OsGAMYBL2 is required for pollen maturation and germination in rice

Yichen Xiao , Yulu Zhou , Jianxin Shi , Dabing Zhang
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Abstract

The reproductive development process of rice is not only important for basic developmental biology research, but also of great benefit to production practice. However, current understanding of the reproductive growth process is relatively limited, especially in the later stages of development. Here we report that OsGAMYBL2, a homologous gene of GAMYB, is required for pollen maturation and germination in rice, and that loss of function of OsGAMYBL2 leads to male sterility in rice. We generated three alleles of OsGAMYBL2 knockout mutants using CRISPR/Cas9 system, all of them lost fertility when grown in the paddy field. Semi-section and electron microscopy observations revealed that there are no visible morphological changes in the outer and inner surfaces of anther and in the surface of mature pollens between mutants and wild type. However, pollens at late developmental stages exhibited much lighter pollen viability staining and some of them shriveled. In vitro and in vivo germination experiments showed that mutant pollens fail to germinate, leading to abnormal pollen tube formation and subsequent defective pollination. Furthermore, we observed that as compared with high temperature at 30 ​°C, low-temperature at 22 ​°C affects less on osgmybl2 fertility. Collectively, we proposed that OsGAMYBL2 is likely involved in the regulation of starch accumulation during later pollen development with a thermal-sensitive way, which improves our understanding on the reproductive development in plants.

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水稻花粉成熟和萌发需要OsGAMYBL2
水稻的生殖发育过程不仅在基础发育生物学研究中具有重要意义,而且在生产实践中具有重要意义。然而,目前对生殖生长过程的了解相对有限,特别是在发育的后期阶段。本文报道了水稻花粉成熟和萌发需要同源基因OsGAMYBL2, OsGAMYBL2功能缺失导致水稻雄性不育。我们利用CRISPR/Cas9系统产生了3个OsGAMYBL2基因敲除突变体等位基因,它们在水田生长时都失去了育性。半切片和电镜观察显示,突变体与野生型的花药内外表面和成熟花粉表面没有明显的形态变化。而发育后期花粉活力染色较轻,部分花粉枯萎。体外和体内萌发实验表明,突变花粉无法萌发,导致花粉管形成异常,进而导致授粉缺陷。此外,我们观察到,与30°C的高温相比,22°C的低温对osgmybl2的育性影响较小。综上所述,我们提出OsGAMYBL2可能以热敏的方式参与了花粉发育后期淀粉积累的调控,从而提高了我们对植物生殖发育的认识。
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