本文選題：劍麻 + APX　； 參考：《海南大學(xué)》2017年碩士論文

【摘要】：劍麻是世界最重要的麻類經(jīng)濟(jì)作物,其應(yīng)用廣泛,綜合價(jià)值高,前景廣闊,但是其受到斑馬紋病的危害。劍麻斑馬紋病是影響劍麻生產(chǎn)種植的最嚴(yán)重的病害之一,該病致病菌為煙草疫霉菌,是一種傳染性病害,會導(dǎo)致劍麻麻區(qū)植株染病,最終導(dǎo)致劍麻產(chǎn)量大量降低,嚴(yán)重地制約劍麻產(chǎn)業(yè)的可持續(xù)發(fā)展,造成巨大的經(jīng)濟(jì)損失。本研究以提高劍麻品種H.11648抗病性為目的,開展以下研究:1.以pCAMBIA1300為基礎(chǔ)載體,構(gòu)建了病原菌誘導(dǎo)型啟動子Hrs203J啟動APX 基因和 hevein-like 的雙價(jià)表達(dá)載體:pCAMBIA1300-Hr203J-Hevein-NOS-Hr203J-APX-NOS,用凍融法將載體導(dǎo)入根癌農(nóng)桿菌EHA105中。2.將從廣東廣西采集的14株劍麻斑馬紋病病原菌進(jìn)行rDNA-ITS序列分析,結(jié)果證明它們都是煙草疫霉菌,并且發(fā)現(xiàn)其基因類群與區(qū)域有一定聯(lián)系,通過14株病菌致病力的分析,發(fā)現(xiàn)其致病力與菌株所在區(qū)域沒有直接聯(lián)系,篩選出致病力較強(qiáng)的2株煙草疫霉菌菌株。3.將植物表達(dá)載體 pCAMBIA1300-Hr203J-Hevein-NOS-Hr203J-APX-NOS通過農(nóng)桿菌介導(dǎo)法對122個(gè)劍麻愈傷組織進(jìn)行遺傳轉(zhuǎn)化,獲得9株劍麻抗性植株,轉(zhuǎn)化率約為7.4%。對獲得的抗性劍麻幼苗進(jìn)行PCR、RT-PCR檢測,證明4株劍麻苗呈陽性,表明APX基因和hevein-like成功導(dǎo)入到劍麻基因組中,陽性轉(zhuǎn)化率為44.4%。并將2株較強(qiáng)的菌株分別接種到4株陽性轉(zhuǎn)基因苗葉片上,其結(jié)果為2號轉(zhuǎn)基因劍麻苗株的抗病為2級高抗病,編號1、3、4劍麻苗抗病性呈中抗為3級抗病性。
[Abstract]:Sisal is the most important economic crop of hemp in the world. It is widely used with high comprehensive value and broad prospect, but it is damaged by zebra stripe disease. Sisal zebra stripe is one of the most serious diseases affecting the production and planting of sisal. The pathogen of the disease is Phytophthora tabacum, which is an infectious disease. Seriously restrict the sustainable development of sisal industry, resulting in huge economic losses. In order to improve the disease resistance of sisal variety H. 11648, the following research was carried out: 1. Using pCAMBIA1300 as the basic vector, the bivalent expression vector: pCAMBIA1300-Hr203J-Hevein-NOS-Hr203J-APX-NOSwas constructed, and the vector was transfected into Agrobacterium tumefaciens EHA105 by freeze-thaw method. The bivalent expression vector: pCAMBIA1300-Hr203J-Hevein-Hr203J-APX-NOSwas constructed. The rDNA-ITS sequence analysis of 14 strains of sisal zebra stripe disease collected from Guangdong and Guangxi was carried out. The results showed that they were all Phytophthora tabacum, and found that their gene groups were related to the region, and the pathogenicity of 14 strains were analyzed. It was found that the pathogenicity was not directly related to the region where the strain was located, and two strains of Phytophthora tabacum with strong pathogenicity were screened out. The plant expression vector pCAMBIA1300-Hr203J-Hevein-NOS-Hr203J-APX-NOS was transformed into 122 calli of sisal by Agrobacterium tumefaciens. Nine sisal resistant plants were obtained and the transformation rate was about 7.4%. The results of RT-PCR showed that four sisal seedlings were positive, which indicated that APX gene and hevein-like were successfully introduced into the genome of sisal, and the positive transformation rate was 44.4%. The two stronger strains were inoculated into the leaves of 4 positive transgenic seedlings respectively. The results showed that the disease resistance of transgenic sisal seedling No. 2 was high in grade 2, and the disease resistance of No.2 transgenic sisal seedling was middle to grade 3.
【學(xué)位授予單位】：海南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S435.63

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本文編號：1997515