Genes from Lycopersicon chmielewskii affecting tomato quality during fruit ripening

Three chromosomal segments from the wild tomato, L. chmielewskii, introgressed into the L. esculentum genome have been previously mapped to the middle and terminal regions of chromosome 7 (7M, 7T respectively), and to the terminal region of chromosome 10 (10T). The present study was designed to investigate the physiological mechanisms controlled by the 7M and 7T segments on tomato soluble solids (SS) and pH, and their genetic regulation during fruit development. The effects of 7M and 7T were studied in 64 BC₂F₅ backcross inbred lines (BILs) developed from a cross between LA 1501 (an L. esculentum line containing the 7M and 7T fragments from L. chmielewskii), and VF145B-7879 (a processing cultivar). BILs were classified into four homozygous genotypes with respect to the introgressed segments based on RFLP analysis, and evaluated for fruit chemical characteristics at different harvest stages. Gene(s) in the 7M fragment reduce fruit water uptake during ripening increasing pH, sugars, and SS concentration. Gene(s) in the 7T fragment were found to be associated with higher mature green fruit starch concentration and red ripe fruit weight. Comparisons between tomatoes ripened on or off the vine suggest that the physiological mechanisms influenced by the L. chmielewskii alleles are dependent on the translocation of photosynthates and water during fruit ripening.     

Use of molecular markers in breeding for soluble solids content in tomato — a re-examination

Summbary Through earlier breeding efforts, portions of the genome of the wild species Lycopersicon chmielewskii have been introgressed into the cultivated tomato (Rick 1974). These introgressed chromosomal segments have been reported to increase soluble solids in fruit of certain tomato varieties (Rick 1974). Recently, two of the introgressed segments have been identified with RFLP markers and tested for effects on soluble solids in a single F2 population (Osborn et al. 1987). Based on results from that experiment, it was determined that one of the detected segments contains gene(s) controlling soluble solids and concluded that tomato varieties could be improved for this character by indirect selection for the linked RFLP marker (Osborn et al. 1987). In this report, we have independently tested the association between RFLP and isozyme markers and genes controlling soluble solids and other characters in the above described material. These experiments differ from the previous ones in that a set of 132 molecular markers (isozymes and DNA clones) of known chromosomal position have been used. Three introgressed chromosomal segments from L. chmielewskii have been identified using these markers. They map to the middle and the end of chromosome 7 (> 40 cM apart) and to the end of chromosome 10. The effects of these segments on soluble solids and other horticultural characters were tested in crosses with three different cultivars over a period of two years. Two of the three segments were found to increase soluble solids, however the effect of one of these was dependent on genetic background. Both segments were found to be associated with deleterious characters including increase in fruit pH, lower yield and small fruit. These results confirm the utility of molecular markers for detecting genes underlying quantitative variation but demonstrate the danger in establishing breeding programs around such linkages until the effects of the quantitative genes have been tested in a variety of genetic backgrounds and for associated effects on other characters of agronomic importance.     

High-resolution linkage analysis and physical characterization of the EIX-responding locus in tomato

An ethylene-inducing xylanase (EIX) from Tricohoderma viride is a potent elicitor of ethylene biosynthesis, localized cell death and other defense responses in specific cultivars of tobacco (Nicotiana tabacum) and tomato (Lycopersicon esculentum). Wild species of tomato, such as Lycopersicon cheesmanii and Lycopersicon pennellii, do not respond to EIX treatment. The F₁ progeny of a L. esculentum×L. cheesmanii and a L. esculentum×L. pennellii cross responded to EIX treatment with an increase in ethylene biosynthesis and the induction of localized cell death. The F₂ progeny of the above mentioned crosses segregated 3:1 (responding:non-responding). We mapped the EIX-responding locus (Eix) to the short arm of chromosome 7 using a population of introgression lines (ILs), containing small RFLP-defined chromosome segments of L. pennellii introgressed into L. esculentum. RFLP analysis of 990 F₂ plants that segregated for the introgressed segment mapped the Eix locus 0.1 cM and 0.9 cM from the flanking markers TG61 and TG131, respectively. Using the marker TG61 we isolated a yeast artificial chromosome (YAC) clone that carries 300-kb DNA segments derived from the Eix region. By mapping the ends of this YAC clone we show that it spans the Eix locus. Thus, positional cloning of the Eix locus appears feasible. Received: 20 March 1999 / Accepted: 30 April 1999     

Effects of the Lycopersicon chmielewskii sucrose accumulator gene (sucr) on fruit yield and quality parameters following introgression into tomato

A gene controlling fruit sucrose accumulation, sucr, was introgressed from the wild tomato species Lycopersicon chmielewskii into the genetic background of a hexose-accumulating cultivated tomato, L. esculentum. During introgression, the size of the L. chmielewskii chromosomal segment containing sucr was reduced by selection for recombination between RFLP markers for the sucr gene and flanking loci. The effects of sucr on soluble solids content, fruit size, yield and other fruit parameters were studied in the genetic background of the processing tomato cultivar Huntl00. In a segregating BC₅F₂ generation, the smallest introgression containing sucr-associated markers was necessary and sufficient to confer high-level sucrose accumulation, the effects of which were completely recessive. Fruit of sucr/sucr genotypes were smaller than those of +/sucr or +/+ genotypes at all stages of development. The timing of sugar accumulation and total sugar concentration were unaffected by sugar composition. No differences in total fruit biomass (fresh weight of red and green fruit) at harvest were observed between the genotypes, and sucrose accumulators produced greater numbers of fruit than hexose accumulators in one family. However, the proportion of ripe fruit at harvest, and hence yield of ripe fruit, as well as average ripe fruit weight and seed set were reduced in sucr/sucr genotypes. Sucrose accumulation was also associated with increased soluble solids content, consistency, serum viscosity, predicted paste yield and acidity, and decreased color rating. In the first backcross to L. chmielewskii, hexose accumulators (+/sucr) had larger fruit than sucrose accumulators (sucr/sucr), while no difference in soluble solids was detected.     

Molecular mapping of chromosome segments introgressed from Solanum lycopersicoides into cultivated tomato (Lycopersicon esculentum)

The wild nightshade Solanum lycopersicoides (accessionLA2951) was backcrossed to the cultivated tomato (Lycopersicon esculentum cv ’VF36’), then inbred through single-seed descent for several generations. Over 300 backcross-inbred families thereby derived were genotyped at 139 marker loci, consisting of RFLPs, allozymes, and monogenic morphological markers, to identify introgressed S. lycopersicoides chromosomes and segments thereof. The pattern of genotypes observed in the lines indicated a high degree of overall synteny between the S. lycopersicoides genome and that of tomato. Two putative single-copy RFLP probes revealed secondary loci in this wide cross. Recovery of the L. esculentum genome was more rapid than expected, with an average value in the BC₂ generation of 97.8%, versus the expected value of 87.5%. This was due to widespread segregation distortion that favored L. esculentum alleles as well as a tendency for plants homozygous for in- trogressed segments to be partially or completely male-sterile, thereby preventing the fixation of S. lycopersicoides markers in many lines. Despite these difficulties, nearly every S. lycopersicoides marker (or approximately 98% of the genome, measured in centi Morgans) was represented in at least 1 backcross-inbred line, with only a region on chromosome 4L missing from the population as a whole. Although the extent of transmission and fixation of introgressed segments varied according to chromosome, overall approximately 66% of the S. lycopersicoides genome was represented by homozygous in- trogressions with sufficient fertility to reproduce by self-pollination. An excess of terminal (vs. interstitial) segments was noted, and putative heterozygous substitutions for chromosomes 6, 7, 8, and 10 were found. Recombination within certain introgressed regions was reduced over 100-fold. These backcross-inbred lines are expected to facilitate the genetic analysis of traits identified in S. lycopersicoides and their transfer into horticultural tomatoes. Received: 16 March 1999 / Accepted: 22 June 1999     

Introgressions fromLycopersicon pennellii can improve the soluble-solids yield of tomato hybrids

RFLP-defined chromosome segments covering the entire tomato genome were introgressed from the wild green-fruited speciesLycopersicon pennellii into the cultivated tomato (L. esculentum cv M82; Eshed et al. 1992). SixL. pennellii chromosome segments were selected for a detailed evaluation based on previous observations of their effects on the two yield components, fresh tomato yield and total soluble-solids content (Brix). Differences in the quantitative traits measured between M82 and the introgression lines, or their hybrids with different inbred parents, can be attributed to the alien chromosome segments. Replicated field trials, grown at wide and dense spacing, identified three quantitative trait loci (QTLs) for solublesolids content on chromosomes 1, 5 and 7. In plants heterozygous for the chromosome-5 locus there was a 50% increase in soluble-solids yield in wide but not in dense spacing. Plants heterozygous for the chromosome-1 QTL/s were tested over a 2-year period, in three genetic backgrounds, and showed a significant 16% elevation in soluble-solids yield only in dense spacing. These results demonstrate that wild tomato germplasm can be used to improve the yield of the cultivated crop.     

Identification of random amplified polymorphic DNA (RAPD) markers linked to the v locus in barley, Hordeum vulgare L.

 Recombinant backcross lines of barley were produced from a cross between Kanto Nakate Gold (KNG; two-rowed) and Azumamugi (AZ; six-rowed) after backcrosses of F₁ plants with AZ as the recurrent parent. Each of these lines had an introgressed segment from chromosome 2 of KNG. Two recombinant backcross lines, L1 and M3-13, were used for an initial screening of polymorphism. After screening a total of 888 oligonucleotides as arbitrary primers, we identified eight random amplified polymorphic DNAs (RAPDs) between backcross lines and AZ. Among the RAPD fragments, CMNA-38₇₀₀ was linked to the v locus with a recombination frequency of zero, while OPJ-09₈₅₀ and OPP-02₇₀₀ were linked to the v locus at a map distance of 1.4 cM. Thus, the three RAPD markers were clustered around the v locus since the lengths of introgressed chromosomal segments in the L1 and M3-13 lines were no less than 38 cM. The other five RAPD fragments that we identified were not linked to the v locus. Received: 14 January 1997 / Accepted: 14 February 1997     

Sink Metabolism in Tomato Fruit : III. Analysis of Carbohydrate Assimilation in a Wild Species

Carbohydrate composition and key enzymes involved in carbohydrate metabolism were assayed throughout development of Lycopersicon esculentum and L. chmielewskii fruit. Translocation and assimilation of asymmetric sucrose and total soluble solids content was also determined in both species. The data showed that L. chmielewskii accumulated less starch than L. esculentum, and this was related to a lower level of ADPglucose pyrophosphorylase and a higher level of phosphorylase in L. chmielewskii. L. chmielewskii accumulated sucrose throughout fruit development rather than glucose and fructose which were accumulated by L. esculentum. A low level of invertase and nondetectable levels of sucrose synthase were associated with the high level of sucrose in L. chmielewskii. Translocation and assimilation of asymmetrically labeled sucrose indicated that sucrose accumulated in L. chmielewskii fruit was imported and stored directly in the fruit without intervening metabolism along the translocation path. In contrast, the relatively low level of radioactive sucrose found in L. esculentum fruit appeared to arise from hydrolysis and resynthesis of sucrose. The possible relationship between the level of soluble solids and differences in carbohydrate metabolism in sink tissue of the two species is discussed.     

Engineering of interstitial foreign chromosome segments containing the K+/Na+ selectivity gene Kna1 by sequential homoeologous recombination in durum wheat

Targeted homoeologous recombination mediated by the absence of the Ph1 locus is currently the most efficient technique by which foreign genes can be introgressed into polyploid wheat species. Because intra-arm homoeologous double cross-overs are rare, introgressed foreign genes are usually on terminal foreign chromosome segments. Since the minimum length of such a segment is determined by the position of a gene in the chromosome, large chromosome segments with undesirable genetic effects are often introgressed. Introgression of foreign genes on short interstitial segments based on two cycles of homoeologous recombination is described here. The utility of the technique is demonstrated by the introgression of the Kna1 locus, which controls K⁺/Na⁺ selectivity in T. aesivum L., on short interstitial segments of chromosome 4D into chromosome 4B of Triticum turgidum L. The level of recombination in a homoeologous segment is not significantly affected by a juxtaposed proximal homologous segment in the absence of the Ph1 locus.     

RFLP analysis of the size of chromosomal segments retained around the Tm-2 locus of tomato during backcross breeding

Summary Genes introduced into cultivated plants by backcross breeding programs are flanked by introgressed segments of DNA derived from the donor parent. This phenomenon is known as linkage drag and is frequently thought to affect traits other than the one originally targeted. The Tm-2 gene of Lycopersicon peruvianum, which confers resistance to tobacco mosaic virus, was introduced into several different tomato cultivars (L. esculentum) by repeated backcrossing. We have measured the sizes of the introgressed segments flanking the Tm-2 locus in several of these cultivars using a high density map of restriction fragment length polymorphic (RFLP) markers. The smallest introgressed segment is estimated to be 4 cM in length, while the longest is over 51 cM in length and contains the entire short arm of chromosome 9. Additionally, RFLP analysis was performed on remnant seed from different intermediate generations corresponding to two different backcross breeding programs for TMV resistance. The results reveal that plants containing desirable recombination near the resistance gene were rarely selected during backcrossing and, as a result, the backcross breeding method was largely ineffective in reducing the size of linked DNA around the resistance gene. We propose that, by monitoring recombination around genes of interest with linked RFLP markers, one can quickly and efficiently reduce the amount of linkage drag associated with introgression. Using such a procedure, it is estimated that an introgressed segment can be obtained in two generations that is as small as that which would otherwise require 100 backcross generations without RFLP selection.     

Evaluation of breeding utility of a chromosomal segment from Lycopersicon chmielewskii that enhances cultivated tomato soluble solids

A chromosomal segment from the wild tomato species, Lycopersicon chmielewskii, when backcrossed into the tomato cultivar VF145B-7879, increases fruit soluble solids concentration. In the investigation reported here, the near-isoline (VF145B-7M) homozygous for the chmielewskii (chm) chromosomal segment was crossed to the cultivar to study allelic interactions and then hybridized to three commercial cultivars (UC204c, E6203, and ChicoIII) to investigate the effect of this segment when heterozygous in different genetic backgrounds. Parents, isogenic hybrids, and unmodified hybrid controls were evaluated in three consecutive years of field study. The chm segment, when either homozygous or heterozygous, significantly increased soluble solids concentration by 13% and 12%, respectively, over VF145B-7879 (esc/esc), suggesting dominant gene action. Averaged across the three isohybrids and one isoline, one dose of this segment significantly increased soluble solids concentration by 6.0% compared to the unmodified hybrids and VF145B-7879. Other than increasing fruit total and soluble solids in mature-green and ripe-red tomatoes, no consistent negative effects of the chmielewskii segment were observed in the various genetic backgrounds on plant yield, fruit weight, or fruit pH. These results favor the use of this segment in breeding programs to develop fresh market and processing tomato cultivars with enhanced quality and reduced processed product dehydration costs. Received: 15 April 2000 / Accepted: 13 January 2001     

Development, utilization of introgression lines using a synthetic wheat as donor

A series of introgression lines (ILs) were generated from repeated backcrossing between the exotic hexaploid wheat genotype Am3 and the common wheat genotype Laizhou953. Am3 was synthesized by crossing Triticum carthlicum with Aegilops tauschii and was used as the donor parent in this study, and Laizhou953 was used as the recurrent parent. Two hundred and five SSR markers showing polymorphism between the two parents were used to identify the introgressed Am3 chromosome segments in 97 BC4F3 ILs. The introgressed segments in each line and the length of the introgressed segments were estimated according to the wheat SSR consensus map. The introgressed segments from Am3 in the 97 lines covers 37.7% of the donor genome. The introgressed segments were most found on 2D, 3B, 6B, and 1D with coverage of 59.8, 59.5, 59.1, and 59% of the chromosomes, respectively. None of the 97 lines tested contained chromosome 4D segments introgressed from Am3. Introgressed segments for each of the chromosomes were mapped using the consensus wheat linkage map. Nine agronomic traits from BC4F3 lines were evaluated and the phenotype showed most lines have the tendency to be more similar to the recurrent parent. There were lines showing better agronomic traits than the recurrent parent, which indicated the introgression of favorable alleles from the exotic hexaploid wheat into the elite cultivar Laizhou953. Marker and phenotype data were used to identify quantitative trait loci (QTLs) controlling these nine traits. In total, 38, 33, and 28 putative QTLs were detected for seven of the nine traits in 2003, 2004, and 2005, respectively. Some of these agronomic important QTLs were detected in more than one season. Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.     

Recombination around the Tm2a and Mi resistance genes in different crosses of Lycopersicon peruvianum

The amount of recombination in three different intraspecific crosses of the wild tomato species Lycopersicon peruvianum was investigated for the short arm of chromosome 6 that harbors the Mi nematode resistance gene and the centromeric region of chromosome 9 that contains the Tm2a virus resistance gene. These two genes have been introgressed into the cultivated tomato and are associated with a significant reduction in recombination in the respective region when crossed to other L. esculentum lines. For both regions and all crosses within L. peruvianum significantly more recombination (up to more than ten fold) was observed in the gametes derived from the female parent than in those from the male parent. In general, the differences were more pronounced for chromosome 6 than for chromosome 9. The amount of recombination in the three intraspecific L. peruvianum crosses was compared with the amount of recombination observed in the standard interspecific cross used for the construction of a saturated genetic map of tomato (L. esculentum x L. pennellii). In two of three cases for each region, more recombination was observed in the intraspecific crosses and in one case for each region significantly less recombination was found in the intraspecific cross when compared to the interspecific cross. Specifically for the Mi-carrying region, crosses within L. peruvianum exhibited up to 15-fold more recombination than crosses between resistant and susceptible L. esculentum lines, and such crosses will allow the fine mapping of this gene for the purpose of map-based cloning.     

The development of lettuce backcross inbred lines (BILs) for exploitation of the<Emphasis Type="Italic"> Lactuca saligna</Emphasis> (wild lettuce) germplasm

Backcross inbred lines (BILs) were developed in which chromosome segments of Lactuca saligna (wild lettuce) were introgressed into L. sativa (lettuce). These lines were developed by four to five backcrosses and one generation of selfing. The first three generations of backcrossing were random. Marker-assisted selection began in the BC₄ generation and continued until the final set of BILs was reached. A set of 28 lines was selected that together contained 96% of the L. saligna genome. Of these lines, 20 had a single homozygous introgression (BILs), four had two homozygous introgressions (doubleBILs) and four lines had a heterozygous single introgression (preBILs). Segregation ratios in backcross generations were compared to distorted segregation ratios in an F₂ population, and the results indicated that most of the distorted segregations can be explained by genetic effects on pollen- or egg-cell fitness. By means of BIL association mapping we were able to map 12 morphological traits and hundreds of additional amplified fragment length polymorphic (AFLP) markers. The total AFLP map now comprises 757 markers. This set of BILs is very useful for future genetic studies.Communicated by F. Salamini     

Introgression into tomato (Lycopersicon esculentum) of the L. chmielewskii sucrose accumulator gene (sucr) controlling fruit sugar composition

High sucrose concentration in fruit of Lycopersicon chmielewskii is governed by the recessive sucrose accumulator gene (sucr) that is situated in the pericentromeric region of chromosome 3. The sucr gene was introgressed into the genetic background of the hexose-accumulating cultivated tomato (L. esculentum cv Hunt 100) by marker-assisted selection using tightly linked RFLP markers and a tomato acid invertase cDNA as probes for sucr. RFLP mapping indicated that the segment containing sucr comprised over 43.2 cM in the BC₁F₂ generation, representing over one-third of the total length of chromosome 3. By selecting for crossovers between sucr and the flanking visual marker r (yellow fruit flesh) and RFLP marker TG288, we were able to reduce the size of the sucr introgression fragment to 0.8–7.1 cM by the BC₅ generation. Smaller recombinant fragments were not obtained despite screening a large BC₆F₂ population. The smallest sucr introgression reduced recombination between the flanking visual markers sy (sunny) and bls (baby lea syndrome) by 38%. To facilitate future introgression and recombination experiments, a PCR-based test for the sucr gene was developed using primers specific to the tomato invertase gene. This assay takes advantage of a small deletion that maps to the second intron of the L. chmielewskii nvertase gene. The assay detected significant allelic variation both within and between hexose- and sucrose-accumulating Lycopersicon spp.     

Inheritance and genetic mapping of fruit sucrose accumulation in Lycopersicon chmielewskii

Fruit of the domestic tomato (Lycopersicon esculentum Mill.) accumulate soluble sugars primarily in the form of the hexoses, glucose and fructose. In contrast, the predominant sugar in fruit of the wild tomato relative, L. chmielewskii, is sucrose. In the present study, the inheritance and linkage relations of sucrose accumulation were examined in interspecific L. esculentum x L. chmielewskii populations. In backcrosses to either the wild or domestic tomato, segregation for sucrose accumulation permitted qualitative analysis of the trait and indicated monogenic recessive control, although deviations from Mendelian inheritance were observed in some populations. This major gene, designated sucr, was mapped in F₂, F₃, and BC₁F₂ populations using a set of 95 informative RFLP and isozyme markers covering the tomato genome. A map location near the centromere of chromosome 3 was established, with tight linkage to the genomic clone TG102. Association of sucrose accumulation with yellow fruit, encoded by an allele of the r gene, permitted alignment with the classical map, thereby confirming the map location of sucr. A linkage map of the region surrounding sucr was obtained by monitoring recombination between flanking markers in the back-crosses to tomato. A cDNA clone of tomato fruit acid invertase, TIV1, was mapped to TG102 and sucr, with no recombination between the two RFLP markers observed in over 1700 meiotic products. Despite the tight linkage, TG102 and TIV1 hybridize to distinct restriction fragments, hence do not represent the same gene. The genetic data strongly suggest that sucr is an allele of the invertase gene and thus support previous biochemical studies that demonstrated low invertase activity in sucrose-accumulating fruit. L. hisutum, another low-invertase, sucrose-accumulating species, was hybridized with L. chmielewskii and the resulting F₁ plants accumulated sucrose, indicating that genetic control of soluble sugar composition is conserved in these two species.     

Inheritance of long staple fiber quality traits of Gossypium barbadense in G. hirsutum background using CSILs

Gossypium hirsutum is a high yield cotton species that exhibits only moderate performance in fiber qualities. A promising but challenging approach to improving its phenotypes is interspecific introgression, the transfer of valuable traits or genes from the germplasm of another species such as G. barbadense, an important cultivated extra long staple cotton species. One set of chromosome segment introgression lines (CSILs) was developed, where TM-1, the genetic standard in G. hirsutum, was used as the recipient parent and the long staple cotton G. barbadense Hai7124 was used as the donor parent by molecular marker-assisted selection (MAS) in BC₅S_1–4 and BC₄S_1–3 generations. After four rounds of MAS, the CSIL population was comprised of 174 lines containing 298 introgressed segments, of which 86 (49.4%) lines had single introgressed segments. The total introgressed segment length covered 2,948.7 cM with an average length of 16.7 cM and represented 83.3% of tetraploid cotton genome. The CSILs were highly varied in major fiber qualities. By integrated analysis of data collected in four environments, a total of 43 additive quantitative trait loci (QTL) and six epistatic QTL associated with fiber qualities were detected by QTL IciMapping 3.0 and multi-QTL joint analysis. Six stable QTL were detected in various environments. The CSILs developed and the analyses presented here will enhance the understanding of the genetics of fiber qualities in long staple G. barbadense and facilitate further molecular breeding to improve fiber quality in Upland cotton.     

An Introgression Line Population of Lycopersicon Pennellii in the Cultivated Tomato Enables the Identification and Fine Mapping of Yield-Associated Qtl

Methodologies for mapping of genes underlying quantitative traits have advanced considerably but have not been accompanied by a parallel development of new population structures. We present a novel population consisting of 50 introgression lines (ILs) originating from a cross between the green-fruited species Lycopersicon pennellii and the cultivated tomato (cv M82). Each of the lines contains a single homozygous restriction fragment length polymorphism-defined L. pennellii chromosome segment, and together the lines provide complete coverage of the genome and a set of lines nearly isogenic to M82. A field trial of the ILs and their hybrids revealed at least 23 quantitative trait loci (QTL) for total soluble solids content and 18 for fruit mass; these estimates are twice as high as previously reported estimates based on traditional mapping populations. For finer mapping of a QTL affecting fruit mass, the introgressed segment was recombined into smaller fragments that allowed the identification of three linked loci. At least 16 QTL for plant weight, 22 for percentage green fruit weight, 11 for total yield and 14 for total soluble solids yield were identified. Gene action for fruit and plant characteristics was mainly additive, while overdominance (or pseudo-overdominance) of wild species introgressions was detected for yield.     

Introgression and DNA marker analysis of Lycopersicon peruvianum, a wild relative of the cultivated tomato, into Lycopersicon esculentum, followed through three successive backcross generations

 Segregation of the Lycopersicon peruvianum genome was followed through three generations of backcrossing to the cultivated tomato L. esculentum cv ‘E6203’ using molecular markers. Thirteen BC₁ plants were genotyped with 113 markers, 67 BC₂ plants with 84 markers, and finally 241 BC₃ plants were genotyped with 177 markers covering the entire genome and a BC₃ map constructed. Several segments of the genome, including parts of chromosomes 3, 4, 6, and 10, quickly became fixed for esculentum alleles, possibly due to sterility problems encountered in the BC₁. Observed overall heterozygosity and chromosome segment lengths at each generation were very near the expected theoretical values. Markers located near the top telomeric region of chromosome 9 showed segregation highly skewed towards the wild allele through all generations, suggesting the presence of a gamete promoter gene. One markers, TG9, mapped to a new position on chromosome 9, implying an intrachromosomal translocation event. Despite the great genetic distance between the two parents, overall recombination was only 25% less than that observed in a previous tomato cross, indicating that L. peruvianum genes may be more readily introgressed into cultivated germplasm than originally believed. Received: 9 April 1997 / Accepted : 20 May 1997     

Mapping and introgression of a tomato yellow leaf curl virus tolerance gene,TY-1

The whitefly-transmitted tomato yellow-leaf curl gemini-virus (TYLCV) is a major pathogen of tomatoes. The wild tomato species Lycopersicon chilense, which is resistant to the virus, was crossed to the cultivated tomato, L. esculentum. The backcross-1 selfed (BC1S1) generation was inoculated and a symptomless plant was selected. This plant was analyzed using 61 molecular markers, which span the tomato genome, to determine which L. chilense chromosome segments were introgressed. A BC2S1 population was cage-inoculated with viroliferous whiteflies (Bemisia tabaci), the natural insect vector of the virus, and subjected to RFLP analysis. Markers on chromosomes 3 and 6 were significantly associated with the level of tolerance; the association of chromosome-6 markers was further substantiated in two additional BC2S1 populations. A tolerant BC2S1 plant which was homozygous for L. chilense introgressions in chromosomes 3, 6 and 7 was crossed to generate a BC3S1 population which was planted in an infested field. A TYLCV-tolerance gene with partial dominance, TY-1, was mapped to chromosome 6; two modifier genes were mapped to chromosomes 3 and 7. Field and whitefly-mediated cage inoculations of nearly-isogenic lines in BC3S3 supported our conclusion that TY-1 is the major TYLCV-tolerance locus.