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.     

Factors Influencing Transmission of Didymella rabiei (Ascochyta Blight) from Inoculated Seed of Chickpea Under Controlled Conditions

Ascochyta blight of chickpea (Cicer arietinum), caused by the fungus Didymella rabiei, has the potential to cause 100% crop loss in severe epiphytotics. Management of this disease often involves reducing sources of inoculum. The influence of sowing depth, host resistance, seed infection level and soil temperature on disease transmission was investigated in a series of glasshouse and growth room trials using seed artificially inoculated with D. rabiei. A positive correlation (R²=0.9992) was observed between rate of seed infection and the incidence of disease on seedlings. Disease transmission to seedlings was not significantly influenced by sowing depth (1, 3 and 6 cm) in separate trials on two cultivars. Susceptibility of the host showed no obvious influence on the frequency of disease transmission in two trials conducted using four cultivars ranging from highly susceptible to moderately susceptible/moderately resistant. Trials conducted in controlled conditions showed that there was no obvious relationship between soil temperature (5, 9, 14 and 19 °C) and the incidence of disease on seedlings.     

Genetics of virulence in Ascochyta rabiei

In order to critically test the hypothesis that virulence variation in the Ascochyta rabiei/chickpea pathosystem is a discrete character under simple genetic control, a genetic cross was made between a highly virulent isolate of A. rabiei from Syria and a less virulent isolate from the USA. Two independent virulence assays conducted by inoculating susceptible and resistant chickpea cultivars under controlled conditions with 77 independent progeny isolates from this cross revealed a continuous distribution of disease phenotypes. Bimodality, as would be predicted for the segregation of virulence under simple genetic control, was not supported by statistical tests of the progeny phenotype distribution. anova revealed highly significant pathogen‐genotype × host‐genotype interactions demonstrating the segregation of genes controlling specialization on the two cultivars tested. These interactions could be localized to two isolates that changed virulence rank on the cultivars. It was concluded that variation in virulence to these two cultivars is under quantitative genetic control. If this conclusion applies to other cultivars, it can be speculated that the discrete categories of virulence variation identified in previous studies were probably the result of incomplete sampling of host resistance or pathogen virulence variation and/or of selection for increased virulence in contemporary A. rabiei populations.     

Determination of agronomic practices for the management of blight of chickpea caused by <Emphasis Type="Italic">Ascochyta rabiei</Emphasis> in Turkey: 1. Appropriate sowing date

In order to determine the most appropriate dates for planting chickpea in central Anatolia, Turkey, six cultivars were planted at three sites that differed in disease pressure. In two of the sites, disease pressure from Ascochyta rabiei was promoted by spreading infected chickpea debris on the soil surface at the time of planting and, at one of these, sprinkle irrigation was applied. In the third site, where conditions were dryer, no artificial inoculum was provided. Plants from seeds sown in early March had the most disease and in the sprinkle irrigated plots the disease severity ranged from 7.8 on the most susceptible cv. Canitez to 3.3 on the least susceptible Gokce as scored on the 1–9 scale where 1 = no disease and 9 represents a plant killed by the fungus. There was an inverse relationship between disease severity and yield, production from blight resistant cultivars of around 2,000 kg ha⁻¹ being more than twice that of susceptible ones. Delaying planting for 3–5 weeks reduced the severity of ascochyta blight but also reduced the yields in four of the six cultivars. In contrast, reduction in disease severity by delayed sowing resulted in yield increases for the susceptible cvs Canitez and Local, although yield level was not as much as those of the less susceptible cvs sown early. Delay of 6–9 weeks almost eliminated ascochyta blight but yields of all cultivars were seriously compromised by drought stress. In consequence, chickpea farmers are recommended to use resistant or tolerant cultivars and sow early in March. For less resistant cultivars, sowing in early April is recommended. Further delay is not recommended unless irrigation is provided and fungicide spraying is recommended where signs of infection are present under conditions conducive to the disease.     

Diagnostics,genetic diversity and pathogenic variation of ascochyta blight of cool season food and feed legumes

Molecular diagnostic techniques have been developed to differentiate the Ascochyta pathogens that infect cool season food and feed legumes, as well as to improve the sensitivity of detecting latent infection in plant tissues. A seed sampling technique was developed to detect a 1% level of infection by Ascochyta rabiei in commercial chickpea seed. The Ascochyta pathogens were shown to be genetically diverse in countries where the pathogen and host have coexisted for a long time. However, where the pathogen was recently introduced, such as A. rabiei to Australia, the level of diversity remained relatively low, even as the pathogen spread to all chickpea-growing areas. Pathogenic variability of A. rabiei and Ascochyta pinodes pathogens in chickpea and field pea respectively, appears to be quantitative, where measures of disease severity were based on aggressiveness (quantitative level of infection) rather than on true qualitative virulence. In contrast, qualitative differences in pathogenicity in lentil and faba bean genotypes indicated the existence of pathotypes of Ascochyta lentis and Ascochyta fabae. Therefore, reports of pathotype discrimination based on quantitative differences in pathogenicity in a set of specific genotypes is questionable for several of the ascochyta-legume pathosystems such as A. rabiei and A. pinodes. This is not surprising since host resistance to these pathogens has been reported to be mainly quantitative, making it difficult for the pathogen to overcome specific resistance genes and form pathotypes. For robust pathogenicity assessment, there needs to be consistency in selection of differential host genotypes, screening conditions and disease evaluation techniques for each of the Ascochyta sp. in legume-growing countries throughout the world. Nevertheless, knowledge of pathotype diversity and aggressiveness within populations is important in the selection of resistant genotypes.     

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.     

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.     

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.     

Alternative hosts and plant tissues for the survival,sporulation and spread of the Ascochyta blight pathogen of chickpea

Various crop and weed species were infected naturally by Didymella rabiei (anamorph: Ascochyta rabiei) in blight-affected chickpea fields in the Palouse region of eastern Washington and northern Idaho, USA. The fungus was isolated from asymptomatic plants of 16 species commonly found in commercial crops in this region. Isolates of the pathogen from crop and weed species were pathogenic to chickpea and indistinguishable in cultural and morphological characteristics from isolates of D. rabiei from chickpea. Both mating types of D. rabiei were isolated from eight naturally infected plant species. Chickpeas were infected by D. rabiei when plants emerged through infested debris of seven crop and weed species. The teleomorph developed on overwintered tissues of seven plant species infected naturally by D. rabiei in a blight screening nursery and on debris of wheat, white sweet clover and pea inoculated with ascospores of D. rabiei or conidia of two compatible isolates of the pathogen. Didymella rabiei naturally infected 31 accessions of 12 Cicer spp. and the teleomorph developed on the overwintered debris of all accessions, including those of three highly resistant perennial species. The fungus developed on the stem and leaf pieces of ten plant species common to southern Spain inoculated with conidia of two compatible isolates of D. rabiei, and formed pseudothecia with asci and viable ascospores on six of ten species and pycnidia with conidia on all plant species.     

Airborne ascospores ofDidymella rabiei as a major primary inoculum for Ascochyta blight epidemics in chickpea crops in southern Spain

The incidence and severity of Ascochyta blight in potted chickpea trap plants exposed for 1-wk periods near infested chickpea debris in Córdoba, Spain, or in chickpea trap crops at least 100 m from infested chickpea debris in several locations in southern Spain were correlated with pseudothecial maturity and ascospore production ofDidymella rabiei from nearby chickpea debris. The period of ascospore availability varied from January to May and depended on rain and maturity of pseudothecia. The airborne concentration of ascospores ofD. rabiei was also monitored in 1988. Ascospores were trapped mostly from the beginning of January to late February; this period coincided with that of maturity of pseudothecia on the chickpea debris. Most ascospores were trapped on rainy days during daylight and 70% were trapped between 12.00 and 18.00 h. Autumn-winter sowings of chickpea were exposed longer to ascospore inoculum than the more traditional spring sowings because the autumn-winter sowings were exposed to the entire period of ascospore production on infested chickpea debris lying on the soil surface.     

Development of the teleomorph of Ascochyta rabiei on culture media

Ascochyta blight caused by Didymella rabiei (anamorph: Ascochyta rabiei) is an important foliar disease of chickpea in many countries. The fungus is heterothallic and requires the pairing of two compatible mating types for the teleomorph to develop. In nature, the teleomorph only develops on chickpea debris that overwinters on the soil surface in the presence of both mating types. When natural and synthetic agar media were seeded with conidial suspensions of compatible isolates of D. rabiei from Spain and the United States and incubated under favourable conditions for teleomorph development, the teleomorph only developed on 2?% water agar amended with powdered chickpea stems or hot water extracts of chickpea stems, but not on 14 other natural or synthetic media. Ascospore isolates of D. rabiei from pseudothecia that developed on agar media were indistinguishable in cultural and morphological characteristics from isolates of the fungus from chickpea. Production of pseudothecia and ascospores on the best culture medium was always lower than on stem pieces of chickpea straw used as a control treatment. Ascospores discharged from pseudothecia that developed on powdered chickpea stem media onto chickpea seedlings were pathogenic, inducing symptoms identical to those caused by ascospores from chickpea stem pieces or conidia from a chickpea isolate of the fungus. This is the first report of the teleomorph of D. rabiei developing on culture media.     

Towards identifying pathogenic determinants of the chickpea pathogen <Emphasis Type="Italic">Ascochyta rabiei</Emphasis>

Ascochyta blight is a serious disease of cool-season grain legumes (chickpea, faba bean, lentil and pea) caused by fungal species of the anamorphic genus Ascochyta and related genera. Despite extensive studies on the biology, ecology, epidemiology and management of the disease, little is known about the pathogenic determinants of these pathogens. This research aims at using Ascochyta rabiei as a model for the genus in investigating genetic factors of pathogenicity, with the ultimate goal of elucidating pathogenic mechanisms. Three advances were made: (1) insertional mutants with altered pathogenicity were identified through in vivo screening, and genomic regions adjacent to the insertion sites in selected mutants were determined; (2) a phage library of A. rabiei genomic DNA was constructed, and the library was estimated to provide complete coverage of the A. rabiei genome. This library was used successfully to recover clones with DNA adjacent to insertional mutation sites and to isolate specific genes; (3) DNA probes specific for an acyl-CoA ligase (cps1) and a polyketide synthase gene (pks1) were developed and library clones containing the corresponding genomic regions were identified from the phage library. These advances provide the foundation and necessary tools for experimentation of ectopic complementation assays and targeted mutagenesis to elucidate the genetic mechanisms of pathogenicity of A. rabiei.     

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.     

Validation of a QTL for resistance to ascochyta blight linked to resistance to fusarium wilt race 5 in chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.)

Ascochyta blight caused by Ascochyta rabiei and fusarium wilt caused by Fusarium oxysporum. f. sp. ciceris are the two most serious diseases of chickpea (Cicer arietinum). Quantitative trait loci (QTL) or genes for ascochyta blight resistance and a cluster of resistance genes for several fusarium wilt races (foc1, foc3, foc4 and foc5) located on LG2 of the chickpea map have been reported independently. In order to validate these results and study the linkage relationship between the loci that confer resistance to blight and wilt, an intraspecific chickpea recombinant inbred lines (RIL) population that segregates for resistance to both diseases was studied. A new LG2 was established using sequence tagged microsatellite sites (STMS) markers selected from other chickpea maps. Resistance to race 5 of F. oxysporum (foc5) was inherited as a single gene and mapped to LG2, flanked by the STMS markers TA110 (6.5 cM apart) and TA59 (8.9 cM apart). A QTL for resistance to ascochyta blight (QTL_AR3) was also detected on LG2 using evaluation data obtained separately in two cropping seasons. This genomic region, where QTL_AR3 is located, was highly saturated with STMS markers. STMS TA194 appeared tightly linked to QTL_AR3 and was flanked by the STMS markers TR58 and TS82 (6.5 cM apart). The genetic distance between foc5 and QTL_AR3 peak was around 24 cM including six markers within this interval. The markers linked to both loci could facilitate the pyramiding of resistance genes for both diseases through MAS.     

Towards the first linkage map of the<Emphasis Type="Italic">Didymella rabiei</Emphasis> genome

A genetic map was developed for the ascomyceteDidymella rabiei (Kovachevski) v. Arx (anamorph:Ascochyta rabiei Pass. Labr.), the causal agent of Ascochyta blight in chickpea (Cicer arietinum L.). The map was generated with 77 F₁ progeny derived from crossing an isolate from the U.S.A. and an isolate from Syria. A total of 232 DAF (DNA Amplification Fingerprinting) primers and 37 STMS (Sequence-Tagged Microsatellite Site) primer pairs were tested for polymorphism between the parental isolates; 50 markers were mapped, 36 DAFs and 14 STMSs. These markers cover 261.4cM in ten linkage groups. Nineteen markers remained unlinked. Significant deviation from the expected 1:1 segregation ratios was observed for only two markers (Prob. of χ²<0.05). The implications of our results on ploidy level of the asexual spores are discussed. http://www.phytoparasitica.org posting Sept. 5, 2002.     

Responses of cucumber cultivars to induction of systemic resistance against anthracnose by plant growth promoting fungi

Initial experiment on the reactions of five Japanese cultivars of cucumber toColletotrichum orbiculare infection in the greenhouse revealed that cv Suyo and Gibai were susceptible and moderately susceptible, respectively, while cv Shogoin fushinari and Sagami hanjiro were resistant to infection byC. orbiculare; cv Ochiai fushinari was moderately resistant. The ability of 16 plant growth promoting fungi (some isolates belonged to species ofPhoma and some non-sporulating isolates) isolated from zoysiagrass rhizospheres to induce systemic resistance in the above five cucumber cultivars was tested by growing plants in potting medium infested with barley grain inocula of PGPF in the greenhouse. The second true leaves of 21-day-old plants were challenge inoculated withC. orbiculare and disease assessed. Nine, out of 16 isolates, caused significant reduction of disease caused byC. orbiculare in at least two cultivars.Phoma isolates (GS8-1 and GS8-2) and non-sporulating isolates (GU21-2, GU23-3, and GU24-3) significantly reduced the disease in all the five cultivars. The disease suppression in cucumber was due to the induction of systemic resistance, since the inducer(s) and the pathogen were separated spatially and that the inducer did not colonize aerial portions. The resistance induced by certain isolates in a susceptible cultivar was less than that in a resistant cultivar. Disease suppression caused by isolate GU21-2 was similar to theC. orbiculare induced control in certain cultivars. The average rate of expansion of lesion diameter on leaves due toC. orbiculare was slower due to induction with the selected plant growth promoting fungi compared to the uninduced control plants. Roots of four cultivars were colonized by only three isolates, however, roots of one cultivar (Suyo) was colonized by five isolates suggesting the cultivar-specific root colonization ability.Abbreviations cv cultivar(s) - PGPF plant growth promoting fungal isolates - PGPR plant growth promoting rhizobacteria     

Genetic and pathogenic diversity within Ascochyta rabiei(Pass.) Lab. populations in Pakistan causing blight of chickpea (Cicer arietinum L.)

Pathogenic and genetic diversity in Ascochyta rabiei populations in Pakistan were evaluated. Biological pathotyping of 130 A. rabiei isolates (obtained from hierarchically collected samples) was conducted on a set of three chickpea differentials, i.e. ILC 1929 (susceptible), ILC 482 (tolerant) and ILC 3279 (resistant), under controlled conditions. Disease severity data were recorded 12 days after inoculation. Statistical analysis grouped the isolates into three pathotype classes. Four isolates belonged to pathotype I (least aggressive), 79 isolates to pathotype II (medium aggressive) and 47 isolates to pathotype-III (highly aggressive).Genetic analysis was performed using RAPDs and oligonucleotide fingerprinting, where Hinf I-digested DNA was hybridized to the³²P-endlabeled oligonucleotide probes (CAA)₅, (GAA)₅, (GA)₈, (CA)₈and (GATA)₄. Dendrograms produced by cluster analysis discriminated 46 genotypes in the A. rabiei population of Pakistan. Genetic distances and relatedness between isolates were calculated. At a genetic distance of 0.3, genotypes were divided into six distinct genotype groups A, B, C, D, E and F containing 16, 11, 2, 5, 5 and 7 isolates, respectively. Most of the genotypes were area specific or predominated in certain areas but did not belong to a distinct pathotype, while most of the aggressive isolates (pathotype III) occurred in Northern Punjab and in the North Western Frontier Province.     

黄淮麦区主栽小麦品种抗茎基腐病评价及茎秆和籽粒中毒素积累分析

为明确黄淮麦区主栽小麦品种对茎基腐病的抗性水平、不同抗性指标的相关性,以及茎杆和籽粒中镰刀菌毒素积累情况,在采用苗期茎基部滴注法和成株期混合播种法进行抗性鉴定的基础上,本研究还利用超高效液相色谱高分辨质谱联用技术,测定了小麦茎杆和籽粒中6种常见镰刀菌毒素的含量.结果表明,供试的20个小麦品种中,苗期抗病、中抗、感病和高...