水稻OsMS5-1/OsMS5-2双突变体花粉形态电镜扫描观察

Electron microscopy scanning observations on rice OsMS5-1/OsMS5-2 double-mutant pollen

【目的】观察水稻MS5同源基因OsMS5-1/OsMS5-2双突变体的形态特征,为进一步研究水稻MS5同源基因OsMS5-1、OsMS5-2在水稻雄性育性中的功能提供参考。【方法】以前期通过RNAi干扰技术创制的两个水稻MS5同源基因OsMS5-1、OsMS5-2的粳稻中花11干扰单突变体T1代为亲本,将其进行正反杂交以创制双突变体,以野生型粳稻中花11成熟花粉粒为参照,对双突变体的花粉粒进行碘—碘化钾染色及电镜扫描观察。【结果】正反交双突变体的花粉粒均未被染色,呈细小、不规则、透明状,其不育度可达100%,而正常可育的野生型花粉粒被染为深蓝色,呈较大而规则的正圆形。双突变体的花粉粒存在发育缺陷,其外形、萌发孔和花粉粒外壁均明显不同于正常可育的野生型中花11的花粉粒。【结论】水稻OsMS5-1及OsMS5-2基因双突变体花粉粒形态特征与野生型可育花粉粒存在一定差异,但两种双突变体不育花粉粒总体上差别不明显,可作为鉴别早期不育与可育花粉的重要指标。

[Objective]The rice MS5 homologous genes OsMS5-1/OsMSS-2 double mutant pollen morphology was ob- served and studied in order to further understand the MS5 homologous genes OsMSS-1, OsMS5-2 in rice and its function- al effects on male rice sterility. [ Method ] Utilizing Zhonghua 11 ＇ s single interference mutant TI, created from the 2 rice MS5 homologous genes OsMSS-1 and OsMSS-2 using RNAi interference, as parent, the gene was reciprocally crossed to create double mutants; then, using wild Zhonghua 11 mature pollen grains as reference, the double mutant pollen grains were examined by I2-KI staining and electron microscopy scanning. [Result]Reciprocal double mutant pollen grains were not stained, small, irregularly shaped, transparent, and measured with a sterile rate up to 100%, while the normal tertile wild pollen grains were dyed dark blue, large, and regularly shaped. The double mutant pollen grains were considered developmental defects; the grains＇ shape, aperture, and exine were significantly different from the normal fertile wild Zhonghua 11 pollen grains. [ Conclusion ]The morphologies of rice OsMSS-I and OsMSS-2 gene mutant pollen grain and the wild fertile pollen grains exhibited certain characteristic differences, while the two double mutant sterile male pollen grains exhibited no significant differences in general ; this result could be used to distinguish early sterile and fertile pollen.