本文選題：普通小麥 + 雄性不育��； 參考：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：小麥337S不育系是對(duì)長(zhǎng)日高溫、短日低溫均敏感的兩極光溫敏小麥雄性不育系,具有不育性穩(wěn)定、制種區(qū)域廣之特點(diǎn)。本實(shí)驗(yàn)室早期以337S光溫敏雄性不育系在短日低溫不育環(huán)境條件及正�？捎h(huán)境條件下的花藥為材料,采用數(shù)字基因表達(dá)譜技術(shù)對(duì)其進(jìn)行RNA-Seq測(cè)序分析,獲得了大量與小麥生殖發(fā)育相關(guān)的差異基因,并對(duì)其中差異表達(dá)顯著的關(guān)鍵基因進(jìn)行了分子克隆。同時(shí)利用Gateway技術(shù)構(gòu)建了候選基因TaMS337S-1和TaMS337S-2的超量表達(dá)載體,通過(guò)基因槍和農(nóng)桿菌介導(dǎo)的轉(zhuǎn)基因技術(shù)轉(zhuǎn)化不同小麥品種進(jìn)行功能驗(yàn)證,探究TaMS337S-1和TaMS337S-2基因在小麥生長(zhǎng)過(guò)程中的主要作用,并為337S光溫敏不育系短日低溫?cái)∮肿訖C(jī)理的研究提供線索�，F(xiàn)已基本完成了這兩個(gè)基因的遺傳轉(zhuǎn)化工作,主要實(shí)驗(yàn)結(jié)果如下:以華麥2152和華麥2566為受體通過(guò)基因槍轟擊法轉(zhuǎn)化TaMS337S-1基因,沒(méi)有得到轉(zhuǎn)基因植株。以華麥2152、華麥2566、鄭麥9023、隴春23、科農(nóng)199和Bob White幼胚為受體材料,通過(guò)基因槍轟擊轉(zhuǎn)化TaMS337S-2基因得到再生小麥51株,分別為4株華麥2566,41株隴春23,1株Bob White。PCR初步檢測(cè)后顯示2株隴春23為陽(yáng)性植株,其余均為陰性再生苗。以華麥2152、華麥2566、隴春23和科農(nóng)199為受體材料,利用農(nóng)桿菌侵染法轉(zhuǎn)化TaMS337S-1基因得到再生植株7株,分別為3株華麥2566,4株隴春23。PCR結(jié)果顯示有2株華麥2566為轉(zhuǎn)基因陽(yáng)性植株,其表型為花絲外露、無(wú)花藥。以華麥2152、華麥2566、隴春23和科農(nóng)199為受體材料通過(guò)農(nóng)桿菌轉(zhuǎn)化TaMS337S-2基因共得到9株再生苗,分別為1株華麥2566,4株科農(nóng)199,4株隴春23,但PCR檢測(cè)沒(méi)有發(fā)現(xiàn)轉(zhuǎn)基因陽(yáng)性苗。
[Abstract]:Wheat 337s male sterile line is sensitive to long day high temperature and short day low temperature sensitive male sterile line of wheat, which has stable sterility and wide seed production area. The anthers of 337S photo-thermo-sensitive male sterile lines were used as materials in short day low temperature sterility environment and normal fertile environment in our laboratory. Digital gene expression technique was used to analyze the RNA-Seq sequence of the male sterile lines. A large number of differentially expressed genes related to wheat reproductive development were obtained and the key genes with significant differentially expressed genes were cloned. At the same time, the overexpression vectors of candidate genes TaMS337S-1 and TaMS337S-2 were constructed by using Gateway technology, and then transformed into different wheat varieties by gene gun and Agrobacterium tumefaciens. To explore the main role of TaMS337S-1 and TaMS337S-2 genes in wheat growth, and to provide clues for the study of the molecular mechanism of short-day and low-temperature abortion of 337S photo-thermosensitive male sterile lines. The main results are as follows: Huamai 2152 and Huamai 2566 were used as receptors to transform TaMS337S-1 gene by bombardment of gene gun, and no transgenic plants were obtained. The immature embryos of Huamai 2152, Huamai 2566, Zhengmai 9023, Longchun 23, Konong 199 and Bob White were used as recipient materials, and 51 regenerated wheat plants were obtained by bombardment of TaMS337S-2 gene with gene gun. The results of Bob White.PCR analysis showed that 2 Longchun 23 plants were positive plants, and the rest were negative regenerated plantlets. Using Huamai 2152, Huamai 2566, Longchun 23 and Konong 199 as receptor materials, 7 regenerated plants were obtained by Agrobacterium tumefaciens infection method. The results showed that 2 Huamai 2566 plants were transgenic positive plants. Its phenotype is filaments exposed, no anther. Using Huamai 2152, Huamai 2566, Longchun 23 and Konong 199 as receptor materials, 9 regenerated seedlings were obtained by Agrobacterium tumefaciens transformation into TaMS337S-2 gene, one of them was 1 Huamai 2566 / 4 Konong 1994 strain Longchun 23, but no transgenic seedlings were found by PCR test.
【學(xué)位授予單位】：華中農(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)】：S512.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 王永琦;楊小振;莫言玲;鄭俊，

本文編號(hào)：1782264