本文選題：伽師瓜 + 植物病毒病��； 參考：《新疆大學》2017年碩士論文

【摘要】：甜瓜(Cucumis melo L.)不僅高產(chǎn)、優(yōu)質(zhì),而且經(jīng)濟效益高,其市場銷售范圍也很廣,在新疆栽培歷史悠久,也是世界重要的經(jīng)濟作物。素有“植物癌癥”之稱的植物病毒病,是影響多種農(nóng)作物減產(chǎn)的主要原因,在瓜類作物中造成的危害尤為嚴重。病毒病一旦大面積發(fā)生,甜瓜就會嚴重減產(chǎn),品質(zhì)下降,甚至絕收。而且病毒在田間還會經(jīng)常發(fā)生復合侵染,這使得病毒病的防治更加困難。近幾年新疆瓜果產(chǎn)業(yè)一直受到植物病毒的影響,而主栽甜瓜品種不抗植物病毒病,所以利用基因工程技術(shù)轉(zhuǎn)化病毒來源的外殼蛋白基因也是一種可行的方法,由此開展實驗并得到以下結(jié)果:1.從新疆吐魯番收集的發(fā)病葉片,分離純化后確定為西瓜花葉病毒�？寺～@得西瓜花葉病毒外殼蛋白(coat protein,CP)基因序列,經(jīng)HindⅢ與XbaⅠ酶切后插入pCAMBIA2300載體中,獲得植物雙元表達載體。2.以厚皮甜瓜伽師為材料,建立以子葉為外植體的快速高效再生體系。甜瓜子葉外植體接種在MSB+1.5 mg·L-1 6-BA誘導分化培養(yǎng)基上,誘導率可達75%;添加不同濃度AgNO3誘導不定芽,以MSB+1.5 mg·L-1 6-BA+2.0 mg·L-1AgNO3最適,誘導率可達65%,產(chǎn)生單芽較多,縮短了再生周期。3.通過農(nóng)桿菌GV3101的介導甜瓜和本氏煙草,得到了許多擬陽性轉(zhuǎn)基因煙草和甜瓜,提取擬陽性植株基因組DNA,經(jīng)過PCR鑒定,得到了11株轉(zhuǎn)基因煙草,4株轉(zhuǎn)基因甜瓜。4.對轉(zhuǎn)基因煙草的SOD、POD和CAT的酶活性進行檢測,發(fā)現(xiàn)這三種酶的活性均高于野生型煙草,初步驗證了轉(zhuǎn)基因煙草對西瓜花葉病毒存在抗性,可作為早期篩選抗性植株的輔助指標。統(tǒng)計T1代幼苗Kan抗性遺傳分離比為3:1的轉(zhuǎn)基因煙草,得到1、4、5、6、9、10、11這七個符合株系。本實驗結(jié)果為進一步開展病毒病與植物抗病性的研究奠定基礎。
[Abstract]:Cucumis melo L. Not only high yield, good quality, but also high economic benefit, its marketing range is also very wide, it has a long history of cultivation in Xinjiang, and is also an important cash crop in the world. Plant virus disease, known as "plant cancer", is the main reason that affects the yield reduction of many crops, especially in melon crops. Once the virus disease occurs in a large area, melon will seriously reduce production, quality decline, or even end-harvest. And the virus often occurs in the field complex infection, which makes it more difficult to prevent and cure the virus disease. In recent years, the melon and fruit industry in Xinjiang has been affected by plant viruses, and the main melon varieties are not resistant to plant virus disease, so it is also a feasible method to use genetic engineering technology to transform the coat protein gene from the virus source. The experiment was carried out and the following results were obtained: 1: 1. Watermelon mosaic virus was isolated and purified from the infected leaves collected from Turpan, Xinjiang. The coat protein (CPP) gene of watermelon mosaic virus was cloned and inserted into pCAMBIA2300 vector by Hind 鈪，

本文編號：2001177