水稻黄绿叶突变体ygl13的鉴定及候选基因分析

doi:10.3864/j.issn.0578-1752.2015.21.001

Abstract

Abstract: 【Objective】The current study was conducted aiming at phenotypic characterization and candidate gene analysis of the yellow-green mutant ygl13, so as to add to our knowledge of the formation and regulation of the molecular mechanisms responsible for leaf-color mutations in rice.【Method】A new rice mutant exhibiting stable inheritance was identified as derived from ethyl methane sulfonate (EMS)-treated restorer line Jinhui10 (Oryza sativa), tentatively named as yellow-green leaf 13 (ygl13). Morphological characteristics, the photosynthetic pigment contents and the agronomic traits were measured systematically. Transmission electron microscopy was conducted to analyze the ultrastructure of the mesophyll cells and chloroplasts in the ygl13 mutant and wide-type plants. The ygl13 was crossed with indica sterile line Xinong1A whose plant and leaves were normally green, and the morphological phenotype and segregation ratio of F₁ and F₂ were used for genetic analysis, F₂ for gene mapping, and putative genes in the fine mapped region were analyzed, and the candidate genes in the mutant and the wild type were sequenced, respectively.【Result】The ygl13 leaves displayed yellow-green compared with the wild type. And the photosynthetic pigment contents of chlorophyll a, chlorophyll b, and carotenoid decreased significantly at the seedling and the booting stages. The results from transmission electron microscope demonstrated that the structure of the chloroplast in the mutant ygl13 developed abnormally with poor thylakoids, less grana stacks and scattered distribution when compared with the wide type. According to the performance of agronomic traits, compared with the wild type Jinhui 10, the grain number per panicle increased by 26.06%, and the pant height and seed setting rate decreased by 12.33% and 18.82%. As for the panicle length, effective panicles per plant, filled grain number per panicle and 1000-grain weight, there was no significant difference between the wild type and ygl13. Genetic analysis demonstrated that the mutant trait was controlled by a single recessive gene as the number of green seedlings verses that of yellow-green seedlings approached 3:1（χ²=2.35＜χ²_0.05=3.84). Genetic mapping of the mutant gene was conducted using 602 recessive individuals from the F₂ segregation population. Finally, YGL13 was mapped on the short arm of chromosome 8 between InDel marker ID43 and ID69, and with an interval of 318 kb. There were 52 genes in this region and the sequencing analysis of these candidate genes between the mutant and its wild type revealed a single base change (G1005A) of the OsSIG1 gene (LOC_Os08g06630) in the encoded product resulted in a premature stop codon and protein truncation with 334 residues not the primary protein with 520 residues in the mutant ygl13. The qRT-PCR results showed that, the expression level of genes associated with pigment metabolism and photosynthesis is in disorder. 【Conclusion】 The mutant ygl13 was controlled by a single recessive gene. The gene YGL13 was allelic to OsSIG1 which was documented as a plastid sigma factor previously in rice.

Key words: rice (Oryza sativa L.), yellow-green leaf mutant, OsSIG1, genetic analysis, candidate gene analysis

WANG Ya-qin, SHI Jun-qiong, ZHANG Ting, LI Yan, ZHANG Tian-quan, ZHANG Xiao-long, SANG Xian-chun, LING Ying-hua, HE Guang-hua. Characterization and Candidate Gene Analysis of Yellow-Green Leaf Mutant ygl13 in Rice (Oryza sativa)[J].Scientia Agricultura Sinica, 2015, 48(21): 4197-4208.

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