發(fā)布時(shí)間：2018-04-04 17:58

  本文選題：近等基因系　切入點(diǎn)：SSR分子標(biāo)記　出處：《山西大學(xué)》2016年碩士論文

【摘要】：油菜是中國(guó)主要種植的油料作物,對(duì)我國(guó)國(guó)民經(jīng)濟(jì)的發(fā)展有著重要的作用,提高油菜的油分含量一直是油菜育種研究的首要任務(wù)。WRKY轉(zhuǎn)錄因子作為一種調(diào)控因子在植物調(diào)控網(wǎng)絡(luò)中占據(jù)著重要地位,參與植物的生長(zhǎng)發(fā)育、抗逆、抗病等一系列的調(diào)控過(guò)程。我們通過(guò)轉(zhuǎn)錄組測(cè)序和分析得到了兩個(gè)與含油量有關(guān)的WRKY轉(zhuǎn)錄因子BnWRKY52、BnWRKY52。本實(shí)驗(yàn)利用高代自交姐妹系構(gòu)建了兩個(gè)高含油量和中低含油量的近等基因系,并以這兩個(gè)近等基因系材料為基礎(chǔ)材料,初步研究了BnWRKY52、BnWRKY16在油菜種子成熟與萌發(fā)過(guò)程中的表達(dá)情況。具體情況如下:1.以NJ9高油品系為基礎(chǔ)材料,利用連續(xù)自交的方法構(gòu)建含油量差異的擬近等基因系。以來(lái)源于蕓薹屬Brassica DB公共數(shù)據(jù)庫(kù)的156對(duì)SSR引物對(duì)9個(gè)擬近等基因系株系材料進(jìn)行擴(kuò)增,篩選出32對(duì)條帶清晰、重復(fù)性好的引物用于近等性分析。基于32對(duì)SSR引物PCR擴(kuò)增結(jié)果,經(jīng)過(guò)遺傳和似性系數(shù)估算以及聚類分析鑒定近等基因系的近等性,結(jié)果顯示9個(gè)候選株系材料遺傳相似性系數(shù)均在0.85到0.94之間,其中YC13-554和YC13-559的遺傳相似性系數(shù)最大為0.94,確定為近等基因系。而后對(duì)這兩個(gè)材料成熟期的農(nóng)藝性狀進(jìn)行調(diào)查,結(jié)果進(jìn)一步確定了YC13-554和YC13-559為高(中低)含油量近等基因系。2.以含油量差異的近等基因系材料YC13-554和YC13-559為基礎(chǔ)材料,利用實(shí)時(shí)定量PCR的方法,檢測(cè)轉(zhuǎn)錄因子BnWRKY52、BnWRKY16在種子成熟過(guò)程中的基因表達(dá)量。結(jié)果顯示,在種子成熟的不同時(shí)期BnWRKY52在油菜YC13-554和YC13-559中的表達(dá)均呈現(xiàn)上升趨勢(shì),且在油菜YC13-559中表達(dá)量比在YC13-554中高;WRKY16在油菜YC13-554和YC13-559的表達(dá)均只出現(xiàn)一個(gè)較大的表達(dá)峰,在油菜YC13-559中的表達(dá)峰出現(xiàn)早于油菜YC13-554中。據(jù)以上結(jié)果表明,轉(zhuǎn)錄因子WKY52和WRKY16與油菜油分合成的調(diào)控有關(guān)。為驗(yàn)證所得結(jié)果,選擇了三個(gè)已確定的與油菜含油量有關(guān)的關(guān)鍵酶的基因二�；视王；D(zhuǎn)移酶(DTAG)、乙酰輔酶A羧化酶、Wril,檢測(cè)其表達(dá)量,并使BnWRKY52、BnWRKY16表達(dá)量與之進(jìn)行對(duì)比。結(jié)果顯示,兩者之間表達(dá)模式相似。3.以近等基因系材料YC13-554和YC13-559為基礎(chǔ)材料,利用實(shí)時(shí)定量PCR技術(shù)檢測(cè)轉(zhuǎn)錄因子BnWRKY52、BnWRKY16在油菜種子萌發(fā)過(guò)程中的基因表達(dá)量。結(jié)果顯示,在種子萌發(fā)的不同時(shí)期,BnWRKY52、BnWRKY16在油菜YC13-554與油菜YC13-559中的表達(dá)均有兩個(gè)表達(dá)峰,分別在幼苗突破種皮與長(zhǎng)出第一片真葉的時(shí)期,且在YC13-559中比YC13-554中起始表達(dá)早。推測(cè)BnWRKY52與BnWRKY16參與油菜種子萌發(fā)過(guò)程中油脂的代謝調(diào)控。
[Abstract]:Rape is the main oil crop planted in China, which plays an important role in the development of our national economy.Improving the oil content of rapeseed has been the primary task of rapeseed breeding. WRKY transcription factor plays an important role in the plant regulatory network as a regulatory factor, and participates in a series of regulation processes such as plant growth and development, stress resistance, disease resistance and so on.Two WRKY transcription factors, BnWRKY52 and BnWRKY52, which are related to oil content, were obtained by sequencing and analysis of transcriptome.The details are as follows: 1.Using NJ9 high oil strain as the basic material, a pseudo isogenic line with different oil content was constructed by continuous self-crossing.Nine pseudo isogenic lines were amplified by 156 pairs of SSR primers from the common database of Brassica Brassica DB. 32 pairs of primers with clear bands and good reproducibility were selected for near-isomorphism analysis.Based on the results of 32 pairs of SSR primer PCR amplification, genetic and similarity coefficients were estimated and cluster analysis was used to identify the near isogenic lines. The results showed that the genetic similarity coefficients of the 9 candidate lines ranged from 0.85 to 0.94.The genetic similarity coefficient between YC13-554 and YC13-559 was 0.94, which was identified as a near isogenic line.Then the agronomic characters of the two materials at maturity were investigated. The results showed that YC13-554 and YC13-559 were high (medium and low) oil content near isogenic lines.The gene expression of transcription factor BnWRKY52 and BnWRKY16 during seed maturation was detected by real-time quantitative PCR based on YC13-554 and YC13-559.緇撴灉鏄劇ず,鍦ㄧ瀛愭垚鐔熺殑涓嶅悓鏃舵湡BnWRKY52鍦ㄦ補(bǔ)鑿淵C13-554鍜孻C13-559涓殑琛ㄨ揪鍧囧憟鐜頒笂鍗囪秼鍔，

本文編號(hào)：1711052