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  本文關(guān)鍵詞：小麥不同胚性愈傷率成熟胚特性比較和PvTB1基因轉(zhuǎn)化研究　出處：《西北農(nóng)林科技大學(xué)》2016年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 小麥(Triticum aestivum L.) 成熟胚 胚性愈傷組織 PvTB1 轉(zhuǎn)基因

【摘要】：小麥?zhǔn)鞘澜绲谝淮蠹Z食作物,對(duì)人類(lèi)生活有著十分重要的意義。由于全球耕地面積減少,人口數(shù)量逐年上升,提高糧食產(chǎn)量刻不容緩。近年來(lái)植物轉(zhuǎn)基因技術(shù)發(fā)展迅速,已在小麥遺傳改良中得到了初步應(yīng)用,為培育高產(chǎn)品種開(kāi)辟了一條新的途徑。目前,在小麥遺傳轉(zhuǎn)化的受體材料中,胚性愈傷組織應(yīng)用最為普遍。成熟胚由于不受生長(zhǎng)季節(jié)限制、可周年取材且供給充足,被認(rèn)為是誘導(dǎo)胚性愈傷組織最為方便的外植體。小麥成熟胚的特征特性與胚性愈傷組織誘導(dǎo)的數(shù)量、質(zhì)量必然具有一定相關(guān)性,迄今國(guó)內(nèi)外尚未有相關(guān)研究的報(bào)道。同時(shí),分蘗是小麥重要的農(nóng)藝性狀之一,對(duì)小麥高產(chǎn)、穩(wěn)產(chǎn)影響顯著,而采用遺傳轉(zhuǎn)化技術(shù)對(duì)小麥分蘗特性進(jìn)行遺傳改良,將具有重要的育種和生產(chǎn)應(yīng)用前景。為了探究胚性愈傷率誘導(dǎo)效果不同的普通小麥成熟胚特性的差異,本研究以胚性愈傷率不同的10種小麥基因型的成熟胚為材料,對(duì)其部分生理生化指標(biāo)進(jìn)行了測(cè)定,并觀察分析了其外部形態(tài)和內(nèi)部微觀結(jié)構(gòu)。同時(shí),利用基因槍法,將克隆自生物能源作物柳枝稷中調(diào)控分蘗性狀的基因PvTB1對(duì)普通小麥進(jìn)行了遺傳轉(zhuǎn)化。取得的主要研究結(jié)果如下:1.以胚性愈傷率不同的10種小麥基因型的成熟胚為材料,對(duì)胚性愈傷率高、低不同的兩類(lèi)小麥成熟胚的可溶性糖、可溶性蛋白、纖維素、谷胱甘肽和游離氨基酸的測(cè)定。結(jié)果顯示,兩類(lèi)小麥成熟胚的可溶性蛋白和纖維素含量存在顯著性差異,而可溶性糖、谷胱甘肽和游離氨基酸的含量在兩類(lèi)小麥成熟胚間的差異不顯著。相關(guān)性分析發(fā)現(xiàn),小麥成熟胚胚性愈傷率與纖維素含量存在極顯著正相關(guān)(P=0.002,P0.01),與可溶性蛋白含量存在顯著負(fù)相關(guān)(P=0.01,P0.05)。2.利用體視顯微鏡對(duì)胚性愈傷率不同的10種小麥基因型的成熟胚外部形態(tài)進(jìn)行觀察,結(jié)果發(fā)現(xiàn)胚性愈傷率高、低不同的兩類(lèi)小麥成熟胚的高度之間存在顯著性差異。利用石蠟切片方法對(duì)成熟胚的內(nèi)部結(jié)構(gòu)進(jìn)行觀察比較,結(jié)果顯示兩類(lèi)小麥成熟胚的內(nèi)部結(jié)構(gòu)之間差異不顯著。3.以SQ26、西農(nóng)928、西農(nóng)1376和鄭麥9023小麥基因型為轉(zhuǎn)化外植體材料,采用基因槍法,將調(diào)控柳枝稷分蘗性狀的基因PvTB1對(duì)普通小麥進(jìn)行遺傳轉(zhuǎn)化,經(jīng)篩選共獲得409株抗性苗,移栽到溫室后存活350株。通過(guò)PCR鑒定分析,在鄭麥9023中獲得1株含有目標(biāo)基因的轉(zhuǎn)基因植株,轉(zhuǎn)化效率約為0.01%。通過(guò)對(duì)轉(zhuǎn)基因植株的分蘗數(shù)、穗長(zhǎng)、株高、旗葉長(zhǎng)、穗下第一節(jié)長(zhǎng)和小穗數(shù)6個(gè)農(nóng)藝性狀進(jìn)行統(tǒng)計(jì)分析,結(jié)果表明轉(zhuǎn)基因植株與對(duì)照植株之間并沒(méi)有顯著性差異,初步說(shuō)明Pv TB1基因在普通小麥中沒(méi)有表達(dá)或表達(dá)量不足。
[Abstract]:Wheat is the largest food crop in the world, which is of great significance to human life. Due to the decrease of global cultivated land, the population has increased year by year. It is urgent to increase grain yield. In recent years, plant transgenic technology has been developed rapidly and has been applied in wheat genetic improvement, which opens a new way for breeding high-yield varieties. Embryogenic callus is the most common recipient material in wheat genetic transformation. Mature embryos can be obtained annually and supplied with sufficient materials because they are not restricted by growing season. It is considered that it is the most convenient explant to induce embryogenic callus. The characteristic characteristics of wheat mature embryo must have certain correlation with the quantity and quality of embryogenic callus induction. At the same time, tillering is one of the important agronomic traits of wheat, which has a significant effect on high yield and stable yield of wheat. Genetic transformation technology is used to improve the tillering characteristics of wheat. In order to explore the difference of mature embryo characteristics of common wheat with different embryogenic callus induction rate. In this study, some physiological and biochemical indexes of mature embryos of 10 wheat genotypes with different embryogenic callus rates were measured, and their external morphology and internal microstructure were observed and analyzed. Using genetic gunshot. The genetic transformation of common wheat was carried out by cloning the gene PvTB1, which was cloned from switchgrass, a bioenergy crop, to regulate tillering traits. The main results were as follows:. 1. The mature embryos of 10 wheat genotypes with different embryogenic calli were used as materials. The determination of soluble sugar, soluble protein, cellulose, glutathione and free amino acid in two kinds of wheat mature embryos with high and low embryogenic callus rate. There was significant difference in soluble protein and cellulose content between the two types of wheat mature embryos, while the contents of soluble sugar, glutathione and free amino acids were not significantly different between the two types of wheat mature embryos. There was a very significant positive correlation between the callus rate of mature embryo and cellulose content, and a significant negative correlation between callus rate and soluble protein content (P0. 01). The external morphology of mature embryos of 10 wheat genotypes with different embryogenic calli was observed by stereoscopic microscope. The results showed that the embryogenic callus rate was high. There were significant differences in the height of mature embryos between the two types of wheat. The internal structure of mature embryos was observed and compared by paraffin section method. The results showed that there was no significant difference in the internal structure of mature embryos between the two types of wheat. The genotypes of SQ26, Xinong 928, Xinong 1376 and Zhengmai 9023 were used as the transformed explants. The gene PvTB1, which regulates the tiller character of switchgrass, was transformed into common wheat by gene gun method, and a total of 409resistant seedlings were obtained by screening. A transgenic plant containing target gene was obtained from Zhengmai 9023 by PCR analysis. The transformation efficiency was about 0.01. The agronomic characters of transgenic plants including tiller number, ear length, plant height, flag leaf length, first node length under ear and spikelet number were analyzed statistically. The results showed that there was no significant difference between transgenic plants and control plants. The results showed that PV TB1 gene was not expressed or was not expressed in common wheat.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：S512.1

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