植物学报 ›› 2022, Vol. 57 ›› Issue (3): 276-287.DOI: 10.11983/CBB22018

• 研究报告 • 上一篇    下一篇

水稻黄绿叶调控基因YGL18的克隆与功能解析

杨凯如1, 贾绮玮1, 金佳怡1, 叶涵斐1, 王盛1, 陈芊羽1, 管易安1, 潘晨阳1, 辛德东1, 方媛1,*(), 王跃星2,*(), 饶玉春1,*()   

  1. 1浙江师范大学化学与生命科学学院, 金华 321004
    2中国水稻研究所水稻生物学国家重点实验室, 杭州 310006
  • 收稿日期:2022-01-19 接受日期:2022-03-18 出版日期:2022-05-01 发布日期:2022-05-18
  • 通讯作者: 方媛,王跃星,饶玉春
  • 作者简介:ryc@zjnu.cn
    wangyuexing@caas.cn;
    * E-mail: fy0579@zjnu.cn;
  • 基金资助:
    国家级大学生创新创业训练计划(202010345067);国家级大学生创新创业训练计划(202110345011);国家自然科学基金(31971921);广西水稻遗传育种重点实验室开放基金(2018-15-Z06-KF12);中国水稻生物学国家重点实验室开放项目(20200102)

Cloning and Functional Analysis of Rice Yellow Green Leaf Regulatory Gene YGL18

Kairu Yang1, Qiwei Jia1, Jiayi Jin1, Hanfei Ye1, Sheng Wang1, Qianyu Chen1, Yian Guan1, Chenyang Pan1, Dedong Xin1, Yuan Fang1,*(), Yuexing Wang2,*(), Yuchun Rao1,*()   

  1. 1College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China
    2State Key Laboratory of Rice Biology, China National Rice Research Institute, Hangzhou 310006, China
  • Received:2022-01-19 Accepted:2022-03-18 Online:2022-05-01 Published:2022-05-18
  • Contact: Yuan Fang,Yuexing Wang,Yuchun Rao

摘要: 叶色突变体往往伴随着叶绿素含量变化及叶绿体结构异常, 是研究叶绿体发育与光合作用相关基因功能的重要材料。该研究通过甲基磺酸乙酯(EMS)诱变籼稻(Oryza sativa subsp. indica)品种华占(HZ)获得黄绿叶突变体, 将其命名为ygl18 (yellow-green leaf 18)。与野生型相比, 黄绿叶突变体ygl18自三叶期起叶片开始变黄且程度不断加深, 同时伴随着光合速率与叶绿素含量下降, 且结实率、千粒重及有效穗数均显著降低。透射电镜观察结果显示, ygl18的叶绿体结构紊乱, 基质片层疏松, 发育受到抑制, 与叶片出现黄绿色表型一致。遗传分析表明, ygl18突变性状受1对隐性等位核基因控制, 这对等位基因位于水稻第3号染色体长臂标记InDel2和InDel3之间115.2 kb范围内。进一步研究发现该突变体表型是编码铁氧还蛋白FdC2的基因LOC_Os03g48040的5'UTR发生突变所致。通过CRISPR转基因实验验证了该基因对表型的控制作用。研究结果揭示了叶色调控网络的遗传基础, 可为今后选育高光效水稻品种提供新线索。

关键词: 水稻, 叶色变异, 遗传分析, 图位克隆, 铁氧还蛋白

Abstract: Leaf color mutants are often accompanied by changes in chlorophyll content and abnormal chloroplast structure, and serve as essential materials for studying the functions of chloroplast development and photosynthesis-related genes. In this study, we obtained a yellow-green leaf mutant named yellow-green leaf 18 (ygl18) from Oryza sativa subsp. indica cv. ‘HZ’ with ethyl methanesulfonate (EMS). Compared with the wild type, the leaves of ygl18 turned yellow at three-leaf stage and the degree of yellowing increased as it grew, accompanied by decreasing photosynthetic rate and chlorophyll content. The seed-setting rate, 1 000-grain weight, and effective panicle number were significantly lower than those of the wild type. We observed disordered chloroplast structure, loose stromal lamellas, and stalled development in the mutant using transmission electron microscopy. Genetic analysis indicated that the mutant feature (or phenotype) of ygl18 is controlled by a pair of recessive nuclear alleles, which were located in a 115.2 kb region between markers InDel2 and InDel3 on the long arm of chromosome 3. We found mutations in the 5′UTR of LOC_Os03g48040 encoding FdC2 (ferredoxin C2). The gene’s function on controlling the mutant phenotype was verified using CRISPR transgenic experiments. Our results revealed a genetic basis for leaf color regulatory network and provide new clues for breeding photosynthetically efficient rice varieties in the future.

Key words: Oryza sativa (rice), leaf color variation, genetic analysis, map-based cloning, ferredoxin