拟南芥雄性育性相关基因的功能分析

发布时间：2018-07-18 20:58

【摘要】：植物繁殖和选择育种的一个重要目标是植株雄性育性的调控。对雄性不育分子机制的深入理解将成为控制雄性育性和杂种繁育一种有效的途径。ms606是本实验室在前期工作中鉴定的一个拟南芥雄性不育突变株系。ms606突变体植株败育,药室内壁次生加厚异常,花药不开裂,表明MS606与药室内壁的木质化有关。MS35/MYB26是木质素生物合成途径上游一个重要的激活因子,为了进一步了解MS606和MYB26在调控花药药室内壁次生加厚过程中的作用,本论文工作以ms35为母本,ms606为父本,进行了人工杂交得到杂交子二代(F2),并对F2代植株的基因型、表型和基因表达进行鉴定,获得了一个纯合的ms35ms606双突变体,这为进一步分析MS606和MS35在调控药室内壁次生加厚中的作用提供了宝贵的实验材料。生长素在雄蕊发育过程中有着重要作用,为了探讨MS606是否通过生长素途径调控雄蕊发育,本论文工作用不同浓度的IAA处理野生型和突变体的花器官;构建并鉴定出ms606背景的DR5:GUS植株,并对其花药进行GUS染色;检测不同时期花蕾中生长素的含量;对花蕾中生长素合成基因(YUC2、YUC6)和运输基因(PIN2、PIN7和AUX1)的表达进行分析。结果显示,不同浓度的IAA处理ms606无法恢复突变体花药开裂缺陷表型。GUS染色表明,野生型和ms606花药中生长素表达的时期相同,均在花发育的第10时期开始表达,12时期后期无GUS信号,但在第11时期,突变体花药中GUS活性明显强于野生型,且对花蕾中生长素的测定发现突变体第10-11时期含量显著高于野生型。利用RT-PCR和qRT-PCR分析发现ms606花蕾中生长素合成基因YUC2和YUC6的表达量显著降低,PIN2和PIN7的表达也发生改变,但输入载体AUX1的表达无差异,这些实验成果均表明MS606基因突变可在一定程度上影响到花蕾中的生长素途径基因的表达。除此之外,突变体中茉莉酸诱导途径基因(MYB21、MYB24、MYB108)和绒毡层发育基因(AMS、ABCG26)的表达均发生变化。由此可见,MS606功能缺失可对多个途径产生影响,但详细的作用机制仍需进一步深入研究。
[Abstract]:One of the important goals of plant breeding and selective breeding is the regulation of plant male fertility. An in-depth understanding of the molecular mechanism of male sterility will be an effective way to control male fertility and hybrid breeding. Ms606 is a sterile mutant of Arabidopsis thaliana. The secondary thickening of the chamber wall was abnormal and the anther was not cracked, which indicated that MS606 was an important activator in the upstream of lignin biosynthesis pathway. In order to further understand the role of MS606 and MYB26 in the regulation of secondary wall thickening of anthers, the second generation of hybrid (F2) was obtained by artificial hybridization with ms35 as the female parent of MS606. A homozygous ms35ms606 double mutant was obtained by identification of phenotype and gene expression, which provides valuable experimental materials for further analysis of the role of MS606 and MS35 in regulating the secondary wall thickening. Auxin plays an important role in stamen development. In order to investigate whether MS606 regulates stamen development through auxin pathway, different concentrations of IAA were used to treat flower organs of wild type and mutant. The plant of DR5: Gus with ms606 background was constructed and identified, and its anthers were stained with Gus. The contents of auxin in buds at different stages were detected, and the expression of auxin synthase genes (YUC6) and transporter genes (PIN2PIN7 and AUX1) in flower buds were analyzed. The results showed that different concentrations of ms606 could not restore the anther dehiscence phenotype. Gus staining showed that the expression of auxin in wild type and ms606 anther was the same period. All of them expressed no Gus signal at the 10th stage of flower development, but at the 11th stage, the Gus activity in the anthers of the mutant was significantly stronger than that in the wild type. The content of auxin in flower bud was significantly higher than that in wild type in 10-11 period. RT-PCR and qRT-PCR analysis showed that the expression of auxin synthesis genes YUC2 and YUC6 in ms606 buds also decreased significantly, but the expression of AUX1 was not different in the input vector AUX1. These results indicated that MS606 gene mutation could affect the expression of auxin pathway genes in flower buds to some extent. In addition, the expression of Jasmonic acid inducible pathway gene (MYB21, MYB24, MYB108) and tapetum development gene (AMS-ABCG26) were all changed in the mutant. It can be seen that the absence of MS606 function can affect many ways, but the detailed mechanism still needs to be further studied.
【学位授予单位】：山西大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q943.2

【参考文献】