RAPD markers associated with salt tolerance in an interspecific cross of tomato (Lycopersicon esculentum×L. pennellii)

This study was conducted to identify randomly amplified polymorphic DNA (RAPD) markers associated with quantitative trait loci (QTLs) conferring salt tolerance during germination in tomato. Germination response of an F₂ population (2000 individuals) of a cross between UCT5 (Lycopersicon esculentum, salt-sensitive) and LA716 (L. pennellii, salt-tolerant) was evaluated at a salt-stress level of 175 mM NaCl+17.5 mM CaCl₂ (water potential ca. –9.5 bars). Germination was scored visually as radicle protrusion at 6-h intervals for 30 consecutive days. Individuals at both extremes of the response distribution (i.e., salt-tolerants and salt-sensitives) were selected. The selected individuals were genotyped for 53 RAPD markers and allele frequencies at each marker locus were determined. The linkage association among the markers was determined using a “Mapmaker” program. Trait-based marker analysis (TBA) identified 13 RAPD markers at eight genomic regions that were associated with QTLs affecting salt tolerance during germination in tomato. Of these genomic regions, five included favorable QTL alleles from LA716, and three included favorable alleles from UCT5. The approximate effects of individual QTLs ranged from 0.46 to 0.82 phenotypic standard deviation. The results support our previous suggestion that salt tolerance during germination in tomato is polygenically controlled. The identification of favorable QTLs in both parents suggests the likelihood of recovering transgressive segregants in progeny derived from these genotypes. Results from this study are discussed in relation to using marker-assisted selection in breeding for salt tolerance. Received: 16 June 1997 / Revision received: 11 August 1997 / Accepted: 2 September 1997     

Mapping QTLs conferring early blight (Alternaria solani) resistance in a Lycopersicon esculentum×L. hirsutum cross by selective genotyping

Most commercial cultivars of tomato, Lycopersicon esculentum Mill., are susceptible to early blight (EB), a devastating fungal (Alternaria solani Sorauer) disease of tomato in the U.S. and elsewhere in the world. Currently, sanitation, long crop rotation, and routine application of fungicides are the most common disease control measures. Although no source of genetic resistance is known within the cultivated species of tomato, resistant resources have been identified within related wild species. The purpose of this study was to identify and validate quantitative trait loci (QTLs) conferring EB resistance in an accession (PI126445) of the tomato wild species L. hirsutum Humb. and Bonpl. by using a selective genotyping approach. A total of 820 BC₁ plants of a cross between an EB susceptible tomato breeding line (NC84173; maternal and recurrent parent) and PI126445 were grown in a greenhouse. During late seedling stage, plants were inoculated with mixed isolates of A. solani and subsequently evaluated for EB symptoms. The most resistant (75 plants = 9.1%) and most susceptible (80 = 9.8%) plants were selected and subsequently transplanted into a field where natural infestation of EB was severe. Plants were grown to maturity and evaluated for final disease severity. From among the 75 resistant plants, 46 (5.6% of the total) that exhibited the highest resistance, and from among the 80 susceptible plants, 30 (3.7% of the total) that exhibited the highest susceptibility, were selected. The 76 selected plants, representing the two extreme tails of the response distribution, were genotyped for 145 restriction fragment length polymorphism (RFLP) markers and 34 resistance gene analogs (RGAs). A genetic linkage map, spanning approximately 1298 cM of the 12 tomato chromosomes with an average marker distance of 7.3 cM, was constructed. A trait-based marker analysis (TBA), which measures differences in marker allele frequencies between extreme tails of a population, detected seven QTLs for EB resistance, one on each of chromosomes 3, 4, 5, 6, 8, 10 and 11. Of these, all but the QTL on chromosome 3 were contributed from the resistant wild parent, PI126445. The standardized effects of the QTLs ranged from 0.45 to 0.81 phenotypic standard deviations. Four of the seven QTLs were previously identified in a study where different populations and mapping strategy were used. The high level of correspondence between the two studies indicated the reliability of the detected QTLs and their potential use for marker-assisted breeding for EB resistance. The location of several RGAs coincided with locations of EB QTLs or known tomato resistance genes (R genes), suggesting that these RGAs could be associated with disease resistance. Furthermore, similar to that for many R gene families, several RGA loci were identified in clusters, suggesting their potential evolutionary relationship with R genes.     

An integrated high-density RFLP-AFLP map of tomato based on two Lycopersicon esculentum×L. pennellii F2 populations

 Two independent F₂ populations of Lycopersicon esculentum×L. pennellii which have previously been investigated in RFLP mapping studies were used for construction of a highly saturated integrated AFLP map. This map spanned 1482 cM and contained 67 RFLP markers, 1078 AFLP markers obtained with 22 EcoRI+MseI primer combinations and 97 AFLP markers obtained with five PstI+MseI primer combinations, 231 AFLP markers being common to both populations. The EcoRI+MseI AFLP markers were not evenly distributed over the chromosomes. Around the centromeric region, 848 EcoRI+ MseI AFLP markers were clustered and covered a genetic distance of 199 cM, corresponding to one EcoRI+ MseI AFLP marker per 0.23 cM; on the distal parts 1283 cM were covered by 230 EcoRI+MseI AFLP markers, corresponding to one marker per 5.6 cM. The PstI/MseI AFLP markers showed a more even distribution with 16 PstI/MseI AFLP markers covering a genetic distance of 199 cM around the centromeric regions and 81 PstI/MseI AFLP markers covering a genetic distance of 1283 cM on the more distal parts, corresponding to one marker per 12 and 16 cM respectively. In both populations a large number of loci showed a significant skewed segregation, but only chromosome 10 loci showed skewness that was similar for both populations. This ultra-dense molecular-marker map provides good perspectives for genetic and breeding purposes and map-based cloning. Received: 3 September 1998 / Accepted: 27 October 1998     

Course of meiosis in F1 interspecific hybrids of Lycopersicon esculentum Mill. × Licopersicon chilense Dun.

A study was conducted into the course of meiosis in F₁ interspecific hybrids of Lycopersicum esculentum Mill (mutant line Mo 638) × Lycopersicum chinense Dul. and its parental forms. An F₁ interspecific hybrid was obtained through the embryo culture technique. A decrease in the chiasma frequency and an increase in the frequency of univalents and meiotic abnormalities compared to their parental forms were detected in hybrid plants. The number of univalents and the percentage of main impairments decreased, as the height of bud tier locations increased. A conclusion was made regarding the connection between the regularity of meiosis in the examined F₁ interspecific hybrids of Lycopersicon esculentum × Lycopersicon chilense, on the one hand, and the hybrid nature of genotypes and the influence of environmental factors, on the other hand.     

Characterisation of a cytoplasmic male-sterile hybrid line between Lycopersicon peruvianum Mill. ×Lycopersicon pennellii Corr. and its crosses with cultivated tomato

 The cytoplasmic male-sterile (CMS) line CMS-pennellii (BC₁₀P₂ L. peruvianum×L. pennellii) and its complex hybrids with L. esculentum were studied. The established sterility was classified as the sporogenous type. As a result of the interaction of the genome of L. pennellii and the cytoplasm of L. peruvianum clear changes were established in the profiles of malic enzyme and esterase. Restriction fragment length polymorphism (RFLP) was detected between the mitochondrial (mt) genomes of CMS-pennellii and the cytoplasm donor, L. peruvianum, for two mtDNA probes: atpA and nad3. The established differences in the isozyme pattern and mt genomes are considered as useful markers to distinguish fertile and sterile plants. A breakthrough in the unilateral incompatibility of CMS-pennellii and the incorporation of the genome of L. esculentum on a CMS background is reported. The analysis of the complex hybrids assumes the interaction of two dominant genes – a maintainer gene from L. pennellii and a restorer gene from cultivated tomato. The hybrids produced with L. esculentum provide the basis for the development of a CMS system in cultivated tomato. Received: 25 May 1998 / Accepted: 26 August 1998     

Tomato chromosome 1: high resolution genetic and physical mapping of the short arm in an interspecific Lycopersicon esculentum×L. peruvianum cross

A detailed map of part of the short arm of chromosome 1 proximal to the Cf-4/Cf-9 gene cluster was generated by using an F₂ population of 314 plants obtained from the cross between the remotely related species Lycopersicon esculentum and L. peruvianum. Six markers that cosegregate in an L. esculentum×L. pennellii F₂ population showed high recombination frequencies in the present interspecific population, spanning an interval of approximately 13?cM. Physical distances between RFLP markers were estimated by pulsed field gel electrophoresis of high-molecular-weight DNA and by identifying YACs that recognized more than one RFLP marker. In this region 1?cM corresponded to 55–110?kb. In comparsion with the value of 730?kb per cM averaged over the entire genome, this reflects the remarkably high recombination frequencies in this region in the hybrid L. esculentum×L. peruvianum progeny population. The present data underline the fact that recombination is not a process that occurs randomly over the entire genome, but can vary dramatically in intensity between chromosomal regions and among populations.     

Induced parthenocarpy—a way of changing the levels of endogenous hormones in tomato fruits (Lycopersicon esculentum Mill.) 1. Extractable hormones

Ethylene, indol-3-acetic acid (IAA), gibberellin-like substances (GAs) and abscisic acid (ABA) were analysed in extracts from normal, seed-containing and parthenocarpic tomato fruits throughout fruit development. Parthenocarpic fruit growth was induced with an auxin (4-CPA), morphactin (CME) or gibberellic acid (GA₃) and compared with that of pollinated control fruits. Fruit growth was only affected by the treatment with GA₃, decreasing size and fresh weight by 60%. The peak sequence of hormones during fruit development was ethylene-GAs-IAA-ABA. Seeded fruits contained the highest levels of IAA and ABA but the lowest levels of GAs. Also, in seeded fruits, a high proportion of IAA and ABA was found in the seeds whereas this was not the case for GAs.Hormone levels of tomato fruits may be successfully, easily and reproducibly altered by inducing parthenocarpic fruit growth and thus eliminating development of seeds which are a major source of hormone synthesis. In spite of markedly changed hormone levels, there was no obvious relationship between fruit growth and extractable hormones per se. However, the results indicate that a high ratio of GAs: auxins is unfavourable for growth of tomato fruits.     

Mapping of QTLs for lycopene and other fruit traits in a Lycopersicon esculentum × L. pimpinellifolium cross and comparison of QTLs across tomato species

Quantitative trait loci (QTLs) for several fruit traits in tomato were mapped and characterized in a backcross population of an interspecific cross between Lycopersicon esculentum fresh-marker breeding line NC84173 and L. pimpinellifolium accession LA722. A molecular linkage map of this cross that was previously constructed based on 119 BC1 individuals and 151 RFLP markers was used for the QTL mapping. The parental lines and 119 BC1S1 families (self-pollinated progeny of BC1 individuals) were grown under field conditions at two locations, Rock Spring, PA, and Davis, CA, and fruits were scored for weight (FW), polar (PD) and equatorial diameters (ED), shape (FS), total soluble solids content (SSC), pH and lycopene content (LYC). For each trait, between 4 and 10 QTLs were identified with individual effects ranging between 4.4% and 32.9% and multilocus QTL effects ranging between 39% and 75% of the total phenotypic variation. Most QTL effects were predictable from the parental phenotypes, and several QTLs were identified that affected more than one trait. A few pairwise epistatic interactions were detected between QTL-linked and QTL-unlinked markers. Despite great differences between PA and CA growing conditions, the majority of FW QTLs (78%) and SSC QTLs (75%) in the two locations shared similar genomic positions. Almost all of the QTLs that were identified in the present study for FW and SSC were previously identified in six other studies that used different interspecific crosses of tomato; this indicates conservation of QTLs for fruit traits across tomato species. Altogether, the seven studies identified at least 28 QTLs for FW and 32 QTLs for SSC on the 12 tomato chromosomes. However, for each trait a few major QTLs were commonly identified in 4 or more studies; such ‘popular’ QTLs should be of considerable interest for breeding purposes as well as basic research towards cloning of QTLs. Notably, a majority of QTLs for increased SSC also contributed to decreased fruit size. Therefore, to significantly increase SSC of the cultivated tomato, some compromise in fruit size may be unavoidable. This revised version was published online in June 2006 with corrections to the Cover Date.     

Inheritance of downy mildew resistance, β-1,3-glucanases and peroxidases in pearl millet [Pennisetum glaucum (L.) R. Br.] crosses

The inheritance of resistance to downy mildew disease and the defense-related enzymes β-1,3-glucanase and peroxidase was studied in crosses of pearl millet using a generation-mean analysis. The study material comprised six generations (susceptible and resistant parents, F₁, F₂, BC₁ and BC₂) in three crosses. Seedlings from these generations were inoculated with the downy mildew pathogen Sclerospora graminicola and disease incidence was recorded. Analysis of constitutive levels of β-1,3-glucanase and peroxidase in the seedlings of different generations indicated that the resistant populations showed higher enzyme activities, while lower activities of the enzymes were recorded in the susceptible populations. In the generation-mean analysis, the significance of scaling tests revealed the existence of non-allelic interactions in the inheritance of resistance to downy mildew as well as with the enzymes. Among the gene effects, both additive and dominant effects were significant. All the non-allelic interaction effects were significant in the crosses. Studies on the isozyme patterns of the enzymes substantiated the results of the disease-incidence experiments in most of the generations. The results indicated that the inheritance of downy mildew disease resistance and the expression of β-1,3-glucanase and peroxidase in pearl millet is not only under the control of additive and dominant genes but are also governed by complex non-allelic interactions. Received: 30 April 2000 / Accepted: 17 October 2000     

Detection of QTLs associated with shoot wilting and root ammonium uptake under chilling temperatures in an interspecific backcross population from Lycopersicon esculentum ×L. hirsutum

The genetic basis for shoot wilting and root ammonium uptake under chilling temperatures was examined in an interspecific backcross (BC₁) population derived from Lycopersicon esculentum Mill. cv T5 and wild Lycopersicon hirsutum f. typicum accession LA1778. The chilling sensitivity of shoot wilting and ammonium uptake was evaluated in four replicated cuttings from each of 196 BC₁ plants. Wilting was evaluated at two different times: 2 hours (wilting 2 h) and 6 hours (wilting 6 h recovery) after root exposure to 4°C. The BC₁ plants were genotyped with 89 polymorphic RFLP markers, and composite interval mapping was used to detect quantitative trait loci (QTLs). Three QTLs, one each on chromosomes 5, 6 and 9, were detected for wilting 2 h. The presence of a L. hirsutum (H) allele at the QTL on chromosomes 5 and 9 decreased wilting, while the H allele at the QTL on chromosome 6 increased wilting. To analyze plant recovery from wilting at 6 h, subsets of the BC₁ population were selected, based on phenotype and genotype, because not all plants wilted at 2 h. The phenotype subset (wilting 6 h-PS) included plants that wilted to a greater degree at 2 h, and the genotype subsets included plants carrying specific allelic compositions at the QTL for wilting 2 h on chromosomes 5 (wilting 6 h-GS-ch5), 6 (wilting 6 h-GS-ch6), and 9 (wilting 6 h-GS-ch9). On chromosome 6, a QTL was located that was associated with three subsets (wilting 6 h-PS, wilting 6 h-GS-ch5 and wilting 6 h-GS-ch9), while on chromosome 7 a QTL was detected with two subsets (wilting 6 h-PS and wilting 6 h-GS-ch5). Three additional QTLs were detected within a single subset: chromosome 1 (wilting 6 h-GS-ch6), chromosome 11 (wilting 6 h-GS-ch5) and chromosome 12 (wilting 6 h-GS-ch9). The presence of the H allele at the QTL on chromosomes 7 and 12 had a positive effect, enhancing recovery from wilting, while the H allele at the other QTL had a negative effect. Three traits were used to evaluate the chilling sensitivity of root ammonium uptake: ammonium uptake before a chilling episode, ammonium uptake after the chilling episode, and the relative inhibition of uptake (difference in uptake rates before and after chilling divided by the rate before chilling). One QTL was detected on chromosome 3 for the rate before chilling and one on chromosome 6 for the relative inhibition of ammonium uptake. Our results demonstrate that shoot wilting and ammonium uptake under chilling are controlled by multiple QTLs. Received: 10 August 1999 / Accepted: 25 March 2000     

Changes in abscisic acid and its β-D-glucopyranosyl ester levels during tomato (Lycopersicon esculentum Mill.) seed development

Summary The role of abscisic acid (ABA) in tomato (Lycopersicon esculentum Mill.) zygotic embryogenesis was analysed. ABA and ABA ß-D-glucopyranosyl ester (ABA-GE) changes were determined in seeds and fruit tissues — placenta and mesocarp — during seed development, which was defined with eight embryo stages: from globular (stage 1) to mature embryo (stage 8). In whole seeds, ABA changes paralleled fresh and dry weight pattern curves and could be characterized by a high increase during embryo growth followed by a decrease as the seed matured and dehydrated. Moreover this dehydration phase led, at stage 8, to a new ABA distribution within the seed, preferentially into integument and embryo. Fruit tissue analyses provided new information about the ABA origin in seeds. ABA-GE levels were also measured and the results suggested different ABA metabolism in seed and fruit tissues.Abbreviations ABA abscisic acid - ABA-GE abscisic acid ß-D-glucopyranosyl ester - ABTS 2,2 — azino — bis (3 — ethylben-zthiazoline — 6 — sulfonic acid) - BHT butylhydroxytoluene - DW dry weight - ELISA Ezyme linked immunosorbent assay - HPLC high performance liquid chromatography     

RAPD and AFLP tagging and mapping of Beta (B) and Beta modifier (MoB), two genes which influence β-carotene accumulation in fruit of tomato (Lycopersicon esculentum Mill.)

The Beta (B) locus in tomato (Lycopersicon esculentum) increases fruit β-carotene content at the expense of lycopene, resulting in orange-pigmented fruit. Expression of B is influenced by the beta-modifier (Mo _B ) gene which segregates independently of B. RAPD and AFLP analyses were performed using near isogenic lines (NILs) unique for B and bulked segregant analysis (BSA) of a L. esculentum×L. cheesmanii-derived F₂ population segregating for B. Using 1018 random primers for RAPD analysis and 64 primer pairs for AFLP analysis, we identified polymorphic products which distinguished the NILs and the two bulked DNA samples constructed for BSA. A single 100 bp AFLP amplification product (E-ACA/M-CTG₁₀₀) which distinguished the NILs cosegregated with Mo _B and was demonstrated to be tightly linked to the locus. E-ACA/M-CTG₁₀₀ exhibited a recombination frequency of 1.7% in the F₂ progeny derived from an initial cross between the isolines. The Mo _B locus was mapped to the long arm of chromosome 6. Two RAPD products (OPAR18₁₁₀₀ and UBC792₈₃₀) of 1100 bp and 830 bp, respectively, were polymorphic between orange- and red-fruited bulks constructed from F₂ individuals in the L. esculentum and L. cheesmanii mating series. OPAR18₁₁₀₀ and UBC792₈₃₀ displayed recombination frequencies of 4.2% and 7.6%, respectively, in F₂ progeny. The B-linked OPAR18₁₁₀₀ marker was also mapped to the long arm of chromosome 6, proximal to Mo _B , and revealed linkage between B and Mo _B . Received: 9 April 1999 / Accepted: 27 April 1999     

Inheritance of Organelle DNA in Rye (Secale cereale L.) with Triticale (×Triticale Thch.) Hybrids

The mode of inheritance of chloroplast and mitochondrial DNA (mtDNA) in rye × triticale intergeneric hybrids has been studied with the use of specific PCR markers for loci 18S/5S and 3rbcL in organelle DNA. In rye × triticale BC₁, mtDNA copies of two types, paternal and maternal, have been found; in BC₂ plants, only paternal mtDNA and chloroplast DNA (cpDNA) have been detected. Mechanisms determining the inheritance and/or differential amplification of organelles of a specific type are discussed.     

Inheritance of resistance to the two-spotted spider mite from L. pimpinellifolium (Jusl.) Mill. accession ‘TO-937’

The two-spotted spider mite (Tetranychus urticae Koch) is an important pest of tomato (Lycopersicon esculentum Mill.) crops in temperate regions as this spider mite has a very large capacity for population increase and causes severe tomato yield losses. There is no described tomato cultivar fully resistant to this pest, although resistant accessions have been reported within the green-fruited tomato wild species L. pennellii (Corr.) D’Arcy and L. hirsutum Humb. & Bonpl. We observed a L. pimpinellifolium (Jusl.) Mill. accession, ‘TO-937’, which seemed to be completely resistant to mite attacks and we crossed it with the susceptible L. esculentum cultivar. ‘Moneymaker’ to obtain a family of generations consisting of the two parents, the F₁, the F₂, the BC₁ to L. esculentum, and the BC₁ to L. pimpinellifolium. This family was evaluated for mite resistance in a polyethylene greenhouse using an experimental design in 60 small complete blocks distributed along 12 double rows. Each block consisted of five F₂ plants in one row and one plant of each of the two parents, the F₁, the BC₁ to L. esculentum, and the BC₁ to L. pimpinellifolium in the adjacent row. Plants at the 10–15 leaf stage were artificially infested by putting on them two pieces of French bean leaf heavily infested with T. urticae. After two months, evaluations of infestation were made by visual observation of mite nets and leaf damage. Plants that were free of signs of mite reproduction on the top half were considered as resistant, plants with silky nets only on their basal leaves, intermediate, and plants with mite reproduction on both basal and top canopies were scored as susceptible. Dominance for resistance appeared because all the ‘To-937’, BC₁ to L. pimpinellifolium, and F₁ plants were resistant. Not all ‘Moneymaker’ plants behaved as susceptible because 35% of plants were intermediate. In the BC₁ to L. pimpinellifolium and the F₂, most plants were scored as resistant, only 7 % BC₁ and 3 % F₂ plants were intermediate, and a single F₂ plant (0.3 %) was susceptible. With these figures, resistance seemed to be controlled by either four or two genes according to whether segregation in the BC₁ or in the F₂, respectively, were considered. These results could in part be explained because of appearance of negative interplot interference due to the high frequency of resistant genotypes within most of the generations. Therefore, the family was evaluated again but using a different experimental design. In the new experiment, 16 ‘TO-937’, 17 ‘Moneymaker’, 17 F₁, 37 BC₁ to L. pimpinellifolium, 38 BC₁ to L. esculentum, and 125 F₂ plants were included. Each of these test plants was grown besides a susceptible ‘Moneymaker’ auxilliary plant that served to keep mite population high and homogeneous in the greenhouse. Negative interplot interference was avoided with this design and all the ‘TO-937’, F₁, and BC₁ to L. pimpinellifolium plants were resistant, all ‘Moneymaker’ test plants were susceptible, and 52 % BC₁ to L. esculentum and 25 % F₂ plants were susceptible, which fitted very well with the expected for resistance governed by a single dominant gene. The simple inheritance mode found will favour sucessful introgression of mite resistance into commercial tomatoes from the very close relative L. pimpinellifolium.     

An integrated interspecific AFLP map of lettuce (Lactuca) based on two L. sativa × L. saligna F2 populations

AFLP markers were obtained with 12 EcoRI/ MseI primer combinations on two independent F₂ populations of Lactuca sativa ×Lactuca saligna. The polymorphism rates of the AFLP products between the two different L. saligna lines was 39%, between the two different L. sativa cultivars 13% and between the L. sativa and L. saligna parents on average 81%. In both F₂ populations segregation distortion was found, but only Chromosome 5 showed skewness that was similar for both populations. Two independent genetic maps of the two F₂ populations were constructed that could be integrated due to the high similarity in marker order and map distances of 124 markers common to both populations. The integrated map consisted of 476 AFLP markers and 12 SSRs on nine linkage groups spanning 854 cM. The AFLP markers on the integrated map were randomly distributed with an average spacing between markers of 1.8 cM and a maximal distance of 16 cM. Furthermore, the AFLP markers did not show severe clustering. This AFLP map provides good opportunities for use in QTL mapping and marker-assisted selection. Received: 13 July 2000 / Accepted: 19 January 2001     

Does root-sourced ABA have a role in mediating growth and stomatal responses to soil compaction in tomato (Lycopersicon esculentum)?

Isogenic wild-type (Ailsa Craig) and abscisic acid (ABA)-deficient mutant (flacca) genotypes of tomato were used to examine the role of root-sourced ABA in mediating growth and stomatal responses to compaction. Plants were grown in uniform soil columns providing low to moderate bulk densities (1.1–1.5 g cm^?3), or in a split-pot system, which allowed the roots to divide between soils of the same or differing bulk density (1.1/1.5 g cm^?3). Root and shoot growth and leaf expansion were reduced when plants were grown in compacted soil (1.5 g cm^?3) but leaf water status was not altered. However, stomatal conductance was affected, suggesting that non-hydraulic signal(s) transported in the transpiration stream were responsible for the observed effects. Xylem sap and foliar ABA concentrations increased with bulk density for 10 and 15 days after emergence (DAE), respectively, but were thereafter poorly correlated with the observed growth responses. Growth was reduced to a similar extent in both genotypes in compacted soil (1.5 g cm^?3), suggesting that ABA is not centrally involved in mediating growth in this severely limiting ‘critical’ compaction stress treatment. Growth performance in the 1.1/1.5 g cm^?3 split-pot treatment of Ailsa Craig was intermediate between the uniform 1.1 and 1.5 g cm^?3 treatments, whereas stomatal conductance was comparable to the compacted 1.5 g cm^?3 treatment. In contrast, shoot dry weight and leaf area in the split-pot treatment of flacca were similar to the 1.5 g cm^?3 treatment, but stomatal conductance was comparable to uncompacted control plants. These results suggest a role for root-sourced ABA in regulating growth and stomatal conductance during ‘sub-critical’ compaction stress, when genotypic differences in response are apparent. The observed genotypic differences are comparable to those previously reported for barley, but occurred at a much lower bulk density, reflecting the greater sensitivity of tomato to compaction. By alleviating the severe growth reductions induced when the entire root system encounters compacted soil, the split-pot approach has important applications for studies of the role of root-sourced signals in compaction-sensitive species such as tomato.     

T-DNA-tagged chromosome 12 in donor Lycopersicon esculentum × L. pennellii is retained in asymmetric somatic hybrids with recipient Solanum lycopersicoides

Summary Asymmetric somatic hybrid plants were recovered after fusing irradiated mesophyll protoplasts of donor Lycopersicon esculentum × L. pennellii (EP) interspecific hybrid with callus-derived protoplasts of recipient Solanum lycopersicoides. EP plant A54 had been previously transformed by an agrobacterium vector, and the T-DNA insert mapped to the L. esculentum chromosome 12. The T-DNA insert conferred kanamycin resistance to EP that was subsequently used to select cell fusion products and recover asymmetric hybrid plants that retained tagged chromosome 12. Doses of 50- and 100-Gy irradiation promoted the elimination of only a few donor chromosomes. At 200 Gy, the regenerated plants had ploidy levels higher than tetraploid. However, the T-DNA tagged chromosome 12 was always retained in the asymmetric hybrid plants tested. Likewise, all plants from the 100-Gy series, with the exception of number 160, were mixoploid in the root-tip cells. Such mixoploid asymmetric somatic hybrids could be stabilized by inducing adventitious shoots on leaf strips cultured on shoot regeneration medium containing kanamycin. The asymmetric hybrid plants did not produce viable seed when self-pollinated or backcrossed to tomato or S. lycopersicoides. Present address: Department of Biology, University College of London, Gower Street, London, UK     

Inheritance and genetic mapping of two nuclear genes involved in nuclear–cytoplasmic incompatibility in peas (Pisum sativum L.)

Genetic analysis was performed to finely map and assess the mode of inheritance of two unlinked nuclear genes Scs1 and Scs2 involved in incompatibility of the nuclear genome of the cultivated pea Pisum sativum subsp. sativum with the cytoplasm of the wild pea of the subspecies P. sativum subsp. elatius, accession VIR320. Based on the segregation of genotypes in the progeny of the test-crosses, we concluded that if the cytoplasm was inherited from the wild pea VIR320, the Scs1 allele from the cultivated pea was gametophyte lethal and sporophyte recessive lethal. The Scs2 allele from the cultivated pea reduced male gametophyte viability. In homozygote, Scs2 from cultivated parent brought about nuclear–cytoplasmic conflict manifested as chlorophyll deficiency, reduction of blade organs, and low pollen fertility of about 20%. In heterozygote, Scs1 and Scs2 genes reduced pollen fertility by ca 50 and 30%, respectively. The Scs1 and Scs2 genes involved in nuclear–cytoplasmic incompatibility were genetically mapped. The distance between the markers bordering Scs1 comprised about 2.5 cM on linkage group III. The map distance between the bordering markers in the neighborhood of Scs2 varied substantially from cross to cross in the range of 2.0–15.1 cM on linkage group V.     

Field response of tomato (Lycopersicon esculentum Mill ‘Pusa Ruby’) to inoculation with a VA mycorrhizal fungusGlomus fasiculatum and withAzotobacter vinelandii

Summary The response of tomato (Lycopersicon esculentum Mill) to inoculation with the vasicular arbuscular mycorrhizal (VAM) fungusGlomus fasiculatum andAzotobacter vinelandii singly and in combination was tested in the field. It was found thatG. fasiculatum as well asA. vinelandii significantly increased leaf area, shoot dry weight, nitrogen content phosphorus content and yield in respect to uninoculated control. While, VAM fungal treatment alone could bring about substantial increase in growth, nitrogen content, phosphorus content and yield, its combination withA. vinelandii produced additional effects on leaf area, shoot dry weight, phosphorus content and yield. Contribution No. 304/83 of Indian Institute of Horticultural Research, Bangalore-89.     

A linkage map of chickpea (Cicer arietinum L.) based on populations from Kabuli × Desi crosses: location of genes for resistance to fusarium wilt race 0

Two recombinant inbred line (RIL) populations derived from intraspecific crosses with a common parental line (JG62) were employed to develop a chickpea genetic map. Molecular markers, flower colour, double podding, seed coat thickness and resistance to fusarium wilt race 0 (FOC-0) were included in the study. Joint segregation analysis involved a total of 160 markers and 159 RILs. Ten linkage groups (LGs) were obtained that included morphological markers and 134 molecular markers (3 ISSRs, 13 STMSs and 118 RAPDs). Flower colour (B/b) and seed coat thickness (Tt/tt) appeared to be linked to STMS (GAA47). The single-/double-podding locus was located on LG9 jointly with two RAPD markers and STMS TA80. LG3 included a gene for resistance to FOC-0 (Foc0₁/foc0₁) flanked by RAPD marker OPJ20₆₀₀ and STMS marker TR59. The association of this LG with FOC-0 resistance was confirmed by QTL analysis in the CA2139 × JG62 RIL population where two genes were involved in the resistance reaction. The STMS markers enabled comparison of LGs with preceding maps.