一个水稻早衰突变体基因的精细定位

doi:10.3864/j.issn.0578-1752.2014.11.001

Abstract

Abstract: 【Objective】The objective of this study is to conduct genetic analysis and gene mapping for the rice early leaf senescence mutant W330.【Method】An early leaf senescence mutant, designated as W330, was isolated from the progeny of 60Co-γ-treated indica rice cv. Zh8015. Phenotypes of the W330 mutant were observed and its main agronomic traits were analyzed under field conditions in Hangzhou and Sanya. After the W330 mutant was crossed with 02428, leaf phenotypes of the F1 progenies，the segregation ratio of normal and early senescence plants in the F2 populations were investigated. An F2 population derived from the cross of cv. 02428 with W330 was used for genetic analysis and gene fine mapping. Candidate gene assay was conducted by gene sequencing, enzyme digestion, gene expression and phylogenetic analysis.【Result】W330 plants showed leaf senescence phenotype from 3-leaf-stage to maturity stage. Compared with wild-type parent Zhonghui8015, the plant height，tiller and blade width of W330 decreased significantly, and the number of productive panicles per plant, the number of spikelets per panicle and seed setting rate were also significantly reduced. All F1 plants generated by crossing early senescence mutant W330 with 02428 showed normal leaf. The segregation ratios of normal plants and early senescence plants in two F2 populations were both 3:1, indicating that the W330 was controlled by a single recessive nuclear gene which was mapped on the short arm of chromosome 3. With developed SSR and Indel markers, the gene was finally narrowed to an interval of 21.5 kb between markers CD-5 and CD-7. Among four ORFs in this region, the LOC_Os03g0131200 encoding a Catalase (OsCATC) was probably the candidate gene. The sequencing analysis and enzyme digestion revealed the variable splicing in the first intron was probably responsible for the early senescence phenotype, which was confirmed by assay of gene expression and enzyme activity. Compared with wild-type parent Zhonghui8015，the content of catalase activity decreased by 47.8% and the content of H2O2 increased by 2.7 times. It was speculated that the W330 mutant gene was allelic to OsCATC gene. Otherwise, phylogenetic analysis revealed that OsCATC was an independent evolution in rice. Expression analysis of catalase genes by real-time PCR showed that OsCATA and OsCATB have a significant increase in the expression of leaves, while the expression of OsCATC has no significant changes. It was a speculated that the three highly homologous genes have complementary mechanisms in rice plants. 【Conclusion】 The W330 mutant gene was allelic to OsCATC gene. A point mutation in introns of OsCATC gene in the W330 mutant makes catalase inactive, resulting in the leaf senescence phenotype.

Key words: Oryza sativa L. , early senescence , fine mapping , variable splicing

ZHAO Chun-De-1, 2 , ZHANG Ying-Xin-1, 2 , LIU Qun-恩1, 2 , YU Ning-1, 2 , CHENG Shi-Hua-1, 2 , CAO Li-Yong-1, 2 . Fine Mapping of an Early Senescence Gene in Rice[J].Scientia Agricultura Sinica, 2014, 47(11): 2069-2077.

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