Ascochyta blight, caused by the fungus Ascochyta rabiei (Pass.) Lab., is one of the most devastating diseases of chickpea (Cicer arietinum L.) worldwide. Research was conducted to map genetic factors for resistance to ascochyta blight using a linkage map constructed with 144 simple sequence repeat markers and 1 morphological marker (fc, flower colour). Stem cutting was used to vegetatively propagate 186 F2 plants derived from a cross between Cicer arietinum L. 'ICCV96029' and 'CDC Frontier'. A total of 556 cutting-derived plants were evaluated for their reaction to ascochyta blight under controlled conditions. Disease reaction of the F1 and F2 plants demonstrated that the resistance was dominantly inherited. A Fain's test based on the means and variances of the ascochyta blight reaction of the F3 families showed that a few genes were segregating in the population. Composite interval mapping identified 3 genomic regions that were associated with the reaction to ascochyta blight. One quantitative trait locus (QTL) on each of LG3, LG4, and LG6 accounted for 13%, 29%, and 12%, respectively, of the total estimated phenotypic variation for the reaction to ascochyta blight. Together, these loci controlled 56% of the total estimated phenotypic variation. The QTL on LG4 and LG6 were in common with the previously reported QTL for ascochyta blight resistance, whereas the QTL on LG3 was unique to the current population.
Random amplified polymorphic DNA, simple sequence repeat, and inter-simple sequence repeat markers were used to estimate the genetic relations among 65 pea varieties (Pisum sativum L.) and 21 accessions from wild Pisum subspecies (subsp.) abyssinicum, asiaticum, elatius, transcaucasicum, and var. arvense. Fifty-one of these varieties are currently available for growers in western Canada. Nei and Li's genetic similarity (GS) estimates calculated using the marker data showed that pair-wise comparison values among the 65 varieties ranged from 0.34 to 1.00. GS analysis on varieties grouped according to their originating breeding programs demonstrated that different levels of diversity were maintained at different breeding programs. Unweighted pair-group method arithmetic average cluster analysis and principal coordinate analysis on the marker-based GS grouped the cultivated varieties separately from the wild accessions. The majority of the food and feed varieties were grouped separately from the silage and specialty varieties, regardless of the originating breeding programs. The analysis also revealed some genetically distinct varieties such as Croma, CDC Handel, 1096M-8, and CDC Acer. The relations among the cultivated varieties, as revealed by molecular-marker-based GS, were not significantly correlated with those based on the agronomic characters, suggesting that the 2 systems give different estimates of genetic relations among the varieties. However, on a smaller scale, a consistent subcluster of genotypes was identified on the basis of agronomic characters and their marker-based GS. Furthermore, a number of variety-specific markers were identified in the current study, which could be useful for variety identification. Breeding strategies to maintain or enhance the genetic diversity of future varieties are proposed.
Lentil anthracnose (Colletotrichum truncatum (Schwein.) Andrus et W.D. Moore is a potential threat in many lentil (Lens culinaris Medik.) production regions of North America. In the lentil germplasm maintained in Germany and North America, 16 lines were reported to have resistance to race Ct1, but none has resistance reported to race Ct0. The objective of this study was to examine accessions of wild Lens species for their resistance to races Ct1 and Ct0 of lentil anthracnose. Five hundred and seventy-four wild accessions of six species and control lines were screened in two replications under both field and greenhouse conditions using a 1-9 scoring scale (1, highly resistant; 2-3, resistant; 4-5, moderately resistant; 6-7, susceptible; and 8-9, highly susceptible). Indianhead and PI 320937 were resistant while Eston and Pardina were susceptible to race Ct1 as expected. However, none of the check lines were resistant to race Ct0. Among the six Lens wild species tested, accessions of Lens ervoides (Brign.) Grande had the highest level of resistance, 3-5 to race Ct1 and Ct0 followed by L. lamottei Czefr. in the field and greenhouse. Lens orientalis (Boiss.), L. odemensis L., L. nigricans (M. Bieb.) Godron and L. tomentosus L. were highly susceptible, 8-9 to race Ct0 in the greenhouse. The highest frequency of resistance, especially in L. ervoides (Brign.) Grande, was found in accessions originating from Syria and Turkey. The usefulness of these L. ervoides (Brign.) Grande accessions as sources of resistance to the more virulent race of anthracnose in a lentil breeding program is discussed.
Lentil, Lens culinaris subsp. culinaris Medic., is an important legume crop on the Canadian prairies. Anthracnose, a fungal disease caused by Colletotrichum truncatum (Schwein.) Andrus & W.D. Moore, is a major barrier to seed yield and quality in lentil. Pathogenicity testing has revealed two races, Ct1 and Ct0, of C. truncatum in western Canada. No cultivar or landrace of cultivated lentil has been reported with resistance to anthracnose race Ct0. A search for Ct0 resistance in the wild species identified a high frequency of resistant accessions in Lens ervoides (Brign.) Grande. To incorporate higher levels of resistance from L. ervoides to the two races of anthracnose, a cross was made between a susceptible L. culinaris cultivar, Eston, and a resistant accession of L. ervoides germplasm, L‐01‐827A, which has both Ct0 and Ct1 resistance. Embryo rescue technique was used to obtain an F1 hybrid. Single‐seed descent was used to advance the individual F2 plants to F7:8 recombinant inbred lines. Evidence of transfer of resistance to both anthracnose races Ct1 and Ct0 from the wild species to cultivated lentil is presented. Chi‐square tests of goodness of fit indicated that resistance to race Ct1 and race Ct0 may be conferred by two recessive genes. However, these results may be skewed due to variable fertility encountered in development of the population. Selection of resistant lines for use in pyramiding genes in breeding programs should result in a more durable level of resistance to anthracnose in lentil.
The inheritance of an inter-simple-sequence-repeat (ISSR) polymorphism was studied in a cross of cultivated chickpea (Cicer arietinum L.) and a closely related wild species (C. reticulatum Lad.) using primers that anneal to a simple repeat of various lengths, sequences and non-repetitive motifs. Dinucleotides were the majority of those tested, and provided all of the useful banding patterns. The ISSR loci showed virtually complete agreement with expected Mendelian ratios. Twenty two primers were used for analysis and yielded a total of 31 segregating loci. Primers based on (GA)n repeats were the most abundant while primers with a (TG)n repeat gave the largest number of polymorphic loci. Nucleotides at the 5' and 3' end of the primers played an important role in detecting polymorphism. All the markers showed dominance. We found an ISSR marker linked to the gene for resistance to fusarium wilt race 4. The marker concerned, UBC-855500, was found to be linked in repulsion with the fusarium wilt resistance gene at a distance of 5.2 cM. It co-segregated with CS-27700, a RAPD marker previously shown to be linked to the gene for resistance to fusarium wilt race 1, and was mapped to linkage group 6 of the Cicer genome. This indicated that genes for resistance to fusarium wilt races 1 and 4 are closely linked. The marker UBC-855500 is located 0.6 cM from CS-27700 and is present on the same side of the wilt resistance gene. To our knowledge this is the first report of the utility of an ISSR marker in gene tagging. These markers may provide valuable information for the development of sequence-tagged microsatellite sites (STMS) at a desired locus.
Ascochyta blight (AB) caused by Ascochyta rabiei (teleomorph, Didymella rabiei) Pass. Lab. is an important fungal disease of chickpea worldwide. Only moderate sources of resistance are available within the cultivated species and we hypothesized that the available sources may carry different genes for resistance, which could be pyramided to improve field resistance to AB. Four divergent moderately resistant cultivars CDC Frontier, CDC Luna, CDC Corinne, and Amit were each crossed to a highly susceptible germplasm ICCV 96029. Parents, F(1) and F(2) generations were evaluated under controlled conditions for their reactions to AB. A total of 144 simple sequence repeat (SSR) markers were first mapped to eight linkage groups (LG) for the CDC Frontier x ICCV 96029 population. Then based on the evidence from this population, 76, 61, and 42 SSR markers were systematically chosen and mapped in CDC Luna, CDC Corinne, and Amit populations, respectively. Frequency distributions of the AB rating in the F(2) generation varied among the four populations. Composite interval mapping revealed five QTLs (QTL1-5), one on each of LG 2, 3, 4, 6, and 8, respectively, distributed across different sources, controlling resistance to AB. CDC Frontier contained QTL2, 3, and 4 that simultaneously accounted for 56% of phenotypic variations. CDC Luna contained QTL 1 and 3. CDC Corinne contained QTL 3 and 5, while only QTL 2 was identified in Amit. Altogether these QTL explained 48, 38, and 14% of the estimated phenotypic variations in CDC Luna, CDC Corinne, and Amit populations, respectively. The results suggested that these QTLs could be combined into a single genotype to enhance field resistance to AB.
Fusarium wilt caused by Fusarium oxysporum Schlechtend.:Fr. f. sp. ciceris (Padwick) Matuo & K. Sato is the most widely spread soilborne disease of chickpea (Cicer arietinum L.). To advance our understanding of the genetics of wilt resistance and aid chickpea breeding programs, we developed a set of F6 recombinant inbred lines (RILs) between Fusarium wilt susceptible (C‐104) and resistant 315) parents. Prior screening of selected F3 plants identified two primers (UBC‐170 and CS‐27) that produced random amplified polymorphic DNA {RAPD) markers associated with Fusarium wilt race 1 resistance. Analysis of the RILs with these primers yielded an estimate of 7% recombination between the two markers and the locus for wilt resistance, and 6% recombination between the loci corresponding to the two RAPD markers. The DNA fragments were cloned and sequenced in order to construct primers that would amplify only the markers of interest. Primer pair CS‐27F/CS‐27R amplified a fragment linked to the allele for susceptibility to race 1 of Fusarium wilt and thus constitute allele specific associated primers (ASAPs), whereas UBC‐170FFLIBC‐170R produced a single band for both resistant and susceptible genotypes, thus demonstrating locus specificity rather than allele specificity. The use of markers generated by the RAPD or ASAP approaches can facilitate the introgression of resistance genes into susceptible lines and expedite the screening of chickpea germplasm resources and will be useful in extending the genetic map of chickpea.
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