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     

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.     

Search in <Emphasis Type="Italic">Solanum</Emphasis> (section <Emphasis Type="Italic">Lycopersicon</Emphasis>) germplasm for sources of broad-spectrum resistance to four <Emphasis Type="Italic">Tospovirus</Emphasis> species

The genus Tospovirus was considered as monotypic with Tomato spotted wilt virus (TSWV) being the only assigned species. However, extensive studies with worldwide isolates revealed that this genus comprises a number of species with distinct virulence profiles. The Neotropical South America is one center of Tospovirus diversity with many endemic species. Groundnut ringspot virus (GRSV), TSWV, Tomato chlorotic spot virus (TCSV), and Chrysanthemum stem necrosis virus (CSNV) are the predominant tomato-infecting species in Brazil. Sources of resistance were found in Solanum (section Lycopersicon) mainly against TSWV isolates from distinct continents, but there is an overall lack of information about resistance to other viral species. One-hundred and five Solanum (section Lycopersicon: Solanaceae) accessions were initially evaluated for their reaction against a GRSV isolate by analysis of symptom expression and systemic virus accumulation using DAS-ELISA. A subgroup comprising the most resistant accessions was re-evaluated in a second assay with TSWV, TCSV, and GRSV isolates and in a third assay with a CSNV isolate. Seven S. peruvianum accessions displayed a broad-spectrum resistance to all viral species with all plants being free of symptoms and systemic infection. Sources of resistance were also found in tomato cultivars with the Sw-5 gene and also in accessions of S. pimpinellifolium, S. chilense, S. arcanum, S. habrochaites, S. corneliomuelleri, and S. lycopersicum. The introgression/incorporation of these genetic factors into cultivated tomato varieties might allow the development of genetic materials with broad-spectrum resistance, as well as with improved levels of phenotypic expression.     

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.     

Characterization of resistance to Globodera rostochiensis pathotype Ro1 in cultivated and wild potato species accessions from the Vavilov Institute of Plant Industry

Potato cyst nematodes (PCN) collected in six localities in the Leningrad region of North West Russia were identified as Globodera rostochiensis pathotype Ro1 and were used for subsequent resistance tests. Seventy‐nine accessions of cultivated and closely related wild potato species from the VIR collection in Russia were screened on resistance to G. rostochiensis pathotype Ro1 and on the presence of molecular markers for H1 and Gro1‐4 resistance genes. No associations were detected between the resistance level of diploid and tetraploid Andean and tetraploid Chilean potato landraces (indigenous cultivars) and their related wild species and their geographical distribution or presence of PCR‐based markers that are associated with the H1 and Gro1‐4 genes. At the same time, all susceptible genotypes lacked such markers. New sources of resistance were found and could be used in breeding.     

High salt tolerance potential in Lycopersicon species during germination

Summary The potential to improve seed germination responses to salinity was evaluated for 13 accessions representing six wild Lycopersicon species and 20 accessions of L. esculentum. Germination response times increased in all accessions at 100 mM NaCl. Analysis indicated that one accession of L. peruvianum (PI126435) germinated faster under high salinity than all other accessions and was closely followed by L. pennellii (LA716). The fastest germinating L. esculentum accession, PI174263, ranked third. Additional wild ecotypes exhibiting rapid germination at 100 mM NaCl were identified among L. pimpinellifolium and L. peruvianum.     

Screening for new sources of resistance to Clavibacter michiganensis subsp. michiganensis (Cmm) in tomato

Bacterial canker of tomato, caused by Clavibacter michiganensis subsp. michiganensis (Cmm), is considered the most serious bacterial threat, resulting in high damages in production areas. Worldwide, Cmm is subjected to quarantine regulations.There is no cultivar in market containing Cmm resistance genes. This project aimed to screen tomatoes or wild relatives of tomato for resistance to Cmm, to be used for starting breeding programs. We have screened 24 different wild accessions of tomato and found several new tolerant sources: Solanum pimpinellifolium GI.1554, S. parviflorum LA735 and S. parviflorum LA2072. We also confirmed the tolerance which was reported previously in S. peruvianum LA2157, S. peruvianum PI127829, S. peruvianum LA385, S. habrochaites LA407 and S. lycopersicum cv. IRAT L3. No immunity was found. Also accessions showing a low disease score still contained high titers of bacteria as determined by a dilution plating method, using tow selective media. These results were confirmed with a TaqMan real time PCR assay, which was developed to determine and quantify Cmm in planta.     

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.     

DNA marker-assisted evaluation of potato genotypes for potential resistance to potato cyst nematode pathotypes not yet invading into Japan

One of major objectives of crop breeding is conferring resistance to diseases and pests. However, large-scale phenotypic evaluation for many diseases and pests is difficult because strict controls are required to prevent their spread. Detection of disease resistance genes by using DNA markers may be an alternative approach to select potentially resistant accessions. Potato (Solanum tuberosum L.) breeders in Japan extensively use resistance gene H1, which confers nearly absolute resistance to potato cyst nematode (Globodera rostochiensis) pathotype Ro1, the only pathotype found in Japan. However, considering the possibility of accidental introduction of the other pathotypes, breeding of resistant varieties is an important strategy to prevent infestation by non-invading pathotypes in Japan. In this study, to evaluate the prevalence of resistance genes in Japanese genetic resources, we developed a multiplex PCR method that simultaneously detects 3 resistance genes, H1, Gpa2 and Gro1-4. We revealed that many Japanese varieties possess not only H1 but Gpa2, which are potentially resistant to other pathotypes of potato cyst nematode. On the other hand, no genotype was found to have the Gro1-4, indicating importance of introduction of varieties having Gro1-4. Our results demonstrate the applicability of DNA-marker assisted evaluation of resistant potato genotypes without phenotypic evaluation.     

Screening wild Lycopersicon species for resistance to powdery mildew (Oidium lycoperiscum)

Since the late 1980s powdery mildew, designated Oidium lycopersicum, frequently invaded the tomato crop in Western Europe. All commercial cultivars are susceptible. To screen for resistance in wild species a reliable and efficient disease test was developed. Young plants with two to three true leaves are inoculated at high relative humidity by spraying with a freshly prepared suspension of 2×10⁴ conidia, ml^–1. Symptoms are periodically evaluated according to a scale based on the percentage of leaf area with mycelium.One hundred and twenty seven accessions, representing eight wild Lycopersicon species, were screened for resistance to O. lycopersicum. A large variation in resistance was found between species. L. hirsutum was the most resistant species; L. pennellii was moderately resistant; species of the subgeneric group of L. esculentum and of the peruvianum-complex were all susceptible. L. parviflorum was classified separately due to a large variation between accessions. Except for this species, a low variation was found between accessions within species. High levels of resistance were observed in four accessions of L. hirsutum, in one of L. parviflorum and in one of L. peruvianum. This resistance is characterized by a very low disease incidence and a strongly restricted mycelium growth and lack of sporulation.     

Response of accessions within tomato wild species,Solanum pimpinellifolium to late blight

Late blight (LB), caused by the oomycete Phytophthora infestans, is one of the most devastating diseases of the cultivated tomato (Solanum lycopersicum) worldwide. Most commercial cultivars of tomato are susceptible to LB. Previously, three major LB resistance genes (Ph‐1, Ph‐2, Ph‐3) were identified and incorporated into a few commercial cultivars of tomato. Reduced effectiveness and potential breakdown of the resistance genes has necessitated identification, characterization and utilization of new sources of resistance. We evaluated the response of 67 accessions of the wild tomato species, S. pimpinellifolium to LB, under multiple field and greenhouse (GH) conditions and compared them with six control genotypes. Sixteen accessions were identified with strong LB resistance in both field and GH experiments. However, 12 accessions exhibited resistance similar to a control line which was homozygous for Ph‐2 + Ph‐3. Genotyping accessions with molecular markers for Ph‐2 and Ph‐3 were not conclusive, indicating that resistance in these accessions could be due to these or other resistance genes. Strong correlations were observed between field and GH disease response and between foliar and stem infection.     

Heritability of late blight resistance in tomato conferred by Solanum pimpinellifolium accession PI 224710

Late blight (LB), caused by the oomycete Phytophthora infestans, is one of the most destructive diseases of tomato and potato worldwide. Identifying and characterizing new sources of resistance is essential given the emergence of new aggressive and fungicide‐resistant P. infestans isolates. Recently, we reported identification of several new sources of LB resistance within the tomato wild species, Solanum pimpinellifolium. In this study, we examined heritability (h²) of LB resistance conferred by the S. pimpinellifolium accession PI 224710 using a parent–offspring regression (correlation) analysis. F₂ and F₃ progeny populations, derived from crosses between PI 224710 and a LB‐susceptible tomato breeding line, were evaluated for response to LB infection. To obtain a better estimate of h², the F₃ progeny were evaluated for LB resistance in two separate replicated experiments. The h² estimates were similar in the two experiments and averaged ~0.87, suggesting that this resistance was highly heritable. Two different methods estimated involvement of one resistance locus. Breeding and mapping efforts are underway to further assess the viability of this newly reported LB resistance.     

Genetic analysis of late blight resistance in Solanum pimpinellifolium accession PI 270441: Heritability and response to selection

Late blight (LB), caused by Phytophthora infestans, is a destructive disease of tomato (Solanum lycopersicum) worldwide. Currently, there are few commercial cultivars of tomato with resistance to LB, and the disease is mainly controlled by heavy use of fungicides. Due to the emergence of fungicide‐resistant pathogen isolates, there is a concerted effort to identify new genetic sources of resistance and breed new resistant cultivars. A recent screening identified several new tomato accessions with strong resistance to LB. Here, we report on the genetic basis of LB resistance in S. pimpinellifolium accession PI 270441, as determined by generation means analysis and analysis of response to selection, using populations derived from crosses with LB‐susceptible breeding line Fla. 8059. Heritability of LB resistance ranged from 0.76 to 0.78, and the minimum number of genes was estimated 1—few. These results suggest that transfer of LB resistance from PI 270441 to the cultivated tomato should be feasible via a traditional backcross breeding approach. Genetic mapping studies are underway to identify molecular markers associated with resistance in this accession.     

Screening recently identified whitefly/spider mite‐resistant wild tomato accessions for resistance to Tuta absoluta

The tomato leaf miner (Tuta absoluta) is a serious pest of tomato (Solanum lycopersicum) in the tropics and subtropics. Previous World Vegetable Center studies identified selected accessions of S. galapagense, S. cheesmaniae and S. pimpinellifolium that were resistant to whitefly (Bemisia tabaci Genn.) and spider mite (Tetranychus urticae Koch). Here, we evaluated these accessions for resistance to T. absoluta based on the number of eggs from choice bioassays, and larval mortality and adults emerged percentages in no‐choice feeding bioassays at WorldVeg Eastern and Southern Africa (WorldVeg) and the International Centre for Insect Physiology and Ecology (icipe). At WorldVeg, S. galapagense VI063177 exhibited high resistance in both choice and no‐choice bioassays. There was strong negative correlation between larval mortality and adults emerged percentages in the no‐choice feeding bioassays. Results from the icipe experiments were consistent with those of the WorldVeg screening, except for S. pimpinellifolium accession VI030462 , which was susceptible at icipe. Tuta absoluta is rapidly spreading and the resistance sources reported here will be valuable in breeding tomato varieties resistant to this insect and others.     

Pyramiding of genes conferring resistance to Tomato yellow leaf curl virus from different wild tomato species

Tomato (Solanum lycopersicum) production in tropical and subtropical regions of the world is limited by the endemic presence of Tomato yellow leaf curl virus (TYLCV). Breeding programmes aimed at producing TYLCV‐resistant tomato cultivars have utilized resistance sources derived from wild tomato species. So far, all reported breeding programmes have introgressed TYLCV resistance from a single wild tomato source. Here, we tested the hypothesis that pyramiding resistances from different wild tomato species might improve the degree of resistance of the domesticated tomato to TYLCV. We have crossed TYLCV‐resistant lines that originated from different wild tomato progenitors, Solanum chilense, Solanum peruvianum, Solanum pimpinellifolium, and Solanum habrochaites. The various parental resistant lines and the F₁ hybrids were inoculated in the greenhouse using viruliferous whiteflies. Control, non‐inoculated plants of the same lines and hybrids were exposed to non‐viruliferous whiteflies. Following inoculation, the plants were scored for disease symptom severity, and transplanted to the field. Resistance was assayed by comparing yield of inoculated plants to those of the control non‐inoculated plants of the same variety. Results showed that the F₁ hybrids between the resistant lines and the susceptible line suffered major yield reduction because of infection, but all hybrids were more resistant than the susceptible parent. All F₁ hybrids resulting from a cross between two resistant parents, showed a relatively high level of resistance, which in most cases was similar to that displayed by the more resistant parent. In some cases, the hybrids displayed better levels of resistance than both parents, but the differences were not statistically significant. The F₁ hybrid between a line with resistance from S. habrochaites and a line with resistance from S. peruvianum (HAB and 72‐PER), exhibited the lowest yield loss and the mildest level of symptoms. Although the resistance level of this F₁ hybrid was not statistically different from the level of resistance displayed by the 72‐PER parent itself, it was statistically better than the level of resistance displayed by the F₁ hybrids between 72‐PER and any other resistant or susceptible line.     

A novel source of resistance to the two-spotted spider mite in Lycopersicon pimpinellifolium (Jusl.) Mill.: its genetics as affected by interplot interference

We have previously found an accession of Lycopersicon pimpinellifolium (Jusl.) Mill. (`TO-937') that appeared to resist attack by the two-spotted spider mite (Tetranychus urticae Koch). L. pimpinellifolium is a very close relative of the cultivated tomato (Lycopersicon esculentum Mill.) and thereby a potential source of desirable traits that could be introgressed to the crop species. The objective of this study was to investigate the genetics of the resistance present in `TO-937'. Resistance to infestation by the spider mite was quantified in 24-plant plots of L. pimpinellifolium accessions `TO-937' and `PE-10', L. pennellii accession `PE-45', L. esculentum cultivars `Moneymaker', `Roma' and `Kalohi' (reported to be partially resistant: Stoner & Stringfellow, 1967), and the interspecific F₁ cross, L. esculentum `Moneymaker' × L. pimpinellifolium `TO-937'. Only `TO-937', the F<sub>1</sub>, and`PE-45' were found to be resistant. Resistance of `TO-937' was complete when evaluated in two small greenhouses completely planted with `TO-937' so as to simulate the genotypic homogeneity usual in commercial crops. Generations (P₁, P₂, F₁, F₂, BC₁P₁, and BC₁P₂) of a P₁ (susceptible) × P₂ (resistant) cross (`Moneymaker' × `TO-937') were studied for resistance in a single-plant per plot design. Resistance of `TO-937' was inherited with complete dominance and appeared to be controlled by either two or four genes according to whether segregation in the F₂ or the BC₁P₁, respectively, were considered. However, calculation of the number of genes involved in the resistance was complicated by negative interplot interference due to the high frequency of resistant genotypes within most of the generations. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic analysis of resistance to tomato late blight in Solanum pimpinellifolium accession PI 163245

Late blight (LB), caused by Phytophthora infestans, is one of the most devastating diseases of tomato (Solanum lycopersicum) worldwide. Due to the emergence of new and aggressive P. infestans isolates, identifying new genetic resistance to LB is a priority in tomato breeding. Recently, we reported the identification of several Solanum pimpinellifolium accessions with strong LB resistance. In this study, we investigated the utility of resistant accession PI 163245 for tomato breeding by examining heritability (h²) of resistance and the response to selection for resistance. Estimates of h² based on F₂ : F₃ and F₃ : F₄ parent : offspring correlation analyses averaged 0.79 and 0.94, respectively, suggesting the heritable nature of LB resistance in PI 163245. Analysis of response to selection for resistance from F₂ to F₄ generations indicated a realized h² of 0.63, confirming the utility of this resistance in tomato breeding. Two methods of estimating the minimum number of loci involved indicated the presence of one major resistance locus. Currently, genetic mapping and breeding efforts are underway to further confirm the viability of this accession for improving tomato LB resistance.     

New sources of resistance to the potato cyst nematodes Globodera pallida and G. rostochiensis in the Commonwealth Potato Collection

The Commonwealth Potato Collection contains 77 species following the incorporation of the personal collection of Professor JG Hawkes. The material originates from several expeditions to the Americas and represents a potentially valuable source of resistance to pests and pathogens, including potato cyst nematodes (PCN). The germplasm from the Hawkes collection screened in this study included taxa mainly from Mexico, Bolivia, Argentina and Peru. Seedlings of 198 accessions, from 63 different species were screened for reaction to the PCN species G. pallida and G. rostochiensis. Of the 198 accessions tested, 56% contained G. pallida-resistant clones and 53% contained clones resistant to G. rostochiensis. Twelve species in this survey are believed to contain novel, previously unreported sources of resistance. The geographical distribution of resistance indicated a contrast between the two PCN species. Although, as in previous studies, resistance to G. rostochiensis was centred on Bolivia and Argentina, G. pallida resistance was found across the geographical range of tuber-bearing Solanum. The possibility of cross-resistance arising from selective pressure from the more northern species G. mexicana is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.