擬南芥雄性育性相關(guān)基因的功能分析
發(fā)布時(shí)間：2018-07-18 20:58
【摘要】：植物繁殖和選擇育種的一個(gè)重要目標(biāo)是植株雄性育性的調(diào)控。對(duì)雄性不育分子機(jī)制的深入理解將成為控制雄性育性和雜種繁育一種有效的途徑。ms606是本實(shí)驗(yàn)室在前期工作中鑒定的一個(gè)擬南芥雄性不育突變株系。ms606突變體植株敗育,藥室內(nèi)壁次生加厚異常,花藥不開裂,表明MS606與藥室內(nèi)壁的木質(zhì)化有關(guān)。MS35/MYB26是木質(zhì)素生物合成途徑上游一個(gè)重要的激活因子,為了進(jìn)一步了解MS606和MYB26在調(diào)控花藥藥室內(nèi)壁次生加厚過(guò)程中的作用,本論文工作以ms35為母本,ms606為父本,進(jìn)行了人工雜交得到雜交子二代(F2),并對(duì)F2代植株的基因型、表型和基因表達(dá)進(jìn)行鑒定,獲得了一個(gè)純合的ms35ms606雙突變體,這為進(jìn)一步分析MS606和MS35在調(diào)控藥室內(nèi)壁次生加厚中的作用提供了寶貴的實(shí)驗(yàn)材料。生長(zhǎng)素在雄蕊發(fā)育過(guò)程中有著重要作用,為了探討MS606是否通過(guò)生長(zhǎng)素途徑調(diào)控雄蕊發(fā)育,本論文工作用不同濃度的IAA處理野生型和突變體的花器官;構(gòu)建并鑒定出ms606背景的DR5:GUS植株,并對(duì)其花藥進(jìn)行GUS染色;檢測(cè)不同時(shí)期花蕾中生長(zhǎng)素的含量;對(duì)花蕾中生長(zhǎng)素合成基因(YUC2、YUC6)和運(yùn)輸基因(PIN2、PIN7和AUX1)的表達(dá)進(jìn)行分析。結(jié)果顯示,不同濃度的IAA處理ms606無(wú)法恢復(fù)突變體花藥開裂缺陷表型。GUS染色表明,野生型和ms606花藥中生長(zhǎng)素表達(dá)的時(shí)期相同,均在花發(fā)育的第10時(shí)期開始表達(dá),12時(shí)期后期無(wú)GUS信號(hào),但在第11時(shí)期,突變體花藥中GUS活性明顯強(qiáng)于野生型,且對(duì)花蕾中生長(zhǎng)素的測(cè)定發(fā)現(xiàn)突變體第10-11時(shí)期含量顯著高于野生型。利用RT-PCR和qRT-PCR分析發(fā)現(xiàn)ms606花蕾中生長(zhǎng)素合成基因YUC2和YUC6的表達(dá)量顯著降低,PIN2和PIN7的表達(dá)也發(fā)生改變,但輸入載體AUX1的表達(dá)無(wú)差異,這些實(shí)驗(yàn)成果均表明MS606基因突變可在一定程度上影響到花蕾中的生長(zhǎng)素途徑基因的表達(dá)。除此之外,突變體中茉莉酸誘導(dǎo)途徑基因(MYB21、MYB24、MYB108)和絨氈層發(fā)育基因(AMS、ABCG26)的表達(dá)均發(fā)生變化。由此可見,MS606功能缺失可對(duì)多個(gè)途徑產(chǎn)生影響,但詳細(xì)的作用機(jī)制仍需進(jìn)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.
【學(xué)位授予單位】：山西大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：Q943.2
本文編號(hào)：2132755
[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.
【學(xué)位授予單位】：山西大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：Q943.2
【參考文獻(xiàn)】
相關(guān)期刊論文 前3條
1 ;A Genetic Pathway for Tapetum Development and Function in Arabidopsis[J];Journal of Integrative Plant Biology;2011年11期
2 ;AtMYB103 is a crucial regulator of several pathways affecting Arabidopsis anther development[J];Science China(Life Sciences);2010年09期
3 毛健民,李俐俐;擬南芥——植物界的“果蠅”[J];生物學(xué)通報(bào);2001年12期
，本文編號(hào)：2132755
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