Breaking breeding barriers in Lycopersicon. 5. The inheritance of the unilateral incompatibility between L. peruvianum (L.) Mill. and L. esculentum Mill. and the genetics of its breakdown

Summary The validity of the current hypotheses on the inheritance of unilateral incompatibility is disputed and the importance of intrapopulational research on the inheritance of unilateral incompatibility is emphasized.Such research has been carried out on the unilateral incompatibility between L. peruvianum and L. esculentum with inbred material of L. peruvianum. The results indicate that in this species the inhibition of L. esculentum pollen tube growth, built up of distinct processes, is based on independent dominant genes, and that the breakdown of the phenomenon is a consequence of segregation of recessive alleles, the expressivity of at least some of these being influenced by environmental factors.Research demonstrating differences between self-incompatibility and interspecific pollen tube growth inhibition is reviewed and it is concluded from this and own research that the two phenomena are distinct.     

Breaking breeding barriers in Lycopersicon. 3. Inheritance of self-compatibility in L. peruvianum (L.) Mill.

Summary A brief survey is given of the genetics of self-compatibilith in species with a one-locus gametophytic system of incompatibility.A study has been made of the genetics of the self-compatibility found in L. peruvianum.From the results of various test crosses and selfings and of cytological research it is concluded that self-compatibility in L. peruvianum can be based on different types of S-allele mutations, on addition of an S-allele bearing chromosome fragment, or on genes modifying the S-allele expressivity.The results further indicate that generation of new S-alleles is a frequently occurring phenomenon in inbred material of L. peruvianum and that pollen with an S-allele mutation or an extra chromosome fragment is less vital than normal pollen.A short notation for incompatibility genotypes is given.     

Breaking breeding barriers in Lycopersicon. 2. Breakdown of self-incompatibility in L. peruvianum (L.) Mill.

Summary Attempts were made to break the self-incompatibility in L. peruvianum by selection of mutated S-alleles through large-scale self-pollination on clones with pollen which was mutagenically treated in different stages of development. Besides self-compatibility was searched for in inbred lines.The self-incompatibility in L. peruvianum was found to be very strict indeed. From more than 22,000 self-pollinations on 5 clones 1527 seeds were obtained, seed set being very erratic. The possible causes of this seed set are discussed. From 1527 seeds 1036 plants were raised and tested for self-compatibility.In the progenies of 4 clones from 823 plants tested, 7 were more or less self-compatible. It is concluded that this self-compatibility was spontaneous and not the result of the mutagenic treatment.In the progeny of the fifth clone plants were found which reacted as self-compatible at a temperature of about 40°C and as self-incompatible at lower temperatures. It is suggested that this character—a high temperature sensitive incompatibility reaction—is governed by one recessive gene.In inbred lines plants were found with a stable form of self-compatibility and also plants with a high temperature sensitive incompatibility reaction.The problems of an incompatibility research on inbred material are discussed. A brief survey is given of some of the characters found in inbred L. peruvianum.     

Self-compatibility in Lycopersicum peruvianum (L.) mill.

Summary Mention is made of self-compatibility found in Lycopersicum peruvianum. This character is of importance for genetic research on this species and opens up the possibility of a better exploitation of species hybrids with L. esculentum.     

Overcoming unilateral breeding barriers between Lycopersicon peruvianum and cultivated tomato,Lycopersicon esculentum

Summary Treatment of Lycopersicon peruvianum stigmas with an artificial medium analogous to stigmatic exudate allowed pollen germination and growth on immature pistils. Growth of Lycopersicon esculentum pollen tubes to L. peruvianum ovules, an otherwise incompatible cross, was achieved following such treated bud-pollinations. No plantlets were recovered, although a few embryos from this cross at the globularity heart stage of development were excised at 22 days after pollination, indicating the presence of crossing failures as severe as in the reciprocal cross. Hybrid plants were obtained from the reciprocal cross, using as pollen parent an L. peruvianum line selected for congruity with L. esculentum. Bud pollinations to L. peruvianum, using these interspecific F₁ hybrids as the pollen parent, allowed viable embryo development and plantlet recovery. Resulting backcross plants, which possess approximately 1/4 L. esculentum genome in a L. peruvianum cytoplasm, may facilitate further introgression of the L. esculentum nuclear genes into in foreign cytoplasm.     

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.     

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.     

Efficient hybridization between Lycopersicon esculentum and L. peruvianum via 'embryo rescue' and in vitro propagation

For the transfer of valuable traits from wild species into the cultivated tomato, excised globular-stage embryos 13 and 15 days after pollination (DAP) were cultured in vitro. Plants were regenerated from interspecific crosses of Lycopersicon esculentum cv. ‘Early pink’×L. peruvianum PI270435, and backcrosses of L. esculentum‘Giban (JF) No. 1’× (‘Early pink’× PI270435). Somatic embryos and single cotyledons emerged on hypocotyl sections of the embryos. Five to nine plantlets per embryo were obtained by clonal propagation. The hybrid nature of the plants is confirmed by comparing hybrids and parents in their ability to regenerate shoots from leaf segments in vitro, by comparing plant morphology and characteristics and by chromosome number. This study describes an efficient ‘embryo rescue’ method, as well as somatic embryogenesis by clonal propagation. A novel and simple method for the characterization of the interspecific hybrids is also reported.     

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.     

Somatic hybridization as a tool for tomato breeding

Protoplast fusion can be used to produce somatic hybrids of species that cannot be obtained by sexual hybridization. The possibility to introgress genes from Solanum species into the cultivated tomato species Lycopersicon esculentum, and to obtain novel cytoplasm-nucleus combinations (cybrids) was considered as an important strategy to extend the genetic variation available for tomato breeding. Somatic hybrids between L. esculentum and other Lycopersicon species, as well as between L. esculentum and Solanum or Nicotiana species, have been produced. Specific mutants, genotypes with antibiotic resistances, and metabolic inhibition by iodoacetate or iodoacetamide and irradiation were used for the selection of hybrids. In addition, the improvement of protoplast culture techniques and the use of the favourable tissue culture traits derived from species such as L. peruvianum, which have been introduced into tomato by classical breeding, allowed the efficient recovery of somatic hybrids. However, the occurrence of somatic incongruity in fusion combinations of L. esculentum and Solanum and even more in L. esculentum and Nicotiana, did not allow the production of true cybrids and/or fertile hybrids, indicating the importance of both cytoplasm-nucleus and nucleus-nucleus interactions in somatic incongruity. Another problem with fusions between distantly related species is the strongly reduced fertility of the hybrids and the very limited homoeologous recombination between chromosomes of the parental species. Partial genome transfer from donor to recipient through microprotoplast (+) protoplast fusion, and the production of monosomic or disomic chromosome addition lines, light overcome some of these problems. In symmetric somatic hybrids between L. esculentum and S. tuberosum the occurrence of limited somatic and meiotic recombination was demonstrated. Fertile progeny plants could be obtained, though at a low frequency, when embryo rescue was performed on a large scale after backcrossing hexaploid somatic tomato (+) potato hybrids with a tetraploid potato genotype. The potential value of genomic in situ hybridization (GISH) and RFLPs for the analysis of the genome/chromosome composition of the hybrids has been demonstrated for intergeneric somatic hybrids between Lycopersicon and Solanum.Abbreviations cpDNA chloroplast DNA - mtDNA mitochondrial DNA     

Trichomes on leaves of Lycopersicon hirsutum,L. esculentum and their hybrids

Summary Density of certain types of trichomes were characterized on leaflets of Lycopersicon hirsutum, L. esculentum and their hybrids. Trichome density covaried with leaflet development, and depending on the species and type of trichome, density also covaried with leaflet surface. Type IV trichomes were not observed on leaflets of L. esculentum and Type V trichomes were not observed on leaflets of L. hirsutum. Type IV and V trichomes were present on F₁ hybrids and densities of these two types of trichomes segregated in the F₂ population. When corrected for differences of leaflet size between L. hirsutum and L. esculentum, densities of Type VI trichomes were more similar between species than uncorrected densities. There was little difference in Type VI density among F₂ individuals. The appearance, lipid and phenol staining properties, and native fluorescence of the multicellular tip of the Type VI trichome differed between L. hirsutum and L. esculentum. F₁ hybrids were intermediate for these characters and these characters segregated in the F₂ generation.The investigation reported in the paper (84-10-94) is in connection with a project of the Kentucky Agricultural Experiment Station and is published with the approval of the Director. This material is based upon work supported, in part, by the U.S. Department of Agriculture, Agreement No. 59-2213-1-1-717-0 (Competitive Research Grants Program).     

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.     

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.     

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     

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.     

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.     

Introgression of in vitro regeneration capability of Lycopersicon pimpinellifolium Mill. into recalcitrant tomato cultivars

The goal of this research was to study the introgression of the high regeneration capacity of Lycopersicon pimpinellifolium Mill line WV-700, in recalcitrant tomato cultivars (L. esculentum Mill cvs. Petomech, Santa Rita and VFN-8) using backcrossing. Hypocotyl explants of in vitro germinated seeds were cultivated in half strength MS medium supplemented with 5mg/l 6-BA to assess their shoot regeneration capacity. The apical shoot of the in vitro germinated seeds were cultured on a separate medium. Apical shoots from genotypes showing high regeneration rates were acclimated in a glasshouse and used as pollen donors for the next backcrossing. After four backcrossings, the material showed a similar mean fruit weight for the cultivated tomato and a high regeneration capacity similar to the wild species. It is shown that L. pimpinellifolium can be used with success as donor parent to introgress in vitro regeneration capacity to recalcitrant tomato cultivars. This revised version was published online in August 2006 with corrections to the Cover Date.     

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.     

Presence of Gene Del in Wild Species of the Genus Lycopersicon Mill.

The carotene composition of the fruits of F₁ hybrids between Lycopersicon esculentum Mill. — cv. 1/D (genotype Del⁺Dd⁺) and wild species of genus Lycopersicon with various fruit pigmentation — red, orange, and green — has been investigated. The results reveal that the red and orange-fruited species carry the recessive allele Del⁺, while the green-fruited species carry a δ-carotene gene which is identical or similar in action to the dominant gene Del. The test of allelism that has been carried out proves that the δ-carotene gene from Lycopersicon pennellii Corr. is allelic to gene Del.     

Attempts to overcome partial incompatibility between Hordeum vulgare L. and H. bulbosum L.

Summary Partial incompatibility has already been reported between Hordeum vulgare and H. bulbosum during the course of doubled haploid production, and in this paper attempts to overcome the breeding barrier are described. The methods which seem to offer most success are those of environmental adjustments and the adoption of new genotypes of H. bulbosum. Further cultivars of H. vulgare exhibiting this phenomenon are also noted.