Seid Ahmed

Genetic improvement of grass pea for low neurotoxin (β-ODAP) content

Grass pea is a promising crop for adaptation under climate change because of its tolerance to drought, water-logging and salinity, and being almost free from insect-pests and diseases. In spite of such virtues, global area under its cultivation has decreased because of ban on its cultivation in many countries. The ban is imposed due to its association with neurolathyrism, a non-reversible neurological disorder in humans and animals due to presence of neurotoxin, β-N-oxalyl-L-α,β-diaminopropionic acid (β-ODAP) in its seedlings and seeds. The traditional varieties of grass pea contain 0.5-2.5% β-ODAP. Exploitable genetic variability for β-ODAP has been observed for development of low ODAP varieties, which along with improved agronomic and detoxification practices can help reduce the risk of lathyrism. Collaborative efforts between ICARDA and NARS have resulted in development of improved varieties such as Wasie in Ethiopia, Ratan, Prateek and Mahateora in India, and BARI Khesari-1 and BARI Khesari-2 in Bangladesh with <0.10% β-ODAP. Soil application of 15-20 kg ha(-1) zinc sulphate, early planting, and soaking seeds in water have shown significant effects on β-ODAP. Because of the often cross-pollination nature, the current breeding procedures being followed in grass pea requires paradigm shift in its approach for a possible genetic breakthrough.

Pathotype IV, a New and Highly Virulent Pathotype of Didymella rabiei, Causing Ascochyta Blight in Chickpea in Syria

The causal agent of Ascochyta blight disease of chickpea (Cicer arietinum L.) is highly variable because of the presence of a sexual phase (Didymella rabiei). There is also selection pressure on the pathogen due to wide adoption of improved resistant chickpea cultivars in some countries. The pathogen is able to produce pathotypes with specific virulence on particular cultivars. Three pathotypes, I, II, and III, have been reported (3). In this study, we confirmed the presence of a new and highly virulent pathotype that we designate as pathotype IV. To test the pathogenicity of the isolates collected and maintained at ICARDA, 10 isolates representing a wide spectrum of pathogenic variation, including those classified by S. M. Udupa et al. (3) and a putatively identified more virulent type, which was collected from a chickpea production field in the Kaljebrine area, Syria, were inoculated onto a set of differential chickpea genotypes. The differential genotypes, ILC 1929, ILC 482, ILC 3279, and ICC 12004, were sown in individual 10-cm-diameter pots containing potting mix and arranged in a randomized block design with three replications in a plastic house maintained at 18 to 20°C. Each differential genotype was inoculated individually with the 10 isolates following the methodology of S. M. Udupa et al. (3). DNA was extracted from single-spored isolates to compare the genotypes of the isolates using three simple sequence repeat (SSR) markers (ArA03T, ArH05T, and ArH06T) (2) and to determine the frequency of mating types (MAT) through the use of MAT-specific PCR primers for MAT1-1 and MAT1-2 (1). Host genotype reactions were measured on a 1 to 9 rating scale (1 = resistant and 9 = plant death). On the basis of the pathogenicity tests, the isolates were classified into four pathotypes: I (least virulent, killed ILC 1929 but not ILC 482, ILC 3279, or ICC12004); II (virulent, killed ILC 1929 and ILC 482 but not ILC 3279 or ICC12004); III (more virulent, killed ILC 1929, ILC 482, and ILC 3279 but not ICC12004); and IV (highly virulent, killed all four host differentials). Of 10 single-spore isolates tested, four showed similar disease reactions unique to pathotype I, four revealed pathotype II reactions, and one isolate each behaved like pathotype III or pathotype IV. SSR fingerprinting of these isolates provided evidence for genetic diversity since SSR ArH05T was highly polymorphic and amplified five bands, including pathotypes III- and IV-specific bands, which need further investigation to discern if this locus has any role to play in the virulence. MAT-type analysis showed that seven isolates were MAT1-1 while the remaining three isolates were MAT1-2. Only pathotype I showed the profile of MAT1-2 and the other three pathotypes were MAT1-1. Initially, a number of chickpea wild relatives were screened to identify sources of resistance to pathotype IV, but none of the accessions tested showed resistance. However, efforts are underway to combine minor and major gene(s) available in the breeding program in addition to a further search of the wild gene pools to control pathotype IV. References: (1) M. P. Barve et al. Fungal Genet. Biol. 39:151, 2003. (2) J. Geistlinger et al. Mol. Ecol. 9:1939, 2000. (3) S.M. Udupa et al. Theor. Appl. Genet. 97:299, 1998.

Effect of methionine supplement on physical responses and neurological symptoms in broiler chicks fed grass pea (Lathyrus sativus)-based starter ration

Starter feeding experiments of broiler chicks with raw grass pea (Lathyrus sativus L.) supplemented with different levels of DL-methionine were undertaken for 4 weeks to assess the toxicity of grass pea-based feed and to correlate it with neurological symptoms. Four hundred fifty day-old broiler chicks were divided into two groups and were given formulations containing 35% (ration I) or 98.5% (ration II) grass pea, respectively. Each ration included controls and treatments with added methionine of four different concentrations. Feed intake, weight gain and feed conversion efficiency (FCE) were much higher in ration I than in ration II and these parameters significantly improved by addition of methionine in both rations. Significant increase of neurological signs with higher grass pea intake and significant reduction of acute neurological signs with addition of methionine were observed. Tolerance for grass pea was enhanced with increasing methionine in the diet and with age. Despite a similarity in the initial intake, a significant (p0.05) increase in the final feed intake by the chicks with methionine addition was found in both rations. These results suggest that methionine can improve a grass pea-based diet for broiler chicks and especially can protect young chicks from neurological symptoms.

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.

Climatic, edaphic and altitudinal factors affecting yield and toxicity of Lathyrus sativus grown at five locations in Ethiopia.

A 2 years (2005-2006) data analysis based on agronomic, qualitative, climatic and edaphic factors was carried out using 10 grass pea (Lathyrus sativus L.) genotypes grown at five eco-divergent locations (Alem Tena, Debre Zeit, Denbi, Akaki, Chefe Donsa) in Ethiopia. Crop yield showed considerable variability among locations, years and genotypes. Path coefficient analysis indicated that rainfall and days to maturity have a large positive influence on yield. High level of micronutrients Mn(2+) and S(2-) negatively affected yield. Path analysis revealed that Zn(2+)/P, days to maturity, yield and K(+) were dominant variables affecting the response variable β-ODAP (β-N-oxalyl-L-α,β-diaminopropionic acid), the neuro-excitatory amino acid in grass pea seeds considered as the cause of neurolathyrism. Linear correlation analysis between β-ODAP and the 35 factors considered showed that β-ODAP level was positively correlated (r > 0.70) with K(+) and sunshine hours (ssh) and negatively correlated (r < 0.70) with soil pH, days to maturity and yield. The strongest correlation of ssh with β-ODAP level was found during the phase of crop maturity. Our results suggest that β-ODAP biosynthesis and its response to environmental stress are maximized during the post-anthesis stage.

Yield loss of faba bean (Vicia faba) due to chocolate spot (Botrytis fabae) in sole and mixed cropping systems in Ethiopia

Two field experiments were conducted during 2004 and 2005 cropping seasons at Adet Agricultural Research Center, Ethiopia to assess yield losses caused by chocolate spot(Botrytis fabae) of faba bean in sole and mixed cropping systems using two cultivars. Cropping systems were sole faba bean (FB), faba bean mixed with field pea (FB: FP), barley (FB: BA) and maize (FB: MA). Mancozeb was sprayed at the rate of 2.5 kg a.i/ha at 7-, 14- and 21-day interval to generate different levels of chocolate spot disease in all the four cropping systems, and unsprayed control was also included. The treatments were arranged in a randomised complete block design (RCBD) with four replications. FB: MA mixed cropping significantly reduced disease severity and the area under disease progress curve (AUDPC) and increased faba bean grain yield. The highest faba bean grain yield among the three mixed croppings under different spray schedules was obtained from FB: MA mixed cropping in both 2004 and 2005 (2.56 and 3.74 t/ha, respectively) cropping seasons. There were highly significant yield differences (P < 0.05) among the spray intervals of mancozeb in both seasons. The highest grain yield (4.9 t/h) was recorded from the 7-day spray interval in 2005. The unsprayed faba bean had a lower grain yield (1.9 t/ha in 2004 and 2.3 t/ha in 2005) compared to the sprayed plots. The highest relative yield loss (67.5%) was calculated in 2005 from FB: FP mixed cropping in unsprayed plots. The relative yield losses in the unsprayed plots were in the range of 35.8–41.5% in 2004 and 52.6–67.5% in the 2005 cropping season. Severity and AUDPC were inversely correlated with faba bean grain yield. Significant differences were recorded in the 100-seed weight and days to maturity (DM). The unsprayed plots had shorter DM ranging from 126 to 128.5 day (except FB: MA mixed cropping) in 2004 and 122–123.9 days in 2005. In the sprayed plots DM was relatively longer than the unsprayed plots. A higher seed weight was recorded in the sole FB (56 g) and FB: MA (55 g) mixed cropping, and the lowest value of 100-seed weight was recorded from FB: FP (53 g) mixed cropping. The productivity of the mixed cropping evaluated by land equivalent ratio (LER) exceeded that of sole cropping. Faba bean grain yield was highly influenced by the severity of chocolate spot. The disease affects the DM, forcing early maturing of the plants.

Evaluation of Local Isolates of Trichoderma Spp. against Black Root Rot (Fusarium solani) on Faba Bean

Faba bean (Vicia fabae L.) is one of the most important pulse crops in Ethiopia and is now cultivated on large areas in many countries. In most growing areas, however, the production of the crop is constrained by several disease infections, including fungal diseases. Black root rot caused by Fusarium solani is the most destructive disease of faba bean. The antagonistic potentials of locally isolated Trichoderma spp. from rhizosphere soils of faba bean plants in the highlands of northeastern Ethiopia were evaluated against F. solani, responsible for black root rot. All isolates of Trichoderma spp. had strong biological control activity against F. solani in vitro as well as in vivo pot experiment. Under dual culture, the percentage of mycelial growth inhibition of F. solani by the Trichoderma ranged from 33.9 to 67.0%. The highest (67.0%) inhibition was obtained from isolate TS036, while the lowest (33.9%) by isolate TS015. Pathogen-inoculated faba bean plants grown in pots that were treated with antagonists had taller plant heights and biomass than the Trichoderma untreated control inoculated with F. solani. The Trichoderma isolates significantly reduced black root rot severity on faba bean seedlings with disease reduction ranging from 64.4 to 74.6% over control. Use of Trichoderma species can be a potential source of biological control agents for the management of black root rot in faba beans grown in the region. Hence, the potential Trichoderma isolates under field condition might used as a components in the integrated management of F. solani that caused faba bean black root rot in the highlands of northeastern Ethiopia.

Pathogenic and genetic diversity of Botrytis fabae Sand. isolates from faba bean fields in different agro-ecological zones of Northern Ethiopia

The identity and genetic diversity of the causal agent of chocolate spot (Botrytis cinerea or Botrytis fabae) in Ethiopia is still poorly defined and this hinders efforts aimed at integrated management of the disease, especially breeding for resistance. Chocolate spot disease epidemics are more severe in the moist, humid western districts of northern Ethiopia (NW) compared to the sub-humid eastern districts (NE) but knowledge of pathogen identity and genetic diversity is needed to facilitate epidemiological studies. Of a total of 120 Botrytis isolates examined, all were found to be B. fabae and none fitted the morphological description of B.cinerea. In pathogenicity tests, representative isolates caused typical chocolate spot symptoms and were re-isolated from infected leaves, indicating that B. fabae is the causal agent of chocolate spot in Ethiopia. Marked differences were found among the isolates in colony morphology and growth rate but the morphology of isolates was unrelated to their pathogenicity. A preliminary virulence (=severity of disease) test with 76 isolates revealed differences in virulence of the B. fabae isolates, with NE isolates being generally more virulent than the NW isolates. Genetic analysis based on 129 amplified fragment length polymorphism (AFLP) markers showed that genetic differentiation among pathogen populations was low (GST = 0.02 to 0.03; θ = 0.02, p = 0.05), indicating limited geographic delimitation and significant gene flow. Total gene diversity (HT = 0.22) was mostly attributable to diversity within populations (HS = 0.21). Thus, only 4.5% of the total variability was attributable to frequency differences between agroecological zones. Genotypic diversity (GD), defined as the probability that two individuals taken at random had different genotypes, was high for populations from NW and NE, and from the moist and subhumid agro-ecological zones (GD = 0.99). However, cluster analysis showed high similarity among many isolates (>75% similarity index), suggesting that such isolates have a familial structure or are clonally related. The phenetic tree revealed groups with low bootstrap values that did not reflect the grouping of isolates based on virulence or agro-ecological zone. The implications of these findings for chocolate spot resistance breeding in Ethiopia are discussed.

Status of chickpea fusarium wilt (Fusarium oxysporum f.sp. ciceris) in northwestern Ethiopia

Surveys for incidence of fusarium wilt of chickpea were carried out in six districts of North and South Gondar, and East Gojam administrative zones, Amhara National Regional State in northwestern Ethiopia in 2006–2007 and 2007–2008 main cropping seasons. In each district, five representative peasant administrations (PAs) were selected based mainly on chickpea-growing area coverage, and, in each PA, disease was assessed in five randomly selected farmers’ fields. In the two cropping seasons, the mean incidences recorded in each district were as follows: Gondar Zuria, 34.16% and 34.11%; Dembia, 37.90% and 35.36%; Libo-Kemkem, 34.74% and 28.81%; Fogera, 34.74% and 28.81%; Dejen, 34.74% and 28.81% and Enemay, 33.34% and 37.64%. The result indicated that fusarium wilt is currently highly distributed in all surveyed chickpea-growing areas of northwestern part of Ethiopia. Therefore, possible management options are vital to alleviate the problem.

New faba bean germplasm with multiple resistances to Ascochyta blight, chocolate spot and rust diseases

Faba bean is one of the oldest crops that originated in the Fertile Crescent and is now distributed around the world and produced under different agro-climatic conditions. It is mainly cultivated in high rainfall and irrigated areas which are favorable for the development of foliar diseases causing severe crop losses. The purpose of this study was to identify new sources of resistance to the three key foliar diseases. A total of 2000 accessions, breeding lines and populations were screened for resistance to chocolate spot, Ascochyta blight and rust diseases from 2005 to 2012, at Lattakia Station, Syria. A total of 30 lines combining Ascochyta blight, chocolate spot and rust resistance were identified from accessions collected from China, Italy, Jordan, Lebanon and Spain. These new sources of resistance will enhance the development of faba bean breeding lines with combined resistance to the three foliar diseases.