Relationships of the Sardinian isolate of tomato yellow leaf curl geminivirus with its whitefly vectorBemisia tabaci Gen

Relationships of an Italian isolate of tomato yellow leaf curl geminivirus from Sardinia (TYLCV-S) with its whitefly vectorBemisia tabaci were studied by means of experimental transmissions from tomato to tomato plants. TYLCV-S was confirmed to be transmitted in a persistent, circulative manner. The minimum latent period in the vector was between 17 and 20 h from the beginning of the acquisition access period (AAP). The maximum retention of infectivity was 8 days from the end of the AAP. Both acquisition and inoculation feeding times influenced the detected proportion of infective insects, with patterns well described by an exponential model. Acquisition was more efficient than inoculation. Males were significantly less efficient vectors than females. Nymphs were as efficient as adults in acquiring the virus. The length of AAP influenced both the retention of infectivity, and the pattern of transmission in serial transfer transmission tests with individual females. No significant difference in transmission efficiency was detected between two colonies ofB. tabaci, one inducing typical silverleaf symptoms on squash, the other inducing only mild symptoms with more than 50 whiteflies per plant. The phenomenon of periodic acquisition was not unequivocally proved for TYLCV-S.Research supported by the National Research Council of Italy, Special Project RAISA, Sub-project N. 2. Paper N. 1961.Supported by a grant from the Istituto Agronomico Mediterraneo, Valenzano (Bari), Italy.     

番茄黄化曲叶病毒与番茄褪绿病毒复合侵染对番茄黄化曲叶病毒传播的影响

番茄黄化曲叶病毒(tomato yellow leaf curl virus, TYLCV)是一种由烟粉虱传播的单链环状DNA病毒, 在田间可与多种病毒发生复合侵染, 如番茄褪绿病毒(tomato chlorosis virus, ToCV)等?本文对比了TYLCV单独侵染和TYLCV与ToCV复合侵染对烟粉虱获取和传播TYLCV的影响?结果表明, 与取食TYLCV单独侵染的番茄相比, 取食复合侵染番茄的烟粉虱对TYLCV的传毒率显著提高, 且番茄植株和烟粉虱体内TYLCV的病毒积累量也显著提高?试验结果说明复合侵染会提高烟粉虱的传毒率, 促进TYLCV的发生与流行?     

Comparison of the reaction of tomato lines to infection by tomato yellow leaf curl begomovirus in Lebanon

Tomato yellow leaf curl begomovirus (TYLCV), transmitted by the whitefly Bemisia tabaci , is epidemic in Africa, the Middle East and South-East Asia. It is also reported in some European countries and the American continent. In Lebanon, it is the major limiting factor for summer and autumn production of tomato. Comparison of the nucleotide sequence in the intergenic region with other reported leaf curl viruses showed the Lebanese TYLCV isolate to be closely related to Egyptian, Israeli and Jamaican isolates (94–96% identity). However, it is not closely related to isolates from Sardinia, Spain and Thailand, or to tomato leaf curl isolates from India, Taiwan and Australia. In field and greenhouse screening tests conducted for 5 years on 67 tomato lines, several were identified as promising. TY-Carla, PSR and RS lines were among the most promising with determinate growth, while S&G 143 and the DR lines were the most promising with semi-determinate and indeterminate growth, respectively. Virus concentrations in most, but not all, tolerant tomato lines were significantly lower than in the susceptible lines. None of the lines tested was immune to the virus. A survey of TYLCV alternative hosts on at least 58 plant species, using nucleic acid hybridization, showed that Amaranthus sp., Malva sp., Sonchus oleraceus , Plantago sp., Solanum melongena , Phaseolus vulgaris and Mercurialis annua may play an important role in the epidemiology of TYLCV in Lebanon. Mercurialis annua is a newly reported host for TYLCV.     

Tomato leaf curl geminivirus in Australia: occurrence, detection, sequence diversity and host range

The occurrence of whitefly transmitted geminiviruses in Australia was studied using a mixed DNA probe capable of detecting a range of distinct geminiviruses. The only geminivirus species detected was Tomato leaf curl virus (TLCV), which is spread across a vast geographical region of far-northern coastal Australia, an area inhabited by the Australasian-Oceania biotype of Bemisia tabaci . The newly introduced silverleaf whitefly, B. tabaci biotype B, forms high population densities in the eastern coastal region of Queensland and is currently located approximately 150 km from the nearest known TLCV-infected area. The viral host range appeared to be narrow and of 58 species of crop plants and weeds inoculated using the B biotype, only 11 became infected with the virus, including five that did not show foliar symptoms. A DNA fragment of 694 nt, including the complete C4 open reading frame (ORF), the overlapping N-terminal part of the C1 ORF and the viral iterons involved in replication, was amplified from 11 TLCV field isolates and sequenced. Sequence analysis revealed an overall sequence variation of up to 14% in this region, as well as the presence of distinct viral iterons.     

陕西省早春茬番茄定植时间与番茄黄化曲叶病毒病发生的关系

为了明确早春茬番茄定植时间与番茄黄化曲叶病毒病发生的关系,采取小区对比试验对不同定植时间下番茄上烟粉虱种群消长动态、番茄黄化曲叶病毒病发病株率、发病程度及番茄产量和经济效益等进行了系统调查。结果表明:番茄田烟粉虱种群消长动态在年度间基本相同,4月上旬始见,5月中旬达到高峰,烟粉虱发生后15 d左右出现番茄黄化曲叶病毒病症状。4月上旬以前定植番茄黄化曲叶病毒病发生较轻,单位面积产量和经济效益较高,4月上旬以后定植发病较重,产量和经济效益较低。定植时间(X_1)与番茄黄化曲叶病毒发病株率(Y_1)及病情指数(Y_2)均呈显著正相关,其相关关系为Y_1=1.470 4X_1+7.947 2,Y_2=0.876 7X_1-6.441 7。生产实践中采取双膜或三膜覆盖栽培等措施适当提早定植,避免番茄感病期与烟粉虱发生高峰期相遇,在不使用任何防治措施下,可有效预防番茄黄化曲叶病毒病暴发成灾。     

北京地区番茄黄化曲叶病毒病的鉴定及防治对策

番茄黄化曲叶病毒病是一种由烟粉虱传播的病毒病,给番茄生产造成严重威胁。2009年在北京郊区调查时发现部分保护地种植的番茄植株表现典型黄化曲叶症状。通过提取典型症状样品总DNA利用粉虱传双生病毒检测简并引物PA/PB,进行PCR扩增到541bp的特异条带。通过测序和核苷酸序列比对表明该序列与番茄黄化曲叶病毒序列相似性最高为99%。分子检测结果表明北京郊区部分保护地种植的番茄已被烟粉虱传播的番茄黄化曲叶病毒侵染危害。     

Pcr-amplification of tomato yellow leaf curl virus (TYLCV) DNA from squashes of plants and whitefly vectors: Application to the study of TYLCV acquisition and transmission

DNA of tomato yellow leaf curl virus (TYLCV), a geminivirus transmitted by the whitefly Bemisia tabaci, was amplified from squashes of infected tomato plants and of viruliferous vectors using the polymerase chain reaction (PCR). Samples of infected tissues as small as 1 mm² were squashed onto a nylon membrane. A 1 × 2 mm strip containing the squash was introduced into a 25 µl PCR reaction mix. The reaction products were subjected to gel electrophoresis, blotted and hybridized with a radiolabeled virus-specific DNA probe. TYLCV DNA was amplified from squashes of leaves, roots, and stem of infected tomato and from individual viruliferous whiteflies. The same squash could be used several times to amplify different virus DNA fragments with various sets of primers. Thus plant and insect squashes can be used as templates for the amplification of geminiviral DNA with no need to prepare tissue extracts or purify nucleic acids. The squash-PCR procedure was applied to study whitefly transmission of TYLCV. Tomato plants were inoculated by placing a single viruliferous insect in the center of a young leaflet. In some plants TYLCV DNA was detected at the site of inoculation as early as 5 min after the beginning of the access feeding and in all plants after 30 min. The squash-PCR procedure also was applied to the study of TYLCV acquisition by the insect vector. TYLCV DNA was detected in the head of whiteflies as early as 5 min after the beginning of the access feeding on infected tomato plants. Viral DNA was detected in the thorax after 10 min and in the abdomen after 25 min.     

Assessment of the effectiveness of Ty genes in tomato against tomato leaf curl Bangalore virus

Tomato leaf curl Bangalore virus (ToLCBaV), a monopartite begomovirus transmitted by the whitefly Bemisia tabaci, has become a major constraint in tomato production in the Indian subcontinent. Earlier breeding efforts in India led to the adoption of tomato cultivars carrying the resistance gene Ty-2. However, it has been observed recently that such cultivars/hybrids are susceptible to begomoviruses. This requires the identification of additional or new sources of resistance against ToLCBaV. The present study was undertaken to assess the effectiveness of several Ty genes in providing resistance against infection by ToLCBaV. The kinetics of virus multiplication in different Ty resistance gene stocks of tomato were estimated and compared using quantitative PCR data. Accumulation of viral genomic units and symptom severity were lower in tomato lines carrying Ty-3 and Ty-2 + Ty-3 compared with those carrying Ty-2 alone. All tested lines carrying Ty-2 showed typical tomato leaf curl disease symptoms. Tomato lines carrying Ty-2 + ty-5 and ty-5 + Ty-6 combinations had a significantly higher disease severity index and viral genomic units compared to those carrying Ty-3 and Ty-2 + Ty-3 at 30 days postinoculation. The accumulation level of ToLCBaV genomic units serves as a good indicator for resistance selection along with other parameters of disease. We present here a comprehensive overview of the effectiveness of Ty-2, Ty-3, and ty-5 alone as well as the combinations Ty-2 + Ty-3 and Ty-2 + ty-5 against ToLCBaV.     

Detection and molecular characterization of viruses associated with tomato yellow leaf curl disease in cucurbit crops in Jordan

In this study, Tomato yellow leaf curl Sardinia virus (TYLCSV) and the strains Israel and Mild of Tomato yellow leaf curl virus (TYLCV-IL, TYLCV-Mld) were detected for the first time in four cucurbit crops in Jordan by nested polymerase chain reaction (nPCR). These viruses cause the tomato yellow leaf curl disease (TYLCD) in tomato. Cucumber, squash, melon and watermelon plants inoculated with TYLCV-IL[JO:Cuc], TYLCV-Mld, TYLCSV-IT[IT:Sar:88] and the Jordanian isolate of TYLCV (TYLCV-JV) did not show disease symptoms. However, virus-specific fragments were detected in uppermost leaves of symptomless plants by nPCR. A whitefly transmission test showed that Bemisia tabaci could transmit TYLCV-Mld from cucumber into tomato and jimsonweed plants. However, all infected tomato plants remained symptomless. In addition, results of semi-quantitative PCR (sqPCR) analysis showed that the relative amount of TYLCV-Mld DNA acquired by B. tabaci from cucumber plants was less than that acquired from tomato plants.     

天津市烟粉虱隐种鉴定及其携带TYLCV、内共生菌情况和抗药性监测

为明确天津市烟粉虱Bemisia tabaci隐种的类别及其寄主适应性、传毒能力、携带内共生菌情况和抗药性,采用mt COI酶切法对从武清、西青、蓟州和宁河4个区的番茄、黄瓜及辣椒3种寄主上采集的12个烟粉虱种群进行隐种鉴定,采用PCR检测其携带番茄黄化曲叶病毒（tomato yellow leaf curl virus,TYLCV）和内共生菌情况,并采用浸叶法测定其对4种常用药剂的抗性。结果表明,采集的烟粉虱种群以MED隐种为主,占所有检测个体的93.33%,有3个种群为MED和MEAM1隐种混合发生。所有检测个体中有36.25%的个体携带TYLCV,在6个种群中检测到TYLCV,其中5个种群有超过50%的个体携带TYLCV。在12个种群中共检测到Hamiltonella、立克次氏体Rickettsia、Cardinium和杀雄菌属Arsenophnus共4种内共生菌,携带个体比例分别为90.63%、48.96%、43.75%和8.33%,进一步对内共生菌协同感染情况进行分析,发现有HARC、HRC、HAC、HR、HC和AC共6个协同感染型,感染率分别为4.17%、28.13%、3....     

Quantitation over time of tomato yellow leaf curl geminivirus DNA in its whitefly vector

ABSTRACT The amount of tomato yellow leaf curl geminivirus (TYLCV) DNA that accumulated in the vector Bemisia tabaci was studied by quantitative chemiluminescent dot-blot assay, using digoxigenin-labeled specific DNA probes. Large groups of female whiteflies were allowed to feed for 4, 12, 24, or 48 h on TYLCV-infected tomato plants and then were transferred to TYLCV-immune cucumber plants. Insects were sampled at different times during and after acquisition access and tested for TYLCV-DNA content. TYLCV-DNA assays were done either on whole insects oron the head plus prothorax (to include salivary glands) and abdomen separately. The maximum amount of TYLCV DNA, averaging from 0.5 to 1.6 ng per insect, was always attained at the end of the acquisition period. The mean amount then decreased by about 1 to 2% per day, remaining clearly detectable up to 20 days after the end of the acquisition period. Only some whiteflies that were TYLCV-positive in the abdomen were positive for head plus prothorax. In both parts of the body, TYLCV DNA remained detectable up to 18 days after the end of the acquisition period, showing that TYLCV DNA remains in insect tissues much longer than infectivity indicates.     

Farmers' perceptions and practices for managing tomato leaf curl virus disease in southern India

A survey of 174 tomato farmers in five districts of Karnataka State, South India, was conducted between March and July 1998 with the objectives of assessing farmers' knowledge and perceptions of tomato leaf curl virus disease (ToLCVD) and examining their current control practices. The findings showed that farmers were familiar with ToLCVD and regarded it as a serious constraint upon production, particularly in the hot season. However, only 2% of farmers were aware that tomato leaf curl virus (ToLCV) was transmitted by a whitefly vector, Bemisia tabaci . A similarly small number of farmers knew that ToLCVD was caused by a virus, whilst 86% of farmers believed that it was caused by high temperatures. Approximately 90% of farmers relied primarily on pesticides for control of ToLCVD. Few ToLCV-resistant varieties were available but one such variety, Avinash II, was grown by 13% of farmers in the hot season when incidence of the disease is highest. Farmers visiting experimental trials at a farmer field day, showed a strong interest in new advanced breeding tomato lines with resistance to ToLCV. Factors affecting the adoption of ToLCV-resistant varieties by tomato farmers are discussed.     

Note:Bemisia tabaci biotype B associated with tomato yellow leaf curl disease epidemics on rhodes Island, Greece

In 2006 an outbreak of tomato yellow leaf curl disease occurred in tomato crops on Rhodes Island, Greece. Diseased plants were found to be infested with the B biotype of theBemisia tabaci (Gennadius) complex and greenhouse and open-field-grown tomato crops were infected withTomato yellow leaf curl virus (TYLCV) introduced from the Middle East. This is the first report of TYLCV and the B biotype ofB. tabaci on Rhodes Island in Greece. http://www.phytoparasitica.org posting Dec. 11, 2007.     

Tomato dwarf leaf curl virus, a new bipartite geminivirus associated with tomatoes and peppers in Jamaica and mixed infection with tomato yellow leaf curl virus

Genomic characterization using nonradioactive probes, polymerase chain reaction with degenerate primers for whitefly transmitted geminiviruses and nucleotide sequencing were used to describe a new bipartite geminivirus, associated with dwarfing and leaf curling of tomatoes and peppers in Jamaica. Partial DNA-A and DNA-B clones were obtained. DNA sequence analysis showed that tomato and pepper samples have a similar geminivirus associated with them. Nucleotide sequence identity > 92% between the common regions of DNA-A and DNA-B confirmed the bipartite nature of the Jamaican geminivirus isolates. Nucleotide sequence comparisons of DNA-A and DNA-B with those of geminiviruses representing the major phylogenetic groups of Western Hemisphere geminiviruses showed the greatest similarity to potato yellow mosaic virus and members of the Abutilon mosaic virus cluster of geminiviruses. This new virus is given the name tomato dwarf leaf curl virus (TDLCV) because of the dwarfing and leaf curling symptoms associated with infected tomato plants. Polymerase chain reaction and Southern hybridization showed mixed infections of TDLCV with tomato yellow leaf curl virus from Israel in 16% of the field samples of tomatoes and peppers.     

侵染广西番茄的番茄黄化曲叶病毒的分子鉴定

番茄黄化曲叶病毒病是番茄生产上的毁灭性病害,严重影响番茄产量及品质。2019年从广西百色市采集疑似番茄黄化曲叶病叶片样品,采用滚环扩增(RCA)及基因克隆等方法,获得了4个烟粉虱传双生病毒的全基因序列,4个分离物的全长序列分别为2 776、2 781、2 752、2 781 bp,均编码6个开放阅读框;核苷酸相似性比较发现,4个分离物彼此间的相似性均在90%以上,与已报道的番茄黄化曲叶病毒Tomato yellow leaf curl virus (TYLCV)各分离物间的相似性也在91%以上;系统进化分析表明,TYLCV广西分离物BS-2和BS-4与TYLCV-Hunan、BS-1和BS-3与TYLCV-YN6553等分离物处于独立的小分支,说明TYLCV广西分离物与TYLCV-Hunan、TYLCV-YN6553具有较近的亲缘关系。本研究首次报道TYLCV在广西发生。     

番茄褪绿病毒与番茄黄化曲叶病毒复合侵染对番茄褪绿病毒传播的影响

番茄褪绿病毒Tomato chlorosis virus(ToCV)是一种由烟粉虱Bemisia tabaci传播的正义单链RNA病毒,在田间常与番茄黄化曲叶病毒Tomato yellow leaf curl virus(TYLCV)复合侵染而造成番茄生产上重大的经济损失。为了明确ToCV与TYLCV的复合侵染对烟粉虱传播ToCV所造成的影响,本文采用RT-PCR以及qRT-PCR检测了复合侵染的番茄对烟粉虱获取和传播ToCV的影响。研究表明,烟粉虱取食复合侵染的番茄后对ToCV的传播效率显著提高,仅25头烟粉虱的传毒率即可达到100%,ToCV在烟粉虱以及番茄体内的累积量均显著提高。说明这种复合侵染促进了烟粉虱对ToCV的传播,在田间应当及时防控烟粉虱,警惕病毒与烟粉虱的蔓延。     

Identification of whitefly transmitted tomato yellow leaf curl geminivirus from Iran and a survey of its distribution with molecular probes

A virus causing a disease of tomato, prevalent in the southern provinces of Iran, with symptoms of leaf-curling, stunting, reduction of leaf size, leaf corrugation, shortening of internodes and severe reduction in fruit yield, was shown to be transmissible to healthy tomato plants by grafting and by whiteflies ( Bemisia tabaci ), but not by sap inoculation. Geminivirus DNA was detected in extracts of diseased tomato plants by dot-blot hybridization assays using as probes full-length cloned DNA of Australian, Italian (Sardinian) or Jordanian strains of tomato yellow leaf curl virus (TYLCV). Geminivirus coat protein was detected in whitefly inoculated plants by dot immunobinding assay using polyclonal antibody raised against Jordanian TYLCV. A limited survey using the dot-blot hybridization assay for virus detection indicated the presence of the virus in tomato-growing provinces of southern but not northern Iran. Whitefly transmission experiments to tomato under controlled greenhouse conditions showed that some isolates of TYLCV-like geminiviruses from different parts of Iran differ in symptomatology.     

南疆温室番茄黄化曲叶病病毒种类的分子鉴定

为明确南疆温室番茄黄化曲叶病的病毒种类,利用双生病毒的兼并引物通过PCR扩增,对采集的20个番茄病株进行了分子检测.从20个病株中均扩增到约500 bp的目标片段,对其中4株进行克隆和测序,其相互间序列同源性为97.1％ ～99.3％,与番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)的同源性较高,为98.6％ ～ 99.5％.随机选取莎车分离物KS2-5进行全基因组的克隆和测序,KS2-5 DNA全长为2781 nt(序列号:JQ807735),具有典型的双生病毒基因组特征,与TYLCV其它分离物同源性达到98.9％～99.5％,而与其它粉虱传双生病毒的序列同源性较低,为68.3％ ～75.5％,表明危害南疆温室番茄的病毒种类为番茄黄化曲叶病毒TYLCV.     

Spread of Tomato yellow leaf curl virus in Sicily: Partial Displacement of Another Geminivirus Originally Present

The geminivirus Tomato yellow leaf curl virus (TYLCV) was reported for the first time in Italy in 2002. We have followed its spread in Sicily, where Tomato yellow leaf curl Sardinia virus (TYLCSV), another tomato-infecting geminivirus, is endemic and has been causing severe crop losses since 1989. The presence of the two viruses was monitored in the main tomato growing area, the Ragusa province, analyzing samples with yellow leaf curling symptoms. At first (spring–summer 2002) both viruses were always found in mixed infections, but in 2003 and 2004 18–35% of plants were found infected by TYLCV alone and 8–28% by TYLCSV alone, with 41–69% carrying both viruses. TYLCV has spread quickly in the area, demonstrating, as in other parts of the world, its high virulence and invasiveness; however it has not, so far, completely displaced TYLCSV. An infectious clone of TYLCV from Sicily (TYLCV-IT) was sequenced. The nucleotide sequence was 97% identical to other TYLCV strains of the ‘severe’ type, found in many countries worldwide.