烟草花叶病毒灭活因子的初步研究
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摘要
本研究选用福建农业大学植物病毒研究所保存的烟草花叶病毒
TMV的强毒株系经大量繁殖并提纯后，用一系列的物理和化学和生
物方法对其进行了灭活处理；选用多种植物进行抗病毒活性物质的提
取，并将提取物对TMV进行了灭活处理；采用半叶法与A蛋白酶联
免疫吸附法（PAS-ELISA）对病毒的灭活结果进行了检测。
检测结果发现：物理因素对病毒的灭活起了重要作用，最重要
的物理因素为温度和光线（包括紫外线）。高温是灭活病毒的一种有效
途径。而且随着温度的上升，TMV的存活率下降。当温度达到80℃
以上时，TMV已经难以存活了。紫外线的照射能迅速有效地灭活病
毒。在近距离内，用30W的紫外灯照射10min就能有效地灭活病毒。
当照射时间达到20min以上时，病毒的存活率就降到了很低的水平。
铜、银等重金属离子和乙醇、甲醛、过氧化氢、SDS等多种有机物
对于TMV有一定程度的灭活作用。此外，多种植物提取物对TMV
也具有一定程度的灭活作用，包括羊蹄甲（Bauhinia variegata）、乌
莓（Cayratia japonica）、蟛蜞菊（Wedelia chinensis）、鬼针草（Bidens
bipinnata）、榕树（Ficus microcarpa）、苦瓜（Monordica charantia）、
心叶烟（Nicotiana glutinosa）、黄槐（Cassia surattensis）、细叶桉
（Eucalyptus tereticornis）等植物叶片的提取液。在这些提取物中苦
瓜蛋白等物质对TMV具有抑制作用。
最后，本文还对植物抗病毒蛋白的作用机理，提取植物抗病毒活
性物质的注意事项以及病毒灭活理论在生产上的一些应用做了讨论。
This thesis chooses the tobacco mosaic virus which is kept
in the institute of plant virology in the Fujian agriculture
university as the study material.After greatly reproducing and
purifying of TMV,a series of physical chemical and biologic
methods were used to inactivate the virus.And the method of
half-leaf method and PAS-ELISA were used to check up the
result.
The final result is that:the physical factors do a great deal
to inactivate the virus,the most important physical factors
contain temperature and light. And with the increasing of the
temperature,the livability of TMV come down.When the
tempterature is higher than 80 C ,TMV can hardly
survive.Ultraviolet radiation can quickly and efficiently
inactivate TMV.In close quarters,10 minutes of UV radiation
produced, by UV lamp of 30W can kill TMV in some
degree.When the irradiation is more than 20 minutes,the
survival of TMV go down to a very low level. However the
chemical factors which have something to do with the
inactivation of water virus may possiblly contain:,heavy metals
(such as copper and silver) and some chemicals (such as
C2H5OH,CH3CHO,H202 and so on)which can change the nucleic
acid and the protein crust of the virus (such as formaldehyde and
benzene) . And some materials purified from some plants may
inactivate the TMV in some degrees.These plant contain Bauhinia
variegata . Cayratia japonica Wedelia chinensis Bidens bipinnata )
2
Ficus micro carpa Monordica charantia Nicotiana glutinosa Cassia
surattensis.. Eucalyptus tereticornis and so on.Among these extractions
balsam pear protein is found to be able to inactivate TMV.
At last,this paper particularly discusses the inactivating
mechanism of plant antivirus protafn ,phonetic notations in
extracting plant antivirus substance and the appliance of the
inactivating theory in yield.
TMV的强毒株系经大量繁殖并提纯后，用一系列的物理和化学和生
物方法对其进行了灭活处理；选用多种植物进行抗病毒活性物质的提
取，并将提取物对TMV进行了灭活处理；采用半叶法与A蛋白酶联
免疫吸附法（PAS-ELISA）对病毒的灭活结果进行了检测。
检测结果发现：物理因素对病毒的灭活起了重要作用，最重要
的物理因素为温度和光线（包括紫外线）。高温是灭活病毒的一种有效
途径。而且随着温度的上升，TMV的存活率下降。当温度达到80℃
以上时，TMV已经难以存活了。紫外线的照射能迅速有效地灭活病
毒。在近距离内，用30W的紫外灯照射10min就能有效地灭活病毒。
当照射时间达到20min以上时，病毒的存活率就降到了很低的水平。
铜、银等重金属离子和乙醇、甲醛、过氧化氢、SDS等多种有机物
对于TMV有一定程度的灭活作用。此外，多种植物提取物对TMV
也具有一定程度的灭活作用，包括羊蹄甲（Bauhinia variegata）、乌
莓（Cayratia japonica）、蟛蜞菊（Wedelia chinensis）、鬼针草（Bidens
bipinnata）、榕树（Ficus microcarpa）、苦瓜（Monordica charantia）、
心叶烟（Nicotiana glutinosa）、黄槐（Cassia surattensis）、细叶桉
（Eucalyptus tereticornis）等植物叶片的提取液。在这些提取物中苦
瓜蛋白等物质对TMV具有抑制作用。
最后，本文还对植物抗病毒蛋白的作用机理，提取植物抗病毒活
性物质的注意事项以及病毒灭活理论在生产上的一些应用做了讨论。
This thesis chooses the tobacco mosaic virus which is kept
in the institute of plant virology in the Fujian agriculture
university as the study material.After greatly reproducing and
purifying of TMV,a series of physical chemical and biologic
methods were used to inactivate the virus.And the method of
half-leaf method and PAS-ELISA were used to check up the
result.
The final result is that:the physical factors do a great deal
to inactivate the virus,the most important physical factors
contain temperature and light. And with the increasing of the
temperature,the livability of TMV come down.When the
tempterature is higher than 80 C ,TMV can hardly
survive.Ultraviolet radiation can quickly and efficiently
inactivate TMV.In close quarters,10 minutes of UV radiation
produced, by UV lamp of 30W can kill TMV in some
degree.When the irradiation is more than 20 minutes,the
survival of TMV go down to a very low level. However the
chemical factors which have something to do with the
inactivation of water virus may possiblly contain:,heavy metals
(such as copper and silver) and some chemicals (such as
C2H5OH,CH3CHO,H202 and so on)which can change the nucleic
acid and the protein crust of the virus (such as formaldehyde and
benzene) . And some materials purified from some plants may
inactivate the TMV in some degrees.These plant contain Bauhinia
variegata . Cayratia japonica Wedelia chinensis Bidens bipinnata )
2
Ficus micro carpa Monordica charantia Nicotiana glutinosa Cassia
surattensis.. Eucalyptus tereticornis and so on.Among these extractions
balsam pear protein is found to be able to inactivate TMV.
At last,this paper particularly discusses the inactivating
mechanism of plant antivirus protafn ,phonetic notations in
extracting plant antivirus substance and the appliance of the
inactivating theory in yield.
引文
王先彬，王启燕．香菇培养物水浸液对烟草花叶病毒(TMV)侵染心叶烟的抑制作用．微生物学报，1986,26(4)：363-365．
田波，裴美云．植物病毒研究方法．科学出版社，1987：190．
安德荣，慕小倩，蔡马等．植物病毒复制抑制物的提取和其抗性测定．西北农业学报，1994,3(1)：95-98．
刘学端，张碧峰．抗植物病毒剂的研究和应用．国外农学-植物保护．1994,7(3-4)：8-11．
严惠琴，蒋蕙惠，李洁等．自来水厂常规加氯处理灭活脊髓灰质炎病毒的效果。病毒学杂志，1987，(2)：55-59．
罗明典．病毒应用于农业的一些进展[J]．病毒学杂志，1986,1(3)：3-12．
郑耀通，林奇英，谢联辉．水体环境中的植物病毒及生态效应．中国病毒学 2000，15(1)：1-7．
赵文彬，周克捷，吴桃林等．脊髓灰质炎减毒活疫苗的热稳定性试验．中国病毒学，1992，7(4)：456-458．
郭明．植物病毒抑制物的研究进展．甘肃农业科技，1992,(7)：29-29．
韩友圻，黄祥瑞，张敏丽等．紫外线对甲型肝炎病毒灭活机理的研究．中国病毒学，1997，12(1)：83-86．
Chen Z C,White R F,Antoniw J F and Lin Q.Effect of pokeweed antiviral protein(PAP) on the infection of plant viruses.Plant Pathology.1991,40:612-620.
Harmon SA;Richards OC,Summers DF et al. The 5'-terminal nucleotides of hepatitis A virus RNA,but not poliovirus RNA. Are required for infectivity.J virol,1991,65:2757.
Rao A L N,Varma.Transmission studies with cucumber green mottle mosaic virus[J].Phytopathol Z, 1984,109(4):325～331.
田波，裴美云．植物病毒研究方法．科学出版社，1987：190．
安德荣，慕小倩，蔡马等．植物病毒复制抑制物的提取和其抗性测定．西北农业学报，1994,3(1)：95-98．
刘学端，张碧峰．抗植物病毒剂的研究和应用．国外农学-植物保护．1994,7(3-4)：8-11．
严惠琴，蒋蕙惠，李洁等．自来水厂常规加氯处理灭活脊髓灰质炎病毒的效果。病毒学杂志，1987，(2)：55-59．
罗明典．病毒应用于农业的一些进展[J]．病毒学杂志，1986,1(3)：3-12．
郑耀通，林奇英，谢联辉．水体环境中的植物病毒及生态效应．中国病毒学 2000，15(1)：1-7．
赵文彬，周克捷，吴桃林等．脊髓灰质炎减毒活疫苗的热稳定性试验．中国病毒学，1992，7(4)：456-458．
郭明．植物病毒抑制物的研究进展．甘肃农业科技，1992,(7)：29-29．
韩友圻，黄祥瑞，张敏丽等．紫外线对甲型肝炎病毒灭活机理的研究．中国病毒学，1997，12(1)：83-86．
Chen Z C,White R F,Antoniw J F and Lin Q.Effect of pokeweed antiviral protein(PAP) on the infection of plant viruses.Plant Pathology.1991,40:612-620.
Harmon SA;Richards OC,Summers DF et al. The 5'-terminal nucleotides of hepatitis A virus RNA,but not poliovirus RNA. Are required for infectivity.J virol,1991,65:2757.
Rao A L N,Varma.Transmission studies with cucumber green mottle mosaic virus[J].Phytopathol Z, 1984,109(4):325～331.