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Acta Agronomica Sinica ›› 2019, Vol. 45 ›› Issue (7): 1050-1058.doi: 10.3724/SP.J.1006.2019.82054

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

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 Online:2019-07-12 Published:2019-03-01
  • Contact: Ding-Yang YUAN,Mei-Juan DUAN E-mail:yuandingyang@hhrrc.ac.cn;duanmeijuan@163.com
  • 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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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

Fig. 1

Phenotype of wild type (WT) R1128 and mutant wsl1 at seedling, tillering, and booting stages A: five-leaf stage. Left: WT; Right: wsl1. B-D: the mutant wsl1 at tillering stage in Changsha (B), tillering stage in Sanya (C), and booting stage in Changsha (D)."

Table 1

Comparison of agronomic traits between wild type R1128 and mutant wsl1"

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**

Fig. 2

Comparison of photosynthetic pigments at seedling stage and tillering stage in the mutant wsl1 and wild type R1128 A: comparison of photosynthetic pigments of the seeding stage; B: comparison of photosynthetic pigments of the tillering stage; Chl a: chlorophyll a; Chl b: chlorophyll b; Total Chl: content of chlorophyll a and chlorophyll b; Car: carotenoids. * represents significant difference between the wsl1 mutant and wild type at the 0.05 probability level; ** represents significant difference between the wsl1 mutant and wild type at the 0.01 probability level."

Fig. 3

Ultrastructures of chloroplasts in the mesophyll cell of the wsl1 mutant and wild type R1128 A: mesophyll cell of the wild type; B: mesophyll cell of the wsl1 mutant’s green leaf; C: mesophyll cell of the wsl1 mutant’s leaf regions with color change from green to white; D: mesophyll cell of the wsl1 mutant’s white leaf. c: chloroplast; v: vacuole."

Fig. 4

Cross-section of wild type R1128 and wsl1 mutant leaf at tillering stage A: cross section of wild type leaf; B: cross section of wsl1 mutant’s leaf. M: motor cell."

Table 2

Genetic analysis of the wsl1 mutant"

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

Fig. 5

Fine mapping of WSL1 gene n: number of group; recombinant: number of recombinant."

Table 3

Sequence of markers linked with WSL1"

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

Table 4

Genes in mapping area of WSL1"

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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