application and Safety Control of Genetically Modified Crops, Southwest University, Chongqing 400716, China
    2 School of Life Sciences, Southwest University, Chongqing 400716, China
  • Received:2017-09-23 Accepted:2018-01-08 Published:2018-05-20 Published online:2018-01-29
  • Contact: Guang-Hua HE
  • Supported by:
    This study was supported by the National Key Research and Development Program of China (2017YFD0100201), Chongqing Science and Technology Commission Project (CSTCCXLJRC201713, cstc2016shms-ztzx0017), and the Fundamental Research Funds for the Central Universities (XDJK2016C111).

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摘要：

叶色突变既可作为形态标记用于杂交稻育种, 又是研究光合系统的结构和功能、叶绿素生物合成及其调控机制的理想材料。EMS (ethyl methane sulfonate)诱变籼稻恢复系“缙恢10号”获得1个稳定遗传的黄绿叶突变体, 暂命名为ygl6 (yellow-green leaf 6)。前期我们通过图位克隆筛选出候选基因Os12g23180, 通过遗传互补实验证实了黄绿叶基因YGL6为Os12g23180, BLASTp分析表明YGL6基因编码NAD(P)-结合的Rossmann折叠超家族蛋白质, 属于短链脱氢酶/还原酶家族, 推断为异黄酮还原酶、糖脱水酶或mRNA结合蛋白。利用qRT-PCR进行表达模式的分析表明YGL6基因仅在绿色组织如心叶、成熟叶、叶鞘和绿色颖壳中表达, 尤其以心叶的表达量最高, 同时YGL6基因表达还受光照的诱导。构建亚细胞定位载体, 转水稻原生质体结果表明YGL6蛋白定位于叶绿体。本研究丰富了水稻突变体库, 为YGL6基因的功能分析奠定了基础。

关键词: 水稻(Oryza sativa), 黄绿叶, YGL6基因, 叶绿体, 表达模式

Abstract:

Leaf color mutants are used not only as morphological markers in hybrid rice breeding, but also as ideal materials in studies on the structure and function of photosystem, chlorophyll biosynthesis and regulation mechanism. A new rice mutant exhibiting stable inheritance was derived from ethyl methane sulfonate (EMS)-treated restorer line Jinhui 10 (Oryza sativa), tentatively named as yellow-green leaf 6 (ygl6). The ygl6 leaf displayed yellow-green at seeding stage, and pale green at jointing stages. The YGL6 complementation experiment implied that the Os12g23180 is the YGL6 gene. The expression pattern analysis indicated that YGL6 was expressed in green tissues including young leaves, mature leaves, sheaths and green glume, with the highest expression level in young leaves. And YGL6 expression was induced by light. Transient expression of the YGL6-GFP protein in rice protoplast showed that YGL6 was localized in chloroplasts. These results provide a foundation for functional analysis of YGL6.

Key words: rice (Oryza sativa), yellow-green leaf 6 (ygl6), YGL6 gene, chloroplast, expression pattern

图1

YGL6基因的互补验证 A: WT、ygl6和转基因互补植株; B: WT、ygl6和转基因互补植株的叶片; C: WT、ygl6和转基因互补植株突变位点的测序结果; D: WT、ygl6和转基因互补植株的色素含量。*表示在0.05水平上差异显著。"

图2

YGL6基因的进化分析"

图3

YGL6基因表达分析 A: 苗期YGL6基因在WT和ygl6的表达分析; B: 孕穗期YGL6基因在WT的表达分析。*表示在0.05水平上差异显著。"

图4

光照对YGL6基因表达的分析 A: 连续黑暗培养后在不同光照时间YGL6基因的表达; B: 连续光照培养后在不同黑暗时间YGL6基因的表达。"

图5

YGL6蛋白的亚细胞定位 A~D: 阴性对照(35S-GFP); E~H: YGL6蛋白定位(35S-YGL6ORF-GFP)。"

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