Sympatric ascochyta complex of wild Cicer judaicum and domesticated chickpea

The aim of this study was to isolate, identify and characterize ascochyta blight pathogens from Cicer judaicum , a wild annual Cicer species which grows in Israel and other Mediterranean countries in sympatric distribution with legume crops, and determine their virulence and aggressiveness to other wild and domesticated legumes. Native C. judaicum plants exhibited symptoms resembling ascochyta diseases of grain legume crops. Two distinct pathogens were isolated and identified as Phoma pinodella and Didymella rabiei using morphological and molecular tools; their infectivity was verified using Koch's postulates. The virulence of these pathogens was examined on 13 legume species, of which P. pinodella was virulent to Pisum sativum , P. fulvum , C. judaicum , C. arietinum , C. reticulatum , C. pinnatifidum and C. bijugum . Didymella rabiei infected all these Cicer species, but not the other legume species tested. Aggressiveness of the pathogens was tested on wild and domesticated chickpea and pea. Didymella rabiei isolated from C. judaicum had significantly higher ( P  < 0·001) aggressiveness than P. pinodella from C. judaicum on both wild and domesticated chickpea. Disease severity on the former species ranged from 62·5% to 70% and on the latter from 41% to 56%. Phoma pinodella isolates from C. judaicum were more aggressive on C. arietinum and P. sativum than on C. judaicum and P. fulvum . Results of the current study suggest that C. judaicum may serve as an alternative host to ascochyta pathogens that endanger chickpea and possibly other crops and wild species growing in close proximity.     

Aggressiveness of eight <Emphasis Type="Italic">Didymella rabiei</Emphasis> isolates from domesticated and wild chickpea native to Turkey and Israel,a case study

Ascochyta blight, caused by Didymella rabiei, affects both domesticated chickpea and its congeneric wild relatives. The aim of this study was to compare the aggressiveness of D. rabiei isolates from wild and domesticated Cicer spp. in Turkey and Israel on wild and domesticated hosts from both countries. A total of eight isolates of D. rabiei sampled from C. pinnatifidum, C. judaicum and C. arietinum in Turkey and Israel was tested on two domesticated chickpea cultivars and two wild Cicer accessions from Turkey and Israel. Using cross-inoculation experiments, we compared pathogen aggressiveness across the different pathogen and host origin combinations. Two measures of aggressiveness were used, incubation period and relative area under the disease progress curve. The eight tested isolates infected all of the host plants, but were more aggressive on their original hosts with one exception; Turkish domesticated isolates were less aggressive on their domesticated host in comparison to the aggressiveness of Israeli domesticated isolates on Turkish domesticated chickpea. C. judaicum plants were highly resistant against all of the isolates from different origins except for their own isolates. Regardless of the country of origin, the wild isolates were highly aggressive on domesticated chickpea while the domesticated isolates were less aggressive on the wild hosts compared with the wild isolates. These results suggest that the aggressiveness pattern of D. rabiei on different hosts could have been shaped by adaptation to the distinct ecological niches of wild vs. domesticated chickpea.     

A comparative study of Turkish and Israeli populations of Didymella rabiei,the ascochyta blight pathogen of chickpea

The Fertile Crescent is the centre of domestication of chickpea (Cicer arietinum) and also the place of origin of its pathogens. Agrosystems provide different environments to natural eco‐systems, thus imposing different types of selection on pathogens. Here, the genetic structure and in vitro temperature growth response of the chickpea pathogen Didymella rabiei from domesticated chickpea (59 isolates from Turkey and 31 from Israel) and wild Cicer spp. (three isolates from Turkish C. pinnatifidum and 35 from Israeli C. judaicum) were studied. Six sequence‐tagged microsatellite site (STMS) primer pairs were used to determine the genetic structure of the 128 D. rabiei isolates. Turkish isolates exhibited the highest genetic diversity (H = 0·69). Turkish and Israeli D. rabiei from domesticated chickpea were genetically closer to each other than isolates from the wild Cicer spp. Analysis of molecular variance showed that 54% of the genetic variation resided between isolates from wild and domesticated origins. EF1‐α sequences distinguished between D. rabiei isolates from domesticated and wild Cicer spp. by four polymorphic sites. Nevertheless, a certain degree of mixing between isolates from wild and domesticated origin was demonstrated using the Bayesian algorithm as well as with principal coordinates analysis. Isolates sampled from domesticated chickpea from both countries were better adapted to temperatures typical of Levantine spring and had a significantly larger colony area at 25°C than at 15°C (typical Levantine winter temperature). These observations were in accordance to the heritability values of the temperature growth response.     

The sympatric Ascochyta pathosystems of Near Eastern legumes,a key for better understanding of pathogen biology

The primary and secondary centres of origin of domesticated plants are often also the places of origin of their pathogens. Therefore, the Near Eastern cradle of agriculture, where crop plants, their wild progenitors, and other con-generic taxa grow sympatrically, may hold some clues on the biology of the pathogens of the respective crops. Unlike the situation in the well-studied Near Eastern cereals and their important diseases, hardly any data are available on basic questions regarding grain legumes. What is the role of genetic diversity at resistance loci of the wild hosts and is it greater compared with the cultigens? Are populations of Ascochyta pathogens infecting wild legumes genetically distinct from populations infecting their domesticated counterparts, and if so, is this differentiation related to differences in host specialization or to adaptation to different ecological conditions? Do isolates originating from wild taxa exhibit a similar level of aggressiveness and have different aggressiveness alleles compared with those originating from domesticated grain legumes? In this review we propose an experimental framework aimed at gaining answers to some of the above questions. The proposed approach includes comparative epidemiology of wild vs. domesticated plant communities, co-evolutionary study of pathogens and their hosts, phenotypic and genetic characterization of fungal isolates from wild and domesticated origins, and genetic analyses of pathogenicity and parasitic fitness among progeny derived from crosses between isolates from wild and domesticated hosts.     

Structure and pathogenic variability in Ascochyta rabiei populations on chickpea in the Canadian prairies

A population study of Ascochyta rabiei from the Canadian prairies was conducted to assess pathogenicity among isolates with the objectives to investigate the existence of a race or pathotype structure and to evaluate whether there had been a shift to higher aggressiveness between 1998 and 2002. Ninety-nine isolates collected in 1998, 2001 and 2002 were inoculated onto seven differential chickpea genotypes. Significant isolate × differential interactions occurred, but accounted for a small proportion of the total variability. It was found that very few interaction effects between all combinations of differentials and isolates were significant and frequency distributions of disease severity of isolates tested on the differentials revealed continuous distributions. These results suggest that no genotype-specific relationship existed between A. rabiei and its host and that Canadian populations of the pathogen cannot be objectively classified into races or pathotypes. Isolates from 2001 and 2002 caused significantly more disease than isolates from 1998, suggesting that disease epidemics encountered since 1999 were in part caused by a shift in the population to higher aggressiveness.     

Characterization of chickpea differentials for pathogenicity assay of ascochyta blight and identification of chickpea accessions resistant to Didymella rabiei

Forty-eight chickpea germplasm lines, including 22 differentials used in previous studies, were characterized for disease phenotypes following inoculation with six isolates of Didymella (anamorph Ascochyta ) rabiei , representing a wide spectrum of pathogenic variation. Representative isolates were also directly compared with six previously identified races on eight chickpea genotypes. Many of the chickpea differentials reacted similarly to inoculation with each isolate of D. rabiei , and several previously identified races caused similar levels of disease on the differentials. This indicates that the number of differentials can be reduced significantly without sacrificing accuracy in describing pathogenic variation of D. rabiei on chickpea. Pathogenic variation among samples of US isolates allowed classification of the isolates into two pathotypes. The distribution of disease phenotypes of the 48 germplasm lines was bimodal after inoculation with pathotype I isolates, whereas the distribution of disease phenotypes was continuous after inoculation with pathotype II isolates. Such distinct distribution patterns suggest that chickpea plants employ different resistance mechanisms to each pathotype and that the two pathotypes may have different genetic mechanisms controlling pathogenicity. The advantages of using the two-pathotype system in assaying pathogenicity of the pathogen and in studying resistance mechanisms of the host are discussed. Three chickpea accessions, PI 559361, PI 559363 and W6 22589, showed a high level of resistance to both pathotypes, and can be employed as resistance sources in chickpea breeding programmes for resistance to ascochyta blight.     

Influence of Temperature and Inoculum Density of Fusarium oxysporum f. sp. ciceris on Suppression of Fusarium Wilt of Chickpea by Rhizosphere Bacteria

The effects of temperature and inoculum density of Fusarium oxysporum f. sp. ciceris race 5 on suppression of Fusarium wilt in chickpea (Cicer arietinum) cv. PV 61 by seed and soil treatments with rhizobacteria isolated from the chickpea rhizosphere were studied in a model system. Disease development over a range of temperatures (20, 25, and 30 degrees C) and inoculum densities (25 to 1,000 chlamydospores per gram of soil) was described by the Gompertz model. The Gompertz relative rate of disease progress and final amount of disease increased exponentially and monomolecularly, respectively, with increasing inoculum densities. Disease development was greater at 25 degrees C compared with 20 and 30 degrees C. At 20 and 30 degrees C, disease development was greater at 250 to 1,000 chlamydospores per gram of soil compared with 25 to 100 chlamydospores per gram of soil. At 25 degrees C, increasing inoculum densities of the pathogen did not influence disease. Nineteen Bacillus, Paenibacillus, Pseudomonas, and Stenotrophomonas spp. out of 23 bacterial isolates tested inhibited F. oxysporum f. sp. ciceris in vitro. Pseudomonas fluorescens RGAF 19 and RG 26, which did not inhibit the pathogen, showed the greatest Fusarium wilt suppression. Disease was suppressed only at 20 or 30 degrees C and at inoculum densities below 250 chlamydospores per gram of soil. Bacterial treatments increased the time to initial symptoms, reduced the Gompertz relative rate of disease progress, and reduced the overall amount of disease developed.     

Ascochyta blight of chickpea in Australia: identification, pathogenicity and mating type

The aetiology of blight of chickpea in South Australia was studied following sporadic disease outbreaks over several years that had been tentatively identified as Phoma blight. Nine fungal isolates from diseased chickpeas were tested for pathogenicity in the glasshouse, of which two caused symptoms resembling those of Ascochyta blight. The two aggressive isolates were identified as Ascochyta rabiei based on morphological characteristics of cultures and RAPD analysis. This was further confirmed by successful mating to international standard isolates, which showed that the two Australian isolates were MAT1-1. These isolates are accessioned as DAR 71767 and DAR 71768, New South Wales Agriculture, Australia. This is the first time that A. rabiei has been positively identified in commercial chickpeas in the southern hemisphere. The pathogen was found (in 1992) in only one of 59 seed samples harvested throughout Australia between 1992 and 1996 and tested using International Seed Testing Association methods. The teleomorph has not been found in Australia and results to date suggest that only one mating type is present. This suggests that quarantine restrictions on imported chickpea seed should be retained to prevent the introduction of the opposite mating type.     

Allelopathic potential of wild onion (Asphodelus tenuifolius) on the germination and seedling growth of chickpea (Cicer arietinum)

Water extracts obtained from the roots, shoots, and fruits of mature wild onion ( Asphodelus tenuifolius ) plants and soil taken from an A. tenuifolius field were used to determine their allelopathic effects on the germination and seedling growth of chickpea ( Cicer arietinum ) in the laboratory. The roots, shoots, and fruits of A. tenuifolius were soaked individually in water in a ratio of 1:20 (w/v) for 24 h to prepare the extracts. Distilled water was used as the control. The germinated seeds were taken out from the Petri dishes and counted every day for 12 days. The seeds of chickpea were also sown in sand and in each of the controlled, normal soil and the soil taken from the A. tenuifolius -infested field in Petri dishes to record the length and weight of the roots and shoots 18 days after sowing. The mean germination time reached the maximum amount for the stem and fruit extracts. The fruit extract caused the most reduction in the germination index and the germination percentage of chickpea. The different wild onion organ extracts significantly reduced the root and shoot length and biomass of the chickpea seedlings compared with the distilled water. The fruit extract of wild onion proved to be the most detrimental to the root length, shoot length, and dry weight of the chickpea seedlings. The soil beneath the A. tenuifolius plants significantly reduced the emergence, root length, shoot length, shoot dry weight, and seedling dry weight but increased the root dry weight of the chickpea seedlings. It is suggested that A. tenuifolius releases phytotoxic compound(s).     

Production of Didymella rabiei Pseudothecia and Dispersal of Ascospores in a Mediterranean Climate

ABSTRACT Temperature and wetness conditions required for development and maturation of Didymella rabiei pseudothecia were determined in a series of experiments conducted in controlled-environmental conditions. Initial stages of pseudothecium formation occurred at temperatures ranging from 5 to 15 degrees C. Incubation at low temperatures was essential for subsequent pseudothecium maturation. This requirement was satisfied for chickpea stem segments incubated at 5 or 10 degrees C for three consecutive weeks or during periods of 3 or 5 days, separated by periods at higher temperatures. Following the low-temperature requirement, subsequent pseudothecium development was independent of temperature in the range tested (5 to 20 degrees C). Wetness was essential for pseudothecium production: pseudothecia formed and matured on stem segments maintained continuously wet but also on those exposed to periods of three or five wet days, separated by dry periods. The dispersal of D. rabiei ascospores was studied using chickpea plants as living traps in the field. Trap plants were infected mainly when exposed during rain but also in rainless periods. Results of this study enabled us to describe the developmental events leading to the production of the teleomorph stage and the dispersal of ascospores by D. rabiei in the Mediterranean climate of Israel.     

Influence of temperature on plant–rhizobacteria interactions related to biocontrol potential for suppression of fusarium wilt of chickpea

Seed and soil treatment with Pseudomonas fluorescens RGAF 19, P. fluorescens RG 26, Bacillus megaterium RGAF 51 and Paenibacillus macerans RGAF 101 can suppress fusarium wilt of chickpea ( Cicer arietinum ), but the extent of disease suppression by these rhizobacteria is modulated by soil temperature. In this work, the effect of temperature on plant–rhizobacteria interactions was assessed in relation to biocontrol potential for suppression of fusarium wilt of chickpea. Seed and soil treatment with those rhizobacteria delayed seedling emergence compared with nontreated controls, and either increased or had no deleterious effect on chickpea growth. Pseudomonas fluorescens isolates significantly increased chickpea shoot dry weight at 20°C and root dry weight at 25 and 30°C. All bacterial isolates colonized the chickpea rhizosphere and internal stem tissues at 20, 25 and 30°C, and there was a positive linear trend between bacterial population size in the rhizosphere and temperature increase. The maximum inhibition of mycelial growth and conidial germination of Fusarium oxysporum f. sp. ciceris race 5 in vitro occurred at a temperature range optimal for bacterial growth and production of inhibitory metabolites. These results demonstrate the need to understand the effects of environmental factors on the biological activities of introduced rhizobacteria of significant importance for plant disease suppression.     

Host Specificity of Ascochyta spp. Infecting Legumes of the Viciae and Cicerae Tribes and Pathogenicity of an Interspecific Hybrid

ABSTRACT Ascochyta spp. (teleomorphs: Didymella spp.) infect a number of legumes, including many economically important species, and the diseases they cause represent serious limitations of legume production worldwide. Ascochyta rabiei, A. fabae, A. pisi, A. lentis, and A. viciae-villosae are pathogens of chickpea (Cicer arietinum), faba bean (Vicia faba), pea (Pisum sativum), lentil (Lens culinaris), and hairy vetch (V. villosa), respectively. Inoculations in the greenhouse and in growth chambers demonstrated that A. fabae, A. lentis, A. pisi, A. rabiei, and A. viciae-villosae were host specific. Isolates caused no visible disease symptoms on "nonhost" plants (plants other than the hosts they were originally isolated from) but were recovered consistently from inoculated, surface-disinfested, nonhost tissues. Interspecific crosses of A. pisi x A. fabae and A. viciae-villosae x A. lentis produced pseudothecia with viable ascospores, and the hybrid status of the ascospore progeny was verified by the segregation of mating type and amplified fragment length polymorphism (AFLP) markers. Interspecific progeny were morphologically normal in culture but exhibited more phenotypic variation compared with progeny from intraspecific crosses. Mating type and the majority of AFLP markers segregated in Mendelian 1:1 ratios in both intraspecific and interspecific crosses. A total of 11 and 7% of AFLP markers showed segregation distortion among progeny from interspecific crosses and intraspecific crosses, respectively; however, this difference was not significant (P = 0.90). Only 30 of 114 progeny isolates from the A. fabae x A. pisi cross inoculated in the greenhouse caused lesions on pea and only 4 caused disease on faba bean. In all, 15 of 110 progeny isolates were pathogenic to pea and none were pathogenic to faba bean under growth chamber conditions. Although no obvious postzygotic, intrinsic isolating barriers were identified in any of the interspecific crosses, it appears that host specialization may act as both a prezygotic, ecological isolating barrier and a postzygotic, extrinsic, ecological isolating barrier in these fungi. Host specificity, coupled with low pathogenic fitness of hybrids, may be an important speciation mechanism contributing to the maintenance of hostspecific, phylogenetic lineages of these fungi.     

Genetic diversity and host differentiation among isolates of Phytophthora infestans from cultivated potato and wild solanaceous hosts in Peru

To test the hypothesis that isolates of Phytophthora infestans attacking wild Solanaceae exhibit specialization for particular host species, 115 isolates of P. infestans were collected from cultivated potatoes, nontuber-bearing Solanum spp. of the Basarthrum section and wild tomatoes from five departments in the northern and central highlands of Peru, and characterized using several neutral markers. All isolates belonged to one of four clonal lineages described previously in Peru: EC-1, US-1, PE-3 and PE-7. There was a strong association of three lineages with host species: PE-3 was only isolated from cultivated potato, while PE-7 and US-1 were only isolated from nontuber-bearing Solanum spp. ( Basarthrum section and wild tomatoes). EC-1 was isolated from all host groups sampled. A subset ( n  = 74) of the isolates was evaluated for metalaxyl resistance. High levels of resistance were found almost exclusively in EC-1 and PE-3, while US-1 and PE-7 isolates were generally sensitive. In a detached-leaf assay for lesion diameter using five EC-1 isolates from S. caripense and seven EC-1 isolates from cultivated potato, there was a significant interaction between isolate origin and inoculated host, caused by higher aggressiveness of EC-1 from cultivated potato on its host of origin. In a comparison of EC-1 (seven isolates from cultivated potato) and US-1 (three isolates from S. caripense ), each pathogen lineage was more aggressive on its original host species, causing a highly significant interaction between isolate origin and inoculated host. Wild tomatoes and nontuber-bearing Solanum spp. harbour several pathogen lineages in Peru and could serve as reservoirs of inoculum that might contribute to epidemics on potato or tomato. Potential risks associated with the use of wild Solanum hosts as sources of resistance to P. infestans are discussed .     

The effect of temperature on hatching and penetration of chickpea roots by Pratylenchus thornei

Egg hatch of Pratylenchus thornei was influenced by temperature. It took place at all temperatures within the range 10–25°C and was optimal at 20°C. Root penetration increased steadily with increasing time of incubation up to the end of the experiment 11 days after inoculation. Temperature affected penetration rate in chickpea ( Cicer arietinum ) cultivars UC 27 and JG 62 but not in line P 2245, being significantly higher at 20–25°C than that at 15°C. At the end of the experiment, roots of line P 2245 held at 15°C contained more P. thornei than cultivars UC 27 and JG 62. No difference in percentage penetration among host genotypes was observed at 20 or 25°C. All migratory stages of P. thornei penetrated roots of chickpea from the first to 11th days after inoculation.     

Mating type, pathotype and RAPDs analysis inDidymella rabiei, the agent of ascochyta blight of chickpea

Eleven pathotype groups (A-K), including five not previously reported, ofDidymella rabiei (anamorphAscochyta rabiei), representing isolates of the pathogen from Ascochyta blight-affected chickpeas mainly from India, Pakistan, Spain and the USA, were characterized using 44 single-spore isolates tested against seven differential chickpea lines. Of 48 isolates tested for mating type, 58% belonged to MAT 1-1 and 42% to MAT 1-2. Thirty-nineD. rabiei isolates, as well as two isolates ofAscochyta pisi and six isolates of unrelated fungi, were analyzed using Randomly Amplified Polymorphic DNAs (RAPDs) employing five primers (P2 at 40°C, and OPA3, OPC1, OPC11 and OPC20 at 35°C). Computer cluster analysis (UPGMA / NTSYS-PC) detected a relatively low level of polymorphism among all theD. rabiei isolates, although atca 7% dissimilarity,ca 10 RAPD groups [I-X] were demarcated, as well as subclustering within the larger groups. By the same criteria, the maximum dissimilarity for the whole population ofD. rabiei isolates wasca 13%. No correlation was found between different RAPD groups, pathotype, or mating type ofD. rabiei, although some evidence of clustering based on geographic origin was detected. The use of RAPDs enabled us to identify specific DNA fragments that may have a potential use as genetic markers in sexual crosses, but none which could be used as virulence markers.     

Aggressiveness of Mycosphaerella graminicola Isolates from Susceptible and Partially Resistant Wheat Cultivars

ABSTRACT The selective effect of quantitative host resistance on pathogen aggressiveness is poorly understood. Because two previous experiments with a small number of bread wheat cultivars and isolates of Mycosphaerella graminicola had indicated that more susceptible hosts selected for more aggressive isolates, we conducted a larger experiment to test that hypothesis. In each of 2 years, six cultivars differing in their levels of partial resistance were planted in field plots, and isolates were collected from each cultivar early and late in the growing season. The isolates were inoculated as populations bulked by cultivar of origin, field replicate, and collection date on seedlings of the same six cultivars in the greenhouse. The selective impact of a cultivar on aggressiveness was measured as the difference in aggressiveness between early and late isolates from that cultivar. Regression of those differences on disease severity in the field yielded significance values of 0.0531 and 0.0037 for the 2 years, with moderately resistant cultivars selecting for more aggressive isolates. In a related experiment, the protectant fungicide chlorothalonil was applied to plots of two susceptible cultivars to retard epidemic development. When tested in the greenhouse, isolates of M. graminicola from those plots were significantly more aggressive than isolates from the same cultivars unprotected by fungicide.     

Aggressiveness of Isolates of Phytophthora infestans from the Columbia Basin of Washington and Oregon

ABSTRACT The aggressiveness of 22 isolates of Phytophthora infestans collected from naturally infected potato plants in the Columbia Basin of Washington and Oregon was determined on detached potato leaflets at 18 degrees C in an incubator. Selected isolates were evaluated on whole plants in a greenhouse. Aggressiveness was measured by using the area under the lesion expansion curve (AULEC), incubation period, latent period, sporulation capacity, and lesion size on detached leaflets and the area under the disease progress curve and sporulation capacity on whole plants. The detached-leaflet assay was useful in that a large number of isolates were tested, several components of aggressiveness were studied, and significant differences were found among isolates. Significant variation for components of aggressiveness was found within and among isolates classified according to genotype. Significant interactions among isolates and cultivars were found for some components of aggressiveness, so results were pooled according to cultivar. On average, US-8 and US-11 isolates had higher AULEC scores, indicating aggressiveness higher than that of US-7, US-6, and US-1 genotypes. One US-8 genotype isolate had a higher standardized sporulation capacity than isolates of the other genotypes. US-6 genotype isolates were the least aggressive group, as indicated by low AULEC, sporulation capacity, and lesion size values. The replacement of the US-1 genotype by the US-8 genotype in the Columbia Basin may be partially explained by the increased aggressiveness of US-8 isolates. Additionally, potato growers may need to shorten intervals between fungicide applications and begin applications earlier.     

Biometric Analyses of the Inheritance of Resistance to Didymella rabiei in Chickpea

ABSTRACT Historically, the response of chickpea (Cicer arietinum L.) to Didymella rabiei (causal agent of Ascochyta blight) has been mainly related to as complete resistance and it was commonly assayed with qualitative (nonparametric) scales. Two reciprocal populations, derived from intra-specific crosses between a moderately resistant late flowering Israeli cultivar and a highly susceptible early flowering Indian accession, were tested at F(3) and F(4) generations in 1998 and 1999, respectively. A quantitative (parametric) assessment (percent disease severity) was used to evaluate the chickpea field response to Ascochyta blight. The transformed relative area under the disease progress curve (tRAUDPC) was calculated for each experimental unit for further analyses. Heritability estimates of the tRAUDPC were relatively high (0.67 to 0.85) in both generations for both reciprocal populations. The frequency distributions of tRAUDPC of the populations were continuous and significantly departed from normality (Shapiro-Wilk W test; P of W < 0.0001), being all platykurtic and skewed toward either the resistant or the susceptible parental lines. The presence of major genes was examined by testing the relationship between the F(3) and F(4) family means and the within-family variances (Fain's test). Analyses of these relationships suggested that segregation of a single (or few) quantitative trait locus with major effect and possibly other minor loci was the predominant mode of inheritance. The correlation estimates between the resistance and days to flower (r = -0.19 to -0.44) were negative and significantly (P = 0.054 to 0.001) different from zero, which represents a breeding constraint in the development of early flowering cultivars with Ascochyta blight resistance.     

Population structure and mating system of Ascochyta rabiei in Tunisia: evidence for the recent introduction of mating type 2

The population structure of Ascochyta rabiei (teleomorph: Didymella rabiei ) in Tunisia was estimated among five populations sampled from the main chickpea growing regions using simple sequence repeat markers (SSR) and a mating type ( MAT ) marker. Mating type 2 isolates ( MAT1-2 ) had reduced genetic and genotypic diversity relative to mating type 1 isolates ( MAT1-1 ). This result, coupled with previous observations of lower overall frequency and restricted geographical distribution of MAT1-2 in Tunisia, and recent (2001) observation of the sexual stage, support the hypothesis of a recent introduction of MAT1-2 . Despite the presence of both mating types in Nabeul, Kef and Jendouba, the hypothesis of random mating was rejected in these locations with multilocus gametic disequilibrium tests. Highly significant genetic differentiation ( θ  = 0·32, G _ST = 0·28, P  < 0·001) was detected among populations and genetic distance and cluster analyses based on pooled allele frequencies revealed that populations from Nabeul and Kef were distinct from those in Beja, Bizerte and Jendouba. More than 70% of total gene diversity ( H _T = 0·55) detected was attributable to variation within populations compared to 28% among populations. This result, coupled with the occurrence of private alleles in each population, suggests that gene flow is currently limited among populations, even those separated by short geographic distances. The presence of two main genetic clusters was confirmed using Bayesian model-based population structure analyses of multilocus genotypes (MLGs) without regard to geographic origin of samples. The presence of MAT1-2 isolates in both clusters suggests at least two independent introductions of MAT1-2 into Tunisia that are likely to be the result of importation and planting of infected chickpea seeds.     

The threat of wild habitat to scab resistant apple cultivars

Evaluations of plant resistance to pathogens are rarely made using isolates from wild habitats, although the heterogeneity of such habitats may generate pathogen diversity which could be a source of new virulence in cultivated habitats. The aim of this study was to investigate whether scab resistance factors, identified and characterized in apples using isolates of Venturia inaequalis from a cultivated habitat, remained effective against isolates from a wild habitat. Three V. inaequalis core collections originating from the cultivated apple Malus × domestica and from two wild species, M. sieversii and M. sylvestris, were established to maximize pathogen diversity. For each core collection, 10 isolates were inoculated in mixtures onto 51 genotypes from an apple progeny segregating for two qualitative resistance genes and six quantitative resistance loci (QRL). On each apple genotype, isolates that contributed to the scab symptoms were identified within the mixture using microsatellite markers. The most frequently detected isolates were inoculated singly to compare their aggressiveness according to their host origin. The results showed that isolates from a wild habitat were able to infect the susceptible apple genotypes. However, these isolates were never more aggressive than isolates from the cultivated habitat on the resistance factors tested. It can therefore be concluded that the resistance factors used in this study, identified with V. inaequalis isolates from a cultivated habitat, remained effective against isolates from M. sylvestris and M. sieversii.