Institute of Molecular Biology, National Chung Hsing University, Taichung, Taiwan.
Biodiversity Research Center, Academia Sinica, Taipei, Taiwan.
PLoS One. 2023 Oct 9;18(10):e0292400. doi: 10.1371/journal.pone.0292400. eCollection 2023.
The rice GA biosynthetic gene OsGA3ox1 has been proposed to regulate pollen development through the gametophytic manner, but cellular characterization of its mutant pollen is lacking. In this study, three heterozygotic biallelic variants, "-3/-19", "-3/-2" and "-3/-10", each containing one null and one 3bp-deletion allele, were obtained by the CRISPR/Cas9 technique for the functional study of OsGA3ox1. The three homozygotes, "-19/-19", "-2/-2" and "-10/-10", derived from heterozygotic variants, did not affect the development of most vegetative and floral organs but showed a significant reduction in seed-setting rate and in pollen viability. Anatomic characterizations of these mutated osga3ox1 pollens revealed defects in starch granule accumulation and pollen wall development. Additional molecular characterization suggests that abnormal pollen development in the osga3ox1 mutants might be linked to the regulation of transcription factors OsGAMYB, OsTDR and OsbHLH142 during late pollen development. In brief, the rice GA3ox1 is a crucial gene that modulates pollen starch granule accumulation and pollen wall development at the gametophytic phase.
水稻 GA 生物合成基因 OsGA3ox1 被提出通过配子体方式调控花粉发育,但缺乏对其突变花粉的细胞特征描述。在这项研究中,通过 CRISPR/Cas9 技术获得了三个杂合双等位变异体,“-3/-19”、“-3/-2”和“-3/-10”,每个变异体都包含一个无效和一个 3bp 缺失等位基因,用于 OsGA3ox1 的功能研究。三个纯合子,“-19/-19”、“-2/-2”和“-10/-10”,来源于杂合变异体,不影响大多数营养器官和花器官的发育,但种子结实率和花粉活力显著降低。对这些突变的 Osga3ox1 花粉进行的解剖特征描述显示,淀粉粒积累和花粉壁发育存在缺陷。额外的分子特征表明,Osga3ox1 突变体中异常的花粉发育可能与晚期花粉发育过程中转录因子 OsGAMYB、OsTDR 和 OsbHLH142 的调节有关。总之,水稻 GA3ox1 是一个关键基因,它在配子体阶段调节花粉淀粉粒积累和花粉壁发育。