【摘要】：作為一種重要的成花誘導(dǎo)途徑,春化作用在大多數(shù)冬性或者二年生植物實(shí)現(xiàn)營(yíng)養(yǎng)生長(zhǎng)向生殖生長(zhǎng)的轉(zhuǎn)變過程中起著不可替代的作用。大白菜屬于典型的種子春化型植物,在種子萌發(fā)期便可感受低溫處理,進(jìn)而提前開花。育種上常利用春化作用來加快大白菜的培育進(jìn)程,但目前大白菜種子春化的內(nèi)在調(diào)控機(jī)理并不明了。因此,本研究運(yùn)用iTRAQ技術(shù)研究了大白菜萌發(fā)種子在春化過程中的蛋白表達(dá)情況,以期為揭示大白菜種子春化的調(diào)控機(jī)制提供一定依據(jù)。主要研究?jī)?nèi)容和結(jié)果如下:1.對(duì)大白菜雙單倍體'FT'萌發(fā)種子進(jìn)行不同時(shí)間春化處理,隨著春化天數(shù)的增加,大白菜所需抽薹開花的時(shí)間會(huì)變短,低溫處理25d可有效促進(jìn)'FT'度過春化。2.利用iTRAQ技術(shù)對(duì)大白菜25℃萌發(fā)24h種子(Ger),及該種子經(jīng)4℃春化25d的種芽(Ver)和25℃生長(zhǎng)60h的種芽(Nor,與Ver種芽等長(zhǎng))進(jìn)行蛋白質(zhì)組分析,共鑒定到4197個(gè)蛋白質(zhì),其中在Nor-VS-Ver、Ger-VS-Ver、Ger-VS-Nor中分別鑒定到641、604、644個(gè)差異表達(dá)蛋白。3.對(duì)Nor-VS-Ver中差異表達(dá)蛋白進(jìn)行GO和KEGG功能富集分析。結(jié)果表明,發(fā)生顯著變化的蛋白質(zhì)主要來源于細(xì)胞質(zhì)和細(xì)胞膜且作為一些蛋白酶或者分子伴侶等在大白菜體內(nèi)的碳水化合物代謝,蛋白質(zhì)代謝、次生物質(zhì)合成以及逆境響應(yīng)等生物過程中起重要作用。4.對(duì)Ger-VS-Ver和Ger-VS-Nor中的差異表達(dá)蛋白進(jìn)行Venn圖譜分析,篩選到329個(gè)Ger-VS-Ver特有差異表達(dá)蛋白,同樣進(jìn)行功能富集分析。結(jié)果表明,這些蛋白也涉及糖類、蛋白質(zhì)、氨基酸和脂類等物質(zhì)代謝,并且伴隨能量代謝和信號(hào)轉(zhuǎn)導(dǎo)等過程。5.采用Western blot技術(shù)檢測(cè)2個(gè)GDSL脂肪酶家族蛋白(顯著富集到類胡蘿卜素代謝通路)的表達(dá)情況,qRT-PCR技術(shù)檢測(cè)12個(gè)差異蛋白對(duì)應(yīng)的基因(植物響應(yīng)逆境、生長(zhǎng)發(fā)育、抗氧化等過程相關(guān))的表達(dá)情況,從蛋白水平和轉(zhuǎn)錄水平上驗(yàn)證了蛋白質(zhì)組定量分析的準(zhǔn)確性。6.對(duì)相同樣品的蛋白質(zhì)組和已有轉(zhuǎn)錄組進(jìn)行聯(lián)合分析,結(jié)果表明,差異表達(dá)蛋白與差異表達(dá)基因之間只有少數(shù)基因被關(guān)聯(lián)到且相關(guān)性較低,也暗示著大白菜種子春化過程中蛋白質(zhì)的表達(dá)存在轉(zhuǎn)錄后調(diào)控作用。
[Abstract]:As an important pathway of flower induction, vernalization plays an irreplaceable role in the transformation of vegetative growth to reproductive growth in most winter or biennial plants. Chinese cabbage (Brassica campestris L.) is a typical spring-seed plant, which can be treated with low temperature during seed germination, and then blossoms ahead of time. In breeding, vernalization is often used to speed up the cultivation of Chinese cabbage, but the internal regulation mechanism of Chinese cabbage seed vernalization is not clear at present. Therefore, iTRAQ technique was used to study the protein expression of germinating seeds of Chinese cabbage in the process of vernalization, in order to provide some basis for revealing the regulation mechanism of vernalization of Chinese cabbage seeds. The main contents and results of the study are as follows: 1. The time of bolting and flowering in Chinese cabbage (Brassica campestris L.) was shortened with the increase of the days of vernalization, and 25 days of low temperature treatment could effectively promote the 'FT' through vernalization. 2. The germination seeds of Chinese cabbage dihaploid' FT' were treated with vernalization for different time. The proteomic analysis of the seeds of Chinese cabbage (Brassica campestris L.) (Ger), at 25 鈩，

本文編號(hào)：2456911