Chinese Journal OF Rice Science >
Map-Based Cloning of OsCAD2 Regulating Golden Hull and Internode in Rice
Received date: 2016-12-23
Revised date: 2017-02-08
Online published: 2017-09-10
【Objective】Rice pigments not only have important physiological effects on their own development, but also have been widely used in rice breeding, by-product improvement, and so on. Phenotypic analysis and gene mapping of pigment-related genes could lay the foundation for the further study of rice pigments metabolism and regulation mechanism. 【Method】A golden hull and internode rice mutant, gh881, was isolated from an elite japonica cultivar Changlijing (CLJ) using ethyl methane sulphonate(EMS) mutagenesis strategy;the main agricultural traits of gh881 and its wild type(WT) were measured at mature stage; we crossed gh881 with WT and ZH8015, respectively, followed by observation of F1 and BC1F1 plants phenotype, and we made genetic analysis using Chi-square test in BC1F2 population and gene mapping using map-based cloning in F2 population; the related-genes expression of young panicles, internodes and flag leaf sheaths of gh881 and WT at various stages were studied by qPCR. 【Result】Compared with wild type, gh881 exhibited golden hull and internode at the mature stage. Except for no significant difference in the number of panicles per plant, there were significant reduction in plant height, the seed-setting rate, the number of spikelets per panicle, the number of grains per panicle and 1000-grain weight between WT and gh881. Genetic analysis and gene fine-mapping results suggested that gh881 was controlled by a single recessive gene and the mutant gene was mapped to a region of 33.2 kb in which four open reading flames (ORFs) existed on the short arm of chromosome 2 between markers FH-13 and RH-25. 【Conclusion】Sequencing analysis revealed that a single base mutation (G to A) occurred at the site 3563 bp of OsCAD2 (Os02g0187800) encoding a Cinnamyl-Alcohol Dehydrogenase (CAD), which led to a G297D mutation (codon GGC to GAC). This implied that gh881 might carry a novel allele of OsCAD2. The quantitative real-time PCR analysis showed that the relative expression level of OsCAD2 decreased significantly in the internode, while it almost increased significantly in the flag leaf sheath and panicle, and the related-genes also almost changed significantly. These results demonstrated that OsCAD2 is an important gene involved in lignin metabolic pathway, and may regulate other related-genes expression feedback.
Hong WANG, Yingxin ZHANG, Lianping SUN, Shuai MENG, Peng XU, Weixun WU, Shihua CHENG, Liyong CAO, Xihong SHEN . Map-Based Cloning of OsCAD2 Regulating Golden Hull and Internode in Rice[J]. Chinese Journal OF Rice Science, 2017 , 31(5) : 465 -474 . DOI: 10.16819/j.1001-7216.2017.6170 465
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