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作物学报 ›› 2019, Vol. 45 ›› Issue (7): 1050-1058.doi: 10.3724/SP.J.1006.2019.82054

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

水稻白条纹叶突变体wsl1的遗传分析及基因精细定位

莫祎1,2,孙志忠2,丁佳2,余东2,孙学武2,盛夏冰2,谭炎宁2,袁贵龙2,袁定阳1,2,3,*(),段美娟1,*()   

  1. 1 湖南农业大学, 湖南长沙410128
    2 湖南杂交水稻研究中心杂交水稻国家重点实验室, 湖南长沙410125
    3 国家南方粮油作物协同创新中心, 湖南长沙410128
  • 收稿日期:2018-11-08 接受日期:2019-01-19 出版日期:2019-07-12 网络出版日期:2019-03-01
  • 通讯作者: 袁定阳,段美娟
  • 作者简介:莫祎, E-mail: 759447001@qq.com|孙志忠, E-mail: szznihaoa@163.com
  • 基金资助:
    本研究由国家自然科学基金项目(3167166);水稻种子良种繁育关键技术研究与示范(2018YFD0100802);香港中文大学合作项目培育超高产杂交水稻组合(TK1711793)(CUHK Joint Project to Develop Super-High Yield Hybrid Rice TK1711793);湖南省农业科学院科技创新项目(2017ZD02);湖南省自然科学青年基金项目(2017JJ3166)

Genetic analysis and fine mapping of white stripe leaf mutant wsl1 in rice

MO Yi1,2,SUN Zhi-Zhong2,DING Jia2,YU Dong2,SUN Xue-Wu2,SHENG Xia-Bing2,TAN Yan-Ning2,YUAN Gui-Long2,YUAN Ding-Yang1,2,3,*(),DUAN Mei-Juan1,*()   

  1. 1 Hunan Agricultural University, Changsha 410128, Hunan, China
    2 State Key Laboratory of Hybrid Rice, Hunan Hybrid Rice Research Center, Changsha 410125, Hunan, China
    3 National South Grain and Oil Crops Collaborative Innovation Center, Changsha 410128, Hunan, China
  • Received:2018-11-08 Accepted:2019-01-19 Published:2019-07-12 Published online:2019-03-01
  • Contact: Ding-Yang YUAN,Mei-Juan DUAN
  • Supported by:
    This study was supported by the National Natural Science Foundation of China(3167166);the Research and Demonstration of Key Technology of Improved Seed Breeding of Rice Seeds(2018YFD0100802);the Chinese University of Hong Kong(CUHK Joint Project to Develop Super-High Yield Hybrid Rice TK1711793);the Scientific and Technological Innovation Project of Hunan Academy of Agricultural Sciences(2017ZD02);the Project of Hunan Natural Science Youth Foundation(2017JJ3166)

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摘要：

从粳稻日本晴和籼稻R1128杂交衍生的重组自交系群体中获得一个稳定遗传的白条纹叶突变体wsl1 (white stripe leaf 1), 世代为F10。与亲本R1128相比, 突变体wsl1表现出白条纹叶, 同时叶脉呈现白化, 该性状在苗期就出现并持续整个生育期; 突变体的株高、每穗总粒数、剑叶长、生育期显著增加, 而结实率显著下降, 其他农艺性状没有显著变化。分蘖期突变体wsl1的叶绿素a、叶绿素b和胡萝卜素含量较杂交亲本R1128显著下降; 透射电镜观察表明, 与野生型相比, 突变体的叶绿体形状异常, 不规则。遗传分析表明, 该突变性状由1对隐性核基因控制。精细定位后发现, 目标基因WSL1位于第1染色体短臂上标记M1-54与标记M1-70之间, 两者相距89.7 kb。生物信息学分析表明候选区间内共有8个开放阅读框, 暂未发现已报道的叶色相关基因; 其中LOC_Os01g02080编码肽基脯氨酰顺反异构酶, GO (Gene Ontology)分类显示其可能与类囊体形成有关, 后续将通过比较测序、qRT-PCR等分子实验来确定候选基因。

关键词: 水稻, 白条纹叶突变体, 遗传分析, 基因定位

Abstract:

A white stripe leaf mutant wsl1 was obtained from the recombinant inbred lines derived from the cross of Oryza sativa var. japonica Nipponbare and Oryza sativa L. subsp. indica R1128. The mutant wsl1 showed white striped leaves and albino veins firstly at the seedling stage and then through the whole growth period. Agronomic traits such as plant height, number of spikelets per panicle, flag leaf length and heading date were significantly increased, while the seed setting rate decreased significantly in the mutant. Compared with wild type R1128, the chlorophyll a, chlorophyll b, and carotene contents of mutant leaves obviously decreased. Microscope observation indicated there were significantly decreased normal chloroplast and a large number of abnormal chloroplasts in mutant. Genetic analysis indicated that the mutant phenotype was controlled by a single recessive nuclear gene. WSL1 was mapped on the short arm of chromosome 1, between markers M1-54 and M1-70, with physical distance of about 89.7 kb. There were eight new open reading frames in the candidate region. Among them LOC_Os01g02080 encodes a peptide-based prolyl cis-trans isomerase, GO (Gene Ontology) classification showed that it might be related to thylakoid formation.

Key words: rice, white stripe leaf mutant, genetic analysis, gene mapping

图1

野生型R1128和突变体wsl1在苗期、分蘖期和抽穗期的表型 A: 五叶期幼苗, 左为野生型, 右为wsl1; B: 长沙突变体分蘖期; C: 三亚突变体分蘖期; D: 长沙突变体孕穗期。"

表1

野生型R1128和突变体wsl1农艺性状"

农艺性状Agronomic trait R1128 wsl1
有效穗数 No. of effective panicles 6.25±2.3 5.9±1.9
穗长 Panicle length (cm) 29.8±1.0 32.9±2.8
千粒重 1000-grain weight (g) 23.3±1.5 22.6±1.1
一次枝梗数 No. of primary branch 19.8±2.4 21.1±1.9
二次枝梗数 No. of secondary branch 49.5±6.8 68.7±8.7**
剑叶宽 Flag leaf width (cm) 3.0±0.4 3.0±0.4
株高 Plant height (cm) 121.9±5.8 181.6±6.9**
每穗总粒数 No. of spikelets per panicle 342.0±61.5 447.1±85.9**
结实率 Seed setting rate (%) 76.3±0.9 57.8±0.9**
剑叶长 Flag leaf length (cm) 40.8±7.7 55.1±9.6**
抽穗期 Heading date (d) 87.0±2.5 102±2.1**

图2

野生型R1128和突变体wsl1苗期和分蘖期叶绿素的含量比较 A: 苗期光合色素含量比较; B: 分蘖期光合色素含量比较。Chl a: 叶绿素a; Chl b: 叶绿素b; Total Chl: 叶绿素a+叶绿素b; Car: 类胡萝卜素。*表示野生型与突变体在0.05水平差异显著; **表示野生型与突变体在0.01水平差异显著。"

图3

突变体wsl1和野生型R1128叶肉细胞中叶绿体显微结构 A: 野生型叶肉细胞; B: wsl1 白条纹叶绿区叶肉细胞; C: wsl1白条纹叶绿转白区叶肉细胞; D: wsl1白条纹叶白区叶肉细胞。c: 叶绿体; v: 液泡。"

图4

野生型和突变体wsl1分蘖期的叶片横切面图 A: 野生型叶片横切面; B: wsl1白条纹叶片横切面。M: 运动细胞。"

表2

突变体wsl1的遗传分析"

杂交组合
Cross		 正常植株数
Number of green plant		 白化叶植株数
Number of albino
plant		 F2群体总株数
Total number of F2
plants		 分离比
Theoretical segregation ratio		 χ2
(χ20.05=3.84)
wsl1/Nipponbare 1247 390 1637 3.20 1.301
R1128/wsl1 2702 878 3580 308 0.431

图5

WSL1的精细定位 n: 群体大小; recombinant: 重组单株数。"

表3

WSL1 基因的连锁标记"

标记
Marker		 正向引物
Forward primer (5′-3′)		 反向引物
Reverse primer (5′-3′)
RM10022 CCTCCATAGAGTAAGGTTTGCATGG CCTCCTCCTCTGTCTTTCTCTGC
M1-40 GACGACGCCCGGTGGCTGTG GCAGTATCACCATGGATCCA
M1-54 ACATAAGTATCATGGGCTTA CCGAAGTTGCGTACTAACCG
M1-70 GTTGCCGCGAAGTTTCACGC ACCACCACCCAAAATGAGCT
M1-76 ACCTGGAGGAGCAAATTAGG CGTCCATTCCAATGATTTCC
M1-80 AAGTTATCTTTGTGACTGCT AGAAATGCGGCAGGAACGAC
RI02428 CTTCGTCACCGCAGATGG CTGTTTCTTTGGACCGGTGT

表4

WSL1 定位区域内基因信息"

基因位点
Gene locus		 基因表达产物
Gene product
LOC_Os01g02000 phosphate transporter 1, putative, expressed
LOC_Os01g02010 expressed protein
LOC_Os01g02020 acetyl-CoA acetyltransferase, cytosolic, putative, expressed
LOC_Os01g02040 retrotransposon protein, putative, unclassified, expressed
LOC_Os01g02050 phosphoenolpyruvate carboxylase, putative, expressed
LOC_Os01g02060 TOO MANY MOUTHS precursor, putative, expressed
LOC_Os01g02070 invertase/pectin methylesterase inhibitor family protein, putative, expressed
LOC_Os01g02080 peptidyl-prolyl cis-trans isomerase, putative, expressed
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