Solanum nigrum: an indigenous weed reservoir for a tomato yellow leaf curl geminivirus in southern Spain

A tomato yellow leaf curl geminivirus (TYLCV-AL), was first identified in tomato plants in Almeria, southern Spain in 1992. This virus is transmitted by the tobacco whitefly, Bemisia tabaci (Gennadius), and is presently infecting tomato crops throughout the south eastern region of Spain. Solanum nigrum, collected from a field in south east Spain and exhibiting leaf curl symptoms, was squash blotted onto nylon membrane and gave a positive signal when hybridised to a TYLCV-Is DNA probe. Laboratory tests showed B. tabaci to transmit TYLCV-AL from infected tomato plants to S. nigrum seedlings. The virus could then be acquired by B. tabaci and transmitted back from infected S. nigrum plants to tomato, inducing typical TYLCV symptoms. These results indicate the importance of S. nigrum as a weed host/reservoir for a TYLCV and its possible role in the spread of this virus within Europe.     

烟粉虱传播的番茄褪绿病毒和番茄黄化曲叶病毒对不同番茄品种的复合侵染

为明确烟粉虱传播的番茄褪绿病毒(Tomato chlorosis virus,ToCV)与番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)对不同番茄品种的复合侵染情况,于2015年11月在山东省寿光市温室内采集13个番茄品种共390份疑似发病植株叶片,对不同番茄品种的TYLCV抗性和2种病毒的复合侵染以及温室内发病番茄植株上烟粉虱成虫的带毒率进行检测。结果表明,采集的13个番茄品种经分子标记检测鉴定均为TYLCV杂合抗性;不同番茄品种ToCV与TYLCV的复合侵染率存在明显差异,大果番茄粉宴和贝瑞上复合侵染率最高可达73.3%,而樱桃番茄八喜上未检测到这2种病毒的复合侵染。此外,在发病番茄植株上采集的烟粉虱成虫体内可检测到2种病毒,其中烟粉虱ToCV带毒率为90.7%,TYLCV带毒率为80.0%,同时检测到ToCV与TYLCV的概率为71.3%。表明ToCV和TYLCV的复合侵染在山东省番茄生产中普遍发生,烟粉虱可同时携带这2种病毒并广泛传播。     

Displacement of Tomato Yellow Leaf Curl Virus (TYLCV)-Sr by TYLCV-Is in Tomato Epidemics in Spain

ABSTRACT A progressive displacement of tomato yellow leaf curl virus (TYLCV)-Sr by TYLCV-Is was observed in tomato epidemics in southern Spain based on incidence data of both virus species obtained during surveys conducted between 1996 and 1998. Ecological factors that might be involved in such a displacement, such as competition of TYLCV-Sr and TYLCV-Is in tomato, transmission by local biotypes (B and Q) of Bemisia tabaci, and presence in weeds and alternate crops, have been analyzed. No selective advantage is observed for TYLCV-Sr or TYLCV-Is in tomato plants either infected via Agrobacterium tumefaciens or via B. tabaci. However, TYLCV-Is is more efficiently vectored by local biotypes of B. tabaci; and common bean, a bridge crop between tomato crops, is a host for TYLCV-Is but not TYLCV-Sr. Therefore, common bean acts as a reservoir for TYLCV-Is. These two factors are probably responsible for the displacement of TYLCV-Sr by TYLCV-Is as the causative agent of epidemics in tomato in southern Spain.     

Comparison of Resistance to Tomato Leaf Curl Virus (India) and Tomato Yellow Leaf Curl Virus (Israel) among Lycopersicon Wild Species, Breeding Lines and Hybrids

The objective of this study was to screen wild and domesticated tomatoes for resistance to Tomato yellow leaf curl virus, Israel (TYLCV-Is) and Tomato leaf curl virus from Bangalore isolate 4, India (ToLCV-[Ban4]) to find sources of resistance to both viruses. A total of 34 tomato genotypes resistant/tolerant to TYLCV-Is were screened for resistance to ToLCV-[Ban4] under glasshouse and field conditions at the University of Agricultural Sciences, Bangalore, India. Resistance was assessed by criteria like disease incidence, symptom severity and squash-blot hybridization. All the tomato genotypes inoculated with ToLCV-[Ban4] by the whitefly vector Bemisia tabaci (Gennadius) produced disease symptoms. In some plants of the lines 902 and 910, however, the virus was not detected by hybridization. The tomato genotypes susceptible to ToLCV-[Ban4] by whitefly-mediated inoculation were also found susceptible to the virus under field conditions. However, there were substantial differences between genotypes in disease incidence, spread, symptom severity and crop yield. Despite early disease incidence, many genotypes produced substantially higher yields than the local hybrid, Avinash-2. Sixteen tomato genotypes from India resistant/tolerant to ToLCV-[Ban4] were also tested for TYLCV-Is resistance at the Hebrew University of Jerusalem, Rehovot, Israel. Accessions of wild species, Lycopersicon hirsutum LA 1777 and PI 390659 were the best sources of resistance to both viruses. Lines 902 and 910, which were, resistant to TYLCV-Is were only tolerant to ToLCV-[Ban4] and accession Lycopersicon peruvianum CMV Sel. INRA, resistant to ToLCV-[Ban4], was only tolerant to TYLCV-Is. Implications of using the resistant lines in breeding programme is discussed.     

Typing of Tomato Yellow Leaf Curl Viruses in Europe

Tomato yellow leaf curl disease is spreading in southern Europe, where it has quickly become a serious problem. In recent years, several virus isolates have been characterised. Although with some genetic variability, all isolates found in Europe belong to one of two species Tomato yellow leaf curl-Sardinia (TYLCV-Sar) or Tomato yellow leaf curl-Israel (TYLCV-Is). Several methods were tested to identify and type TYLCV isolates from field samples: (1) RFLP of a DNA fragment amplified from the coat protein gene; (2) PAGE of a fragment amplified from the C2 gene; (3) dot-blot hybridisation. All methods enabled the detection of the TYLCVs and provided good indications for attributing them to one species or the other. However, for typing purposes, the RFLP method was the most reliable, due to the easily recognisable pattern produced by the two virus species present in Europe. Dot-blot hybridisation is less expensive for identifying TYLCVs in large numbers of samples, particularly when a mixture of two probes is used. PAGE of the C2 fragment is the fastest of the methods tested.     

Detection and typing of tomato yellow leaf curl viruses

The denomination Tomato yellow leaf curl virus (TYLCV) comprises several viruses that cause severe damage to tomato crops in warm and temperate regions worldwide. TYLCV viruses are widespread in the Mediterranean Basin, in which two species have been reported: Tomato yellow leaf curl Sardinia virus (TYLCSV) and Tomato yellow leaf curl virus (TYLCV, previously TYLCV-Is). The availability of methods convenient for the diagnosis of these viruses is essential. We have investigated several alternatives for reliable detection and differentiation of TYLCSV and TYLCV. Triple-antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) proved to be very useful for large-scale diagnosis in field situations, but lacked discriminating capacity and sensitivity in the stages of infection in which low virus titre is present. The DNA-based methods are suited to laboratory operations and plant disease clinics, where accuracy of detection and discrimination of viruses is required. Polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) was the most reliable method to discriminate between TYLCSV and TYLCV, but is not suited to high sample turnover. For large-scale testing, tissue print hybridization assay provides a reliable and sensitive alternative to PCR.     

Differentiation of three whitefly-transmitted geminiviruses from the Republic of Yemen

Three viruses collected in southern Yemen in 1990, infecting watermelon, tobacco and tomato were shown to be transmitted by the whiteflyBemisia tabaci and to have particle morphologies typical of geminiviruses. Colonies ofB. tabaci collected from different locations and from different hosts were used in virus transmission tests with the same host range of plants. Colonies established from both watermelon and cotton in the Yemen were identified as the squash silverleaf-inducing B biotype. The culture host of the colony did not influence virus acquisition and transmission efficiencies to and from other hosts. The tobacco and tomato geminiviruses had a similar host range, but differed in their severity in some hosts. Both these viruses differed from the watermelon geminivirus in host range and symptoms.Datura stramonium, an alternative host for all three viruses, could be co-infected by the watermelon and tobacco viruses.B. tabaci was able to acquire both viruses from the co-infectedD. stramonium and infect seedlings of either original host plant species with their respective viruses orD. stramonium with both. The viruses were identified as watermelon chlorotic stunt virus, tobacco leaf curl virus and tomato yellow leaf curl virus and were distinguished by cross hybridisation.     

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。     

Biomolecular relationships among isolates of<Emphasis Type="Italic">Tomato Yellow Leaf Curl Tanzania Virus</Emphasis>

An investigation of the biological properties of the virus causing tomato yellow leaf curl disease in Tanzania was initiated to compare it with other known tomato yellow leaf curl viruses. Properties relating to acquisition and inoculation feeding time, persistence, mechanical inoculation, seed transmission and host range were studied. Results obtained indicate that the virus was transmitted persistently byBemisia tabaci Genn., but it was not mechanically, sap- or seed-transmissible. Minimum acquisition and inoculation feeding time was 30 min.Capsicum annuum, Datura stramonium, Nicotiana glutinosa, N. tabacum andLycopersicon esculentum were found to be hosts of the virus among the plant species tested, whereasPhaseolus vulgaris was not. It is concluded that the properties of the agent causing yellow leaf curl symptoms in tomato plants from different regions in Tanzania are similar to those ofTomato yellow leaf curl Sardinia virus species studied elsewhere. http://www.phytoparasitica.org posting Feb. 20, 2003.     

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.     

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.     

Pepper (Capsicum annuum) Is a Dead-End Host for Tomato yellow leaf curl virus

ABSTRACT Tomato yellow leaf curl (TYLC) is one of the most devastating pathogens affecting tomato (Lycopersicon esculentum) worldwide. The disease is caused by a complex of begomovirus species, two of which, Tomato yellow leaf curl Sardinia virus (TYLCSV) and Tomato yellow leaf curl virus (TYLCV), are responsible for epidemics in Southern Spain. TYLCV also has been reported to cause severe damage to common bean (Phaseolus vulgaris) crops. Pepper (Capsicum annuum) plants collected from commercial crops were found to be infected by isolates of two TYLCV strains: TYLCV-Mld[ES01/99], an isolate of the mild strain similar to other TYLCVs isolated from tomato crops in Spain, and TYLCV-[Alm], an isolate of the more virulent TYLCV type strain, not previously reported in the Iberian Peninsula. In this work, pepper, Nicotiana benthamiana, common bean, and tomato were tested for susceptibility to TYLCV-Mld[ES01/99]and TYLCV-[Alm] by Agrobacterium tumefaciens infiltration, biolistic bombardment, or Bemisia tabaci inoculation. Results indicate that both strains are able to infect plants of these species, including pepper. This is the first time that infection of pepper plants with TYLCV clones has been shown. Implications of pepper infection for the epidemiology of TYLCV are discussed.     

关中地区温室越冬茬番茄黄化曲叶病毒病分子鉴定及流行规律

为了明确关中地区越冬茬番茄黄化曲叶病毒病发生和流行规律,通过分析该病发生与番茄品种、定植期及传播介体烟粉虱之间的关系,并采用PCR技术对田间病原进行分子鉴定。结果表明,番茄黄化曲叶病毒病在8月中下旬至11月上中旬开始侵染,翌年3月中下旬发生再侵染,秋季病情减轻;烟粉虱种群数量与病害发生程度呈线性正相关;不同番茄品种对番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)的抗性差异显著,其中大番茄品种布鲁尼1288和DRW7728,小番茄品种千禧和美红对该病表现为免疫;分子检测结果表明,4个样品中均扩增出543 bp的特异片段,与NCBI数据库Gen Bank的TYLCV序列(登录号为GU084381、KC138544.1、KC138543.1和JX456642.1)的相似性达99%。研究表明,关中地区番茄病毒病为番茄黄化曲叶病毒病,番茄品种、定植期及烟粉虱发生动态是影响该病发生的主要因素。     

A new approach to evaluate relative resistance and tolerance of tomato cultivars to begomoviruses causing the tomato yellow leaf curl disease in Spain

A simple procedure to evaluate relative resistance and tolerance of tomato cultivars to the begomoviruses causing tomato yellow leaf curl (TYLC) disease in Spain was developed. To estimate the resistance and tolerance levels of a cultivar, several formulae were developed based on the ratio of infected plants, virus titre (estimated by tissue–print hybridization) and symptom intensity. The formulae were applied to five commercial tomato cultivars (Amoretto, Birloque, Royesta, Tovigreen and Ulises) naturally infected by TYLC viruses. The analyses showed that Ulises, Birloque and Tovigreen exhibited a moderate resistance, and Ulises was also highly tolerant. There was a positive correlation between symptom intensity and virus titre in infected plants, suggesting that the hybridization technique could also be used as an early estimator of tolerance. Finally, molecular hybridization and nucleotide sequence analyses of the begomovirus intergenic region showed that the local TYLC virus population consisted of a single species, Tomato yellow leaf curl virus (TYLCV, formerly TYLCV-Israel), with low genetic variation (nucleotide identity between isolates higher than 97%).     

Pymetrozine interferes with transmission of<Emphasis Type="Italic">Tomato yellow leaf curl virus</Emphasis> by the whitefly<Emphasis Type="Italic">Bemisia tabaci</Emphasis>

Pymetrozine, a novel compound belonging to the class pyridine-azomethines, is a feeding inhibitor labeled for use against plant pests in the order Hemiptera. Pymetrozine was evaluated for its ability to interfere with whitefly transmission of the begomovirusTomato yellow leaf curl virus (TYLCV). Pymetrozine was applied as Fulfill ^TM 50 WG at two rates (0.291 and 0.582 g formulationl ⁻¹) to tomato seedlings with four to six true leaves. Viruliferous whiteflies (three to five per plant) were added 1, 4, 7 and 11 d after a single application of pymetrozine, and transmission rates were determined 4 wk after the addition of whiteflies. Pymetrozine provided protection against transmission of TYLCV by viruliferous whiteflies for up to 1 wk after a single apliation. No phytotoxicity was observed on tomato transplants. These results indicate that pymetrozine could be an effective tool for tomato transplant producers to protect susceptible transplants from infection by begomoviruses, such as TYLCV. Pymetrozine might also work well as part of an integrated approach to begomovirus management in greenhouse tomato fruit production. http://www.phytoparasitica.org positing Oct. 20, 2003.     

Three distinct groups of isolates of <Emphasis Type="Italic">Tomato yellow leaf curl virus</Emphasis> in Japan and construction of an infectious clone

Complete nucleotide sequences of eight Japanese isolates of Tomato yellow leaf curl virus (TYLCV) were determined and compared with four TYLCV isolates already reported. These isolates separated into three groups – Shizuoka (Sz), Aichi (Ai), Nagasaki (Ng) – and had 99% identities within the groups. Full-length molecules of DNA-A of group Sz consist of 2791nt and those of group Ai contain 2787nt. Both were closely related to TYLCV-Is.M, although those of group Ng had 2793nt and were more closely related to TYLCV-Is. Comparison of common sequences of isolates belonging to groups Sz and Ai had substitutions of 4nt in the intergenic region and nonsynonymous substitutions at open reading frames between the groups. None of the isolates tested had DNA molecules. Agroinfection of four plant species with a DNA-A dimeric infectious clone of TYLCV-SzY, a member of group Sz, resulted in systemic infection. Tomato plants then developed typical yellow leaf curl symptoms.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession numbers AB116629, AB116630, AB116631, AB116632, AB116633, AB116634, AB116635, and AB116636     

湖南省番茄和牵牛花上双生病毒的分子鉴定

双生病毒是一类在全世界范围内广泛发生的单链环状DNA病毒。本文对从湖南采集到的6例(洋姜、番茄、萝卜、赛葵、甘薯、牵牛花)疑似双生病毒侵染的植物叶片进行了分子鉴定。利用滚环扩增技术(RCA)对样品DNA进行扩增,分别对其RCA产物进行酶切,并将酶切得到的片段测序后进行BLAST比对,结果显示番茄样品中的病毒分离物与番茄黄化曲叶病毒相似性最高(99%),牵牛花样品中的病毒分离物与甘薯卷叶病毒相似性最高(99%),证明这两个分离物是单组分DNA-A双生病毒。这是在湖南省首次发现并报道双生病毒的全核酸序列。     

Arabidopsis thaliana,an experimental host for tomato yellow leaf curl disease‐associated begomoviruses by agroinoculation and whitefly transmission

Tomato yellow leaf curl disease is one of the most devastating viral diseases affecting tomato crops worldwide. This disease is caused by several begomoviruses (genus Begomovirus, family Geminiviridae), such as Tomato yellow leaf curl virus (TYLCV), that are transmitted in nature by the whitefly vector Bemisia tabaci. An efficient control of this vector‐transmitted disease requires a thorough knowledge of the plant–virus–vector triple interaction. The possibility of using Arabidopsis thaliana as an experimental host would provide the opportunity to use a wide variety of genetic resources and tools to understand interactions that are not feasible in agronomically important hosts. In this study, it is demonstrated that isolates of two strains (Israel, IL and Mild, Mld) of TYLCV can replicate and systemically infect A. thaliana ecotype Columbia plants either by Agrobacterium tumefaciens‐mediated inoculation or through the natural vector Bemisia tabaci. The virus can also be acquired from A. thaliana‐infected plants by B. tabaci and transmitted to either A. thaliana or tomato plants. Therefore, A. thaliana is a suitable host for TYLCV–insect vector–plant host interaction studies. Interestingly, an isolate of the Spain (ES) strain of a related begomovirus, Tomato yellow leaf curl Sardinia virus (TYLCSV‐ES), is unable to infect this ecotype of A. thaliana efficiently. Using infectious chimeric viral clones between TYLCV‐Mld and TYLCSV‐ES, candidate viral factors involved in an efficient infection of A. thaliana were identified.     

京郊番茄主要病毒病毒原鉴定

近年来, 番茄病毒病, 特别是番茄黄化曲叶病毒（TYLCV）病在北京地区空前暴发, 给番茄生产造成严重威胁, 使番茄的产量和品质显著降低。2012-2013年在北京周边7个区县, 采集疑似感染病毒的番茄植株样品325份, 分别针对TYLCV、烟草花叶病毒（TMV）、黄瓜花叶病毒（CMV）3种病毒进行了PCR或ELISA 检测。检测结果表明, 目前在北京地区大棚或温室内发生的番茄病毒病以TYLCV为主; 露地番茄病毒复合侵染现象比较普遍, 病毒检出率100%, 其中TYLCV检出率达到75%以上。在TYLCV侵染的样品中, TYLCV和CMV复合侵染占20%左右, TYLCV和TMV复合侵染占15%左右。部分样品检测到TYLCV、CMV 和TMV 3种病毒复合侵染的现象。