本文選題：馬鈴薯 + 病毒��； 參考：《草業(yè)學(xué)報(bào)》2017年12期

【摘要】：馬鈴薯病毒積累引起的種薯退化是馬鈴薯生產(chǎn)中造成產(chǎn)量和品質(zhì)下降的重要原因之一。本研究以馬鈴薯病毒病攜帶植株葉片c DNA為試驗(yàn)組(Tester)、脫毒種苗葉片c DNA為驅(qū)動(dòng)組(Driver),采用抑制消減雜交(suppression subtractive hybridization,SSH)技術(shù)構(gòu)建了馬鈴薯病毒誘導(dǎo)應(yīng)答基因的c DNA文庫;為驗(yàn)證文庫構(gòu)建效果,從文庫中隨機(jī)挑取了98個(gè)陽性克隆經(jīng)PCR驗(yàn)證后測序,獲得了45條高質(zhì)量的有效非重復(fù)序列;經(jīng)與Gen Bank進(jìn)行同源比對后發(fā)現(xiàn),其中14條非重復(fù)序列屬于馬鈴薯病毒基因序列,22條與已知基因序列同源性較高,9條無同源參考基因;選取文庫中出現(xiàn)頻率較高的2個(gè)ESTs(expressed sequence tag,表達(dá)序列標(biāo)簽)用qRT-PCR技術(shù)分析發(fā)現(xiàn),其表達(dá)量受馬鈴薯病毒侵染的誘導(dǎo)。結(jié)果表明,該SSH文庫構(gòu)建較為成功,為進(jìn)一步篩選與馬鈴薯病毒致病、防御相關(guān)的應(yīng)答基因,解析馬鈴薯與病毒互作的分子機(jī)理,利用生物技術(shù)手段培育抗病毒馬鈴薯奠定了基礎(chǔ)。
[Abstract]:Seed potato degradation caused by the accumulation of potato virus is one of the important reasons for the decline of yield and quality in potato production. In this study, the cDNA library of potato virus-induced response gene was constructed by suppression subtractive hybridization (suppression subtractive) hybridization technique, using the leaves of potato virus carrying plants as test group (Tester) and virus-free seedling leaves as driving group (driver). In order to verify the effect of library construction, 98 positive clones were randomly selected from the library and sequenced by PCR. Among them, 14 non-repeat sequences belong to 22 sequences of potato virus genes with high homology with known genes and 9 non-homologous reference genes. Two ests (expressed sequence tag, expressed sequence tags with high frequency in the library were selected. The results were analyzed by qRT-PCR. Its expression was induced by potato virus infection. The results showed that the SSH library was successfully constructed. In order to further screen the responsive genes related to the pathogenesis and defense of potato virus, the molecular mechanism of the interaction between potato and virus was analyzed. Using biotechnology to cultivate anti-virus potato laid the foundation.
【作者單位】： 甘肅省農(nóng)業(yè)科學(xué)院生物技術(shù)研究所;
【基金】：甘肅省農(nóng)科院青年創(chuàng)新基金(2013GAAS29) 甘肅省農(nóng)業(yè)科學(xué)院創(chuàng)新團(tuán)隊(duì)(2015GAAS02);甘肅省農(nóng)業(yè)科學(xué)院農(nóng)業(yè)科技創(chuàng)新重大項(xiàng)目(2016GAAS59-02) 國家自然科學(xué)基金項(xiàng)目(31460388)資助
【分類號】：S435.32

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