Model plants are facilitating the genetic characterization and comparative mapping of a number of traditional crops. Medicago truncatula has been widely accepted as a model plant to this end as it provides the essential tools for multiple aspects of legume genetics and genomics. A large set of markers from highly conserved M. truncatula gene regions is being created and used to establish a worldwide framework for comparative genomic studies in legumes. We have investigated the potential for cross-species amplification of 209 expressed sequence tag (EST)-based and 33 bacterial artificial chromosome (BAC)-based microsatellites from M. truncatula in the three most important European legume pulses-pea, faba bean and chickpea-that might facilitate future comparative mapping. Our results revealed significant transferability of M. truncatula microsatellites to the three pulses (40% in faba bean, 36.3% in chickpea and 37.6% in pea). The percentage of M. truncatula EST-SSRs (simple sequence repeats) amplified in the three crops (39-43%) was twofold higher than that of the genomic SSRs (21-24%). Sequence analysis determined that the level of conservation in the microsatellite motif was very low, while the flanking regions were generally well conserved. The variations in the sequences were mainly due to changes in the number of repeat motifs in the microsatellite region combined with indel and base substitutions. None of the functional microsatellites showed direct polymorphism among the parental genotypes tested, consequently preventing their immediate use for mapping purposes.

Fondevilla, S., Carver, T. L. W., Moreno, M. T., Rubiales, D. (2006). Macroscopic and histological characterisation of genes er1 and er2 for powdery mildew resistance in pea. European Journal of Plant Pathology, 115, (3), 309-321 Keywords: grain legumes - haustorium - histology - hypersensitive resistanceIn pea, two single recessive genes, er1 and er2, have been identified for resistance to powdery mildew caused by Erysiphe pisi, but little is known about their mode of action. Pea accessions carrying the genes er1 or er2 and other accessions displaying resistance to powdery mildew in the field were studied. In accessions carrying gene er1, epidermal cell penetration was prevented and very few haustoria or colonies were formed. Under controlled conditions, er1 conferred complete or almost complete resistance to the fungal isolates used and this resistance was not associated with macroscopically visible necrosis. Under field conditions these accessions developed a low level of disease. Resistance in line JI2480 (carrying er2) increased with temperature and leaf age, and complete resistance was expressed only at high temperature (25 ?C) or in mature leaves. This resistance was based mainly on post-penetration cell death, complemented by a reduction of percentage penetration success in mature leaves. Combining the resistance provided by gene er1 and by line JI2480 into new cultivars is likely to increase their level of resistance and enhance durability of the protection.Peer reviewe

Over 4000 species of angiosperms are able to directly invade and parasitise other plants, but only very few are weedy and parasitise cultivated plants. Together with the witchweeds (Striga spp.) and dodders (Cuscuta spp.), the broomrapes (Orobanche spp.) affect important crops causing complete yield loses with severe infestations. Genetic resistance to parasitisation remains as one of the most desirable components in an integrated control strategy. However, breeding for resistance is a difficult task and many aspects of the host/parasite interaction remain unknown. In the present work, we review the cytological and cytochemical studies investigating the mechanisms of resistance involved in the process leading to an incompatible host-parasite interaction. We have attempted to identify the main gaps and problems related to such studies with parasitic plants and present an in-depth review of histochemical techniques used to assess mechanisms of resistance against these parasitic plants. Furthermore, we discuss future research directions and novel techniques and finally, how such techniques can be applied and incorporated within a breeding programme.

The antinutritional factors (ANFs) present in Vicia spp. seeds are a major constraint to the wider utilization of these crops as grain legumes. In the case of faba bean (Vicia faba L.), a breeding priority is the absence vicine and convicine (v-c); responsible for favism in humans and for the reduced animal performance or low egg production in laying hens. The discovery of a spontaneous mutant allele named vc-, which induces a 10-20 fold reduction of v-c contents, may facilitate the process. However, the high cost and difficulty of the chemical detection of v-c seriously restricts the advances in breeding-selection. To identify random amplified polymorphic DNA (RAPD) markers linked to this gene, we have analysed an F(2 )population derived from a cross between a line with high v-c content (Vf6) and the vc- genotype (line 1268). Quantification of v-c was done by spectrophotometry on the parents and the F(2 )population (n = 136). By using bulked segregant analysis (BSA), two RAPD markers linked in coupling and repulsion phase to the allele vc- were identified and further converted into sequence characterized amplified regions (SCARs). Amplification of SCARS was more consistent, although the initial polymorphism between pools was lost. To recover the polymorphisms several approaches were explored. Restriction digestion with HhaI (for SCAR SCH01(620)) and RsaI (for SCAR SCAB12(850)) revealed clear differences between the parental lines. The simultaneous use of the two cleavage amplified polymorphism (CAP) markers will allow the correct fingerprinting of faba bean plants and can be efficiently used in breeding selection to track the introgression of the vc- allele to develop cultivars with low v-c content and improved nutritional value.

Crenate broomrape (Orobanche crenata) is a root parasitic weed that represents a major constraint for grain legume production in Mediterranean and West Asian countries. Medicago truncatula has emerged as an important model plant species for structural and functional genomics. The close phylogenic relationship of M. truncatula with crop legumes increases its value as a resource for understanding resistance against Orobanche spp. Different cytological methods were used to study the mechanisms of resistance against crenate broomrape of two accessions of M. truncatula, showing early and late acting resistance. In the early resistance accession (SA27774) we found that the parasite died before a tubercle had formed. In the late resistance accession (SA4327) the parasite became attached without apparent problems to the host roots but most of the established tubercles turned dark and died before emergence. The results suggest that there are defensive mechanisms acting in both accessions but with a time gap that is crucial for a higher success avoiding parasite infection.Crenate broomrape (Orobanche crenata) is one of the most important parasitic plants attacking legume crops in Mediterranean area, devastating crops and making unusable infested land (Rubiales, 2001(Rubiales, , 2003Rubiales et al., 2002). Being a broomrape (Orobanche sp.), crenate broomrape is an obligate root holoparasite lacking in chlorophyll and depending entirely on the host for its supply of nutrients (Joel et al., 2007). The knowledge of the mechanisms of resistance against the parasite is crucial to develop strategies of control, like breeding for resistance. With this purpose, we have chosen Medicago truncatula as a crenate broomrape host model plant due to its characteristics.M. truncatula is an annual forage legume in the Mediterranean area. Contrary to other legume crops, M. truncatula is an autogamous self-fertile plant with a small and diploid genome, a short life cycle, and a prolific seed production (Blondon et al., 1994). Its simple genetics, the development of new tools and methods for molecular and genetic analysis, and the complete genome sequence (http://www.medicago.org) provide researchers with a valuable data set and making it interesting as a legume model species for laboratory studies (Cook et al., 1997;Cook, 1999) and also in pathogenic interactions (Ellwood et al., 2007; Pérez-de-Luque et al., 2007a).Nowadays, the most numerous and important works about parasitic plants were focused on the development in susceptible host, as Orobanche spp. (Joel and Losner-Goshen, 1994;Neumann et al., 1999), Striga spp. (Dörr, 1997;Reiss and Bailey, 1998), Cuscuta spp. (Vaughn, 2002(Vaughn, , 2003, Viscum spp. (Heide-Jørgensen, 1987), and others (Heide-Jørgensen and Kuijt, 1993, 1995). But little is known about the basis of host resistance to these parasites, just finding the work of Joel et al. (1996) introducing this subject. In the last years only some histological studies of the resistant interactions have been undertaken (Dörr et al., 199...

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Bulk segregant analysis was used to identify random amplified polymorphic DNA (RAPD) markers linked to a gene determining hypersensitive resistance in Vicia faba line 2N52 against race 1 of the rust fungus Uromyces viciae-fabae. The monogenic nature of the resistance was determined by analyzing the F(2) population from a cross between resistant line 2N52 and susceptible line VF-176, and further confirmed in the F(2:3)-derived families. Linkage of the RAPD markers was confirmed by screening 55 F(2) plants segregating for resistance. Three RAPD markers (OPD13(736), OPL18(1032) and OPI20(900)) were mapped in coupling phase to the resistance gene for race 1 ( Uvf-1). No recombinants between OPI20(900) and Uvf-1 were detected. Two additional markers (OPP02(1172) and OPR07(930)) were linked to the gene in repulsion phase at a distance of 9.9 and 11.5 cM, respectively. The application of marker-assisted selection to develop new faba bean varieties with rust resistance genes is discussed.

Broomrapes (Orobanche spp.) are parasitic angiosperms, which attach to the roots of the hosts to take water and nutrients from them. No complete control measures are available to date, but breeding for resistance remains as one of the most feasible and environmentally friendly methods. However, the mechanisms governing the interaction between these parasites and the host are not yet well understood. We studied the cellular changes associated with the resistance to Orobanche crenata in faba bean as mechanisms involved or responsible for resistance. Two cultivars of faba bean, resistant and susceptible to O. crenata infection, were used. The evolution of the infection and the changes in the cell and tissue organisation and wall components of the host cells were followed and evaluated in both genotypes. Samples of compatible and incompatible interactions were fixed and sectioned, and specific cytochemical methods for different cell components were applied, results being analysed under light and epifluorescence microscopy. A higher proportion of O. crenata seedlings unable to penetrate the root was found on the resistant genotype. Reinforcement of cell walls by callose deposition hampers parasite penetration through the cortex. Lignification of endodermal cells prevents further penetration of the parasite into the central cylinder.