本文關(guān)鍵詞： 大白菜 抽薹 突變體 基因定位 克隆　出處：《沈陽農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：大白菜[Brassica campestris ssp.pekinensis(Lour)Olsson]是東亞地區(qū)重要的蔬菜作物,未熟抽薹會(huì)大大降低其商品價(jià)值,因此,抽薹性狀遺傳機(jī)理的研究有重要意義。鑒于突變體是研究植物性狀遺傳機(jī)理的理想材料,本研究以大白菜DH系'FT'為材料,利用EMS誘變與游離小孢子培養(yǎng)技術(shù)相結(jié)合的方法創(chuàng)制出一個(gè)早抽薹突變體ebml,構(gòu)建了突變基因ebml的遺傳連鎖圖譜,并預(yù)測(cè)出了候選基因。主要研究結(jié)果如下:1.突變體表型鑒定結(jié)果表明,與野生型'FT'相比,早抽薹突變體ebml表現(xiàn)為極早抽薹,同時(shí)伴有葉緣上卷。經(jīng)流式細(xì)胞儀檢測(cè),其為二倍體;經(jīng)TTC染色鑒定,其花粉有正常活性。2.以突變體ebml及其野生型'FT'為親本,配制F1、F2和BC1。遺傳分析結(jié)果表明,早抽薹突變性狀為單隱性核基因遺傳。3.以突變體ebml和青梗菜DH系'13A516'為親本構(gòu)建F2分離群體,選取1502株表型為突變性狀的單株為定位群體,采用SSR標(biāo)記方法,將早抽薹突變基因ebwI定位于A04染色體的SSRh1-53和SSRh1-61兩個(gè)標(biāo)記之間,物理距離為73.4kb,定位區(qū)間內(nèi)有7個(gè)基因。4.通過對(duì)目的區(qū)域內(nèi)7個(gè)基因的編碼序列進(jìn)行克隆測(cè)序,發(fā)現(xiàn)在突變體中只有Bra032169基因的cDNA序列與野生型存在差異。根據(jù)基因注釋信息,Bra032169編碼組蛋白甲基轉(zhuǎn)移酶,為擬南芥卷葉基因(CURLYLEAF,CLF)。CLF能夠調(diào)控與開花相關(guān)基因(如AG、FLC、AGL19、FT)的表達(dá)。預(yù)測(cè)Bra032169為ebml的候選基因。5.基因序列結(jié)構(gòu)分析表明,突變體Bra032169基因的第12個(gè)外顯子中插入了一個(gè)53bp片段,可導(dǎo)致翻譯提前終止�；虮磉_(dá)模式分析證明,此插入突變并未影響自身的轉(zhuǎn)錄,推測(cè)可能產(chǎn)生了喪失原有功能的蛋白質(zhì),影響其對(duì)下游基因的調(diào)控作用。
[Abstract]:Chinese cabbage. [Brassica campestris ssp.pekinensis(Lour)Olsson is an important vegetable crop in East Asia. Therefore, it is important to study the genetic mechanism of sprouting characters. In view of the mutant is an ideal material to study the genetic mechanism of plant traits. In this study, an early sprouting mutant ebml was created by combining EMS mutagenesis with free microspore culture with DH strain FT' of Chinese cabbage as the material. The genetic linkage map of mutant gene ebml was constructed and candidate genes were predicted. The main results were as follows: 1. The ebml of early sprouting mutant was very early and accompanied by leaf edge curl, and it was diploid by flow cytometry. The pollen had normal activity by TTC staining. The mutant ebml and its wild type FT 'were used as parents to prepare F _ 1 F _ 2 and BC _ 1.The results of genetic analysis showed that F _ 1 F _ 2 and BC _ 1 were used as parents. The mutant character of early heading and moss was a single recessive nuclear gene. The F _ 2 segregation population was constructed by using the mutant ebml and the DH line of Dioscorea sinensis as parents. 1502 individuals with mutant phenotypes were selected as localizing population and SSR markers were used. The ebwI gene was located between the SSRh1-53 and SSRh1-61 markers of chromosome A04, and the physical distance was 73.4 kb. There were 7 genes. 4. The coding sequences of 7 genes in the target region were cloned and sequenced. Only the cDNA sequence of the Bra032169 gene was different from that of the wild type in the mutant. Bra032169 encodes histone methyltransferase according to the gene annotation information. It is suggested that CURLYLEAF CLFN. CLF can regulate genes related to flowering (such as AGFLCL AGL19) in Arabidopsis thaliana. Bra032169 was predicted to be a candidate gene for ebml. A 53bp fragment was inserted into the 12th exon of the mutant Bra032169 gene, which led to the early termination of translation. The insertion mutation does not affect the transcription of the gene itself, and it may produce a protein that has lost its original function and affect the regulation of the downstream gene.
【學(xué)位授予單位】：沈陽農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S634.1

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