Evaluation of tomato entries with different combinations of resistance genes to tomato yellow leaf curl disease in Tunisia

Tomato yellow leaf curl virus (TYLCV) is one of the most widespread begomoviruses transmitted by the whitefly Bemisia tabaci that cause tomato yellow leaf curl virus diseases (TYLCDs). TYLCD losses can be especially severe in open‐field tomato (Solanum lycopersicum L.) crops. TYLCV is particularly well known and widespread in the Mediterranean (Med) countries where TYLCD control is mostly based on insecticidal control of B. tabaci populations. Unfortunately, Tunisian B. tabaci populations include Middle East‐Asia Minor I and the Med species that have developed resistance to many classes of insecticides. Therefore, TYLCD‐resistant cultivars are essential for sustainable disease management. Six TYLCD resistance genes (Ty) have been introgressed from wild species into cultivated tomato and are available for breeding. Information on the Ty genes or gene combinations is useful for breeding resistant cultivars. To this end, 14 tomato lines carrying different Ty gene combinations and two susceptible tomato entries were evaluated for TYLCD incidence and severity in two field trials during late season in Tunisia. Entries with Ty‐1/Ty‐3 + Ty‐2 offered the highest levels of resistance in Tunisia.     

Reaction of Lycopersicon cultivars and wild accessions to tomato leaf curl virus

Summary 1201 tomato cultivars, breeding lines and accessions of Lycopersicon species were screened for tomato leaf curl virus (TLCV) under field and laboratory conditions during summer seasons of 1986 to 1989. Two lines of L. hirsutum (PI 390658 and PI 390659) and 2 lines of L. peruvianum (PI 127830 and PI 127831) were resistant to TLCV infection. These accessions did not produce any leaf curl symptoms either in field or after inoculation by whitefly Bemisia tabaci with TLCV. Adult whiteflies died within 3 days after releasing on resistant accessions (PI 390658, PI 390659, PI 127830), whereas the whiteflies survived upto 25 days on susceptible tomato cultivars. Under field conditions 0–4 and 5–25 adult whiteflies were observed on resistant and susceptible cultivars respectively. Hybridization was effected using the popular tomato cultivars Arka Sourabh, and Arka Vikas, as the female parents and the resistant Lycopersicon wild species as the pollen parents, to incorporate the resistant gene(s) into the edible tomatoes.     

Evaluation of whitefly-mediated inoculation techniques to screen Lycopersicon esculentum and wild relatives for resistance to Tomato yellow leaf curl virus

For two consecutive years nine hybrids and three varieties of tomato, four Lycopersicon peruvianum and four Lycopersicon chilense accessions were screened for Tomato yellow leaf curl virus (TYLCV) resistance. Three inoculation techniques using Bemisia tabaci, the vector of TYLCV, were compared: (1) artificial mass inoculation-simultaneous infection of cultivated and wild material in greenhouses; (2) artificial cage inoculation-individual infection in insect-proof cages; (3) natural field infection. Artificial inoculations led to higher levels of infection, but different patterns of response to each inoculation technique were found depending on the resistance level. Tomato varieties showed an important fruit set reduction after both artificial and natural inoculations. In contrast, field infection was milder in tomato hybrids, in which yield was barely affected. These hybrids showed a wide range of reactions with the two artificial inoculation techniques, but infection was always more severe after mass inoculation. Extreme severity of mass infection made it difficult to differentiate among variable degrees of resistance that were more reliably detected with cage inoculation. The hybrids F3524, F3522, Fiona, and Tyking showed the highest level of resistance. F3524 and F3522 had an acceptable yield in field and cage assays, but their resistance collapsed under massive conditions of infection. Tyking and Fiona exhibited the best response in all conditions, although their yield was moderately reduced in mass assays. Mass inoculation was not adequate for the screening of wild Lycopersicon. Some susceptible plants escaped infection, probably as a consequence of non-preference mechanisms and loss of vector infectivity. Individual inoculation in cages prevented the risk of non-infection, ensuring 100% disease incidence. This technique allowed the selection of highly resistant wild sources. L. chilense LA 1969 and LA 1963 had the highest level of resistance with the three inoculation techniques. L. peruvianum PI-126944 and L. chilense LA 1932, which were only tested in mass and field conditions, also exhibited a promising response. The results proved that the inoculation technique influences the response of tomato and wild Lycopersicon spp to TYLCV. It is concluded that artificial cage inoculation, although more time-consuming, is the most efficient, adequate, and reliable technique to screen both cultivated and wild Lycopersicon species for resistance to TYLCV. This revised version was published online in July 2006 with corrections to the Cover Date.     

Inheritance of tomato leaf curl virus resistance in Lycopersicon hirsutum f. glabratum

Summary Inheritance of resistance to tomato leaf curl virus (TLCV) was studied in the progenies derived from interspecific crosses between TLCV resistant Lycopersicon hirsutum f. glabratum line B 6013 and five susceptible cultivars (HS 101, HS 102, HS 110, Pusa Ruby and Punjab Chhuhara) of L. esculentum. P₁, P₂, F₁, F₂, B₁ and B₂ progenies of the five crosses were artificially inoculated with local strains of TLCV by means of the vector whitefly, Bemisia tabaci (Genn.). and the disease reaction was studied in all the crosses. Reaction of parents, F₁, F₂ and backcrosses suggests that resistance derived from L. hirsutum f. glabratum B 6013 is based on two epistatic genes, one from the wild parent and one from the cultivated one, resulting in a 13:3 segragation in the F₂.     

Developing tomato breeding lines resistant to tomato yellow leaf curl virus

Using controlled whitefly-mediated inoculation techniques, seven Lycopersicon chilense accessions, highly resistant to isolates of tomato yellow leaf curl virus(TYLCV) from Southern Europe, TYLCV-Sr, were selected. All exhibited similar levels of partial resistance, being symptomless and with low levels of viral DNA accumulation. However, a differential response to infection was found in interspecific hybrids with tomato and inbred lines derived from different L. chilense accessions, allowing a precise discrimination among them. This selection procedure which considers the expression of the resistance genes in the tomato genetic background led to the selection of two highly resistant F₁ hybrids derived from L. chilense LA 1932 and LA 1938. A backcrossing programme was initiated, selecting for horticultural characteristics and TYLCV resistance, in field and controlled inoculation conditions. As a result of this programme, six advanced breeding lines (UPV Ty 1, 3, 6, 9, 17 and 53), exhibiting a high level of resistance to TYLCV-Sr, were obtained. Under high inoculum pressure conditions these lines suffered only 30-40% yield loss relative to non-infected control plants, and compared with 90-95% yield loss in susceptible controls. These lines also have horticultural characteristics appropriate for the fresh market tomato cultivation system in this area, and are a good base material for obtaining commercial hybrids highly resistant to different isolates of TYLCV.     

Identification of sources of resistance in Lycopersicon species to Tomato leaf curl geminivirus (ToLCV) by agroinoculation

A total of 90 genotypes of Lycopersicon species were tested forresistance to the Tomato leaf curlgeminivirus (ToLCV) by agroinoculation andthe vector whitefly (Bemisia tabaciGenn.) inoculation techniques underinsect-proof glasshouse conditions. Therate of infection in the inoculated plantswas determined by detection of the viralDNA in individual plants by the nucleicacid spot hybridization (NASH). Of the 38cultivars and 11 breeding lines of L.esculentum Mill. tested, none was highlyresistant or resistant while three andseven were moderately resistant whenexposed to the cloned virus DNAs byagroinoculation and whitefly inoculationrespectively. On the other hand, among the38 commercial cultivars screened, 16(42.1%) were highly susceptible in vectorinoculations and 31 (81.6%) inagroinoculation. Among the exoticcollection (EC) accessions six were highlyresistant, eleven resistant to whiteflyinoculation and none was highly susceptiblein either of the two tests, indicating thepresence of resistance among the ECaccessions. A higher degree of resistancewas observed in other species of Lycopersicon. While only one accession ofL. cheesmanii Riley was tested, itcould not be infected by either of the twomethods. L. pimpinellifolium (Jusl.)Mill. genotype EC 251580 was similarlyresistant. In L. peruvianum (L.)Mill., five EC accessions could not beinfected by whitefly inoculation, withthree of these being resistant and twomoderately resistant in agroinoculation.This study demonstrates the importance ofthe agroinoculation technique in the virusresistance screening programs andidentifies several good sources ofresistance to the Tomato leaf curlvirus in Lycopersicon species.     

黄化曲叶病毒病胁迫下番茄生化参数光谱响应特性

在黄化曲叶病毒病条件下,分析比较番茄的感病品种和抗病品种叶片生理生化参数变化趋势,并进行了生化参数的显著性检验,发现木质素、PPO、PAL、TAL、总酚的活性都随着取样时间增加。分析特征光谱反射率、光谱植被指数的变化,显著性检验分析表明除了760 nm处的反射率之外,480 nm、552 nm、680 nm以及光谱指数NDVI、GNDVI、BNDVI、GRNDVI、GBNDVI、RBNDVI、PNDVI都具有显著差异。进而运用归一化方法分析光谱指数,特征光谱反射率与生理生化参数之间的相关性,研究结果表明,木质素与光谱指数和特征光谱反射率间拟合度最高,并且木质素与特征波段480 nm、552 nm光谱指数RVI、NDVI、GRNDVI、GNDVI、GBNDVI、RBNDVI、PNDVI具有较好的拟合效果。     

Response of tolerant breeding lines of tomato, Lycopersicon esculentum, originating from three different sources (L. peruvianum, L. pimpinellifolium and L. chilense) to early controlled inoculation by tomato yellow leaf curl virus (TYLCV)

Selection of tomato plants supposedly tolerant to tomato yellow leaf curl virus (TYLCV), based solely on the absence of symptoms in an infested field can be misleading. An inoculation routine was therefore established to avoid escapes and to overcome difficulties associated with the age of the plant at the time of infection. The inoculation routine was applied to a selection of resistant/tolerant individuals generated through a diallel F¹ cross and to F² segregating populations originating from three wild tomato species described as tolerant to TYLCV: Lycopersicon peruvianum EC 104395, Lycopersicon pimpinellifolium Hirsute and Lycopersicon chilense LA 1969. Clear differences were observed between susceptible symptomatic and tolerant symptomless tomato genotypes, indicating that the uncertainty resulting from escapes, from different levels of inoculum, and from the time of inoculation, can be eliminated. The genes involved in tolerance provided different levels of protection; combinations of various tolerant sources and levels in a single genotype gave a higher level of tolerance. Differences in level of protection were found between genes from the same source and between sources; none of the sources tested had complete dominance. The results obtained with the F² segregating population showed that tolerance from L. pimpinellifolium is controlled by one major gene, that from L. chilense by two genes, and that from L. peruvianum by three genes with no dominant effect. The combination of sources for resistance can thus have positive or negative synergistic effects, or no effect. We suggest that a maximal level of tolerance can be obtained by the additive effect of the partly dominant genes from L. pimpinellifolium and L. chilense.     

Evaluation of seven Lycopersicon species for resistance to tomato spotted wilt virus (TSWV)

Summary One hundred eighty-eight accessions of Lycopersicon cheesmanii, L. chilense, L. chmielewskii, L. hirsutum, L. parviflorum, L. pennellii, and L. peruvianum were screened for resistance to three isolates of tomato spotted wilt virus (TSWV). All plants in an accession were initially screened for resistance to TSWV using isolate 85–9 from Arkansas. Visual symptoms were used to cull obviously infected plants, followed by enzyme-linked immunosorbent assay (ELISA) to identify uninfected plants. Cuttings were taken from uninfected plants in the first screening and the resulting plants were inoculated with isolates Glox and T-2 from Texas and Hawaii, respectively. No resistance was identified in L. cheesmanii, L. chmielewskii, L. hirsutum, L. parviflorum, and L. pennellii. However, 33 of 63 L. chilense accession produced 91 of 1268 plants that were uninfected with isolate 85–9 and 20 accessions that produced 40 of 257 plants that were not infected with any of the isolates. After screening with isolate 85–9 9 of 12 L. peruvianum accessions tested had 38 plants uninfected and 8 accessions had 25 plants that were not infected with any of the isolates.     

Resistance in two Lycopersicon species to an Arkansas isolate of tomato spotted wilt virus

Summary Tomato spotted wilt virus (TSWV) was obtained from infected tomatoes in commercial fields in Arkansas in 1985. A greenhouse screening procedure for identifying tomatoes resistant to TSWV was established using an enzyme-linked immunosorbent assay (ELISA) to detect infected plants. Symptom expression was variable and symptom expression was not reliable for identifying infected plants. Germplasm evaluated for resistance to one typical Arkansas isolate (85–9) of TSWV included: twenty cultivars and breeding lines of Lycopersicon esculentum Mill, 52 accessions of L. pimpinellifolium (Jusl.) Mill and 8 accessions of L. peruvianum (L.) Mill. All cultivated accessions and breeding lines evaluated were susceptible. Some individual plants in several accessions of L. pimpinellifolium were resistant and nearly all plants of the L. peruvianum accessions that were evaluated were resistant to isolate 85–9.Dept. of Plant Pathology     

Mapping of QTLs in tomato line FLA456 associated with resistance to a virus causing tomato yellow leaf curl disease

Tomato (yellow) leaf curl disease (TYLCD) is a serious threat to tomato production in the tropics and subtropics. The genetics of resistance to Tomato yellow leaf curl Thailand virus Taiwan strain (TYLCTHV-[TW]) in a highly resistant tomato line FLA456 was studied through quantitative trait loci (QTL) analysis. Four QTLs named qTy4.1, qTy6.1, qTy10.1 and qTy11.1 were detected on chromosomes 4, 6, 10, and 11, respectively, through evaluation of an F₆ recombinant inbred line (RIL) population derived from a cross between FLA456 (resistant) and CLN1621L (susceptible). Gene action of all QTLs was recessive based on disease reaction of the F₁. The markers SINAC1 and SLM4-34 flanked qTy4.1 on chromosome 4, and SLM11-12 and SLM11-17 defined qTy11.1, which co-located with the previously identified Ty-5 and Ty-2 loci, respectively. qTy6.1 was flanked by the markers SLM6-55 and TES-0014, and qTy10.1 by the markers SLM10-80-SLM10-46 on chromosomes 6 and 10. The LOD values of the putative QTLs ranged from 2.79 to 13.76. The phenotypic variance explained by each QTL ranged from 7.1 to 31.9 %. The four QTLs collectively contributed about 60.5 % of the phenotypic variation in resistance against TYLCTHV-[TW]. Group mean severity scores of those RILs possessing three or four qTy were generally lower than RIL groups with only one or no qTy. Given the diversity of begomoviruses that cause TYLCD across the regions, the new QTLs from FLA456 would be valuable in tomato breeding for developing varieties with durable resistance. Two QTL intervals (qTy4.1 and qTy10.1) contained virus resistance candidate genes such as CTV.22 and eukaryotic translation initiation factor 4E.     

Development of tomato hybrids resistant to tomato leaf curl virus disease in South India

Resistance to tomato leaf curl virus disease (ToLCVD) in tomato (Solanum lycopersicum) is scarce but was developed recently in three open pollinated (OP) varieties (Sankranthi, Nandi and Vybhav), which are now available for cultivation in South India. Hybrids with superior yield capabilities, however, are a preferred choice of cultivation by farmers but hybrids are highly susceptible to ToLCVD. In order to develop virus resistant tomato hybrids, the three OPs were crossed with 12 tomato genotypes with superior agronomic characteristics. From the crosses, 20 hybrids were selected (named BLRH-1 to BLRH-20, Bangalore leaf curl virus-resistant hybrid) and evaluated for their resistance to ToLCVD at the University of Agricultural Sciences, Bengaluru, South India, both through whitefly-mediated inoculations in the glasshouse and natural infection in the field during summer 2005. Their growth and fruit yield parameters were compared against the popularly grown hybrids and OPs. Differences occurred between hybrids in disease incidence, spread, symptom severity and fruit yield. Of the 20 hybrids evaluated, 11 were found resistant to ToLCVD in the field, but only three (BLRH-3, BLRH-9 and BLRH-16) remained resistant when challenged with high virus inoculum pressure in the glasshouse through whitefly-mediated inoculations. None of the six commercially available hybrids tested was found resistant to ToLCVD. An examination of the extent of heterosis in resistant hybrids for both quantitative and qualitative characters over the popularly grown hybrid, US-618, indicated a major improvement in virus resistance, yield and fruit quality. All the twenty hybrids yielded significantly more than US-618, and of these, BLRH-3 and BLRH-16 were considered the best with yields over ca. 60 t/ha and resistant to virus both in the glasshouse and field.     

Resistance to tomato spotted wilt virus in transgenic tomato hybrids

Summary Tomato spotted wilt virus (TSWV) causes significant economic losses in the commercial culture of tomato (Lycopersicon esculentum Mill.). Culture practices and introgression of natural sources of resistance to TSWV have only been marginally effective in controlling the TSWV disease. Recently however, high levels of protection against TSWV have been obtained by transforming tobacco with a chimaeric gene cassette comprising the TSWV nucleoprotein gene. This report demonstrates the successful application of this newly-created TSWV resistance gene in cultivated tomato. Transformation of an inbred tomato line with the TSWV nucleoprotein gene cassette resulted in high levels of resistance to TSWV that were maintained in hybrids derived from the parental tomato line. Therefore, transformant lines carrying the synthetic TSWV resistance gene make suitable progenitors for TSWV resistance to be incorporated into the breeding programmes of tomato.     

New sources for high resistance of tomato to the tomato spotted wilt virus from Lycopersicon peruvianum

Mechanical inoculation and transmission by thrips in a growth chamber were used in order to screen Lycopersicon peruvianumand Lycopersicon chilense germplasm for tomato spotted wilt virus (TSWV) resistance. Two highly aggressive Spanish TSWV isolates (HA-931100 and T-941117), having different restrictotypes were used. L. peruvianum accessions PI-126935, PI-126944, CIAPAN 16, PE-18 and CIAPAN 17 showed high resistance to both isolates in mechanical and thrips transmission. Their resistance appears useful in breeding programmes.     

Novel molecular marker associated with Tm2a gene conferring resistance to tomato mosaic virus in tomato

Tomato mosaic virus (ToMV) is an important Tobamovirus that causes significant crop losses. Resistance to the ToMV is conferred by the genes Tm1, Tm2 and Tm2^a. Among these three genes, Tm2^a confers resistance to most strains of the ToMV. Screening of genetic lines under field conditions based on phenotype is time‐consuming and challenging due to concerns associated with stability of the virus and its potential transmission to other plants. Tightly linked molecular markers associated with resistance genes can improve selection efficiency and avoid these problems. This study developed a PCR‐based marker based on restriction site differences from Tm2^a locus‐specific sequences, which was found to be useful in identifying the resistant and susceptible genotypes and was consistent with phenotypic data. The marker is a codominant cleaved amplified polymorphic sequence (CAPS) marker producing 270‐ and 600‐bp DNA fragments from resistant genotypes and an 870‐bp fragment from susceptible genotypes when digested with HaeIII restriction enzyme. This novel marker can be useful for tomato breeders to screen progeny from segregating populations for ToMV resistance.     

Pyramiding genes for resistance to bean common mosaic virus

Summary Gene pyramiding in Phaseolus vulgaris is being utilized to develop more effective resistance to the temperature-insensitive-necrosis-inducing (TINI) strains of Bean Common Mosaic Virus (BCMV) present in the USA. Our data indicate that contrary to previous work, the bc-3 gene is effective against these strains in the absence of the strain unspecific bc-u gene in genotypes possessing the I gene. The epistatic bc-3 gene interferes with traditional efforts to pyramid the other recessive resistance genes by masking their activity. Indirect selection based on markers linked to the other recessive resistance genes would likewise be ineffective without the ability to also select for the bc-u gene which is required for expression of the bc-2 ² gene in germplasm carrying the I gene. Because the most resistant genotype (I, bc-u, bc-I ², bc-2², bc-3) can only be introduced into a wide range of germplasm through the use of molecular markers linked to the different resistance genes, the search for a marker linked to the strain unspecific bc-u gene should also be given priority.     

Linkage relationship between two genes conferring resistance to peanut stripe virus and soybean mosaic

Summary The possible linkage relationship between hypocotyl color, leaflet shape, resistance to soybean mosaic virus (SMV) and to a soybean isolate of peanut stripe virus (PStV-isolate PN) was examined in two soybean lines AGS 129 and Ankur.Hypocotyl color, leaf shape and reactions to SMV-G1 and PStV were found to be inherited monogenically, with purple hypocotyl color, ovate leaf shape and resistance to both of the viruses being dominant. The reactions to SMV and PStV were conditioned by genes with 9 ± 2.4 percent recombination as coupling phase. They were inherited independently from hypocotyl color and leaf shape.     

Sources of resistance to tomato-yellow-leaf-curl-virus (TYLCV) in Lycopersicon species

Summary Sources of resistance to tomato-yellow-leaf-curl-virus (TYLCV) were investigated in 16 accessions of three Lycopersicon species and 55 commercial tomato hybrids and cultivars. All commercial hybrids and cultivars were highly susceptible. Accessions of L. hirsutum, L. hirsutum f. glabaratum and L. pimpinellifolium showed a wide range of reactions. Those of L. peruvianum, LA 385 of L. peruvianum f. humifusum exhibited very high levels of resistance indicating their potential use in local breeding programs. In contrast to earlier findings, back indexing showed that all symptomless genotypes in this investigation were carriers of the TYLCV.     

Major genes for resistance to beet necrotic yellow vein virus (BNYVV) in Beta vulgaris

Summary Inheritance of resistance to beet necrotic yellow vein virus (BNYVV) was studied in segregating F2 and backcross families obtained from crosses between resistant plants of the sugar beet selection Holly-1-4 or the wild beet accession Beta vulgaris subsp. maritima WB42 and susceptible parents. Greenhouse tests were carried out, in which seedlings were grown in a mixture of sand and infested soil. Virus concentrations of BNYVV in the rootlets were estimated by ELISA. To discriminate resistant and susceptible plants, mixtures of normal distributions were fitted to log₁₀ virus concentrations, estimated for segregating F1, F2 and BC populations of both accessions. The hypothesis that Holly-1-4 contained one single dominant major gene was accepted. For WB42, results fitted with the hypotheses that resistance was based on either one (or more) dominant major gene(s) showing distorted segregation, or two complementary dominant genes, which are both required for resistance. Resistance from WB42 appeared to be more effective against BNYVV than resistance from Holly-1-4.This research was carried out as part of a PhD study at the Graduate School Experimental Plant Sciences (EPS), Department of Virology, Wageningen, The Netherlands