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Use of RNAi With OsMYB76R as a Reporter for Candidate Genes Can Efficiently Create and Verify Gametophytic Male Sterility in Rice

文献类型： 外文期刊

作者： Chen, Yun 1 ; Zhu, Wenping 1 ; Shi, Shudan 1 ; Wu, Lina 2 ; Du, Shuanglin 1 ; Jin, Liangshen 1 ; Yang, Kuan 1 ; Zhao, Wenjia 1 ; Yang, Jiaxin 1 ; Guo, Longbiao 3 ; Wang, Zhongwei 4 ; Zhang, Yi 1 ;

作者机构： 1.Yunnan Univ, Sch Agr, State Key Lab Conservat & Utilizat Bioresources Y, Res Ctr Perennial Rice Engn & Technol Yunnan, Kunming, Yunnan, Peoples R China

2.Southwest Univ, Coll Agron & Biotechnol, Chongqing, Peoples R China

3.China Natl Rice Res Inst, State Key Lab Rice Biol, Hangzhou, Peoples R China

4.Chongqing Acad Agr Sci, Biotechnol Res Ctr, Chongqing, Peoples R China

关键词： Oryza sativa; gametophytic male sterility; OsMYB76R; RNAi; GMS preparation and identification

期刊名称：FRONTIERS IN PLANT SCIENCE （ 影响因子：5.754； 五年影响因子：6.612 ）

ISSN： 1664-462X

年卷期： 2021 年 12 卷

页码：

收录情况： SCI

摘要： Gametophytic male sterility (GMS) plays an important role in the study of pollen development and seed propagation of recessive nuclear male sterile lines insensitive to the environmental conditions in hybrid rice breeding. Since the inherent phenotypic and genetic characteristics of GMS, it is very difficult to find and identify the GMS mutants. However, due to the abundance of gene transcription data, a large number of pollen-specific genes have been found, and most of them may be associated with GMS. To promote the study of these genes in pollen development and heterosis utilization, in this study, an easy and efficient method of creating and identifying GMS was established using RNAi and OsMYB76R as a reporter. First, the OsC1/OsMYB76 gene involved in anthocyanin synthesis was modified, and we have validated that the modified OsMYB76R is workable as the same as the pre-modified OsMYB76 gene. Then, the ascorbic acid oxidase gene OsPTD1 was downregulated using RNAi, driven by its own promoter that resulted in abnormal pollen tube growth. Finally, the RNAi elements were linked with OsMYB76R and transformed into an osmyb76 mutant, and the distortion of purple color segregation was found in T-1 and F-1 generations. This indicates that the OsPTD1 GMS was prepared successfully. Compared to current methods, there are several advantages to this method. First, time is saved in material preparation, as one generation less needs to be compared than in the conventional method, and mutation screening can be avoided. In addition, for identification, the cost is lower; PCR, electrophoresis, and other processes are not needed; and no expensive chemicals or instruments are required. Finally, the results are more accurate, with much lower background effects, and no damage to the plant. The result is an easy, efficient, low-cost, and accurate method of preparing and identifying GMS genes.

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