Chinese Journal OF Rice Science >
Identification and Gene Cloning of DSP2 in Rice (Oryza sativa L.)
Received date: 2021-06-17
Revised date: 2021-09-14
Online published: 2022-03-11
【Objective】 Plant height and panicle length play important roles in rice production. Identifying and cloning genes related to rice plant height and panicle architecture will enrich the molecular mechanism of the regulation of rice plant height and panicle development, and will provide a theoretical foundation and genetic resources for molecular design breeding. 【Method】 A dwarf and small panicle mutant dsp2-D(dwarf and small panicle2-Dominant) was obtained from T-DNA insertion population of Nipponbare. The main agronomic traits of dsp2-D and the wild-type were characterized under conventional planting conditions in field. An F2 mapping population was generated by crossing the dsp2-D mutant with Longtefu B (an indica rice variety) and DSP2 was identified by map-based cloning and T-DNA tag method. The candidate gene was determined by the relative expression analysis of three genes located upstream and downstream of the T-DNA insertion site, respectively, which was carried out by real-time RT-PCR. The function of DSP2 was confirmed by overexpressing DSP2 (DSP2-OE) in wild-type Nipponbare.【Result】 The plant height, panicle length, the length of primary and secondary inflorescence branches of dsp2-D was dramatically reduced compared with the wild type. Genetic analysis indicated that the phenotypes of dsp2-D were regulated by an incomplete dominant gene. The DSP2 gene was mapped to a region between RM208 and RM7337 on chromosome 2, and co-segregated with RM3850. Subsequently, T-DNA insertion site was found to be co-segregated with dsp2-D phenotype, and T-DNA insertion sequence was separated by TAIL-PCR, which showed that T-DNA was inserted into the two genes between RM208 and RM7337. Real-time PCR analysis indicated that the expression level of LOC_Os02g57490, which located down-stream of T-DNA site, was dramatically up-regulated. While the expression levels of the rest five detected genes were not significantly changed in dsp2-D. DSP2 (DSP2-OE) overexpressing transgenic plants showed semi-dwarf and small panicle,which were similar with the phenotypes in dsp2-D. The results confirmed that DSP2 gene was the target gene controlling dsp2-D.【Conclusion】 The overexpression of DSP2 caused the mutant phenotypes of dsp2-D. And we found that DSP2 encodes a LOB family transcript factor and negatively regulates plant height and panicle length in rice. Our findings will open new perspectives on DSP2-based improvement of plant height and panicle type of rice.
Key words： rice; dwarf; panicle length; mutant; gene cloning
QIU Linlin, LIU Qiao, FU Yaping, LIU Wenzhen, HU Guocheng, ZHAI Yufeng, PANG Bo, WANG Dekai . Identification and Gene Cloning of DSP2 in Rice (Oryza sativa L.)[J]. Chinese Journal OF Rice Science, 2022 , 36(2) : 150 -158 . DOI: 10.16819/j.1001-7216.2022.210606
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