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.     

Tomato yellow leaf curl virus in the Dominican Republic: Characterization of an Infectious Clone, Virus Monitoring in Whiteflies, and Identification of Reservoir Hosts

ABSTRACT Epidemics of tomato yellow leaf curl disease (TYLCD) in the Dominican Republic in the early to mid-1990s resulted in catastrophic losses to processing tomato production. As part of an integrated management approach to TYLCD, the complete nucleotide sequence of a full-length infectious clone of an isolate of Tomato yellow leaf curl virus (TYLCV) from the Dominican Republic (TYLCV-[DO]) was determined. The TYLCV-[DO] genome was nearly identical in sequence (>97%) and genome organization to TYLCV isolates from Israel and Cuba. This established that TYLCV-[DO] is a bonafide TYLCV isolate (rather than a recombinant virus, such as isolates from Israel [Mild], Portugal, Japan, and Iran), and provided further evidence for the introduction of the virus from the eastern Mediterranean. A reduction in the incidence of TYLCV in the northern and southern processing tomato production areas of the Dominican Republic has been associated with the implementation of a mandatory 3-month whitefly host-free period (including tomato, common bean, cucurbits, eggplant, and pepper). Monitoring TYLCV levels in whiteflies, by polymerase chain reaction with TYLCV-specific primers, established that the incidence of TYLCV decreased markedly during the host-free period, and then gradually increased during the tomato-growing season. In contrast, TYLCV persisted in whiteflies and tomato plants in an area in which the host-free period was not implemented. Surveys for TYLCV reservoir hosts, conducted to identify where TYLCV persists during the host-free period, revealed symptomless infections in a number of weed species. The implications of these findings for TYLCV management in the Dominican Republic are discussed.     

Evidence of a new Tomato yellow leaf curl virus in Japan and its detection using PCR

A new isolate of Tomato yellow leaf curl virus (TYLCV) has been identified from tomato plants in Kochi Prefecture in Japan and designated TYLCV-[Tosa]. The complete nucleotide sequence of the isolate was determined and found to consist of 2781 nt. In phylogenetic analyses of entire nucleotide sequences, TYLCV-[Tosa] was delineated as a single branch and was more closely related to TYLCV-[Almeria] than TYLCV isolates Ng, Sz, or Ai reported in Japan, which had spread since 1996. Isolate TYLCV-[Tosa] is suggested to be a newly introduced, novel isolate of TYLCV that dispersed into Kochi Prefecture. In addition, a rapid method using the polymerase chain reaction to separate TYLCV isolates into four genetic groups was established. This method would be useful for reliable diagnosis based on genetic differences among isolates of TYLCV.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession numbers AB192965 and AB192966     

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。     

Capsicum Species: Symptomless Hosts and Reservoirs of Tomato yellow leaf curl virus

ABSTRACT Five Capsicum species were tested for susceptibility to Tomato yellow leaf curl virus (TYLCV) and the mild strain of TYLCV (TYLCV-Mld). TYLCV was able to infect 30 of 55 genotypes of C. annuum, one of six genotypes of C. chinense, one of two genotypes of C. baccatum, and the only genotype of C. frutescens tested but was unable to infect the one genotype of C. pubescens tested. This is the first evidence for the susceptibility of C. baccatum, C. chinense, and C. frutescens to TYLCV. Unlike TYLCV isolates, TYLCV-Mld was unable to infect C. chinense. No host differences were observed between the Israeli and Florida isolates of TYLCV. None of the Capsicum species showed symptoms after infection with TYLCV or TYLCV-Mld. TYLCV was detected in fruits of C. annuum, but whiteflies were unable to transmit virus from fruits to plants. White-flies were able to transmit both TYLCV and TYLCV-Mld from infected pepper plants to tomato plants. Pepper plants in research plots were found infected with TYLCV at rates as much as 100%. These data demonstrate the ability of some genotypes of pepper to serve as reservoirs for the acquisition and transmission of TYLCV and TYLCV-Mld.     

Virus transmission studies and diagnostic detection of four begomovirus isolates in selected crop hosts and in Bemisia tabaci biotype B*

Virus transmission studies were conducted under glasshouse conditions using the vector Bemisia tabaci biotype B to determine how effectively isolates of the begomoviruses Tomato yellow leaf curl virus (TYLCV) and Tomato leaf curl Bangalore virus (ToLCBV) could be transmitted to phaseolus bean, capsicum and tomato test plants, the latter host used as a positive control for transmission. Diagnostic detection of viruses in these host crops and vector was also evaluated. Polymerase chain reaction (PCR) detection of TYLCV in bean cv. Wade and capsicum cv. Bellboy was achieved 4 weeks after fumigation in asymptomatic plants. Detection of TYLCV in tomato controls was achieved 2 weeks after fumigation with improved frequency of detection at 4 weeks. PCR was found to be a more sensitive method than triple‐antibody sandwich enzyme‐linked immunosorbent assay (TAS‐ELISA) for the detection of TYLCV isolates in all hosts. ToLCBV was detected by PCR and TAS‐ELISA in bean. TYLCV was also detected by PCR in the vector, with a novel internal positive control. This work was carried out to facilitate the development of a diagnostic protocol for the begomoviruses causing tomato yellow leaf curl under the EU SMT programme project –‘Diagnostic protocols for organisms harmful to plants’ (DIAGPRO).     

Current situation of Tomato yellow leaf curl virus in Portugal

The name Tomato yellow leaf curl virus (TYLCV) has been applied to a group of virus species of the genus Begomovirus in the family Geminiviridae that cause a similar tomato disease worldwide. In 1995, TYLCV was first reported in Algarve (southern Portugal) as responsible for an epidemic outbreak of a severe tomato disease. Molecular data have shown that this Portuguese TYLCV isolate was distinct from those previously reported in Europe, as it belonged to the TYLCV-Israel species ¹ . Since then, TYLCV epidemics have occurred annually, being a limiting factor mainly for autumn/winter glasshouse tomato crops. In 1998, TYLCV was also found associated with the emergence of a novel disease of Phaseolus vulgaris in Algarve. The affected bean plants were severely stunted and gave no marketable yield. However, the disease occurs only sporadically, even in conditions of high TYLCV infection pressure. Recently, Tomato chlorosis virus (ToCV), a whitefly-transmitted bipartite closterovirus (genus Crinivirus , family Closteroviridae ), was found associated with an unusual tomato yellow leaf syndrome, in single or mixed infection with TYLCV. The impact of this new pathosystem on tomato production has yet to be determined. Surveys are in progress in mixed cropping systems infested with whiteflies. So far, TYLCV and ToCV diseases are limited to the Algarve region.     

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.     

Experimental transmission of the mild strain of Tomato yellow leaf curl virus (TYLCV) to Amaranthus dubius by Bemisia tabaci

Tomato yellow leaf curl virus (TYLCV) is a monopartite begomovirus from the Old World. The mild strain of this virus (TYLCV-Mld) was described for South America in Venezuela in 2007. Due to the potential risks of establishment of this virus in the field, six common weeds were evaluated for susceptibility to an isolate of TYLCV-Mld by using adults of Bemisia tabaci (Hemiptera: Aleyrodidae) to mediate viral transmission. In this work, detection based on PCR amplification with TYLCV specific primers showed Amaranthus dubius (Amaranthaceae) as the only infected weed. In A. dubius, viral symptoms were observed from 11.0?±?1.3?days post-inoculation and the transmission rate of TYLCV-Mld to this plant species was 83.3%. The successful back-transmission of TYLCV-Mld from A. dubius to tomato plants was demonstrated.     

侵染甜椒的番茄黄化曲叶病毒的分子鉴定和全序列分析

<正>番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)在世界范围内可危害多种作物,造成植株矮化、叶片皱缩变形、局部黄化等症状。该病毒自1964年首次报道以来已蔓延至世界多地。在我国2006年上海首次报道该病毒[1],随后江苏、山东、安徽、北京、河北、天津等地相继报道,危害严重。TYLCV为双生病毒科(Geminiviridae)菜豆金色花叶病毒属(Begomovirus)成员,基因组为单组     

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.     

番茄黄化曲叶病毒在湖北首次报道

2014年春季,在湖北省武汉市发现种植的番茄表现植株矮缩,叶片上卷,叶缘黄化等症状。PCR检测结果显示,所有采集的病样中均存在菜豆金色花叶病毒属病毒。进一步通过滚环扩增方法获得了该病毒的湖北分离物HB01的全基因组。基因克隆及序列分析结果表明,该病毒基因组全长为2 781nt,与已报道的番茄黄化曲叶病毒(TYLCV)各分离物同源性在89.0%以上,而与来自中国不同地区的TYLCV分离物的同源率均在97.0%以上。因此,HB01属于TYLCV的一个分离物。     

Resistance to Monopartite Begomoviruses Associated with the Bean Leaf Crumple Disease in Phaseolus vulgaris Controlled by a Single Dominant Gene

ABSTRACT Tomato yellow leaf curl virus (TYLCV) and Tomato yellow leaf curl Málaga virus are monopartite begomoviruses (genus Begomovirus, family Geminiviridae) that infect common bean (Phaseolus vulgaris), causing bean leaf crumple disease (BLCD). This disease was found to be widespread in southern Spain and causes stunted growth, flower abortion, and leaf and pod deformation in common bean plants. Commercial yield losses of up to 100% occur. In the present study, we have identified and characterized a resistance trait to BLCD-associated viruses in the common bean breeding line GG12. This resistance resulted in a complete absence of BLCD symptoms under field conditions or after experimental inoculation. Our analysis showed that virus replication was not inhibited. However, a severe restriction to systemic virus accumulation occurred in resistant plants, suggesting that cell-to-cell or long-distance movement were impaired. In addition, recovery from virus infection was observed in resistant plants. The reaction of P. vulgaris lines GG12 (resistant) and GG14 (susceptible), and of F(1), F(2), and backcross populations derived from them, to TYLCV inoculation suggested that a single dominant gene conferred the BLCD resistance described here.     

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.     

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.     

截形叶螨抗哒螨灵种群抗性遗传方式的研究

通过对室内选育出的截形叶螨敏感品系和抗哒螨灵品系的杂交和回交试验表明:所测得的截形叶螨正交F1SR(SS♀×RR♂)和反交F1RS(RR♀×SS♂)代的显性度DSR和DRS分别为0.40和0.60,表明抗性由不完全显性基因控制;两个D值(DRS和DSR)95%置信限有重叠,并且经t检验,两个D值不存在显著差异(P0.05),证明截形叶螨对哒螨灵的抗性遗传为常染色体控制;回交F2代(BC1RS和BC1SR)的实际死亡率和期望值经χ2检验,无显著性差异(χ2=9.72,df=9,P0.05),表明抗性遗传由单基因控制。     

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.     

The Natural Occurrence of Two Distinct Begomoviruses Associated with DNAbeta and a Recombinant DNA in a Tomato Plant from Indonesia

ABSTRACT Two begomoviruses (Java virus-1 and Java virus-2), two satellite DNAs (DNAbeta01 and DNAbeta02), and a recombinant DNA (recDNA) were cloned from a single tomato plant from Indonesia with leaf curl symptoms, and the role of these satellite DNAs in the etiology of begomovirus disease was investigated. The genome organizations of the two viruses were similar to those of other Old World monopartite begomoviruses. Comparison of the sequences with other begomoviruses revealed that Java virus-1 was a newly described virus for which the name Tomato leaf curl Java virus (ToLCJAV) is proposed. Java virus-2 was a strain of Ageratum yellow vein virus (AYVV) (AYVV-[Java]). ToLCJAV or AYVV-[Java] alone did not induce leaf curl symptoms in tomato plants. However, in the presence of DNAbeta02, both ToLCJAV and AYVV-[Java] induced leaf curl symptoms in tomato plants. In the presence of DNAbeta01, these viruses induced mild leaf curl symptoms in tomato plants. The recDNA had a chimeric sequence, which arose from recombination among ToLCJAV, AYVV-[Java], DNAbeta01, and DNAbeta02; it was replicated only in the presence of AYVV-[Java] in tomato plants.     

Detection of Tomato yellow leaf curl Sardinia virus in Tunisia

Surveys were undertaken for tomato yellow leaf curl viruses in the main Tunisian tomato-growing areas in fields and plastic houses. Symptoms included yellowing, leaf curling and stunting. Collected samples were submitted to molecular analysis using two approaches: (1) hybridization tests with two DNA probes corresponding to an intergenic region derived from a cloned dimer of an Egyptian full-length TYLCV and to the coat region of a Tunisian TYLCV isolate; (2) PCR amplification coupled to RFLP allowing both identification and clustering of Tunisian isolates. Only Tomato yellow leaf curl Sardinia virus was detected in Tunisia.     

Biological characterization of Tomato leaf curl New Delhi virus from Spain

Tomato leaf curl New Delhi virus (ToLCNDV; family Geminiviridae, genus Begomovirus) is an emerging virus in horticulture crops in Asia, and has recently been introduced in Spain, Tunisia and Italy. No betasatellite DNA was detected in infected tomato and zucchini squash samples from Spain, and agroinoculated viral DNA‐A and DNA‐B were sufficient to reproduce symptoms in plants of both crop species. Infected tomato and zucchini squash plants also served as inoculum sources for efficient transmission either mechanically or using Bemisia tabaci whiteflies. Cucumber, melon, watermelon, zucchini squash, tomato, eggplant and pepper, but not common bean, were readily infected using viruliferous whiteflies and expressed symptoms 8–15 days post‐inoculation. New full‐length sequences from zucchini squash and tomato indicated a high genetic homogeneity (>99% sequence identity) in the ToLCNDV populations in Spain, pointing to a single recent introduction event.