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柑橘杂色褪绿病PCR检测技术和黄龙病传毒率及其在春梢中的分布的研究.pdf53页
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柑橘杂色褪绿病PCR检测技术和黄龙病传毒率及其在春梢中的分布的研究.pdf
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西南大学 硕士学位论文
柑橘杂色褪绿病PCR检测技术与黄龙病传毒率及在春梢中的分布 研究 姓名：鲁昕 申请学位级别：硕士 专业：植物病理学 指导教师：周常勇 座机***号码 茼要 摘要 柑橘杂色褪绿病 CitrusVariegated 引起的一种严重病害．已知分布在巴西、阿根廷、乌拉圭等南美国家。CVC自上世纪八十年 末在巴西发现以来。已经对巴西柑橘产业造成严重打击。 我国尚未发现有CVC，但是随着引种频率加大，存在CVC传入我国的风险，因此严格 的检疫是限制CVC传入我国的有效途径，其效果取决于CVC检测方法的建立和完善·1虱91-
对CVC检测技术研究较深入，其中PCR方法是目前检测CVC最为灵敏、快捷的方法。本研
究选取了国外已发表的五对引物对CVC进行PCR扩增，通过优化各反应条件，建立了五种检
测CVC的常规PCR体系和一种巢式PCR体系，比较了六种体系的灵敏性和特异性；将PCR 的扩增产物在NCBI中使用BLAST与CVC的标准序列进行了比对；并将所建立的体系应用
于实际检测中，检测了来自巴西的疑似CVC样品，主要结论如下： 1． 本研究选取了五对引物建立了检测CVC的五种常规PCR体系和一种巢式PCR体系，五 bp、745bp和700bp，灵敏度
种常规PCR体系扩增出的片断长度分别为137bp、500bp、603
2．PCR产物的核苷酸序列与已知CVC标准序列的比对结果表明，五种常规PCR体系的扩
增产物与CVC标准序列的同源性高达98％以上，扩增条带都为CVC目的条带·
按照微量快速抽提法。对来自巴西圣保罗不同地点的四个农场中的疑似CVC样品进行总
种方法检测结果一致，35个样品中有8个检测到了CVC病原。
性痛害。我国对HLB的研究较深入，本研究对HLB通过不同嫁接接种方法 皮接和芽接 的传毒率情况和
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