温州地区番茄黄化曲叶病毒病流行规律与综合防治
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摘要
双生病毒是植物病毒中一类具有孪生颗粒形态的植物单链DNA病毒,在自然条件下全部由烟粉虱(Bemisia tabaci)传播,因此称粉虱传双生病毒(Whitefly-tansmitted geminiviruses, WTG)。近年来,随着B型烟粉虱全面入侵和日趋严重,2006年秋冬季温州地区各大设施蔬菜生产基地番茄黄化曲叶病毒病暴发流行,发生面积约达300公顷,经济损失达1000万元,广大农民损失惨重。由于番茄双生病毒病是目前国内发生的新病害,因此生产上急需解决以下三个技术问题：①粉虱传双生病毒病诊断技术；②掌握该病害流行规律,进行有效地监测;③组建一套有效的综合防治技术以指导生产
本研究得到浙江省、温州市农业科技重大项目资助(编号：2007C12054与N20070021)。通过病毒PCR分子鉴定、室内传毒试验、田间病情发生调查和大田防效示范推广等试验,在3年内主要取得以下研究结果：(1)、结合病害症状和分子检测,鉴定并明确了温州地区番茄黄化曲叶病毒病的病原是台湾番茄曲叶病毒(ToLCTWV),并建立了一套诊断技术。(2)、掌握了番茄黄化曲叶病毒病侵染循环规律,进而确定适当推迟播种期可以减轻病害发生。(3)、研究明确了传毒媒介B型烟粉虱传播台湾番茄曲叶病毒(ToLCTWV)的能力,确定了介体虫口密度、苗龄及温度对传毒效率的影响。(4)、监测了温州地区烟粉虱的发生动态,明确了苗期烟粉虱的虫口防治指标,并建立了发病的预测模型。(5)、构建了“清洁苗床(铲除中间寄主植物)、防虫网覆盖隔离、黄板预警诱杀、苗期喷施低毒高效农药”相结合的综合防治技术,防治效果达95%以上,有效地解决了番茄黄化曲叶病毒病防治难题。
3年来,本研究成果在温州地区各大蔬菜基地推广应用,面积达5400公顷,平均每季减少农药使用2-3次,减少农药使用量40%以上,节省防治成本492.48万元,挽回产量损失7.83万吨,增收节支达1.45亿元,社会、经济、生态效益显著。
Geminiviruses are plant viruses characterized by their unique twinned particles, which encapsidate a circular single-stranded DNA genome. Under natural conditions, all of them are spreaded by the whitefly (Bemisia tabaci), and they are called whitefly-tansmitted geminiviruses (WTG). With serious invasion of the whitefly (B whitefly) in recent years, an epidemic outbreak of Tomato yellow leaf curl virus (TYLCV) disease in greenhouses took place in Wenzhou areas over 350 ha in 2006, which caused an economic loss of over 10 million yuan (RMB). As a brand-new disease for domestic emergence, it is urgently needed to following technical issures be figured out for agricultural production:(1) diagnostic techniques on Whitefly-transmitted geminiviruses; (2) understanding on prevalent regularity of the disease with effective monitoring; (3) preventive and controlling guidance on production by comprehensive techniques.
This study was supported by Major Fundings of Agricultural Science and Technology in both Wenzhou and Zhejiang (No.2007C12054 and N20070021, respectively). Molecular identification of virus polymerase chain reaction (PCR), lab experiments on transmission of viruses, investigation in fields, and demonstrative expansion on controlling effects in fields. were all studied in the past three years. The results showed that:(1) the plant disease pathogen is Tomato leaf curl Taiwan virus (ToLCTWV) in Wenzhou, and a set of diagnostic techniques through symptoms and molecular identification were established, (2) the cycling rules of invasion and infection of ToLCTWV is comprehensible, which figured out that appropriate delay in the host seedtime may helpful for alleviate damage. (3) the spreading ability of ToLCTWV by B. tabaci (B whitefly) was clarified, and. influences on efficiency of virus transmission were clear on whitefly population density, host seedling age. and temperature. (4) population dynamics of B. tabaci in Wenzhou were monitored, as control index was set up during seedling age, with establishment of model on population forecast. (5) integrated control, such as seedbed cleaning (eradicating the intermediate host plants), isolation by insect-proof masks, yellow board traps, and spraying low-toxic pesticide during seedling, were all used against viruses effectively. The controlling effect appeared to be over 95%.
In the last three years, the proposed integrated control technology was extensively used in Wenzhou. The total area of farms using this technology was about 5400 ha, reducing the usage of pesticide about 2-3 times quarterly, and reducing the pesticide dose by 40%. The control costs were saved about 4.9 million yuan. Besides, the loss of tomato yield was reduced by 7.8300 tons, increasing earnings of 145 million yuan. The results indicated that the social, economic and ecological benefits of the technology were significant.
本研究得到浙江省、温州市农业科技重大项目资助(编号：2007C12054与N20070021)。通过病毒PCR分子鉴定、室内传毒试验、田间病情发生调查和大田防效示范推广等试验,在3年内主要取得以下研究结果：(1)、结合病害症状和分子检测,鉴定并明确了温州地区番茄黄化曲叶病毒病的病原是台湾番茄曲叶病毒(ToLCTWV),并建立了一套诊断技术。(2)、掌握了番茄黄化曲叶病毒病侵染循环规律,进而确定适当推迟播种期可以减轻病害发生。(3)、研究明确了传毒媒介B型烟粉虱传播台湾番茄曲叶病毒(ToLCTWV)的能力,确定了介体虫口密度、苗龄及温度对传毒效率的影响。(4)、监测了温州地区烟粉虱的发生动态,明确了苗期烟粉虱的虫口防治指标,并建立了发病的预测模型。(5)、构建了“清洁苗床(铲除中间寄主植物)、防虫网覆盖隔离、黄板预警诱杀、苗期喷施低毒高效农药”相结合的综合防治技术,防治效果达95%以上,有效地解决了番茄黄化曲叶病毒病防治难题。
3年来,本研究成果在温州地区各大蔬菜基地推广应用,面积达5400公顷,平均每季减少农药使用2-3次,减少农药使用量40%以上,节省防治成本492.48万元,挽回产量损失7.83万吨,增收节支达1.45亿元,社会、经济、生态效益显著。
Geminiviruses are plant viruses characterized by their unique twinned particles, which encapsidate a circular single-stranded DNA genome. Under natural conditions, all of them are spreaded by the whitefly (Bemisia tabaci), and they are called whitefly-tansmitted geminiviruses (WTG). With serious invasion of the whitefly (B whitefly) in recent years, an epidemic outbreak of Tomato yellow leaf curl virus (TYLCV) disease in greenhouses took place in Wenzhou areas over 350 ha in 2006, which caused an economic loss of over 10 million yuan (RMB). As a brand-new disease for domestic emergence, it is urgently needed to following technical issures be figured out for agricultural production:(1) diagnostic techniques on Whitefly-transmitted geminiviruses; (2) understanding on prevalent regularity of the disease with effective monitoring; (3) preventive and controlling guidance on production by comprehensive techniques.
This study was supported by Major Fundings of Agricultural Science and Technology in both Wenzhou and Zhejiang (No.2007C12054 and N20070021, respectively). Molecular identification of virus polymerase chain reaction (PCR), lab experiments on transmission of viruses, investigation in fields, and demonstrative expansion on controlling effects in fields. were all studied in the past three years. The results showed that:(1) the plant disease pathogen is Tomato leaf curl Taiwan virus (ToLCTWV) in Wenzhou, and a set of diagnostic techniques through symptoms and molecular identification were established, (2) the cycling rules of invasion and infection of ToLCTWV is comprehensible, which figured out that appropriate delay in the host seedtime may helpful for alleviate damage. (3) the spreading ability of ToLCTWV by B. tabaci (B whitefly) was clarified, and. influences on efficiency of virus transmission were clear on whitefly population density, host seedling age. and temperature. (4) population dynamics of B. tabaci in Wenzhou were monitored, as control index was set up during seedling age, with establishment of model on population forecast. (5) integrated control, such as seedbed cleaning (eradicating the intermediate host plants), isolation by insect-proof masks, yellow board traps, and spraying low-toxic pesticide during seedling, were all used against viruses effectively. The controlling effect appeared to be over 95%.
In the last three years, the proposed integrated control technology was extensively used in Wenzhou. The total area of farms using this technology was about 5400 ha, reducing the usage of pesticide about 2-3 times quarterly, and reducing the pesticide dose by 40%. The control costs were saved about 4.9 million yuan. Besides, the loss of tomato yield was reduced by 7.8300 tons, increasing earnings of 145 million yuan. The results indicated that the social, economic and ecological benefits of the technology were significant.
引文
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Czosnek H and Laterrot H. A worldwide survey of tomato yellow leaf curl viruses. Archives of Virology.1997.142:1391-1406
Harrison BD. Barker H. Bock KR, Guthrie EJ, Meredith G,and Atkinson M. Plant viruses with circular single-stranded DNA. Nature,1997.270:760-762
Li ZH. Zhou XE. Zhang X. and Xie Y. Molecular characterization of tomato-infecting begomoviruses in Yunnan, China. Archives of Virology,2004,149:1721-1732
Li M. Hu J. Xu FC. and Liu SS. Transmission of Tomato yellow leaf curl virus by two invasive biotypes and a Chinese indigenous biotype of the whitefly Bemisia tabaci. International Journal of Pest Management.2010.56:275-280
Liu J, Li M. Li JM, Huang CJ. Zhou XP, Xu FC and Liu SS. Viral infection of tobacco plants improves performance of Bemisia tabaci but more so for an invasive than for an indigenous biotype of the whitefly. Journal of Zhejiang University SCIENCE B,2010.11:30-40
Mugiira RB, Liu SS and Zhou XP. Tomato yellow leaf curl virus and Tomato leaf curl Taiwan virus invade south-east coast of China. Journal of Phytopathology,2008.156:217-221
Polston JE and Anderson PK. The emergence of whitefly-transmitted geminiviruses in tomato in the western hemisphere. Plant Diseases.1997.81:1358-1369
Rybicki EP and Pietersen G. Plant virus disease problem in the developing world. Advances in Virus Resrearch,1999.53:127-175
Rojas MR. Gilbertson RL. Russell DR and Maxwell DP. Use of degenerate primers in the polymerase chain reaction to detect whitefly-transmitted geminiviruses. Plant Diseases, 1993.77:340-347
Wyatt SD and Brown JK. Detection of subgroup Ⅲ geminivirus isolates in leaf extracts by degenerate primers and polymerase chain reaction. Phytopathology,1996，86:1288-1293
Xie Y and Zhou XP. Molecular characterization of squash leaf curl Yunnan virus, a new begomovirus and evidence for recombination. Archives of Virology,2003,148:2047-2054
Zhou XP, Liu YL, Calvert L. Munoz C. OtimNape GW， Robinson DJ and Harrison BD. Evidence that DNA-A of a geminivirus associated with severe cassava mosaic disease in Uganda has arisen by interspecific recombination. Journal of Genenal Virology,1997,78:2101-2111
Zhou XP, Xie Y, Tao XR, Zhang ZK, Li ZH and Fauquet CM. Characterization of DNA beta associated with begomoviruses in China and evidence for CO-evolution with their cognate viral DNA-A. Journal of General Virology.2003,84:237-247
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Bedford ID， Briddon RW，Jonse P, Alraff N. and Markham PG. Differentiations of three whitefly-transmitted geminiviruses from the Republic of Yemen. European Journal of Plant Pathology.1994.100:243-257
Czosnek H and Laterrot H. A worldwide survey of tomato yellow leaf curl viruses. Archives of Virology.1997.142:1391-1406
Harrison BD. Barker H. Bock KR, Guthrie EJ, Meredith G,and Atkinson M. Plant viruses with circular single-stranded DNA. Nature,1997.270:760-762
Li ZH. Zhou XE. Zhang X. and Xie Y. Molecular characterization of tomato-infecting begomoviruses in Yunnan, China. Archives of Virology,2004,149:1721-1732
Li M. Hu J. Xu FC. and Liu SS. Transmission of Tomato yellow leaf curl virus by two invasive biotypes and a Chinese indigenous biotype of the whitefly Bemisia tabaci. International Journal of Pest Management.2010.56:275-280
Liu J, Li M. Li JM, Huang CJ. Zhou XP, Xu FC and Liu SS. Viral infection of tobacco plants improves performance of Bemisia tabaci but more so for an invasive than for an indigenous biotype of the whitefly. Journal of Zhejiang University SCIENCE B,2010.11:30-40
Mugiira RB, Liu SS and Zhou XP. Tomato yellow leaf curl virus and Tomato leaf curl Taiwan virus invade south-east coast of China. Journal of Phytopathology,2008.156:217-221
Polston JE and Anderson PK. The emergence of whitefly-transmitted geminiviruses in tomato in the western hemisphere. Plant Diseases.1997.81:1358-1369
Rybicki EP and Pietersen G. Plant virus disease problem in the developing world. Advances in Virus Resrearch,1999.53:127-175
Rojas MR. Gilbertson RL. Russell DR and Maxwell DP. Use of degenerate primers in the polymerase chain reaction to detect whitefly-transmitted geminiviruses. Plant Diseases, 1993.77:340-347
Wyatt SD and Brown JK. Detection of subgroup Ⅲ geminivirus isolates in leaf extracts by degenerate primers and polymerase chain reaction. Phytopathology,1996，86:1288-1293
Xie Y and Zhou XP. Molecular characterization of squash leaf curl Yunnan virus, a new begomovirus and evidence for recombination. Archives of Virology,2003,148:2047-2054
Zhou XP, Liu YL, Calvert L. Munoz C. OtimNape GW， Robinson DJ and Harrison BD. Evidence that DNA-A of a geminivirus associated with severe cassava mosaic disease in Uganda has arisen by interspecific recombination. Journal of Genenal Virology,1997,78:2101-2111
Zhou XP, Xie Y, Tao XR, Zhang ZK, Li ZH and Fauquet CM. Characterization of DNA beta associated with begomoviruses in China and evidence for CO-evolution with their cognate viral DNA-A. Journal of General Virology.2003,84:237-247
车海彦,罗大全一,杨旭光,符瑞益,叶莎冰.海南岛番木瓜和扶桑上粉虱传双生病毒的检测及序列分析[J].中国病毒学,2005,20(5)：543-545
蔡健和,秦碧霞,朱桂宁,黄福新,陈永惠,李煜华.番茄黄化曲叶病毒病在广西爆发的原因和防治策略[J].中国蔬菜,2006(7)：47-48
车海彦,罗大全,杨旭光,符瑞益,叶莎冰.海南菜豆上粉虱传双生病毒的分子检测[J].西北农林科技大学学报(自然科学版),2007,35(8)：139-142
车海彦,刘先宝,杨旭光,符瑞益,罗大全.长蒴母草黄脉病毒一个双生病毒的新种[J].植物病理学报,2007.37(6)：578-583
丁铭,杨程,罗延青,岳宁,方琦,苏晓霞,张仲凯.侵染曼陀罗的中国番茄黄化曲叶病毒及其卫星DNA13全基因组结构特征[J].植物病理学报,2008.38(4)：364-369
董迪,何自福,柴兆祥.广东黄秋葵黄脉曲叶病样中检测到烟粉虱传双生病毒[J].植物保护.2010,36(1)：65-68
范三薇,周雪平.从海南省杂草胜红蓟和假马鞭上检测到粉虱传双生病毒[J].植物病理学报,2003.33(6)：513-516
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王冬生,匡开源.张穗,戴富民,李琳一,瞿培荣,程银坤,王次马.上海温室番茄黄化曲叶病毒病的发生与防治[J].长江蔬菜.2006,(10)：25-26
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张穗,王冬生,瞿培荣,温广月,匡开源.上海市番茄黄化曲叶病毒(TYLCV)病的初步鉴定[J].上海农业学报.2006.22(3)：126-132
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