Creating a Fragrant Environment-insensitive Recessive Genic Male Sterile Germplasm Lacking Coleoptile Purple Lines by CRISPR/Cas9 Gene Editing

doi:10.16819/j.1001-7216.2025.240501

Abstract

Abstract: 【Objective】Hybrid rice has developed into the third generation in which environment-insensitive recessive genic male sterility (EI-RGMS) is the essential trait. In many rice breeding programs, fragrance improvement is an important target. Additionally, the purple line in coleoptile is a clearly identifiable anthocyanin trait at the very early germination stage, which facilitates simple, rapid, and accurate identification of hybrid purity. The purpose of this study is to develop a fragrant EI-RGMS material lacking the coleoptile purple line based on maintainer II-32B, providing a new core germplasm for the third-generation hybrid rice.【Methods】 Using CRISPR/Cas9, we knocked out the pollen development gene OsbHLH141, fragrance gene OsBadh2, and coleoptile purple line gene OsMYB76 in the elite rice parent II-32B.【Results】 Mutant lines with all three genes knocked out showed loss of anthocyanin traits, a distinct aroma, and male sterility.【Conclusion】 This study successfully created a fragrant EI-RGMS germplasm without purple line in coleoptile in the II-32B background.

Key words: environment-insensitive, recessive genic male sterility, fragrance, purple line in coleoptile, CRISPR/Cas9

摘要：

【目的】水稻杂种优势利用技术已发展到第三代，环境非敏感型隐性核雄性不育是第三代杂交稻的必需性状，而香味是许多水稻育种计划的重要目标。另外，水稻芽鞘紫线是极早期表现的一种明显稳定的花青素性状，有利于杂交种纯度的简单、快速和准确鉴定。本研究目的是将II-32B制备成香型、无芽鞘紫线的环境非敏感型隐性核雄性不育材料，为研发方便纯度鉴定的香型第三代杂交稻创造核心种质。【方法】利用CRISPR/Cas9技术，同时对优良水稻亲本II-32B的花粉发育相关基因OsbHLH141、香味基因OsBadh2以及芽鞘紫线基因OsMYB76进行敲除。【结果】获得了三个基因同时发生突变的材料，突变体表现为芽鞘紫线等花青素性状缺失，稻米具有香味且雄性不育。【结论】本研究成功获得了以II-32B为背景的香型、无芽鞘紫线的环境非敏感型隐性核雄性不育种质。

关键词:

text-indent:0cm, "> 环境非敏感；隐性核雄性不育；香味；芽鞘紫线；CRISPR/Cas9

YANG Jiaxin, GUAN Yusheng, DU Run, LI Xianyong, CAI Zuokun, WANG Chutao, YANG Qiyang, HE Yongxin, ZHU Zichao, ZHANG Yi . Creating a Fragrant Environment-insensitive Recessive Genic Male Sterile Germplasm Lacking Coleoptile Purple Lines by CRISPR/Cas9 Gene Editing[J]. Chinese Journal OF Rice Science, 2025, 39(5): 643-649.

杨佳欣, 管玉圣, 杜润, 李贤勇, 蔡座坤, 王楚桃, 阳启样, 何永歆, 朱子超, 张毅. 利用CRISPR/Cas9技术创制无芽鞘紫线的香型环境非敏感隐性核雄性不育种质[J]. 中国水稻科学, 2025, 39(5): 643-649.

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