Molecular genetic characterisation of the Asc locus of tomato conferring resistance to the fungal pathogen Alternaria alternata f. sp. lycopersici

The Alternaria stem canker disease of tomato is caused by the fungal pathogen Alternaria alternata f. sp. lycopersici and its host-selective AAL-toxins. Resistance to the pathogen and insensitivity to the toxins are conferred by the Asc locus on chromosome 3L. Sensitivity to AAL-toxins is a relative character; the toxins inhibit development of all tested tomato tissues but susceptible cultivars are much more sensitive than resistant cultivars. In addition to tomato, some other plant and animal species are sensitive to the toxins as well. The likely mode of action of AAL-toxins is interference with sphingolipid biosynthesis by specific inhibition of ceramide synthase activity. To molecularly isolate Asc, transposon tagging and positional cloning strategies are applied. As a first step, transposon insertions and restriction fragment length polymorphism (RFLP) markers are identified in proximity of the Asc locus. Subsequently, the transposons are used to inactivate Asc by insertion mutagenesis, and the RFLP markers are used to identify yeast artificial chromosomes (YACs) with tomato DNA inserts. Once an Asc-insertion mutant and/or a YAC encompassing Asc has been obtained, physical isolation and characterisation of Asc will be conceivable. Elucidation of the molecular role of Asc will illuminate the specificity of host recognition by Alternaria alternata f. sp. lycopersici.Abbreviations AAL-toxin Alternaria alternata lycopersici-toxin - A. a. lycopersici Alternaria alternata f. sp. lycopersici - Asc Alternaria stem canker - HST host-selective toxin     

Resistance at the TM-2 locus in the tomato to tomato mosaic virus

Summary There are three known tomato mosaic virus (TMV) resistance factors, Tm-1, Tm-2 and Tm-2 ², in the tomato. Tm-2 ² is currently the most widely utilised in glasshouse cultivars. Both Tm-2 and Tm-2² can induce systemic necroses in response to virus infection. These are considered to be hypersensitive resistance reactions in view of the low virus concentrations in affected plants and because sub-inoculation usually fails to infect all plants possessing the same resistance gene. The literature relating to TMV resistance at the Tm-2 locus in the tomato is reviewed.Virulent strains may readily establish when Tm-1 or Tm-2 are used, but Tm-2 ² confers more effective resistance. The possible development of aggressive isolates capable of affecting Tm-2 ²/Tm-2² plants is discussed. The establishment of virus types which cause systemic necrosis at normal growing temperatures is considered more likely than widespread infection from fully virulent strain 2² mutants. However, the growing of crops isolated from the TMV reservoirs in the soil considerably reduces the likelihood of even this occurring.     

Introduction of aromatic fragrance into cultivated tomato from the peruvianum complex'

This study was performed to introduce the distinct aromatic fragrance of Lycopersicon peruvianum LA 1554 into the cultivated tomato, Lycopersicon esculentum. The strong breeding barriers existing between these two distantly related species were circumvented by the ovule selection and culture method. A large BC₁F₁ population was developed and among 127 plants, 36 were self‐compatible and yielded fruits. Fruits of some of these selected plants were found to be enriched with a sweet aromatic flavour. Sensory evaluation of the fruit aroma of these selected plants was performed by a panel of 12 members against one of the best consumer‐rated Japanese commercial tomato cultivars, ‘Momotaro’. Although extensive variation was observed in fruit‐aroma in the BC₁F₁ population, panel opinion on ‘flavour‐desirability’ significantly favoured the BC₁F₁ fruits of some selected plants over the cv. ‘Momotaro’. Therefore, it can be concluded that the aromatic fragrance of a ‘L. peruvianum’ accession has successfully been introduced into the cultivated tomato gene pool.     

Transfer of Tomato Leaf Curl Virus Resistance from Lycopersicon hirsutum f. glabratum to L. esculentum

Six lines, i.e., H-2, H-11, H-17, H-23, H-24, and H-36, resistant to Tomato Leaf Curl Virus (TLCV) have been developed with controlled introgression of L. hirsutum f. glabratum into Lycopersicon esculentum. The disease incidence, 120 days after inoculation, of those lines derived from L. hirsutum f. glabratum ranged from 8.3 to 35.0 %, whereas in susceptible varieties it ranged from 95.0 to 100 %. The coefficient of infection (CI) values in the resistant lines were very low, ranging from 0.25 to 4.55, whereas in susceptible varieties CI values ranged from 60.56 to 88.96. Line H-2 had the highest resistance by showing the least disease incidence and CI values. The fruit size and days to maturity in resistant lines were close to those of cultivated susceptible varieties. These lines have the scope for being used as varieties in the TLCV infested areas or as foundation lines for further genetic improvement.     

Performance of Solanum habrochaites LA1777 introgression line hybrids for marketable tomato fruit yield in Asia

Wild relatives of the cultivated tomato (Solanum lycopersicum L.) are major sources of new genetic diversity for tomato improvement. Introgression lines (IL) are near-isogenic lines homozygous for one or several mapped wild DNA fragments in a common recurrent parent. A set of ILs developed by Cornell University from Solanum habrochaites accession LA1777 and recurrent parent E6203 has been made publicly available through the Tomato Genetics Resource Center of the University of California-Davis. Our objective was to identify LA1777 introgressions with potential to increase the marketable fruit yield of tomato grown in the tropics. A subset of ILs were each crossed to CLN2498E (resistant to bacterial wilt and some begomoviruses) to create IL hybrids (ILH). ILH, IL recurrent parent E6203, CLN2498E, and CLN2498E × E6203 (Hchk) were evaluated in replicated trials in Thailand, India, and Taiwan during two dry seasons. Highly significant effects for marketable fruit yield were detected in Thailand and Taiwan. ILH heterozygous for S. habrochaites segments at the bottom of chromosome 1 yielded about 20% than the Hchk at Thailand and Taiwan. Our results agree with previous results from Cornell University researchers who found a segment of S. habrochaites DNA located between TG158 and TG27 associated with increased total fruit yield in previous trials conducted in upper state New York. Yield improvement due to this S. habrochaites introgression can occur over a wide range of environments.     

Pistillate-parental differences and the ability to produce interspecific hybrids between Lycopersicon esculentum and 'peruvianum-complex' species (L. peruvianum and L. chilense)

To overcome the cross-breeding barriers between the cultivated tomato Lycopersicon esculentum and the ‘peruvianum-complex’, hybrid production ability (HPA) and pistillate-parental differences were investigated. As a criterion of HPA, the number of germinated ovules per fruit (GPF) was used. GPF was expressed as GPF = OPF × GPO, where OPF is the number of ovules per fruit, and GPO is the proportion of germinated ovules to total ovules obtained. The interspecific crossing between nine varieties and three ‘peruvianum-complex’ accessions revealed that the cultivars ‘Sekaiichi’,‘Ponde Rooza’ and ‘Early Pink’ showed quite high and stable GPF over the years, but the cv.‘Best of All’ produced no hybrids. Variance analysis for GPF, OPF and GPO, and their correlation with seven sexual organ morphological traits and three fruit morphological traits were performed. These results indicated that choosing both the pistillate parents with wider reproductive organs for high OPF and appropriate environmental conditions for high GPO might be significant for enhancing GPF in interspecific crossing.     

In vitro selection of tomato transformants at the immature embryo stage

Summary A method is described for selecting resistant transformed tomato genotypes in vitro at the stage of immature embryos. The utilization of HLH nutrient medium with the selective agent kanamycin is proposed. Normal development of seedlings from immature embryos, which do not form callus, is a good and true indicator for isolation of resistant genotypes. The immature embryos do not germinate and develop on MS selective medium.     

Diallel analysis over two environments for blossom-end scar size in tomato

Summary To investigate genetic regulation of blossom-end scar size in tomato (Lycopersicon esculentum Mill.), a half diallel cross including 10 parents was grown under warm fall conditions in Bradenton, Florida, and cool winter conditions in Hazeva, Israel. The parents were a random sample representing all available fresh market tomato breeding lines commercially grown under subtropical field conditions. A randomized, complete block design with three replications was used and the blossom-end scar index (BSI), a measure for scar size relative to truit size, was measured on 40 and 25 fruits per plot in Bradenton and Hazeva, respectively. Analysis of variance for BSI indicated highly significant (P=0.001) variation among parents and among F1's in both environments. In Bradenton and Hazeva both, the difference in average BSI between parents and F1's was not significant suggesting insignificant overall dominance effects. Further partitioning of variation within F1's indicated that general combining ability (GCA) effects were highly significant in both environments, whereas specific combining ability (SCA) effects were significant at P=0.001 in Bradenton, but only at P=0.05 in Hazeva. Estimated variance components for GCA and SCA effects indicated that BSI inherited mainly additively in both environments. Analysis combined over environments indicated that variation in sensitivity to environments was 5-fold higher among parents than among F1's. The genotype x environment variance component was not large enough to justify testing over more than one environment for population improvement purposes and early testing of hybrids. Evaluation of inbred lines, however, may have to be done in more than one environment, especially for Suncoast-derived material. Under temperatures in Hazeva, genotype differences were more pronounced and heritabilities higher than under high temperatures in Bradenton. The estimated overall single plot heritability was 0.63. Breeding lines with a pointed blossom-end morphology (e.g. NC 8276 and NC 140 in this study) generally had small blossom-end scars, and intercrossing of these lines or crossing with non-pointed, moderately smooth breeding lines generally resulted in smooth hybrids.     

Low-temperature-related growth and photosynthetic performance of alloplasmic tomato (Lycopersicon esculentum Mill.) with chloroplasts from L. hirsutum Humb. & Bonpl.

Growth and photosynthetic performance were analyzed in alloplasmic tomato at a high- (25/17 ^°C; HTR) and low-temperature regime (12/6 ^°C; LTR) in order to establish the role of cytoplasmic variation on low-temperature tolerance of tomato (Lycopersicon esculentum Mill.). Four alloplasmic tomato lines, containing the nuclear genome of tomato and the plastome of L. hirsutum LA 1777 Humb. & Bonpl., an accession collected at high-altitude in Peru, were reciprocally crossed with 11 tomato entries with a high inbreeding level and a wide genetic variation, resulting in a set of 44 reciprocal crosses. Irrespective of growth temperature, alloplasmic families with alien chloroplasts of L. hirsutum (h) were on average characterized by a high shoot biomass, a large leaf area, and a low specific leaf area in comparison with their euplasmic counterparts. These results do not directly point to an advantageous effect of h-chloroplasts on biomass accumulation at low temperature but rather towards a small general beneficial effect on growth and/or distribution of assimilates. Significant chloroplast-related differences in photosynthetic performance, however, were not detected at both temperature regimes, indicating that h-chloroplasts can properly function in a variable nuclear background of L. esculentum. It is concluded that chloroplast substitution is not an effective method for breeding tomato plants with improved low-temperature tolerance. This revised version was published online in August 2006 with corrections to the Cover Date.     

A CAPS marker TAO1902 diagnostic for the I-2 gene conferring resistance to Fusarium oxysporum f. sp. lycopersici race 2 in tomato

Fusarium oxysporum f. sp. lycopersici inhabits most tomato-growing regions worldwide, causing tomato production yield losses. A molecular marker linked to resistance would be useful for tomato improvement programmes. Thus, a cleaved amplified polymorphic sequence (CAPS) marker TAO1₉₀₂ was developed to identify tomato genotypes possessing the I-2 gene, which confers resistance to F. o. lycopersici race 2. The Rsa I or Fok I restriction fragments corresponded to the presence or absence of the I-2 allele in a segregating 100 F₂ progeny, tomato cultivars, 16 resistant and 20 susceptible to Fusarium wilt, respectively, lines and F₁ hybrids, representing various tomato gene pools. TAO1₉₀₂ may be helpful for selection of F. o. lycopersici -resistant tomato germplasm.