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     

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.     

Variation of resistance to broomrape (Orobanche ramosa) in tomatoes

Summary Twenty five tomato cultivars and one accession of wild tomato were evaluated for their resistance to branched broomrape (Orobanche ramosa) infestation. Tomato cultivars were found different in their resistance to Orobanche based on different growth parameters. Total Orobanche number, haustoria development and number of emerged Orobanche shoots were all different between tomato cultivars. Differences in the growth and fruit yield among tomato cultivars were also found in response to Orobanche infestation. Ranking tomato cultivars for Orobanche resistance indicates the existence of different resistance mechanisms in these cultivars. Relatively high to moderate levels of resistance were obtained in Tiny Tim, Acora, Castler, Pomodora, Orient, Red Alert and the accession LA 1478 of L. pimpinellifolium. The cultivar Tiny Tim showed the highest level of resistance for all measured parameters and in all experiments.     

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.     

Genetics of the viability of pollen grain produced at low temperatures inLycopersicon Mill.

Summary The different viabilities of pollen produced at low temperatures in intra- and inter-specific crosses of tomato were studied. Cultivars Red Top, Moneymaker, and Marroqui were crossed with cultivar E-15 and these four cultivars were hybridized with lines of the wild speciesL. pimpinellifolium PE-13,L. parviflorum PE-52,L. pennellii PE-47, andL. hirsutum PE-37 and PE-41. A six-generation family of the Moneymaker x PE-47 cross was obtained to carry out a more detailed genetical study of pollen grain viability at low temperatures. Pollen grain viability was evaluated during the winter via acetocarmine staining. When the parents were compared with their F₁, the intra-specific tomato crosses showed dominance to better-quality pollen, theL. esculentum x L. pimpinellifolium inter-specific crosses showed positive heterosis, while the crosses ofL. esculentum with the tolerant speciesL. pennellii andL. hirsutum showed intermediate inheritance. However, in theL. esculentum x L. pennellii family, the dominance and the non-allelic interactions (homozygosis x homozygosis) were the most important factors, so that dominance to better viability at low temperatures appeared to be the general mode of inheritance. Genetical control of pollen grain viability at low temperatures seemed to be polygenetic.     

Inheritance of resistance to fruit rot (Phytophthora parasiticaDast.) in tomato (Lycopersicon esculentum Mill.)

Summary Fruit rot disease caused by Phytophthora parasiticaDast. is a limiting factor in tomato production in Himachal Pradesh. 30 to 60 per cent fruits are damaged by this disease. Crosses were made between EC 54725 (Lycopersicon pimpinellifolium), a small tyuited type, resistant to fruit rot and four highly susceptible tomato commercial cultivars (Gola, Sioux, S12, and Lalmani). Studies of F₁'s, F₂'s and back crosses indicated that EC 54725 carries a dominant gene imparting resistance to fruit rot.     

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.     

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.     

Qualitative resistance to powdery mildew in hybrid sweet peas

Lathyrus odoratus L. × Lathyrus belinensis L. hybrids were produced using L. belinensis as the pollen parent, with fertile seed produced by the L. odoratus parent. The F1 hybrid plants were completely self-sterile, but produced viable seeds when backcrossed to L. odoratus. The plants produced by backcrossing resembled L. odoratus, the flower colour being purple/magenta, and were self-fertile. Both hybrid plants and those produced by back crossing to L. odoratus were resistant to Erysiphe pisi DC that causes powdery mildew in sweet peas. Continued backcrossing resulted in hybrid plants, that closely resembled the L. odoratus parent, but segregated for complete resistance/susceptibility to E. pisi,with a ratio of 2.46:1 resistant to sensitive plants. This suggests the presence of a single dominant gene that confers resistance. When resistant and sensitive plants were inoculated with E. pisi and their leaf surfaces examined,using a Scanning Electron Microscope, it was found that although spores germinated on the leaves of both resistant and sensitive plants, spores present on resistant plants collapsed shortly after germination. Possible reasons for this are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Assessment of conditions affecting Agrobacterium-mediated soybean transformation using the cotyledonary node explant

Conditions affecting Agrobacterium-mediated transformation of soybean [Glycine max (L.) Merr.], including seed vigor of explant source, selection system, and cocultivation conditions, were investigated. A negative correlation between seed sterilization duration and seed vigor, and a positive correlation between seed vigor and regenerability of explants were observed in the study, suggesting that use of high vigor seed and minimum seed sterilization duration can further improve transformation efficiency. Selection schemes using glufosinate or bialaphos as selective agents in vitro were assessed. Glufosinate selection enhanced soybean transformation as compared to bialaphos. The use of 6 mg L^-1 glufosinate during shoot induction and shoot elongation stages yielded higher final transformation efficiency ranging from 2.0% to 6.3% while bialaphos at 4 to 6 mg L^-1 gave 0% to 2.1% efficiency. Including cysteine and DTT during cocultivation increased the transformation efficiency from 0.2–0.9% to 0.6–2.9%. This treatment also improved T-DNA transfer as indicated by enhanced transient GUS expression. Shoot regeneration and Agrobacterium infection were attained in twelve soybean cultivars belonging to maturity groups I-VI. These cultivars maybe amenable to genetic transformation and may provide a valuable tool in soybean improvement programs. This revised version was published online in July 2006 with corrections to the Cover Date.     

Factors affecting in vitro flowering and fruiting of green pea (Pisum sativum L.)

Multiple shoots were efficiently regenerated from cotyledonary node and shoot tip explants of Pisum sativum within 15 days on MS medium containing B₅ vitamins and supplelmented with 2.0 mgl^-1 6-benzylaminopurine. The elongated shoots produced on the same medium were excised and transferred to MS medium containing half strength ammonium nitrate (8.25 gml^-1) and supplemented with auxins (indole-3-butyric acid or naphthalene acetic acid) either alone or in combinations with gibberellic acid. Rooting and flowering were observed on the 7th and 15th day after their transfer to rooting medium. The flowers self-fertilised in vitro and produced mature pods within 25 days of rooting. These seeds were germinable both in vitro and in vivo. In vitro seeds sown in pots under field conditions developed into flowering plants, and subsequently produced pods with viable seeds. This revised version was published online in July 2006 with corrections to the Cover Date.     

Production of hybrids between Cajanus acutifolius and C. cajan

There are many wild species of pigeonpea which are endemic to Australia. These wild species are cross incompatible with cultivated species of Indian origin. Cajanus acutifolius is one such species which does not easily cross with cultivated pigeonpea. Interspecific pollinations lead to hybrid seeds which were semi-shrivelled. Very few seeds germinated to give rise to F1 plants. Backcrossing the hybrid plants to C. cajan, the male parent, gave rise to aborting seeds which did not germinate in vivo hence BC1 plants are obtained after saving the aborting embryos in vitro. BC₁ plants showed normal meiotic pairing, but had low pollen fertility. The reasons for embryo abortion and low pollen fertility in spite of normal meiosis could be due to the effect of wild species cytoplasm. This revised version was published online in August 2006 with corrections to the Cover Date.     

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.     

Hybridization Studies of Five Species of the Solanum Nigrum Complex Found in South Africa and two Cocktail Tomato Cultivars

Summary Species from the Solanum nigrum complex are not as popular as potatoes and peppers but are economically important fruit or leafy vegetables in some parts of the world. The aim of this study was to look at the hybridization potential of genotypes from the Solanum nigrum complex, and two cocktail tomato cultivars, to facilitate a future breeding program. Solanum americanum Mill., S. burbankii Bitter, S. chenopodioides Lam., S. retroflexum Dun. and S. scabrum L. are some of the species found in Southern Africa. Crosses, including reciprocals, were made between these five species and two cocktail tomato cultivars. Fifty percent of the crosses made, resulted in fruit set, but only 22.5% of the produced seeds germinated to yield progeny. Cytological analyses of the chromosome numbers showed no variation from the normal euploid series. The occurrence of polyploidy between the species is probably the most efficient barrier to natural hybridization between the species.     

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.     

Plant regeneration via organogenesis and somatic embryogenesis in callus cultures derived from mature zygotic embryos of leek (Allium ampeloprasum L.)

Summary A high frequency plant regeneration system via organogenesis and somatic embryogenesis was established with callus cultures derived from mature zygotic embryos of different leek genotypes (Allium ampeloprasum L.). Four different callus types with varying morphogenetic potential were obtained. Relatively high concentrations of the auxin 2,4-dichlorophenoxy-acetic acid reduced callus weight and subsequent shoot regeneration and primordia formation of the callus. Shoot regeneration and primordia formation of the callus decreased after prolonged subculture on media containing 2,4-dichlorophenoxy acetic acid. A callus growth period of six weeks on Murashige and Skoog medium with 0.25–0.5 mg l^-1 2,4-dichlorophenoxy acetic acid showed the highest rate of shoot regeneration after transfer of callus to regeneration medium with 1 mg l^-1 kinetin.Differences between leek genotypes in callus type, callus weight, shoot regeneration and primordia formation were observed. Histological observations showed that plant regeneration took place, both via the pathway of somatic embryogenesis and organogenesis.Abbreviation 2,4-D 2,4-dichlorophenoxy acetic acid - MS Murashige and Skoog (1962) medium     

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     

Plant regeneration from in vitro cultures of cotyledon explants and anthers of Sinapis alba and its implications on breeding of crucifers

Summary Protocols for plant regeneration from cotyledon explant and anther cultures of Sinapis alba have been developed for creating doubled-haploids and somaclonal variation. Among the several cultivars tested in this study, only Arda responded well to in vitro plant regeneration both from anther-as well as cotyledoncultures. Multiple shoot formation in cotyledon explants, which always followed a brief callusing phase, was found to be the best on MS medium with ZEA (1.0mg/l) and NAA (0.1mg/l). Regeneration frequency declined sharply in the absence of auxin or presence of other cytokinins and/or auxin. The frequency of shoot regeneration also declined with reduction in the photoperiod to 16h. On MS + BAP (1.0mg/l) + NAA (1.0mg/l) medium, cotyledonary explants showed profuse callusing, which could regenerate shoots on high ZEA + low NAA/IAA medium. However, it declined with progressing time in culture. Anthers, excised from fresh as well as cold pretreated buds, cultured on 10% sucrose containing MS media with different hormonal constitution, developed calli and/or embryos. Initial culture temperature was important with embryogenesis occurring only in anthers cultured at 30°C for 3 weeks. A high temperature (35°C) treatment was lethal for both callus as well as embryo formation. While BAP + NAA and ZEA + NAA/IAA supported embryogenesis, further plant regeneration from anther-or embryo-callus could be achieved in ZEA + NAA/IAA media. Some of the regenerants flowered already in vitro and had small and sterile flowers. Cytological examination of some of the root differentiating calli indicated the presence of haploid as well as diploid cells. Shoots were rooted during prolonged incubation on the same medium or on transfer to MS (reduced)/ B5 + ZEA + NAA media.     

Transgenic fertile plants of soybean [Glycine max (L.) Merrill] obtained from bombarded embryogenic tissue

This work describes the production of transgenic, fertile plants of soybean [Glycine max (L.) Merrill]. The transformation method combines the advantages of somatic embryo genesis with the efficiency of particle bombardment of tissues that have a great capacity for in vitro proliferation and regeneration. The results described here represent the first report of transformation of soybean cultivars recommended for commercial growing in South Brazil using somatic embryogenesis, and may open the field for the improvement of this crop in this country by genetic engineering. This revised version was published online in August 2006 with corrections to the Cover Date.     

Interspecific hybrid of Helianthus annuus × H. simulans: Characterization and utilization in improvement of cultivated sunflower (H. annuus L.)

The first success at interspecific hybridization between cultivated sunflower(H. annuus) and a diploid perennial species, H. simulans is reported. The F₁s from both direct and reciprocal directions exhibited dominance of the wild species phenotype and were pollen sterile. Meiosis was irregular in the F₁ hybrids and both univalents and multivalents were observed. Multiplication of the interspecific hybrids was achieved through in vitro culture of nodal sections on Murashige & Skoog medium supplemented with 0.5 mg l^-1 benzyladenine. Fertility of the interspecific hybrids was improved by subjecting the in vitro proliferating shoots to 0.001% colchicine incorporated in the shoot multiplication medium. The amphiploids serve as fertile bridges and facilitate interspecific gene transfer through conventional breeding. This revised version was published online in July 2006 with corrections to the Cover Date.