Genetic Analysis of S5 Regulating the Hybrid Sterility between Indica and Japonica Subspecies in Rice

Author:

Guo Jie123,Zhou Xinqiao123,Chen Ke123ORCID,Ye Chanjuan123ORCID,Liu Juan123,Sun Kangli4,Tang Guang4,Wang Shaokui4ORCID,Zhang Guiquan4,Chen Youding123,Chen Dagang123,Liu Chuanguang123

Affiliation:

1. Key Laboratory of Genetics and Breeding of High Quality Rice in Southern China (Co-Construction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Guangzhou 510640, China

2. Guangdong Key Laboratory of New Technology in Rice Breeding, Guangzhou 510640, China

3. Guangdong Rice Engineering Laboratory/Rice Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China

4. Guangdong Provincial Key Laboratory of Plant Molecular Breeding, South China Agricultural University, Guangzhou 510640, China

Abstract

Hybrid sterility is the major obstacle to the utilization of inter-subspecific heterosis in hybrid rice breeding. The S5 locus, composed of three adjacent genes ORF3, ORF4, and ORF5, plays a crucial role in regulating indica/japonica hybrids’ female sterility. Through a series of crosses involving 38 parents, three alleles of S5, ORF3+ORF4−ORF5n, ORF3+ORF4+ORF5n, and ORF3−/ORF4−/ORF5n, all could be regarded as wide-compatibility alleles, and when crossed with indica or japonica rice, they all showed significantly high fertility. Then, in order to explore the genes’ function, we further knocked out genes by using CRISPR/Cas9-based genome editing. Our results demonstrate that the ORF3+ was not just the protector in the killer-protector system, and knocking out ORF3 of the indica allele seriously affected the rice’s normal development. We observed the concrete enhancing hybrid spikelet fertility from the crosses between the ORF4+ knockout japonica materials with indica varieties. By conducting the comparative RNA-Seq analysis of young spikelets, we found that the ORF4+/ORF4− could modulate the hybrid fertility by affecting the expressions of genes related to the function of the Golgi apparatus. This study indicated that knocking out the ORF4+ of the japonica allele or using the alleles carrying ORF5n would provide effective approaches to overcome indica/japonica hybrid female sterility in rice breeding.

Funder

National Natural Science Foundation of China

Natural Science Foundation of Guangdong Province, China

Special fund for scientific innovation strategy-construction of high-level Academy of Agriculture Science

Guangdong Key Laboratory of New Technology in Rice Breeding

Publisher

MDPI AG

Subject

Agronomy and Crop Science

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