本文選題：甘薯羽狀斑駁病毒 + 甘薯褪綠矮化病毒��； 參考：《廣西大學(xué)》2016年碩士論文

【摘要】：甘薯病毒病是廣西甘薯的主要病害之一。從廣西南寧、崇左、北海、玉林的甘薯種植區(qū)采集疑似甘薯病毒病樣品,通過RT-PCR法進(jìn)行甘薯羽狀斑駁病毒(Sweet potato feathery mottle virus, SPFMV)與甘薯褪綠矮化病毒(Sweet potato chlorotic stunt virus, SPCSV)的檢測,發(fā)現(xiàn)SPFMV、SPCSV在廣西普遍存在,當(dāng)甘薯單獨感染SPFMV與SPCSV時,表現(xiàn)的病毒病癥狀輕微,但甘薯同時感染SPFMV和SPCSV時,表現(xiàn)出植株矮化、葉片皺縮等嚴(yán)重癥狀。設(shè)計擴(kuò)增SPFMV與SPCSV的CP基因的引物,通過克隆并測序分別得到18個SPFMV廣西分離物的CP基因序列和14個SPCSV廣西分離物的CP基因序列。SPFMV廣西分離物CP基因的核苷酸序列945bp,編碼315個氨基酸；SPCSV廣西分離物CP基因的核苷酸序列774bp,編碼257個氨基酸。其CP基因的序列分析結(jié)果表明：SPFMV廣西分離物存在O株系、RC株系、EA株系,不存在C株系,其中O株系是SPFMV廣西分離物的優(yōu)勢株系；SPCSV廣西分離物只有WA株系,無EA株系。將RT-PCR擴(kuò)增得到SPCSV的CP基因克隆到pET30a(+)載體上,轉(zhuǎn)化于大腸桿菌BL21(DE3)pLysS, IPTG誘導(dǎo)表達(dá)得到與預(yù)期大小一致的SPCSV CP的目的蛋白。經(jīng)純化回收后免疫兔子,獲得了SPCSV CP的抗血清。通過ELISA和Western Blot檢測,抗血清的效價達(dá)到1：64000，，特異性好。SPCSV CP抗血清為生產(chǎn)上監(jiān)測SPCSV的侵染提供技術(shù)條件。
[Abstract]:Sweet potato virus disease is one of the major diseases of sweet potato in Guangxi. Samples of suspected sweet potato virus were collected from sweet potato planting areas in Nanning, Chongzuo, Beihai and Yulin, Guangxi. Sweet potato feather mottle virus (Sweet potato feathery mottle virus, SPFMV) and sweet potato chlorotic dwarf virus (Sweet potato chlorotic stunt virus, SPCSV) were detected by RT-PCR method. It was found that SPFMVN SPCSV was common in Guangxi. When sweet potato was infected with SPFMV and SPCSV alone, it showed mild viral symptoms, but when sweet potato was infected with SPFMV and SPCSV, it showed plant dwarfing, leaf shrinkage and other serious symptoms. A primer was designed to amplify the CP gene of SPFMV and SPCSV. The CP gene sequences of 18 SPFMV Guangxi isolates and 14 SPCSV Guangxi isolates were obtained by cloning and sequencing. The nucleotide sequence of CP gene of SPFMV Guangxi isolate was 945bp, encoding 315 amino acids. The nucleotide sequence of the gene was 774 BP, encoding 257 amino acids. The results of sequence analysis of CP gene showed that there were O lines, RC lines, and no lines C, and O lines were the dominant line of SPFMV Guangxi isolate, only WA line and no EA line were found in the Guangxi isolate. The CP gene of SPCSV was cloned into pET30a () vector by RT-PCR amplification and transformed into E. coli BL21DE3pLysS. the target protein of SPCSV CP was induced by IPTG. The antiserum of SPCSV CP was obtained by immunizing rabbits after purification and recovery. The titer of antiserum reached 1: 64000 by ELISA and Western Blot detection. The antiserum of SPCSV with good specificity provided technical conditions for monitoring the infection of SPCSV in production.
【學(xué)位授予單位】：廣西大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S435.661

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