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作物学报 ›› 2022, Vol. 48 ›› Issue (12): 3120-3129.doi: 10.3724/SP.J.1006.2022.12077

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

水稻叶脉黄化突变体yml的遗传分析及基因定位

郭均瑶1,2(), 刘斌美1,2, 杨惠杰1,2, 秦超琦2, 任艳2, 姜鸿瑞2, 陶亮之2, 叶亚峰2, 吴跃进1,2()   

  1. 1安徽大学物质科学与信息技术研究院, 安徽合肥 230031
    2中国科学院合肥物质科学研究院, 安徽合肥 230031
  • 收稿日期:2021-11-06 接受日期:2022-03-25 出版日期:2022-12-12 网络出版日期:2022-04-19
  • 通讯作者: 吴跃进
  • 作者简介:E-mail: 154519697@qq.com
  • 基金资助:
    安徽省自然科学基金项目(2108085MC99);合肥市“借转补”科技专项(J2020G45);安徽省科技重大专项(202003c08020006)

Genetic analysis and gene mapping of the yellow midrib leaf mutant (yml) in rice (Oryza sativa L.)

GUO Jun-Yao1,2(), LIU Bin-Mei1,2, YANG Hui-Jie1,2, QIN Chao-Qi2, REN Yan2, JIANG Hong-Rui2, TAO Liang-Zhi2, YE Ya-Feng2, WU Yue-Jin1,2()   

  1. 1Institutes of Physical Science and Information Technology, Anhui University, Hefei 230031, Anhui, China
    2Hefei Institutes of Physical Science, Chinese Academy of Science, Hefei 230031, Anhui, China
  • Received:2021-11-06 Accepted:2022-03-25 Published:2022-12-12 Published online:2022-04-19
  • Contact: WU Yue-Jin
  • Supported by:
    Natural Science Foundation of Anhui Province(2108085MC99);Hefei Science and Technology Project(J2020G45);Anhui Science and Technology Major Project(202003c08020006)

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摘要：

水稻(Oryza sativa L.)是重要的粮食作物之一, 其产量一直备受关注。水稻叶片是光合作用的重要场所, 叶脉是叶片中具有支撑和运输功能的组织。同时, 叶脉中存在的光合色素也能提供一定的光合作用。本研究采用重离子束诱变籼稻9311获得的叶脉黄化突变体yml进行遗传分析及基因定位研究。该突变体在抽穗期开花后5 d左右, 开始出现叶脉黄化表型, 并于抽穗后期黄化性状明显, 并持续至成熟期。在分蘖期, 突变体与野生型相比, 光合色素含量无显著差异。但在抽穗后期, 突变体叶片及叶脉中光合色素含量显著降低, 导致光合效率明显降低, 净光合速率仅为野生型的50.37%。成熟期突变体的株高、穗长、有效穗数、每穗实粒数、结实率、千粒重均显著低于野生型。通过遗传分析, 确定该突变性状由一对隐性基因控制。利用图位克隆技术, 将该基因定位于6号染色体。进一步通过简单重复序列(SSR)及插入缺失(InDel)标记, 将该基因位置缩小至InDel5与RM3431之间, 物理距离约为700 kb。本研究为后续突变体基因的克隆和功能分析提供了研究基础。

关键词: 水稻, 叶脉, 黄化, 光合特性, 基因定位

Abstract:

Rice (Oryza sativa L.) is an important food crop, and its yield has been concerned for a long time. Rice leaves are the essential sites for photosynthesis, and leaf midrib is the tissue with the function of supporting and transporting. Meanwhile, the photosynthetic pigments present in leaf midrib can also provide a certain amount of photosynthesis. In this study, the yellow midrib leaf (yml) mutant obtained by the heavy ion beam implantation on indica rice 9311 was applied for genetic analysis and gene mapping. The mutant began to had the yellowing phenotype of leaf midrib about 5 days after flowering at heading stage, and the yellowing character was obvious at the late heading stage, and this change could last until the mature stage. At tillering stage, there was no significant difference in photosynthetic pigment content between the mutant and the wild type. However, compared with wild type at the late heading stage, the photosynthetic pigment content in the leaves and midribs of the mutant was significantly lower. Consequently, the photosynthetic efficiency of the mutant was substantially reduced, and the net photosynthetic rate was only 50.37% of wild type. At mature stage, plant height, panicle length, effective panicle, filled grain number per panicle, seed setting rate, and the 1000-grain weight of mutants were significantly lower than wild type. Genetic analysis revealed that this mutant character was controlled by a pair of recessive genes. By using map-based cloning technique, this gene was located on chromosome 6. Moreover, the gene was further located between InDel5 and RM3431 with a physical distance of approximately 700 kb by using simple repeat sequence (SSR) and insertional deletion (InDel) markers. This study provides a research basis for the subsequent cloning and functional analyses of the mutant gene.

Key words: Oryza sativa L., midrib, yellow, photosynthetic, gene mapping

图1

野生型(9311)与突变体yml的表型特征 A: 抽穗期野生型(9311)与突变体yml植株, 标尺为10 cm; B: 抽穗期野生型(9311)与突变体yml的剑叶, 标尺为5 cm; C: 野生型(9311)与突变体yml的穗, 标尺为5 cm。"

表1

野生型(9311)和突变体(yml)农艺性状分析"

材料
Material name		 株高
Plant height
(cm)		 穗长
Panicle length
(cm)		 有效穗数
Effective
panicle		 每穗实粒数
Filled grain number
per panicle		 结实率
Seed setting rate (%)		 千粒重
1000-grain weight (g)
9311 127.33±4.29 22.42±2.17 9.67±1.53 158.62±27.98 71.16±9.55 28.06±2.16
yml 97.41±4.15** 20.67±2.79* 4.67±0.58** 105.00±30.58** 43.20±9.02** 23.10±0.41**

图2

不同时期野生型(9311)与突变体yml叶片与叶中脉光合色素含量以及各光合色素含量比值 A: 分蘖期野生型(9311)与突变体yml叶片光合色素含量; B: 分蘖期野生型(9311)与突变体yml叶中脉光合色素含量; C: 抽穗期野生型(9311)与突变体yml叶片光合色素含量; D: 抽穗期野生型(9311)与突变体yml叶中脉光合色素含量; E: 分蘖期野生型(9311)与突变体yml叶片中类胡萝卜素与总叶绿素比值; F: 抽穗期野生型(9311)与突变体yml叶片中类胡萝卜素与总叶绿素比值。*: P < 0.05; **: P < 0.01。"

表2

野生型(9311)和突变体(yml)叶片的光合特性"

材料
Material		 净光合速率
Photosynthetic rate
(μmol CO2 m-2 s-1)		 蒸腾速率
Transpiration rate
(mol H2O m-2 s-1)		 气孔导度
Stomatal conductance
(mmol H2O m-2 s-1)		 胞间二氧化碳浓度
Intercellular CO2 concentration
(μmol CO2 L-1)
9311 14.95 ±0.74 3.96±0.33 279.97±3.72 338.00±7.21
yml 7.53±0.15** 5.96±0.34** 227.47±15.88** 390.00±9.54**

图3

yml基因初定位电泳图 M: 9311; W: 日本晴; M: DL2000 marker."

图4

yml基因在6号染色体上连锁图谱"

表3

根据籼稻品种9311与粳稻品种日本晴的序列差异新设计的InDel标记"

引物名称
Prime name		 引物序列
Prime sequence
(5'-3')		 物理位置
Physical position
(bp)		 9311 PCR
产物大小
PCR product size of 9311
(bp)		 日本晴PCR
产物大小
PCR product size of Nipponbare
(bp)		 中花11 PCR
产物大小
PCR product size of Zhonghua 11 (bp)
InDel 3 F: CTGGCGGTCGAATTTCTTTA
R: GGCAAATTTCAGCCACAGAT		 7,245,119-7,245,342 160 124 124
InDel 5 F: CAATCAGAATCTGCTTGCCA
R: ACACACATGGCAGGTCACAT		 8,059,757-8,059,997 255 241 241
InDel 13 F: CAGCCACACATTTGGAACCA
R: CTGCAAAGGATCTTGATCCG		 7,813,844-7,814,076 206 233 233

表4

定位区间内相关基因及功能预测"

基因名称
Gene name		 推测功能
Putative function
LOC_Os06g14420 Hydrolase, NUDIX family, domain containing protein, expressed
LOC_Os06g14650 Zinc finger, C3HC4 type domain containing protein, expressed
LOC_Os06g14670 ODORANT1, putative, expressed
LOC_Os06g14700 Myb-like DNA-binding domain containing protein, expressed
LOC_Os06g14710 Myb-like DNA-binding domain containing protein, expressed
LOC_Os06g14750 Phosphatidylinositol-4-phosphate 5-kinase family protein, putative, expressed
LOC_Os06g15380 Acylphosphatase, putative, expressed
LOC_Os06g15390 Acylphosphatase, putative, expressed
LOC_Os06g15410 Angel, putative, expressed
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