A bottleneck in lentil: widening its genetic base in South Asia

This paper introduces the concept of a bottleneck in lentil (Lens culinaris Medikus) in South Asia and then reviews its rupture. South Asia is the largest lentil growing region in the world and where indigenous lentils show a marked lack of variability. This results from its introduction from Afghanistan around 2000 B.C. and it limits breeding progress. Three approaches to widening the genetic base in the region have been tried, namely plant introduction, hybridization and mutation breeding. Introductions from West Asia flower as indigenous material matures. The asynchrony in flowering has isolated the local pilosae ecotype reproductively. However, the introduction of ILL 4605, an early, large-seeded line, has resulted in its release as ‘Manserha 89’ for wetter areas of Pakistan and its widespread use as a parent in breeding programs in the region. Hybridization between pilosae and exotic germplasm, primarily at International Center for Agricultural Research in the Dry Areas (ICARDA) followed by selection in the sub-continent has resulted in cultivars with improved disease resistance and yield in Bangladesh and Pakistan. Mutation breeding has given new morphological markers and several promising lines. These examples illustrate not only the widening of the genetic base of the lentil in South Asia, but also the evolution of a breeding program of an international center and national programs targeted toward specific adaptation. This revised version was published online in July 2006 with corrections to the Cover Date.     

Heritability estimates for winter hardiness in lentil under natural and controlled conditions

Heritability analysis for cold tolerance in lentil was conducted using parental, F₂ and F₃ populations at two locations in Balochistan, Pakistan and one controlled environment in Fort Collins, Colorado, USA. Populations of parental and F₂ families were grown over 2 years (1991‐92 and 1992‐93) at Quetta, Balochistan, Pakistan. In 1992‐93, parental and F₃ families were studied at Quetta, and Kalat, Balochistan, Pakistan. Evaluation for normality using non‐transformed and log‐transformed data failed. Attempts to use analysis of variance were then abandoned in favour of parent‐offspring regression for narrow‐sense heritability. Estimates of narrow‐sense heritability ranged from 0.31±0.06 to 0.71 ± 0.06 under field conditions. Under controlled conditions, the estimated heritability was maximized at 1.00 ± 0.17 using 6‐ to 8‐week‐old lentils. Significant transgressive segregants were found in five of the six populations in the F₃ generation. Transgressive segregants appeared in the controlled F₃ generation but were not observed in field environments. This indicates that cold tolerance is under additive gene control and is environmentally sensitive in gene expression.     

The use of Rapd markers for lentil genetic mapping and the evaluation of distorted F2 segregation

To maximize the extent of polymorphism within a mapping population wide crosses are often made, frequently resulting in distorted segregation. Two parents used in the crosses in this study contained ca 50% from wild lentil genome (Lens culinaris ssp. Orientalis). We investigated the use of random amplified polymorphic DNA (RAPD) in the lentil (Lens culinaris Med.), for genetic mapping and testing for segregation distortion in F₂ populations. In cross 1, 83% of the RAPD markers showed segregation distortion, which was also observed for isozyme and morphological loci. By contrast, in cross 2, there was little (10%) segregation distortion. Out of 390 primers tested, 116 primers (29.7%) yielded 192 polymorphic fragments between parents of cross 2. This polymorphism was confirmed as reproducible. Seventy-eight segregating loci were analyzed for linkage, at a LOD score > 3.0, resulted in 28 RAPD, one RFLP, one morphological and three oligonucleotide markers, which were assigned to 9 linkage groups spanning 206 cM. Clearly, in lentil RAPD markers were valuable for genetic mapping and evaluation of segregation distortion. This revised version was published online in July 2006 with corrections to the Cover Date.     

Application of biotechnology in breeding lentil for resistance to biotic and abiotic stress

Summary Lentil is a self-pollinating diploid (2n = 14 chromosomes) annual cool season legume crop that is produced throughout the world and is highly valued as a high protein food. Several abiotic stresses are important to lentil yields world wide and include drought, heat, salt susceptibility and iron deficiency. The biotic stresses are numerous and include: susceptibility to Ascochyta blight, caused by Ascochyta lentis; Anthracnose, caused by Colletotrichum truncatum; Fusarium wilt, caused by Fusarium oxysporum; Sclerotinia white mold, caused by Sclerotinia sclerotiorum; rust, caused by Uromyces fabae; and numerous aphid transmitted viruses. Lentil is also highly susceptible to several species of Orabanche prevalent in the Mediterranean region, for which there does not appear to be much resistance in the germplasm. Plant breeders and geneticists have addressed these stresses by identifying resistant/tolerant germplasm, determining the genetics involved and the genetic map positions of the resistant genes. To this end progress has been made in mapping the lentil genome and several genetic maps are available that eventually will lead to the development of a consensus map for lentil. Marker density has been limited in the published genetic maps and there is a distinct lack of co-dominant markers that would facilitate comparisons of the available genetic maps and efficient identification of markers closely linked to genes of interest. Molecular breeding of lentil for disease resistance genes using marker assisted selection, particularly for resistance to Ascochyta blight and Anthracnose, is underway in Australia and Canada and promising results have been obtained. Comparative genomics and synteny analyses with closely related legumes promises to further advance the knowledge of the lentil genome and provide lentil breeders with additional genes and selectable markers for use in marker assisted selection. Genomic tools such as macro and micro arrays, reverse genetics and genetic transformation are emerging technologies that may eventually be available for use in lentil crop improvement.     

Variation between and within lentil landraces from Yemen Arab Republic

Summary The variation in 31 landraces of lentil collected from Yemen Arab Republic was partitioned to estimate the variation between populations and the variation between families within populations by studying a total of 568 progenies of randomly selected plants. There was significant variation between populations and between families within populations in seedling pigmentation, time to flower, time to maturity and plant height, but the predominant source of variation was between populations. This variation was unrelated to altitude of collection. The landraces were equally variable across characters. The implications of this analysis of variation on lentil breeding and germplasm collection are discussed.     

Single plant selection for yield in lentil

Summary Individual plant selection for yield in lentil is problematic at a commercial crop density primarily because of inter-plant entanglement by tendrils. Visual plant selection for yield was compared with random selection in the F₅ at three plant densities (66, 133 and 200 seeds/m²) by an evaluation of F₇ progeny yields over two seasons in two populations of lentil. Random plant sampling was as effective as visual plant selection in isolating high-yielding F₇ lines. The plant density of the selection environment did not affect the response to selection. The correlations between the seed number of selected F₅ plants and the mean yield of their F₇ progenies were r=+–0.26 and –0.06 in two populations, indicating the lack of positive response to plant selection for seed number. The results show that 1) random sampling is the most economic of the methods tested of plant selection for yield and 2) the plant density of the environment for plant selection can be low enough to avoid inter-plant entanglement by tendrils, allowing a focus in plant selection on characters, other than yield, of importance to the breeding program and with a higher heritability than yield.     

Genetics of seed coat colour in lentil

Summary Four grey mottled seed coat colour lentil lines/cultivars were crossed to one brown seed coat colour cultivar. The F₁ hybrids were brown seeded in all the crosses. Segregation pattern for seed coat colour in F₂ and F₃ generations revealed that it is under control of a single dominant gene, which is present in the parent UPL 175 while a recessive gene is responsible for grey mottled seed coat colour in Pant L 406, Pant L 639, LG 120 and Rau 101.     

Salinity Effects on the Growth and Ion Contents of Some Chickpea (Cicer arietinum L.) and Lentil (Lens culinaris Medic.) Varieties

The effect of salinity on seed germination, plant yield parameters, and plant Na, Cl and K concentrations of chickpea and lentil varieties was studied. Results showed that in both crops percentage emergence was significantly reduced by increasing NaCl levels (0–8dSm^?1). From the plant growth studies it was found that differences existed among chickpea and lentil varieties in their response to NaCl application. In chickpea, the variety Mariye showed the comparatively lowest germination percentage and the lowest seedling shoot dry weight in response to salinity and was also among the two varieties which had the lowest relative plant height, shoot and root dry weight and grain yield at maturity. Similarly, variety DZ-10-16-2, which was the second best in germination percentage and the highest in terms of seedling shoot dry weight, also had the highest relative plant height, shoot and root dry weights, and grain yield at maturity. In lentil, however, such relationships were less pronounced. Chloride concentration (mg g^?1) in the plant parts at salt levels other than the control was about 2–5 times that of Na. K concentration in the plants was significantly reduced by increasing NaCl levels. Chickpea was generally more sensitive to NaCl salinity than lentil. While no seeds were produced at salinity levels beyond 2dSm^?1 in chickpea (no seeds were produced at this salt level in the most sensitive variety, Mariye), most lentil varieties could produce some seeds up to the highest level of NaCl application. Overall, varieties R-186 (lentil) and Mariye (chickpea) were the most sensitive of all varieties. On the other hand, lentil variety NEL-2704 and chickpea variety DZ-10-16-2 gave comparatively higher mean relative shoot and root dry weights, and grain yield, thus showing some degree of superiority over the others. The observed variations among the varieties may be useful indications for screening varieties of both crops for salt tolerance.     

Fusarium vascular wilt in lentil: Inheritance and identification of DNA markers for resistance

The inheritance of resistance to lentil (Lens culinaris Medik.) vascular wilt caused by Fusarium oxysporum f.sp. lentis was investigated in a cross between resistant (ILL5588) and susceptible (L692–16-l(s)) lines. F_2:4 progenies and F_6:8, F_6:9 recombinant inbred line (RIL) populations were assessed for their wilt reaction for three seasons in a well-established wilt-sick plot. Resistance to wilt was conditioned by a single dominant gene in the populations studied. The map location of the Fw locus was identified for the first time through linkage to a random amplified polymorphic DNA (RAPD) marker (OPK-15₉₀₀) at 10.8 cM. Two other RAPD markers (OP-BH₈₀₀ and OP-DI5₅₀₀) identified by bulked segregant analysis were associated in the coupling phase with the resistance trait, and another marker (OP-C04₆₅₀) was associated with repulsion. The DNA markers reported here will provide a starting point in marker-assisted selection for vascular wilt resistance in lentil.     

Genetics of allozyme variants and linkage groups in lentil

Summary The genetics of 8 electrophoretically detectable enzymes in lentil was examined. The enzyme systems glutamic-oxaloacetic transaminase, malic enzyme, phosphoglucomutase, alcohol dehydrogenase, 6-phosphogluconate dehydrogenase, shikimic dehydrogenase and isocitrate dehydrogenase were assayed. The allozymes at each of the studied loci behaved in a codominant manner and segregated in the expected Mendelian fashion. Linkage tests between these loci and an additional morphological trait revealed two linkage groups that involved 5 loci, the rest were independent of each other.     

Geographic distribution of variation in quantitative traits in a world lentil collection

Summary In a world lentil collection the distribution of variation amongst accessions from 13 major lentil-producing countries was examined on the basis of nine quantitative morphological characters by discriminant analysis and canonical analysis. Stepwise discriminant analysis revealed major differences between accessions from different countries. Three major regional groups were apparent: 1) a levantine group (Egypt, Jordan, Lebanon and Syria, 2) a more northern group composed of Greece, Iran, Turkey, and USSR, and 3) accessions from India and Ethiopia with strikingly similar quantitative morphological characters. Misclassifications of individuals within groups were frequent. Characters useful in discriminating between accessions from different countries were in descending order of importance: time to maturity, lowest pod height and 100-seed weight. The regional grouping indicates the importance of local adaptation through clusters of associated characters with phenological adaptation to the ecological environment as the major evolutionary force in the species.     

Inheritance of new genetic markers in lentil (Lens Miller)

Summary We report on the inheritance of 11 morphological markers and 17 isozymes in lentil (Lens culinaris). The monogenic inheritance of 11 morphological markers and 11 isozymes is confirmed. The inheritance of six isozymes (Aco-2, Enp, Est-3, Est-4, Lap-3, and Mdh-m) is reported for the first time in lentil. This brings the total number of described genes in lentil to 78. Cases of disturbed segregation were more frequent than expected by chance. It is suggested that disturbed segregation was in most cases caused by linkage with a piece of chromosome that showed preferential elimination in crosses between Lens culinaris ssp. odemensis and other subspecies. The prevalence of disturbed segregation in crosses with Lens culinaris ssp. odemensis could limit the usefulness of this subspecies in genetic and linkage studies.     

Comparison of random bulk population and single-seed-descent methods for lentil breeding

Summary Three lentil (Lens culinaris Medic.) populations were advanced from the F₂ to the F₄ generation by singleseed-descent (SSD) and bulk-population (BP) breeding methods and used to compare the relative efficiency of the methods for maintaining genetic variation and selection opportunities.SSD maintained more genetic variation (% MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0dh9WrFfpC0xh9vqqj-hEeeu0xXdbba9frFj0-OqFf% ea0dXdd9vqaq-JfrVkFHe9pgea0dXdar-Jb9hs0dXdbPYxe9vr0-vr% 0-vqpWqaaeaabaGaaiaacaqabeaadaqaaqaaaOqaaiqbeo8aZzaaja% Waa0baaSqaaiaadEgaaeaacaaIYaaaaaaa!3B04!\[\hat \sigma _g^2 \]) in 15 of 21 comparisons of characters that were made. Genetic variances were significantly higher with SSD for plant height, days to maturity and yield in population 1; height of lowest pod in population 2; and days to blooming, height of lowest pod, plant type, and yield in population 3. SSD-derived populations had 10, 9, and 13% more erect lines in the three populations, respectively, when compared to the same populations advanced by BP. The BP method maintained 14, 2, and 4% more taller types in the three populations, respectively, and 16 and 33% more segregants that carried their pods higher from the ground. This indicated a reduced frequency of short plants with low flowers as a result of natural selection operating within BP against less competitive short types. The SSD method is an efficient cost-saving method of advancing lentil populations and is recommended for lentil breeding.Scientific Paper No. 5478.     

Inheritance of black testa colour in lentil (Lens culinaris Medik.)

The inheritance of testa (seed coat) colour and interaction of cotyledon and testa colours were studied in seven crosses of lentil (Lens culinaris Medik.) involving parents with black, brown, tan or green testa and with orange, yellow or dark green cotyledons. Analysis of F₂ and F₃ seed harvested from F₁ and F₂ plants, respectively, revealed that although black testa is dominant over nonblack testa, its penetrance is not complete since both F₁ plants and heterozygous F₂ plants produced varying proportions of seeds with either black or nonblack testa. The F₂ populations of the crosses between parents with brown and tan, as well as brown and green, testa segregated in the ratio of 3 brown : 1 tan and 3 brown : 1 green, respectively, indicating monogenic dominance of brown testa colour over tan or green. The expression of testa colour was influenced by cotyledon colour when parents with brown or green testa are crossed with those having orange or green cotyledons. Thus F₂ seeds from these crosses with a green testa always had green cotyledons and never orange cotyledons. F₂ seeds from these crosses with a brown testa always had orange cotyledons and never green cotyledons. These results suggest diffusion of a soluble pigment from the cotyledons to the testa. This revised version was published online in July 2006 with corrections to the Cover Date.     

QTL mapping of resistance in lentil (Lens culinaris ssp. culinaris) to ascochyta blight (Ascochyta lentis)

Quantitative trait locus (QTL) analysis of ascochyta blight resistance in lentil was conducted using genomic maps developed from two F₂ populations, viz. ILL5588/ILL7537 and ILL7537/ILL6002. Five QTLs for ascochyta blight resistance were identified by composite interval mapping (CIM) across four linkage groups (LG) in population ILL5588/ILL7537. Three QTLs were identified by CIM in population ILL7537/ILL6002 (two in close proximity on LGI and one on LGII). Two of these coincided with regions identified using multiple interval mapping (MIM) and were shown to be conditioned by dominant and partial dominant gene action. Together, they accounted for approximately 50% of the phenotypic variance of disease severity. Comparison between the two populations revealed a potentially common QTL and several common regions that contained markers significantly associated with resistance. This study demonstrated the transferability of QTLs among populations and identified markers closely linked to the major QTL that may be useful for future marker‐assisted selection for disease resistance.     

Genetic variance of six agronomic characters in three lentil (Lens culinaris Medic) crosses

Summary Three lentil (Lens culinaris Medic) crosses (cross 1 = Chilean × PI 297784; cross 2 = Tekoa × PI 212611; cross 3 = Precoz × PI 212611) were advanced in a hierarchal design from the F₂ generation to the F₅ generation and tested in replicated field experiments. The purpose was to estimate the genotypic variances and covariances for several characters and to partition these into components due to additive, dominance, and additive × additive variances.Additive genetic variance was the major component of variance in cross 2 for all characters, except plant height and seed weight. Unexpectedly, estimates of dominance variance appeared to be high in crosses 1 and 3. Estimates of the additive × additive component seemed very small in all three populations. The dominance variance component estimates were consistently high for plant height in the three crosses and for seed weight in crosses 2 and 3. Where parental means were close, additive variance estimates were often low and in many cases negative. The unexpected high ratio of dominance variance to additive variance indicated that dominance variance was important in the crosses we studied.Cooperative investigations of Agricultural Research Service, US Department of Agriculture and the College of Agriculture, Washington State University, Pullman, Washington 99164, Work sponsored by Fellowship from Univ. of Jordan, Amman, Jordan. Scientific paper no. 5825.Formerly: Department of Agronomy and Soils, Washington State University, Pullman, WA., USA. Presently: Department of Plant Production and Plant Protection, University of Jordan, Amman, Jordan.Statistical Services, Washington State University, Pullman, WA 99164, USA.US Department of Agriculture, Agricultural Research Service, Washington State University, Pullman WA 99164, USA.     

RAPD and SCAR markers for resistance to acochyta blight in lentil

Resistance to ascochyta blight of lentil (Lens culinaris Medikus),caused by the fungus Ascochyta lentis, is determined by a single recessive gene, ral ₂, in the lentil cultivar Indian head. Sixty F₂ individuals from a cross between Eston (susceptible) and Indian head (resistant) lentil were analyzed for the presence of random amplified polymorphic DNA (RAPD) markers linked to the ral ₂gene, using bulked segregant analysis (BSA). Out of 800 decanucleotide primers screened, two produced polymorphic markers that co-segregated with the resistance locus. These two RAPD markers, UBC227₁₂₉₀and OPD-10₈₇₀, flanked and were linked in repulsion phase to the gene ral ₂ at 12 cm and 16 cm, respectively. The RAPD fragments were converted to SCAR markers. The SCAR marker developed from UBC227₁₂₉₀ could not detect any polymorphism between the two parents or in the F₂. The SCAR marker developed from OPD-10₈₇₀ retained its polymorphism. The polymorphic RAPD marker UBC227₁₂₉₀ and the SCAR marker developed from OPD-10₈₇₀ can be used together in a marker assisted selection program for ascochyta blight resistance in lentil. This revised version was published online in July 2006 with corrections to the Cover Date.     

Association of isozyme markers with quantitative trait loci in random single seed descent derived lines of lentil (Lens culinaris Medik.)

Summary Polymorphism at isozyme loci was used to locate factors responsible for variation in quantitative traits of lentil. Eight sets of random single seed descent (RSSD) derived lines were developed by advancing individual F₃ plants of interspecific (L. culinaris Medik. × L. orientalis Boiss.) hybrids to the F₆. The RSSD lines in each of the eight sets differed for alleles at 2–8 isozyme loci. In each set, association of isozyme loci with variation in seven quantitative traits (days to flower, days to mature, plant height, biomass, seed yield, harvest index, seed weight) was determined for each pairwise combination of a quantitative trait with a marker locus. Loci affecting variation in all seven quantitative traits were detected by their association with 14 isozyme markers (Aat-c, Aat-m, Aat-p, Adh-1, Fk, Gal-1, Gal-2, Lap-1, Lap-2, Pgd-p, Pgi, Pgm-c, Pgm-p, Skdh). The known position of 10 the 14 isozyme loci on the lentil genetic map was used to mark the genomic regions for possible location of associated quantitative trait loci (QTL). Detected QTL were found to be located in six of the seven linkage groups on lentil genetic map. Regions of the genome represented by linkage groups, 1, 5 and 7 appeared to affect a greater number of traits than other genomic regions represented by linkage groups 2, 3 and 4. Results indicated that the mean expression of quantitative traits at segregating marker locus classes can be used to locate the genetic factors in lentil which influence the behavior of economically important traits.     

Resistance to bean common mosaic potyvirus strains and its inheritance in some Indian land races of common bean

Evaluation of 397 common bean accessions of diverse origin revealed the presence of effective sources of resistance against bean common mosaic potyvirus strains prevalent in Himachal Pradesh, a north-western Himalayan state of India. 21 accessions viz., KR 7, KR 225, KR 295, KRC 4, KRC 7, KRC 11, KRC 12, KRC 13, KRC 16, KRC 22, Amanda, Black Turtle Soup, Contender, Hans, Great Northern UI 123, Improved Tender Green 40031, Jubila, Kentucky wonder, Monroe, Premier and Sanilac, were found resistant to NL-1n and NL-7n strains. Some of the genotypes showed strain specific resistance. Pattern of inheritance studied in Hans, Contender, KRC 4 and KRC 22 of Indian origin against strain NL-1n revealed that single dominant gene governed the resistance in Hans and Contender whereas single recessive gene determined the resistance in KRC 22 and KRC 4. SCAR marker SW13₆₉₀ amplified a product of 690 bp in Hans and Contender, confirmed the presence of I gene in these cultivars. The implications of above genes in breeding for programme for BCMV resistance are discussed.     

Screening lentil for resistance to fusarium wilt: methodology and sources of resistance

Host-plant resistance is the best means to control the key disease of lentil-vascular wilt, caused by Fusarium oxysporum Schlecht. emend. Snyder & Hansen f.sp. lentis Vasudeva and Srinivasan. Systematic screening for resistance to lentil wilt was initiated in the field in 1993, in a wilt-sick plot in North Syria, with a core collection of 577 germplasm accessions from 33 countries. A subset (88 accessions) of mostly resistant accessions was re-screened in 1994. The most resistant accessions came from Chile, Egypt, India, Iran and Romania. Variation among accessions in the temporal pattern of wilting was analyzed. The limited wilting in resistant accessions followed a linear model through time, whereas the pattern for susceptible accessions was better described with an exponential model. This temporal variation emphasizes the need for repeated scoring during screening for resistance to lentil vascular wilt to identify ‘late-wilters’. To overcome spatial variation in inoculum density, the efficacy of using wilt scores from a systematically-repeated susceptible control in covariate analysis was tested. Covariance analysis significantly improved overall screening by 3% in 1993, but the improvement was non-significant in 1994. The results emphasize the relative uniformity of disease pressure in the wilt-sick plot and suggest that covariance analysis of a systematically arranged control will be of greater benefit in land which is less uniformly infected. This revised version was published online in July 2006 with corrections to the Cover Date.