拟南芥At5g58100基因T-DNA纯合插入突变体PCR鉴定及表型观察

PCR identification and phenotype observation of At5g58100 gene T-DNA homozygous insertion mutants of Arabidopsis thaliana

以拟南芥SPOT1功能缺失突变体spot 1和野生型(WT)植株为材料,比较研究了它们在花粉发育和结实率方面的差异.spot 1-1,spot 1-2和spot 1-3突变体都能形成可育的三核花粉,但用扫描电子显微镜(SEM)观察发现其花粉粒皱缩,花粉外壁存在纹理缺陷.用荧光显微镜观察了这些花粉粒的树脂半薄切片,发现花粉内壁无纤维素的自发荧光,spot 1突变体成熟果荚中的种子数量显著少于野生型.对spot 1-3突变体花药发育过程进行观察后发现,从花粉母细胞时期开始,突变体细胞壁组成与野生型已出现差异.10μmol·L^-1(物质的量浓度)茉莉酮酸既不能抑制黄化spot 1幼苗的顶钩反应,又不能抑制1×10^-6(质量分数)乙烯利引起的夸张顶钩反应.上述结果提示SPOT 1基因的功能与小孢子初生外壁、花粉内壁的发育,以及花丝伸长(长度)有密切关系,SPOT1可能与茉莉酮酸信号传导通路相关联.

The differences between the spot 1 mutants and wild type(WT)in their pollen development and spikelet fertility were compared in the paper.The pollen grains with spot 1 mutants(spot 1-1,spot 1-2 and spot 1-3)were shrunken and spotty in exine pattern by scan electron microscopy(SEM)observation,although they can all develop into male fertile pollens.There was no blue fluorescence in pollen wall at trans-section pollen grains by fluorescence microscope analysis.The average number of seeds in mature fruit pods with mutants is significant less than that of wild type because of insufficient pollination.The earliest change in the cell wall component occurs at the microspore mother cell stage of anther development by observation the sequential transverse sections of the spot 1-3 mutant flower inflorescence.10μmol·L^-1 jasmonic acid treatment can neither inhibit the curvature of apical hook of etiolated spot 1 mutant seedling,nor inhibit ethylene from promoting the formation of apical hook of etiolated spot 1 mutant seedling.The results indicated that the function of SPOT1 gene might involve in the construction development of primexine and intine,and in filament elongation.SPOT1 gene might be related to jasmonic acid signal transduction pathway.