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.     

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.     

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.     

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.     

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.     

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.     

Inheritance of resistance to potato cyst-eelworm (Heterodera rostochiensis Woll.) in the genus Lycopersicon

Summary The presence of resistance to potato cyst-eelworm in wild Lycopersicon species was confirmed.The resistance of two plant accessions, L. pimpinellifolium, B6173, and L. peruvianum, B6001, was compared in screening tests against three eelworm isolates, using the susceptible L. esculentum Ailsa Craig as a control. Both accessions were shown to possess high levels of resistance to the Wren isolate (pathotype A) of H. rostochiensis, and resistance to the Castle Donington (pathotypes A, B, and E) and Hare Lane (pathotype B) isolates. The level of resistance in L. pimpinellifolium was greater than that in L. peruvianum. Because of its greater resistance, and the ease with which it can be hybridised with tomato cultivars, L. pimpinellifolium was chosen as a donor parent for breeding programmes designed to incorporate potato cyst-eelworm resistance into a commercial tomato variety.The resistance of L. pimpinellifolium, B6173, to the Wren isolate of H. rostochiensis was shown to be controlled by a single dominant gene for which the symbol Hero is proposed.     

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.     

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.     

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.     

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.     

Inheritance of resistance to two isolates of sterility mosaic pathogen in pigeonpea (Cajanus cajan (L.) Millsp.)

The resistant reaction to tomato spotted wilt virus (TSWV) was found to be determined by a single dominant gene in three Capsicum chinense Jacq. accessions (‘PI 152225’, ‘PI 159236’, ‘7204’). Allelism studies indicated that all C. chinense lines bear the same allele located at the Tsw locus. All the inoculated plants in the allelism tests displayed a resistant hypersensitive phenotype characterized by necrotic local lesions followed by abscission of the inoculated organ. However, a small proportion of them showed late systemic infection. Nine TSWV isolates obtained from these individual plants with systemic symptoms were backinoculated to the three resistant parents. All isolates were able to infect systemically all the resistant accessions without inducing local necrotic lesions. Serological analysis confirmed that these nine viral isolates belong to the TSWV species (serogroup I). Consequently, the susceptible plants in the allelism tests could not be interpreted as possessing a recombinant genotype because of the virulence change in the viral strain. Hobbs et al. (1994) already reported the existence of TSWV pathotypes overcoming the resistance of C. chinense resistant accessions. Practical consequences for pepper breeding associated with the emergence of these resistance-breaking isolates are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Resistance in Lycopersicon species to black leaf mold caused by Pseudocercospora Fuligena

Over 540 accessions of wild Lycopersicon species or their crosses with L. esculentum were screened for resistance in a series of trials. Forty-six accessions were selected for the final screening trial based on lower disease ratings in previous trials. Of these, L. hirsutum had the greatest number of resistant accessions, followed by L. esculentum and L. peruvianum. Twenty accessions were quantified for their levels of resistance based on leaf area infected, area under disease progress curve (AUDPC), and the degree of sporulation. There was a significant positive correlation between the AUDPC calculated from 20 accessions evaluated under growth room and field conditions. Five L. hirsutum accessions had no sporulation associated with leaf lesions, whereas L. esculentum accessions had an average of 1.6×10⁴ conidia/cm² of leaf tissue. There was significant positive correlation between the AUDPC values and the number of conidia per cm² of leaf tissue.     

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.     

Evaluation of French Brassica oleracea landraces for resistance to Plasmodiophora brassicae

A total of 240 kale, 38 cabbage and 126 winter cauliflower French landraces from the B. oleracea genepool of INRA were assessed for resistance to clubroot caused by Plasmodiophora brassicaeWoron. Two French isolates of the pathogen (K and SJ) were used in the experiments under controlled conditions. The reaction of the 126 cauliflower accessions to naturally occurring clubroot was also evaluated in field trials. Kales exhibited considerable variation for expression of disease resistance and high levels of resistance were found in several accessions. In this group, single resistant plants were observed in most of the morphological types and from quite different geographical origins. Cabbage accessions were moderately to highly susceptible to both isolates. All cauliflower populations proved to be highly susceptible to K isolate and moderately susceptible to SJ isolate. In field trials, cauliflowers were also severely infected. Two lines selected from a resistant kale population were highly resistant against a large range of pathotypes of the pathogen. These lines presented a sufficient level of resistance to be directly useful in the breeding program in order to develop cauliflower and broccoli hybrids resistant to clubroot. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Inheritance of resistance to Pyrenochaeta lycopersici in tomato

Summary Tomato accessions (Lycopersicon sp.), along with commercial cultivars and breeding lines were grown in a field infested with the brown root rot (BRR) organism, Pyrenochaeta lycopersici and evaluated for resistance. Three L. esculentum Mill. accessions, P.I. 260397, P.I. 262906 and P.I. 203231, were resistant and were used as male parents in crosses designed to transfer resistance to tomatoes of fresh market type. Through analysis of parental generations and F₁ and F₂ progenies from three crosses the heritability of resistance in the broad sense was estimated to range from 25 to 43 percent. The minimum number of genes influencing resistance was estimated to be from 4 to 8.Florida Agricultural Experiment Stations Journal Series Paper no. 317.     

Tomato Spotted Wilt Virus (TSWV) resistance in tomato derived from Lycopersicon chilense Dun. LA 1938

Tomato spotted wilt virus (TSWV) resistance wasidentified in Y118 (Fla 925-2), an F₁BC₁S₆ tomato line(Lycopersicon esculentum Mill.), derived from a crosswith L. chilense Dun. (LA 1938). This line waspreviously selected for tomato mottle virus (ToMoV)resistance in Florida. Progeny from crosses betweenFla 925-2 and three different TSWV susceptible L.esculentum parents were used in TSWV resistancestudies. A total of 75 F₁ and 596 F₂ plants from allthree crosses were screened for TSWV resistance. ForF₂ plants free of TSWV symptoms, evaluations were madeusing enzyme-linked immunosorbent assay (ELISA). TenF₃ populations used for further greenhouse and fieldscreenings were selected from F₂ plants found to befree of the virus using visual and ELISA criteria ateach evaluation. One F₁ and four F₃ lines werestudied under field conditions (Stellenbosch, SouthAfrica) in which 100% of the `Flora-Dade' susceptiblecontrols were severely infected with TSWV. Theresults of the field study clearly establish that TSWVfield resistance is present in the Fla 925-2 (Y118)derived lines. Studies conducted on these linesrevealed that this resistance has the distinctcharacteristic of often `recovering' from initiallyhigh levels of virus titer in the tissue to levelsbelow detection with ELISA.     

Low temperature seed germination of Lycopersicon species evaluated by survival analysis

Summary Low temperature germination responses were evaluated for 18 high altitude accessions representing five wild Lycopersicon species and 19 accessions of L. esculentum which have reputed ability to germinate in the cold. Survival analysis indicated that one accession of L. chilense germinates better at 10°C than PI 120256, the fastest-germinating L. esculentum genotype, and that PI 120256 germinates as well as PI 126435 (L. peruvianum). Additional wild ecotypes exhibiting rapid germination at 10°C were identified from L. peruvianum and L. hirsutum. These ecotypes may possess genetic potential for introgressing cold germination ability into L. esculentum cultivars.     

Monogenic resistance in tomato to Fusarium oxysporum f. sp. lycopersici race 3

Summary Resistance to fusarium wilt, incited by Fusarium oxysporum (Schlecht.) f. sp. lycopersici (Sacc.) Snyder & Hansen race 3 in tomato (Lycopersicon esculentum Mill.) was discovered in LA 716, a L. pennellii accession. A resistant BC₁F₃ breeding line, E427, was developed from LA 716. E427 was crossed with the susceptible cv. Suncoast and F₁, BCP₁, BCP₂ (to Fla 7155, a susceptible parent) F₂, F₃, and BCP₂S₁ seeds were obtained. Segregation for resistance following root dip inoculation over three experiments indicated a single dominant gene controlled resistance. Five of the 12 BCP₁S₁'s segregated more susceptible plants, whereas one of the 12 segregated more resistant plants than expected (P<0.05). Three of 23 F₃ lines segregated more susceptible plants than expected while 1 of the 23 had more resistant plants than expected (P<0.05). Segregation in all other lines fit expected ratios. Five of the 23 F₃'s were homozygous resistant which was an acceptable fit to expectations (P=0.1–0.5). The gene symbol I ₃ is proposed for resistance to race 3 of the wilt pathogen. Deviations from expected ratios in data reported here and for other breeding lines indicate an effect of modifier genes and/or incomplete penetrance. Plant age at inoculation and seed dormancy did not affect results.Florida Agricultural Experiment Station Journal Series No. 8101.