Selective genotyping to identify late blight resistance genes in an accession of the tomato wild species <Emphasis Type="Italic">Solanum pimpinellifolium</Emphasis>

Late blight (LB), caused by the oomycete Phytophtohra infestans, is one of the most destructive diseases of tomato (Solanum lycopersicum) and other Solanaceae species. Current disease control and prevention strategies are not sufficient to control the disease in tomato. Recent germplasm screening experiments led to the identification of a new source of resistance (PI 270443) in the tomato wild species S. pimpinellifolium. This study was conducted to identify genomic regions associated with LB resistance in this accession. A large F₂ population (n = 986) derived from a cross between PI 270443 and a LB-susceptible tomato breeding line (NCEBR-2) was screened for LB resistance using a highly aggressive isolate of P. infestans. Twenty-five of the most resistant and 29 of the most susceptible, but surviving F₂ individuals were identified based on disease evaluations conducted in the F₂ and F₃ progeny populations. The selected individuals were genotyped with 153 DNA markers located across the 12 tomato chromosomes. A selective genotyping approach led to the identification of two genomic regions on tomato chromosomes 1 and 10 associated with LB resistance in PI 270443. Identification of two genomic regions associated with resistance was consistent with a previous estimate of the number of LB resistance genes in this accession. Research is currently underway to fine map the two resistance genes and incorporate them into new tomato breeding lines and hybrid cultivars.     

Hybridization between pascal celery and parsley

Summary Pascal celery cv. Golden Spartan was hybridized with parsley which is immune to celery blight (Septoria apiicola). Three F₁ hybrids were obtained from over a thousand seedlings grown from the pascal celery parent. The F₁ plants were intermediate for most characters. Segregation for petiole color in the F₂ generation demonstrated that a hybrid actually occurred. Segregation for late blight resistance occurred independently from petiole color.Received for publication 6 December 1979. Michigan Agricultural Experiment Station Journal Article No. 9222.Department of Botany and Plant Pathology, Michigan State University.     

Linkage and quantitative trait locus mapping of foliage late blight resistance in the wild species Solanum vernei

The global cultivation of potato (Solanum tuberosum) is threatened by epidemics caused by new variants of the late blight pathogen, Phytophthora infestans. New sources of durable late blight resistance are urgently needed and these may be found in wild Solanum species. The diploid wild species, S. vernei, has not previously been subjected to mapping of quantitative trait loci (QTLs) for late blight resistance. Two populations designated HGIHJS and HGG, originating from a cross between a clone of S. vernei and two different S. tuberosum clones were evaluated in field trials for late blight infestation. The relative area under the disease progress curve (RAUDPC) was estimated and used for QTL mapping. A linkage map of S. vernei, comprising 11 linkage groups, nine of which could be assigned to chromosomes, was constructed. Results indicated that the resistance in S. vernei was quantitatively inherited. Significant QTLs for late blight resistance were identified on chromosomes VIII (HGG), VI and IX (HGIHJS). In addition, potential QTLs were detected on chromosomes VII (HGIHJS) and IX (HGG). A putative and a significant QTL for tuber yield were found on chromosomes VI and VII in HGG, but no linkage between yield and resistance was indicated. The QTL for late blight resistance, which mapped to chromosome IX, could be useful for late blight resistance breeding as it was located close to the microsatellite marker STM1051 in both populations.     

Apium graveolens PI 181714 is a source of resistance to Fusarium oxysporum f. sp. apii race 4 in celery (A. graveolens var. dulce)

Celery has little genetic diversity and is highly susceptible to the new fungal pathogen Fusarium oxysporum f. sp. apii (Foa) race 4. After screening an Apium graveolens germplasm collection for resistance to Foa race 4, we crossed celery cv. 'Challenger', which is Foa race 2-resistant but Foa race 4-susceptible and A. graveolens PI 181714, which is Foa races 2- and 4-resistant but non-celery type. After selfing F₁s, we screened the F₁S₁ for race 4-resistance and celery-type and then selfed selected F₁S₁. Greenhouse and field trials indicate that three selected F₁S₂ families (76–8-4, 76–8-27 and 76–8-36) are suitable as germplasm for celery breeders for resistance to Foa race 4. A F1S3 76–8–36-124 is either fixed or nearly so for resistance to Foa races 4 and 2. Furthermore, quantitative PCR indicates that PI 181714 is resistant, rather than tolerant, to Foa races 4 and 2, and that this resistance has been introgressed into F1S3 76–8–36-124.     

Mapping of a novel,major late blight resistance locus in the diploid (1EBN) Mexican Solanum pinnatisectum Dunal on chromosome VII

Late blight caused by the oomycete Phytophthora infestans (Mont.) de Bary (Pi) is the most important foliar disease of potato worldwide. An intraspecific hybrid between individuals of a resistant and a susceptible S. pinnatisectum accession was backcrossed to the susceptible parent to generate a segregating population for late blight resistance consisting of 84 plants. In detached‐leaflet assays, reaction to late blight segregated in a 1r:1s manner in BC₁ progeny indicating the presence of a single dominant resistance gene. A genetic map was constructed based on 1,583 DArT/SSR markers which were allocated to 12 linkage groups, covering 1,793.5 cM with an average marker distance of 1.1 cM. The late blight resistance locus derived from S. pinnatisectum was mapped on chromosome VII. In comparison with the previously reported resistance genes Rpi1 and Rpi2, the new target resistance locus most likely is located on the opposite arm of chromosome VII. Results of this study will serve as a basis for future fine mapping of the late blight resistance locus and the development of locus‐specific markers for marker‐assisted selection.     

Identification of Brassica oleracea germplasm with improved resistance to Verticillium wilt

Two hundred and ninety‐nine accessions representing 11 cultivar groups of Brassica oleracea and eight additional accessions of the wild species B. cretica, B. incana, B. insularis and B. villosa were screened for resistance to Verticillium wilt. A disease index (DI) was calculated for each accession, and a correction of the DI was carried out to compensate for a fluctuating infection level between 11 independent trials. A total of 235 , or 77% of the accessions tested, had a DI_corr less or equal to the oilseed rape cv. ‘Express’ (DI_corr= 2.81), the reference cultivar. Only one accession of the wild species, B. incana, showed an enhanced level of resistance (DI_corr= 2.01). Twenty‐four accessions, distributed over eight cultivar groups of B. oleracea were selected for subsequent crosses involving B. rapa. Hybrid plants with 14 accessions were resultant and seed was obtained from crosses where the cultivar groups acephala, alboglabra, botrytis, capitata, gemmifera, italica and sabellica were used as female parents. When progeny of the produced resynthesized rapeseed lines were evaluated for Verticillium wilt resistance, three lines showed a significantly lower disease index (P ≥ 0.01) compared with the cv. ‘Express’. This source of resistance is now being crossed to advanced breeding material of oilseed rape.     

Genetics of resistance to early blight (Alternaria solani Sorauer) in tomato (Lycopersicon esculentum L.)

The genetic nature of early blight resistance in tomato was studied in three crosses at seedling and adult plant stages. A six generation mean analysis of the cross Arka Saurabh (susceptible) × IHR1939 (resistance) and its reciprocal cross revealed that the resistance to early blight was conferred by recessive polygenes at both seedling and adult plant stages. This polygenic early blight resistance revealed the importance of additive and additive × additive gene effects at seedling stage and higher magnitude of dominance and dominance× dominance gene effects at adult plant stage. Evaluation of parents, F₁, F₂ and backcross generations of IHR1816 (resistance) × IHR1939 (resistance) revealed that the early blight resistance genes in IHR1816 (Lycopersicon esculentum NCEBR-1) and IHR1939 (Lycopersicon pimpinellifolium L4394) are independent. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification and mapping of quantitative resistance to late blight (<Emphasis Type="Italic">Phytophthora infestans</Emphasis>) in <Emphasis Type="Italic">Solanum habrochaites</Emphasis> LA1777

Late blight (Phytophthora infestans) can have devastating effects on tomato production over the whole world. Most of the commercial cultivars of tomato, Solanum lycopersicum, are susceptible. Qualitative and quantitative resistance has been described in wild relatives of tomato. In general qualitative resistance can more easily be overcome by newly evolved isolates. Screening of three S. habrochaites accessions (LA1033, LA2099 and LA1777) through a whole plant assay showed that accession LA1777 had a good level of resistance to several isolates of P. infestans. To explore the potential in this wild species, an introgression line (IL) population of S. habrochaites LA1777 was used to screen individual chromosome regions of the wild species by a detached leaf assay. Two major isolates (T_1,2 and T_1,2,4) were used and two parameters were measured: lesion size (LS), and disease incidence (DI). Substantial variation was observed between the individual lines. QTLs were identified for LS but not for DI. The presence of five QTLs derived from LA1777 (Rlbq4a, Rlbq4b, Rlbq7, Rlbq8 and Rlbq12) results in unambiguous higher levels of resistance. All QTLs co-localized with previously described QTLs from S. habrochaites LA2099 except QTL Rlbq4b, which is therefore a novel QTL.     

Selection for <Emphasis Type="Italic">Phytophthora</Emphasis> field resistance in the F<Subscript>2</Subscript> generation of organic outdoor tomatoes

Tomatoes are the most important vegetable, globally as well as in Germany. Outdoor tomato production is seriously impaired due to increasing infections with evolving late blight (Phytophthora infestans) populations. Within organic agriculture, research is being conducted to develop regionally adapted and open pollinated cultivars of outdoor tomatoes with late blight field resistance. In the present experiment, three crosses, including wild, cocktail, and beefsteak tomatoes, were selected for field resistance against late blight in F₂ at one location per cross. The comparison of positive and negative selection in F₃ revealed the selection of single F₂ plants to be efficient in all three crosses. F₂ selection has proved to be a robust and efficient tool for breeding programs. The correlated response to selection in other traits, including yield, fruit weight, days to maturity, harvest period, and plant height, depended on the cross. It was evident that selection for desired traits combined with field resistance against late blight is promising, even in wide crosses. The most undesired attribute of wild tomatoes is the formation of shoots on leaves and in inflorescences. No correlation was observed between field resistance and shoot formation, allowing the selection of genotypes with improved field resistance and yield, but without morphological disadvantages.     

Pyramiding of genes conferring resistance to Tomato yellow leaf curl virus from different wild tomato species

Tomato (Solanum lycopersicum) production in tropical and subtropical regions of the world is limited by the endemic presence of Tomato yellow leaf curl virus (TYLCV). Breeding programmes aimed at producing TYLCV‐resistant tomato cultivars have utilized resistance sources derived from wild tomato species. So far, all reported breeding programmes have introgressed TYLCV resistance from a single wild tomato source. Here, we tested the hypothesis that pyramiding resistances from different wild tomato species might improve the degree of resistance of the domesticated tomato to TYLCV. We have crossed TYLCV‐resistant lines that originated from different wild tomato progenitors, Solanum chilense, Solanum peruvianum, Solanum pimpinellifolium, and Solanum habrochaites. The various parental resistant lines and the F₁ hybrids were inoculated in the greenhouse using viruliferous whiteflies. Control, non‐inoculated plants of the same lines and hybrids were exposed to non‐viruliferous whiteflies. Following inoculation, the plants were scored for disease symptom severity, and transplanted to the field. Resistance was assayed by comparing yield of inoculated plants to those of the control non‐inoculated plants of the same variety. Results showed that the F₁ hybrids between the resistant lines and the susceptible line suffered major yield reduction because of infection, but all hybrids were more resistant than the susceptible parent. All F₁ hybrids resulting from a cross between two resistant parents, showed a relatively high level of resistance, which in most cases was similar to that displayed by the more resistant parent. In some cases, the hybrids displayed better levels of resistance than both parents, but the differences were not statistically significant. The F₁ hybrid between a line with resistance from S. habrochaites and a line with resistance from S. peruvianum (HAB and 72‐PER), exhibited the lowest yield loss and the mildest level of symptoms. Although the resistance level of this F₁ hybrid was not statistically different from the level of resistance displayed by the 72‐PER parent itself, it was statistically better than the level of resistance displayed by the F₁ hybrids between 72‐PER and any other resistant or susceptible line.     

STS and microsatellite marker-assisted selection for bacterial blight resistance and waxy genes in rice, Oryza sativa L.

DNA marker-assisted selection was employed to select Xa-21 bacterial blight resistance and waxy (Wx) genes. Genotypes with both genes were selected from four F₂ populations involving indica × indica, indica × intermediate and japonica × indica crosses. With the assistance of PCR marker, 13 true breeding lines carrying Xa-21 were identified from F₂ generation of IRBB 21 × G 11353 cross. Similarly ten, eleven and fifty two plants having Xa-21 gene were isolated from G 3005-4-1 × IRBB 21, IRBB 21 × HJX 74 and IRBB 21 × SY 2crosses respectively. The lines with Wx gene in both homozygous and heterozygous state were also scored from the above crosses. Twenty plants having both Xa-21 and Wx loci in homozygous state were identified. DNA-based progeny testing was conducted to ensure the selection of homozygous lines for Xa-21 and Wx genes. Finally,twenty true breeding lines with high amylose content and Xa-21 gene were isolated from four crosses. These homozygous lines are phenotypically superior and resistant to Chinese race 5 of the bacterial blight pathogen. Fifty-six germplasm sources were surveyed for PCR polymorphism in order to facilitate future PCR-based marker assisted transfer of bacterial blight resistance genes xa-5, xa-13 and Xa-21 to any desired varietal background which will be useful for selection of parents in breeding programmes. This revised version was published online in August 2006 with corrections to the Cover Date.     

Late blight resistance in a diploid full-sib potato family

Late blight, caused by Phytophthora infestans (Mont.) de Bary, is the most destructive disease of potato worldwide. As this pathogen can rapidly overcome major race‐specific resistance genes, identifying the basis for enhanced quantitative resistance has become a crucial element for implementing advanced breeding strategies. A population of 230 full‐sib progeny derived from a cross between two diploid hybrid Solanum phureja × S. stenotomum clones was evaluated for foliage resistance against late blight in replicated trials at multiple locations in Pennsylvania between 1999 and 2002. In field experiments, plants were evaluated visually for per cent defoliation, and area under the disease progress curve (AUDPC) was determined. The two parents and three control cultivars (‘Atlantic’, ‘Kennebec’ and ‘Katahdin’) were included in all trials. In all three experiments, the presence of a significant number of clones exhibiting transgressive segregation were observed. There were significant differences among environments as well as among clones, and the clone × environment interaction was also significant. Stability analysis revealed that 37 clones made a significant contribution to the overall environment × clone interaction. Broad‐sense heritability for resistance, measured as AUDPC, was estimated as 0.67. The overall results indicate the presence in this potato family of a high level of field resistance against late blight. This segregating diploid family appears to be a good candidate for quantitative trait loci mapping to identify and characterize the genetic components of partial late blight resistance.     

Selection of wild Cicer accessions for the generation of mapping populations segregating for resistance to ascochyta blight

In order to determine genetically diverse parents for the generation of mapping populations segregating for resistance to ascochyta blight in wild Cicer species, the genetic diversity between a selection of resistant and susceptible accessions was assessed using molecular markers. Twenty Cicer accessions — comprising eight C. reticulatum accessions, six C. echinospermum accessions, five C. bijugum accessions, and one C. arietinum accession — were compared using a combination of seven RAPD primers and seven ISSR primers. A total of 231 polymorphic bands were scored and used to determine the genetic distances between accessions using Jaccard similarity coefficients. The most genetically diverse parents for the generation of intraspecific and interspecific populations segregating for resistance to ascochyta blight are reported. This revised version was published online in July 2006 with corrections to the Cover Date.     

Hybridization between Camelina sativa (L.) Crantz (false flax) and North American Camelina species

The potential for gene flow between Camelina sativa, a promising edible and industrial oil crop in Canada, and its wild North American relatives C. alyssum, C. microcarpa and C. rumelica subsp. rumelica, was investigated. Sequence analysis of the nuclear ribosomal internal transcribed spacer region (ITS) was used to differentiate Camelina species and/or accession‐specific nucleotide markers and identify interspecific F₁ hybrids. ITS analysis identified hybrids in progeny of C. alyssum × C. sativa, C. microcarpa accession 36010 × C. sativa and C. sativa × C. rumelica subsp. rumelica. Seed set on C. alyssum × C. sativa F₁ and F₂ progeny was similar to the parents; few seeds were produced on hybrid progeny of C. microcarpa accession 36010 × C. sativa and C. sativa × C. rumelica subsp. rumelica. The study provided evidence that should the species have sympatric distributions and overlapping flowering periods, gene flow between C. sativa and its wild North American relatives is possible and that it would most likely occur with C. alyssum given the high fertility of the F₁ hybrids recovered.     

Effects of glandular trichomes on the development of Phytophthora infestans infection in potato (S. tuberosum)

S. berthaulltii Hawkes, a wild potato species, possesses two types of glandular trichomes on its leaf surface: types A and B wich confer resistance to insects. Preliminary results indicated that the presence of glandular trichomes also confers resistance to Phytophthora infestans, the causal agent of late blight. In this paper we report the evaluation of trichome characteristics (density and exudate activities) performed on 8 lines of potato, including the control varieties Desirée and Tropicana, 5 hybrid lines derived from intercrosses and backcrosses between S. tuberosum and S. berthaultii and one accession of S. berthaultii. Whole plants of the 8 genotypes were inoculated with a spore suspension of Phytophthora infestans in order to evaluate their resistance to late blight. The resultant disease incidence correlated negatively with type A trichome density and with polyphenol-oxidase (PPO) activity of type A trichome glands. The removal of type B trichomes exudate by leaf dipping in ethanol did not influence the disease development after artificial inoculation of detached leaves. Type B trichome exudate was extracted and the fraction corresponding to Fatty Acids Sucrose Esters (FASE) was recovered after Thin Layer Chromatography and tested on the fungal mycelium in vitro. The FASE exhibited a slight inhibitory effect on mycelium growth. The results of the experiments are discussed, and several hypotheses regarding the possible role of types A and B trichomes are formulated. This revised version was published online in July 2006 with corrections to the Cover Date.     

Diallel analyses reveal the genetic control of resistance to ascochyta blight in diverse chickpea and wild Cicer species

Ascochyta blight is a major fungal disease affecting chickpea production worldwide. The genetics of ascochyta blight resistance was studied in five 5 × 5 half-diallel cross sets involving seven genotypes of chickpea (ICC 3996, Almaz, Lasseter, Kaniva, 24B-Isoline, IG 9337 and Kimberley Large), three accessions of Cicer reticulatum (ILWC 118, ILWC 139 and ILWC 184) and one accession of C. echinospermum (ILWC 181) under field conditions. Both F₁ and F₂ generations were used in the diallel analysis. The disease was rated in the field using a 1–9 scale. Almaz, ICC 3996 and ILWC 118 were the most resistant (rated 3–4) and all other genotypes were susceptible (rated 6–9) to ascochyta blight. Estimates of genetic parameters, following Hayman’s method, showed significant additive and dominant gene actions. The analysis also revealed the involvement of both major and minor genes. Susceptibility was dominant over resistance to ascochyta blight. The recessive alleles were concentrated in the two resistant chickpea parents ICC 3996 and Almaz, and one C. reticulatum genotype ILWC 118. The wild Cicer accessions may have different major or minor resistant genes compared to the cultivated chickpea. High narrow-sense heritability (ranging from 82% to 86% for F₁ generations, and 43% to 63% for F₂ generations) indicates that additive gene effects were more important than non-additive gene effects in the inheritance of the trait and greater genetic gain can be achieved in the breeding of resistant chickpea cultivars by using carefully selected parental genotypes.     

Genetics of Hordeum chilense– Current Status

Hordeum chilense, a South-American wild barley, is being used in cereal breeding because of its good crossability with both wild and cultivated Triticeae species. Amphiploids and chromosome addition lines of H. chilense in common and durum wheats have been obtained as a means to transfer desirable agronomic characters from wild barley to wheat. No allosyndetic pairing has been detected in diverse hybrids involving H. chilense. Several biochemical markers have been associated with the seven chilense chromosome pairs. New approaches are necessary to transfer genes from H. chilense to cereals.     

Reaction of tritordeum to Fusarium culmorum and Septoria nodorum

Summary Hordeum chilense is a wild barley extensively used in wide crosses in the Triticeae. It could be a valuable source of resistance to Fusarium culmorum and Septoria nodorum. Some H. chilense x Triticum spp. amphiploids, named tritordeums, were more resistant than the parental wheat line to these diseases, others were not. Average contents of ergosterol and deoxynivalenol (DON) suggested that resistance to colonization by Fusarium was the highest for Hordeum chilense, followed by tritordeum and wheat in decreasing order. In particular, the H. chilense genotypes H7 and H17 enhanced the wheat resistance to F. culmorum in its tritordeum offsprings. Resistance to S. nodorum in tritordeum was not associated with tall plant height. There is sufficient genetic variation for resistance to F. culmorum and S. nodorum among tritordeum to allow the breeding of lines combining short straw and resistance to both diseases.     

Heritability of resistance to foliar late blight in a diploid hybrid potato population of Solanum phureja×Solanum stenotomum

The emergence of new races of Phytophthora infestans has necessitated the search for additional sources of potato germplasm with resistance to late blight. This study examined 281 clones, derived from 72 families of a diploid random-mated hybrid population of Solanum phureja×Solanum stenotomum. The clones were evaluated in a replicated field trial for 2 years with the control cultivar ‘Atlantic’ in Pennsylvania, USA. The P. infestans US-8 A2 mating type culture was used to inoculate spreader rows of susceptible S. tuberosum cv.‘Russet Burbank’. Percent defoliation caused by the late blight fungus was estimated visually in each plot three times near the end of the growing season. Area under the disease progress curve (AUDPC) was estimated and showed that late blight was more severe in 1997 than in 1996. Genetic differences among clones and significant, but small, clone–environment interaction were detected for AUDPC. Broad-sense and narrow-sense heritability estimates, over years, were 0.79 ± 0.05 (P = 0.05) and 0.78 ± 0.29, respectively. Seventy-five percent of the diploid clones had a significantly lower mean AUDPC than Atlantic. These results support the idea that this diploid population is worthy of use in breeding for late blight resistance in tetraploid potato cultivars.     

Introgression of disease resistance genes from Arachis kempff-mercadoi into cultivated groundnut

Arachis kempff‐mercadoi is a wild species from the section Arachis. All kempff‐mercadoi accessions originate from the Santa Cruz province of Bolivia and they represent Arachis species with the A genome. From molecular analysis it was found that although cultivated A. hypogaea is made up of A and B genomes, A. kempff‐mercadoi may not be as closely related to it as are some of the other A genome species. Arachis kempff‐mercadoi is of interest because it has multiple disease resistance. It was crossed with a Spanish A. hypogaea cultivar which is susceptible to foliar diseases and to the insect pest Spodoptera litura. The success rate of the cross A. hypogaea (2n = 40) ×A. kempff‐mercadoi (2n = 20) was very low, but it could be increased by culturing immature seeds in vitro. Although the hybrids were triploids, a few fertile pollen grains were obtained due to the formation of restitution nuclei in the F₁ plants. Interspecific derivatives at the BC₂F₂ generation were scored for early leaf spot, late leaf spot and to Spodoptera damage. Screening results showed that 29% of the derivatives had both early and late leaf spot resistance and that less than 5% of the derivatives had resistance to both the foliar diseases and to Spodoptera.