湖南省烟草马铃薯Y病毒株系的初步鉴定

湖南省烟草马铃薯Y病毒株系的初步鉴定(毕业论文10200字)
摘  要：马铃薯Y病毒（PVY）现广泛分布于我国各烟草区，已成为我国烟草上的主要病害之一。近年来湖南省烟草马铃薯Y病毒病危害逐年加重，严重影响了我省烟草的产量和品质。目前国际上普遍公认De Bokx等的株系划分方法，认为有3个株系：普通型（PVYO）、叶脉坏死型（PVYN）、点条斑型（PVYC）。本文建立了RT-PCR法对湖南省烟草马铃薯Y病毒进行株系鉴定，结果表明N株系为优势株系，并发现O株系和NTN株系(N株系与O株系的重组变异株系)侵染，有株系混合侵染情况。现需加强监控防治，避免NTN株系的扩散，降低PVY对湖南省烟草产业的威胁。
关键字：马铃薯Y病毒株系；烟草；RT-PCR；

Charaterization of Potato Y virus strains in tobacco of Hunan province
    Abstract: Potato Y virus (PVY) is widely distributed in each Chinese tobacco area as one of the main tobacco diseases. In recent years, the harm of Hunan tobacco PVY disease has aggravated year by year, seriously impacted on the production and quality of tobacco. Now, the internationally accepted De Bokx’s division method of PVY strains considers that PVY strains are divided ordinary type (PVYO), veins necrosis type (PVYN), scab streak type(PVYC). This paper has established the RT-PCR method of hunan province tobacco PVY to strain identification and the results show that N strain is an advantage strain and O strain and NTN strain (restructuring variation strain of N strain and O strain) are found, and mixed infection situation is found. We need to strengthen monitoring control, avoid the spread of NTN strain in hunan province, reduce the threat of tobacco industry.

Key words: PVY strain; tobacco; RT-PCR

目 录
摘要    1
关键字    1
1 前言    2
1.1 烟草马铃薯Y病毒病的分布及危害状况    2
1.1.1 烟草马铃薯Y病毒病的分布    2
1.1.2 烟草马铃薯Y病毒病的危害    2
1.2 PVY株系的分化    3
1.3 PVY的检测方法    4
2 材料    5
2.1 样品采集    5
2.2 主要试剂    5
2.3 主要仪器    5
2.4 引物设计    5
3 实验方法    7
3.1 器皿的处理    7
3.2 总RNA的提取    7
3.3 RNA提取质量的检测    8
3.4 RT-PCR检测    8
3.4.1 cDNA的合成    8

3.4.2 PCR扩增    8
3.5 PCR产物的回收    9
3.6 感受态细胞的制备    9
3.7 回收产物的克隆    10
3.7.1 目的片段与pGM-T载体连接    10
3.7.2 连接产物向大肠杆菌的转化    10
3.8 阳性克隆菌落的PCR鉴定    10
3.9 序列的测定    11
4 结果与分析    11
4.1 样品采集    11
4.2 RNA提取质量检测    11
4.2.1 琼脂糖凝胶电泳检测    11
4.2.2 RNA溶液吸光度检测    12
4.3 RT-PCR检测    12
4.4 回收转化克隆    14
4.5 测序结果    15
5 结论与讨论    15
参考文献    17
致谢    18
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