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.     

Identification of a CAPS marker tightly linked to the Tomato yellow leaf curl disease resistance gene <Emphasis Type="Italic">Ty-1</Emphasis> in tomato

During the process of breeding programmes, several resistance genes have been introgressed into tomato (Solanum lycopersicum) cultivars from different wild tomato relatives. A number of these resistance genes have been mapped to chromosome 6. Among them, Ty-1 and Mi, which confer resistance to Tomato yellow leaf curl disease and to Meloidogyne spp., respectively, are in most cases incorporated in commercial hybrids. Several molecular markers tightly linked to Mi have been identified. This study was conducted in order to find an informative molecular marker linked to Ty-1. Six markers mapped in the same region as Ty-1 were analysed in plant material carrying different combinations of Ty-1 and Mi alleles. Three of the six markers revealed polymorphism among the assayed accessions. One allele of JB-1 marker showed association with Ty-1. Furthermore, the presence of Mi did not interfere with the results. The analysis of several accessions of wild tomato relatives with the three polymorphic markers allowed the establishment of the origin of the alleles found in cultivated plant material, showing that introgressions from S. lycopersicum, S. pimpinellifolium and S. habrochaites will not interfere with the results of this marker which tags Ty-1. Furthermore this analysis enabled the location of CT21, the RFLP marker from which JB-1 was designed.     

Nucleotide sequencing, whitefly transmission, and screening tomato for resistance against two newly described begomoviruses in bangladesh

ABSTRACT The molecular diversity of Tomato leaf curl viruses (ToLCVs), from the two main tomato growing areas of Jessore and Joydebpur, Bangladesh, was investigated. The viral DNA was amplified from tomato plants exhibiting mild and severe symptoms by polymerase chain reaction, and the complete genomes of the ToLCVs were sequenced. An isolate of the bipartite Tomato leaf curl New Delhi virus-Severe (ToLCNDV-Svr) was associated with the severe symptom phenotype from Jessore (ToLCNDV-Svr[Jes]). A previously undescribed monopartite virus, designated Tomato leaf curl Joydebpur virus-Mild (ToLCJV-Mld), was sequenced from plants showing mild symptoms. ToLCNDV-Svr[Jes] was most closely related to ToLCNDV-[Lucknow] at 95.7% nucleotide (nt) identity and Tomato leaf curl Gujarat virus-[Varanasi] at 90.6% nt identity, based on DNA-A and -B component sequences. ToLCJV-Mld was similar to Pepper leaf curl Bangladesh virus at 87.1% DNA-A nt identity. Identification of ToLCNDV-Svr[Jes] and ToLCJV-Mld was in addition to the previously described Tomato leaf curl Bangladesh virus, with which they shared 73.2 and 86.0% DNA-A nt identities, thus demonstrating the existence of at least three distinct viruses infecting tomato in Bangladesh. Nucleotide identities and placement in phylogenetic trees suggested that the three ToLCVs may have had different evolutionary pathways. The whitefly, Bemisia tabaci, transmitted the viruses of this study equally efficiently. Four tomato cultivars (TLB111, TLB130, TLB133, and TLB182) resistant/ tolerant to South Indian ToLCV were screened against the Bangladesh ToLCVs in 2003-04. Although challenged by diverse viruses and potentially mixed infections, disease incidence remained low (6 to 45%) in the resistant cultivars compared with local cultivars (68 to 100%).     

Tomato leaf curl Karnataka virus from Bangalore, India, Appears to be a Recombinant Begomovirus

ABSTRACT The genome of Tomato leaf curl virus (ToLCV) from Bangalore, India, a whitefly-transmitted geminivirus, was cloned (pIND9) and sequenced. The circular DNA of 2,759 nucleotides (U38239) is organized similarly to that of other begomoviruses with monopartite genomes. Comparison of the nucleotide sequence of pIND9 with other tomato-associated begomoviruses from India (Tomato leaf curl Bangalore virus [ToLCBV, Z48182]) and Tomato leaf curl New Delhi virus-Severe (ToLCNdV-Svr, U15015) showed moderate DNA sequence identities (82 to 87%) between capsid protein (CP) genes but low identities (66 to 67%) for the intergenic regions and the replication-associated protein (Rep) genes (75 to 81% identity). Phylogenetic trees generated with nucleotide sequences of the Rep and CP genes of 26 begomoviruses indicated that this ToLCV is distinct from other begomoviruses and that it may be a recombinant virus derived from at least three different viral lineages. Tomatoes (Lycopersicon esculentum) inoculated with the cloned DNA monomer of ToLCV (pIND9) via particle bombardment developed leaf curling and yellowing symptoms. The virus was transmitted by Bemisia tabaci biotype B from tomatoes infected via particle bombardment to healthy tomatoes and by sap inoculation from infected tomatoes to tomato, Nicotiana benthamiana and N. tabacum. This ToLCV is a distinct member of the genus Begomovirus from India that differs from the previously characterized Tomato leaf curl Sadasivanagar virus isolate Bangalore 1 (L12739), ToLCBV (Z48182), ToLCBV isolate Bangalore 4 (AF165098), and the bipartite ToLCNdV (U15015, U15016). Thus, this ToLCV is named Tomato leaf curl Karnataka virus (ToLCKV).     

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.     

A simple, efficient agroinoculation soaking procedure for Tomato yellow leaf curl virus

Tomato yellow leaf curl virus (TYLCV, genus Begomovirus, family Geminiviridae), poses a serious threat to tomato crops in tropical and subtropical regions. We developed a simple agroinoculation method with an infectious clone of TYLCV. Dipping of excised sections of susceptible tomato shoots in an Agrobacterium suspension successfully introduced the replicating virus with high efficiency. An additional vacuum treatment for 5 min ensured uniform infection without escapes, allowing evaluation of differences in TYLCV resistance among tomato cultivars. The method can be used in laboratory experiments for virological studies and in breeding programs for resistant cultivars.     

Panel of real‐time PCRs for the multiplexed detection of two tomato‐infecting begomoviruses and their cognate whitefly vector species

A new approach for the simultaneous identification of the viruses and vectors responsible for tomato yellow leaf curl disease (TYLCD) epidemics is presented. A panel of quantitative multiplexed real‐time PCR assays was developed for the sensitive and reliable detection of Tomato yellow leaf curl virus‐Israel (TYLCV‐IL), Tomato leaf curl virus (ToLCV), Bemisia tabaci Middle East Asia Minor 1 species (MEAM1, B biotype) and B. tabaci Mediterranean species (MED, Q biotype) from either plant or whitefly samples. For quality‐assurance purposes, two internal control assays were included in the assay panel for the co‐amplification of solanaceous plant DNA or B. tabaci DNA. All assays were shown to be specific and reproducible. The multiplexed assays were able to reliably detect as few as 10 plasmid copies of TYLCV‐IL, 100 plasmid copies of ToLCV, 500 fg B. tabaci MEAM1 and 300 fg B. tabaci MED DNA. Evaluated methods for routine testing of field‐collected whiteflies are presented, including protocols for processing B. tabaci captured on yellow sticky traps and for bulking of multiple B. tabaci individuals prior to DNA extraction. This work assembles all of the essential features of a validated and quality‐assured diagnostic method for the identification and discrimination of tomato‐infecting begomovirus and B. tabaci vector species in Australia. This flexible panel of assays will facilitate improved quarantine, biosecurity and disease‐management programmes both in Australia and worldwide.     

Characterisation of a Quantitative Resistance to Vector Transmission of Tomato yellow leaf curl virus in Lycopersicon pimpinellifolium

Two wild genotypes from the same species Lycopersicon pimpinellifolium, WVA106 (susceptible) and INRA-Hirsute (so-called ‘resistant’), were compared with respect to their reaction to Tomato yellow leaf curl virus isolate Réunion (TYLCV-Mld[RE]), using both whitefly-mediated inoculation and graft inoculation. Disease incidence and symptom severity were scored. Presence and quantification of viral DNA were assessed by dot blot hybridisation. Upon insect inoculation, accession INRA-Hirsute showed a moderate resistance against TYLCV that was overcome by a high inoculation pressure obtained by increasing the cumulative number of inoculative whiteflies. Temporal analyses of the disease progress in relation to this criterion exhibited that the protection was quantitative, mainly reducing the TYLCV-Mld[RE] incidence by at maximum 50% at low inoculation pressure. When graft inoculated, the final TYLCV-Mld[RE] disease incidence was 100% in both susceptible and resistant genotypes with severe symptoms, suggesting a reduction of virus transmission by a vector resistance as a possible mechanism. Implications of using such type of resistance in breeding programmes are discussed.     

Response of Cucurbita spp. to tomato leaf curl New Delhi virus inoculation and identification of a dominant source of resistance in Cucurbita moschata

Tomato leaf curl New Delhi virus is an emerging whitefly-borne species of begomovirus in Mediterranean regions that poses a severe threat to cucurbit crops of the genus Cucurbita. Until now, only two sources of resistance have been identified in Cucurbita spp., these being PI604506 (cv. Large Cheese) and PI381814 (Indian landrace), both of C. moschata. The resistance of cv. Large Cheese is conferred by a single recessive gene located on chromosome 8. The objective of the present investigation was to screen for tomato leaf curl New Delhi virus (ToLCNDV) resistance among 105 accessions drawn from five species of Cucurbita and, if high resistance was found in any of them, determine the mode of inheritance. Screening was conducted using whitefly-mediated inoculation on all 105 accessions. The accessions showing some resistance were further screened by mechanical inoculation as well as by quantitative PCR-based diagnostics. The results showed that, overall, the accessions of C. pepo and C. maxima were the most susceptible, those of C. argyrosperma and C. ecuadorensis intermediate, and those of C. moschata most resistant to ToLCNDV. Only one accession of C. moschata, BSUAL-252, originating from Japan, was highly resistant to ToLCNDV, showing no symptoms after either method of inoculation, and absence of virus accumulation. Upon crossing BSUAL-252 with a susceptible accession of C. moschata, BSUAL-265, the resistance was observed to be conferred by a single dominant gene. This gene is not linked to the genomic region on chromosome 8 where the locus of the previously identified recessive gene for ToLCNDV resistance resides.     

不同番茄品种对5种植物双生病毒的抗病性鉴定

 由粉虱传双生病毒引起的番茄曲叶病［1］在我国最初仅分布在海南、云南、广东和广西,自2006年上海市和浙江省先后在番茄上发现番茄黄化曲叶病毒（Tomato yellow leaf curl virus,TYLCV）以来,该病害蔓延迅速,在多个省份的番茄上暴发成灾［2］。引起番茄曲叶病害的病原较复杂,在我国其主要病原为TYLCV、中国番木瓜曲叶病毒（Papaya leaf curl China virus, PaLCuCNV）、中国番茄黄化曲叶病毒(Tomato yellow leaf curl China virus, TYLCCNV)、泰国番茄黄化曲叶病毒（Tomato yellow leaf curl Thailand virus, TYLCTHV）和台湾番茄曲叶病毒（Tomato leaf curl Taiwan virus, ToLCTWV）［2~5］,而浙江省的主要病原为TYLCV和ToLCTWV。选育抗病品种是防治番茄黄化曲叶病最有效的手段。了解番茄品种对不同双生病毒的抗性,对因地制宜布局抗病品种具有重要意义。浙杂502、浙粉701、浙粉702是浙江省大规模种植的番茄品种,为了解这些品种对上述5种病毒的抗性,本研究利用5种病毒的侵染性克隆,在人工接种条件下,综合评定分析这3个番茄品种的抗病指标。     

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。     

Methods of controlling Tomato yellow leaf curl virus (TYLCV) and its vector Bemisia tabaci in the Maltese Islands*

Bemisia tabaci and Tomato yellow leaf curl virus (TYLCV) were first observed in Malta in the early 1990s and caused serious damage to glasshouse and outdoor tomato crops. Chemical, physical and biological control methods have been developed, but the effective method has been the use of virus‐tolerant cultivars.     

Resistance of <Emphasis Type="Italic">Solanum</Emphasis> species to <Emphasis Type="Italic">Cucumber mosaic virus</Emphasis> subgroup IA and its vector <Emphasis Type="Italic">Myzus persicae</Emphasis>

Sixty-nine tomato genotypes representing nine Solanum species were evaluated for resistance to Cucumber mosaic virus (CMV) subgroup IA and its aphid vector Myzus persicae. Resistance was assessed by visual scoring of symptoms in the field under natural conditions, and in the greenhouse by artificial inoculations through aphid M. persicae and mechanical transmissions in the year 2007 and 2009. Considerable variation in responses was observed among the evaluation methods used. Field evaluations were found liable to errors as different levels were observed for the same genotypes in the different years, however mechanical inoculation was found to be the most useful in identifying CMV subgroup IA resistance, in contrast aphid transmission was most useful in identifying insect transmission resistance. All genotypes observed as highly resistant to CMV subgroup IA in the field or through vector transmission became systemically infected through mechanical inoculations. Using mechanical inoculation, six genotypes (TMS-1 of S. lycopersicum, LA1963 and L06049 of S. chilense, LA1353, L06145 and L06223 of S. habrochaites) were found resistant and another six (L06188 and L06238 of S. neorickii, L06219 of S. habrochaites, L05763, L05776 and L06240 of S. pennellii) were found tolerant showing mild symptoms with severity index (SI) ranging 1-2 and with delayed disease development after a latent period (LP) of 18–30 days. However, these genotypes were found to be resistant to highly resistant in the field and through inoculation by M. persicae; and they also supported low population levels of M. persicae except TMS-1. Another nine genotypes (LA2184 of S. pimpinellifolium L., LA2727 of S. neorickii, LA0111, L06221, L06127 and L06231 of S. peruvianum L., LA1306, L06057 and L06208 of S. chmielewskii) showing a susceptible response after mechanical inoculation were highly resistant, resistant and tolerant after M. persicae transmission. The resistant genotypes, identified in the present study can be exploited in the breeding programmes aimed at developing tomato varieties resistant to CMV subgroup IA and broadening the genetic base of CMV-resistant germplasm. The differences observed between mechanical and aphid transmission suggests that one should consider both evaluation methods for tomato germplasm screening against CMV subgroup IA.     

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.     

Evaluation of cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis>) cultivars grown in Eastern Europe and progress in breeding for resistance to angular leaf spot (<Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">lachrymans</Emphasis>)

Increased occurrence of cucumber angular leaf spot, Pseudomonas syringae pv. lachrymans, has caused significant losses in cucumber, Cucumis sativus, yield in Poland in recent years. These losses necessitated evaluation of the level of resistance in cucumber cultivars of mainly Polish breeding, cultivated in Eastern Europe, and initiation of a breeding programme for resistance to this disease. Screening for resistance was performed on 84 cucumber accessions under growth chamber conditions using a highly aggressive strain of P. syringae pv. lachrymans. Most of the screened accessions were either susceptible or displayed intermediate resistance. The screening resulted in the identification of five F₁ hybrid cultivars moderately resistant to angular leaf spot. The identified F₁ hybrids were self-pollinated up to the F₄ generation. Individuals resistant to angular leaf spot were identified. These individuals can be used as a source of resistance to angular leaf spot in future breeding efforts.     

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.     

不同番茄品种对番茄黄化曲叶病毒的抗病性鉴定

为评估生产上常用番茄栽培品种对番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)的抗性水平,采用田间大棚自然传毒的方式,通过对各品种的发病时间、发病率及病情指数等参数的比较,结合PCR及ELISA对TYLCV的检测结果,综合分析了20个番茄品种对番茄黄化曲叶病毒的抗病性。不同品种对番茄黄化曲叶病毒的抗性差异较大,试验筛选出仙客6号和金棚1号2份感病材料,发病率和病情指数在90%和60.9以上;佳红8号、10-秋展47和红罗曼2号3份高抗材料,发病率和病情指数均为0;其它表现不同程度抗、耐病水平的材料15份,发病率和病情指数分别在10.0%～85.0%和1.0～40.6之间。     

Tomato leaf curl Guangxi virus is a distinct monopartite begomovirus species

Three begomovirus isolates were obtained from tomato plants showing leaf curl symptoms in Guangxi province of China. Typical begomovirus DNA components representing the three isolates (GX-1, GX-2 and GX-3) were cloned and their full-length sequences were determined to be 2752 nucleotides. Nucleotide identities among the three viral sequences were 98.9–99.7%, but all shared <86.7% nucleotide sequence identity with other reported begomoviruses. The sequence data indicated that GX-1, GX-2 and GX-3 are isolates of a distinct begomovirus species for which the name Tomato leaf curl Guangxi virus (ToLCGXV) is proposed. Further analysis indicated that ToLCGXV probably originated through recombination among viruses related to Ageratum yellow vein virus, Tomato leaf curl China virus and Euphorbia leaf curl virus. PCR and Southern blot analyses demonstrated that isolates GX-1 and GX-2 were associated with DNAβ components, but not isolate GX-3. Sequence comparisons revealed that GX-1 and GX-2 DNAβ components shared the highest sequence identity (86.2%) with that of Tomato yellow leaf curl China virus (TYLCCNV). An infectious construct of ToLCGXV isolate GX-1 (ToLCGXV-GX) was produced and determined to be highly infectious in Nicotiana benthamiana, N. glutinosa, tobacco cvs. Samsun and Xanthi, tomato and Petunia hybrida plants inducing leaf curl and stunting symptoms. Co-inoculation of tomato plants with ToLCGXV-GX and TYLCCNV DNAβ resulted in disease symptoms similar to that caused by ToLCGXV-GX alone or that observed in infected field tomato plants.     

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%).