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.     

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.     

Resistance to root-lesion nematode Pratylenchus neglectus identified in a new collection of two wild chickpea species (Cicer reticulatum and C. echinospermum) from Turkey

Chickpea (Cicer arietinum) is a major legume crop, with Australia being the second largest producer worldwide. Pratylenchus neglectus is a root-lesion nematode that invades, feeds and reproduces in roots of pulse and cereal crops. In Australia, chickpea and wheat (Triticum aestivum) are commonly grown in rotation and annual damage by P. neglectus accounts for large economic losses to both crops. Cultivated chickpea has narrow genetic diversity that limits the potential for improvement in resistance breeding. New collections of wild chickpea species, C. reticulatum and C. echinospermum, have substantially increased the previously limited world collection of wild Cicer germplasm and offer potential to widen the genetic diversity of cultivated chickpea through the identification of accessions with good resistance. This research assessed 243 C. reticulatum and 86 C. echinospermum accessions for response to P. neglectus in replicated experiments under controlled glasshouse conditions from 2013 and 2014 collection missions that were received, tested and analysed in two experimental sets. Multi-experiment analyses showed lower P. neglectus population densities in both sets of wild Cicer accessions tested than Australia's elite breeding cultivar PBA HatTrick at the significance level p < 0.05. Provisional resistance ratings were given to all genotypes tested in both experimental sets, with C. reticulatum accessions CudiB_008B and Kayat_066 rated as resistant in both Set 1 and Set 2. New sources of resistance to P. neglectus observed in this study can be introgressed into commercial chickpea cultivars to improve their resistance to this nematode.     

Different ecological affinities and aggressiveness patterns among Didymella rabiei isolates from sympatric domesticated chickpea and wild Cicer judaicum

Domesticated chickpea (Cicer arietinum) and its wild relative C. judaicum grow in sympatric distribution in Israel and both are susceptible to Ascochyta blight caused by Didymella rabiei. C. arietinum was grown for millennia in drier and hotter Levantine spring conditions while C. judaicum grows in the wetter and milder winters. Accordingly, it is possible that D. rabiei isolates originated from C. arietinum are adjusted to the less favorable spring conditions. Here, 60 isolates from both origins were tested in vitro for their hyphal growth at 15 and 25 degrees C. Isolates from C. arietinum had a significantly larger colony area at 25 degrees C than at 15 degrees C (P < 0.001) while no such differences were detected between isolates from C. judaicum. D. rabiei isolates from wild and domesticated origins were used to inoculate nine C. judaicum accessions and two domesticated chickpea cultivars and their aggressiveness patterns were determined using five measures. On domesticated chickpea, isolates from domesticated origin were significantly more aggressive in four out of the five aggressiveness measures than isolates from wild origin. On C. judaicum, isolates from wild origin were generally more aggressive than isolates from domesticated origin. The results suggest that the habitat segregation between wild and domesticated Cicer influences the pathogens ecological affinities and their aggressiveness patterns.     

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).     

Sources of resistance to ascochyta blight in wild Cicer species

Ascochyta blight [Ascochyta rabiei (Pass.) Lab.] is the major foliar disease of chickpea (Cicer arietinum L.). In search of better sources of resistance to ascochyta blight, 201 accessions of 8 annual wildCicer species were evaluated in field and greenhouse for 3 years (1988 to 1991) at Tel Hadya, Syria. One accession each ofC. judaicum Boiss (ILWC 165) andC. pinnatifidum Jaub. & Spach. (ILWC 159) were consistently rated resistant in both field and greenhouse evaluations. Another three accessions ofC. judaicum (ILWC 61, ILWC 154, ILWC 199) and six accessions ofC. pinnatifidum (ILWC 78, ILWC 88, ILWC 155, ILWC 160, ILWC 162, ILWC 203) were resistant or moderately resistant. The blight-resistant accessions ofC. judaicum originated from Jordan, Lebanon, Syria, and Turkey; and those ofC. pinnatifidum from Syria and Turkey. None of the accessions ofC. bijugum, C. chorassanicum, C. cuneatum, C. echinospermum, C. reticulatum andC. yamashitae were resistant to blight.     

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.     

Life history traits and trade-offs between two species of the ascochyta blight disease complex of pea

Plant disease complexes are a playground to investigate coinfections in natural or cultivated systems. Pathogens of such complexes may affect each other through direct and/or indirect interactions and lead to changes in virulence or aggressiveness, offspring production, and transmission. As coinfections by sympatric host-pathogens can strongly influence pathogen dynamics and their evolutionary trajectories, new insights into the mechanisms of their coexistence is thus of critical importance. In order to characterize differences in ecological niches liable to explain species coexistence on the same host, the inter- and intraspecific diversity of the life history traits in natural collections of the two main pathogens (Peyronellaea (formerly Didymella) pinodes [Dp] and Phoma medicaginis var. pinodella [Pmp]) of the ascochyta blight disease complex of pea was evaluated under controlled conditions. Dp strains developed 1.3 times faster and produced longer, mainly bicellular spores and in lower amounts (3.7 times less) than Pmp strains. Pmp strains were separated into two groups, one producing more pycnidiospores, mainly bicellular, with less resources, and the other with mainly unicellular ones. These three groups can be interpreted as three distinct life history strategies: pioneer colonizer (Dp), scavenger (large-spored Pmp), and intermediate (small-spored Pmp), allowing differentiation in access to and use of resources. While this experimental work provides new insight into coexistence of two species of the ascochyta blight disease complex of pea, it also raises the question of the benefit of having two distinct life history strategies for one species (Pmp).     

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 chickpea genotype and Bacillus sp. on protection from Fusarium wilt by seed treatment with nonpathogenic Fusarium oxysporum

Seeds of kabuli chickpea cultivars ICCV 4 and PV 61 were treated with conidia of nonpathogenic Fusarium oxysporum isolate Fo 90105 suspended in methylcellulose (3 × 10⁶ conidia.seed^-1), or with methylcellulose alone, and sown in soil artificially infested with 500 or 1,000 chlamydospores.g^-1 of F. oxysporum f. sp. ciceris race 5. At an inoculum concentration of 500 chlamydospores.g^-1, seed treatment with Fo 90105 significantly increased the incubation period of the disease by 11 (ICCV 4) or 25 (PV 61) days, and reduced the final disease incidence, disease intensity and the standardized area under the curve of disease intensity over time. This protection from disease was higher and more consistent in PV 61 than in ICCV 4. However, it was annulled with an inoculum concentration of 1,000 chlamydospores.g^-1, except for the incubation period in PV 61 which was increased by 10 days. When ICCV 4 seeds were treated with Fo 90105 (3 × 10⁶ conidia.seed^-1) and/or Bacillus sp. isolate RGAF 51 (1 × 10⁷ cfu.seed^-1), then sown in infested soil, there was no influence by the Bacillus isolate on protection conferred by Fo 90105. However, the degree of protection by the nonpathogenic F. oxysporum was higher and more consistent when plants from treated seeds were grown in sterile sand for 6 days, then transplanted into infested soil.     

Cucumber mosaic virus infection of chickpea stands: temporal and spatial patterns of spread and yield‐limiting potential

Patterns of spread and yield losses were examined when migrant aphid vectors transmitted Cucumber mosaic virus (CMV) within chickpea (Cicer arietinum) plots. When numbers of chickpea plant infection foci were varied to provide initial infection incidences of 0·3–2%, rate of virus spread and its final incidence increased in proportion to initial virus incidence and pathogen progress curves reflected a polycyclic pattern of spread. Chickpea seed yields decreased by 44–45% when CMV incidence reached 61–74% at final assessment in plots with simulated 1–2% initial incidence. In chickpea plots with or without introduced lupin plant infection foci, cumulative spatial data for diseased plants were assessed using spatial analysis by distance indices (sadie ). When CMV spread within a plot, infection was concentrated in large patch clusters and was consistent with comprehensive localized spread around primary infection foci. When introduced infection foci were absent, there was more diffuse spread with many isolated clusters. In individual plants that developed CMV symptoms at different growth stages, shoot and pod dry weight were reduced by 60–65% and 77–79% (early infection) and 44 and 66% (late infection), respectively. Seed yield losses were 78–80% (early) and 65–67% (late), and reduction in 50‐seed weight was 20–25% regardless of time of infection, so seed number was reduced more by early than late infection. Infection also reduced seed quality as, in addition to smaller seed size, seed coats were discoloured and malformed: the proportions of malformed seeds were 9–11% (early), 3–6% (late) and 0·5% (healthy).     

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.     

Orobanche crenata resistance and avoidance in pea (Pisum spp.) operate at different developmental stages of the parasite

Orobanche crenata (broomrape) is an important constraint to pea (Pisum sativum) cultivation in the Mediterranean area, because little resistance is available in commercial crop varieties. Field experiments have demonstrated that some resistance is present in a number of P. sativum and P. fulvum accessions. The goal of this work was to characterize such resistance. The Pisum–O. crenata interaction and the resistance symptoms were studied under controlled conditions by using Petri dish and polyethylene bag assays. The content of phenolics and peroxidase activity in host tissue from infected and non-infected plants were also measured. Resistance and avoidance mechanisms, acting at different developmental stages of the parasite, have been identified, including low stimulation of O. crenata seed germination, unsuccessful penetration of host roots, delay in post-attachment tubercle development and necrosis of the attached tubercles. Infection caused an increase in the content of total soluble phenolics in some Pisum genotypes. Peroxidase activity was higher in resistant than in susceptible accessions. Results obtained with different Pisum genotypes showed that resistance is the result of several mechanisms acting at different stages of the infection process. Resistance is also related to increased levels of peroxidase activity in host roots.     

Development of ascochyta blight (Ascochyta rabiei) in chickpea as affected by host resistance and plant age

Ascochyta blight caused by Ascochyta rabiei, is the most destructive disease in many chickpea growing countries. Disease development varies with the growth stage and host resistance. Hence, disease development was studied in cvs ICCX 810800 (resistant), ICCV 90201 (moderately resistant), C 235 (moderately susceptible), ICCV 96029 and Pb 7 (susceptible) under controlled environment (ICRISAT, Patencheru) and field conditions (Dhaulakuan, Himachal Pradesh) at seedling, post-seedling, vegetative, flowering and podding stages. Under controlled environment, the incubation period and terminal disease reaction (TDR) did not vary significantly at different growth stages against virulent isolate AB 4. Cultivars ICCX 810800, ICCV 90201 and C 235 showed a significantly longer incubation period than the susceptible cv. Pb 7. Cultivar ICCX 810800 showed slow disease progress and the least TDR. Field experiments were conducted during the 2003–2004 and 2004–2005 growing seasons. During 2003–2004, TDR was higher in plants inoculated at podding and the flowering stage and the lowest disease reaction was recorded in ICCX 810800. A severe epidemic during 2004–2005 was attributed to the favourable temperature, humidity and well distributed high rainfall. TDR did not differ significantly at any of the growth stages in susceptible cvs ICCV 96029 and Pb 7. With respect to seeding date and cultivar, the highest yield was recorded in the early-sown crop (1,276.7 kg ha⁻¹) and in ICCV 90201 (1,799.3 kg ha⁻¹), respectively. The yields were greatly reduced in all the cultivars during 2004–2005 and the highest yield was recorded in ICCX 810800 (524.7 kg ha⁻¹). Integrated disease management using resistant cultivars, optimum sowing period and foliar application of fungicides will improve chickpea production. The experiment under controlled environment and field conditions (during the epidemic year) showed a similar disease development.     

Characterization of Ascochyta isolates and susceptibility of pea cultivars to the ascochyta disease complex in Alberta

The relative virulence of 109 Ascochyta isolates collected from pea fields in Alberta from 1996 to 1998 were evaluated on 10-day-old seedlings by the excised leaf-assay technique. Twenty-eight isolates were avirulent, while the others produced lesions of various sizes on pea leaves. DNA samples from 86 isolates were amplified by the RAPD technique using PCR with single primers. One dominant genotype of Ascochyta pisi was identified throughout Alberta, but variations in virulence were not clearly differentiated by the RAPD technique. Five Ascochyta isolates, four virulent and one avirulent, were used to assess the susceptibility of 20 field pea cultivars available in Alberta, including 13 yellow types and seven green types. Based on symptom development, the yellow-type cultivars Swing, Eiffel and Delta, and the green-type cultivar Orb, were the most susceptible. Of yellow-type cultivars, Voyageur, Carneval and Montana were most resistant to A scochyta infection.     

Pathogenic variability in Ethiopian isolates of Fusarium oxysporum f. sp. ciceris and reaction of chickpea improved varieties to the isolates

Twenty-four isolates of Fusarium oxysporum f. sp. ciceris were isolated from wilted chickpea plants obtained from different districts and ‘wilt sickplots’ of central Ethiopia to assess variability in pathogenecity of the populations. Each isolate was tested on 10 different chickpea lines and eight improved chickpea varieties. Isolates showed highly significant variation in wilt severity on the differential lines and improved varieties. Based on the reaction types induced on differential lines, isolates were grouped into four corresponding races. Of the 24 isolates, F13, F20 and F22 were the most virulent. Isolates of race 3 were found in all of the districts and ‘wilt sickplots’ studied. Improved chickpea varieties also showed differential reactions to the isolates. All varieties were resistant to isolates of race 3, while varieties Arerti and DZ-10-4 were resistant to all isolates tested, showing the lowest mean wilt severity. Varieties DZ-10-11 and Maryie were susceptible to isolates F13, F20 and F22 and showed the highest mean wilt severity. Identification of races can be useful in breeding chickpea varieties resistant to wilt. The differential reactions of the improved varieties against different races might be important in managing chickpea wilt through gene deployment.     

Plant defence reactions against fusarium wilt in chickpea induced by incompatible race 0 of Fusarium oxysporum f.sp. ciceris and nonhost isolates of F. oxysporum

Germinated seeds of 'kabuli' chickpea cv. ICCV 4 were inoculated with a conidial suspension of the incompatible race 0 of Fusarium oxysporum f.sp. ciceris (Foc) or of nonhost F. oxysporum resistance 'inducers', and 3 days later were challenged by root dip with a conidial suspension of highly virulent Foc race 5. Prior inoculation with inducers delayed the onset of symptoms and/or significantly reduced the final amount of fusarium wilt caused by race 5. However, the extent of disease suppression varied with the nature of the inducing agent; the nonhost isolates of F. oxysporum were more effective at disease suppression than the incompatible Foc race 0. Inoculation with the inducers gave rise to synthesis of maackiain and medicarpin phytoalexins in inoculated seedlings; these did not accumulate in plant tissues but were released into the inoculum suspension. Inoculation with inducers also resulted in accumulation of chitinase, β-1,3-glucanase and peroxidase activities in plant roots. These defence-related responses were induced more consistently and intensely by nonhost isolates of F. oxysporum than by incompatible Foc race 0. The phytoalexins and, to a lesser extent, the antifungal hydrolases, were also induced after challenge inoculation with Foc race 5. However, in this case the defence responses were induced in both preinduced and noninduced plants infected by the pathogen. It is concluded that the suppression of fusarium wilt in this study possibly involved an inhibitory effect on the pathogen of preinduced plant defences, rather than an increase in the expression of defence mechanisms of preinduced plants following a subsequent challenge inoculation.     

准东煤电煤化工产业开发建设对卡拉麦里山野生动物的影响

准东煤电煤化工产业开发建设在促进当地经济社会发展的同时,给卡拉麦里山的野生动物带来了一定的影响。通过建立野生动物的阻滞增长模型,以蒙古野驴(Equus hemionus)为例进行了建模计算,计算结果表明开发前10年间其种群的固有增长率为4.3026;而开发以后的10年间其固有增长率为0.6。煤电煤化工产业开发建设对其种群的数量增长带来了较大的影响,是影响卡拉麦里山蒙古野驴数量增长的诸多因素中的主要因素。     

Host‐range studies,genetic diversity and evolutionary relationships of ACLSV isolates from ornamental,wild and cultivated Rosaceous species

A large‐scale survey was carried out to study the host range and genetic diversity of Apple chlorotic leaf spot virus (ACLSV) in various Rosaceae species, with a special emphasis on ornamentals and wild shrubs. Samples were tested by DAS‐ELISA using two different antisera, and RT‐PCR amplification of part of the CP gene. There was generally a poor correlation between the results obtained with the two sets of serological reagents and between serological and molecular detection assays. Using a nested RT‐PCR assay developed here, ACLSV was found to be widespread among cultivated, ornamental and wild species of the Rosaceae. The virus was detected for the first time in plum, wild cherry, Crataegus monogyna, Prunus spinosa and Prunus cerasifera in Greece. Sequences of a part of the CP encoding gene and the 3′ untranslated region from ACLSV isolates originating from various wild species and ornamentals were compared to those of isolates from cultivated hosts, showing similar divergence levels. Further phylogenetic analysis using the sequenced region indicated that the isolates from wild or ornamental hosts were not more closely related to each other than to isolates from cultivated hosts. The possible role of different factors in the spread of ACLSV on cultivated, ornamental and wild species is discussed.