Exploitation of wild annual Cicer species for widening the gene pool of chickpea cultivars

Introgression of unadapted genes from the wild Cicer species could contribute to the widening of genetic base of important traits such as yield, yield attributes and resistance to major biotic and abiotic stresses. An attempt was made successfully to intercross two wild annual Cicer species with three cultivated chickpea cultivars. Four interspecific cross‐combinations were made, and their true hybridity was ascertained through morphological and molecular markers. These cross‐combinations were also studied for some important quantitative traits under real field conditions. The range, mean and coefficient of variation of agro‐morphological traits were assessed in the parental lines, their F₁ and F₂ generations to determine the extent of variability generated in cultivated chickpea varieties. A high level of heterosis was recorded for number of pods/plant and seed yield/plant in F₁ generation. Three cross‐combinations of ‘Pusa 1103’ × ILWC 46, ‘Pusa 256’ × ILWC 46 and ‘Pusa 256’ × ILWC 239 exhibited substantially higher variability for important yield‐related traits. The present research findings indicate that these wild annual Cicer species can be easily exploited to broaden the genetic base of cultivated gene pool for improving seed yield as well as adaptation.     

Genetic diversity estimates in Cicer using AFLP analysis

Amplified fragment length polymorphism (AFLP) analysis was used to evaluate the genetic variation among cultivated chickpea and wild Cicer relatives. In total, 214 marker loci were assessed, of which 211 were polymorphic (98.6%) across the 95 accessions that represented 17 species of Cicer. The genetic variation within a species was highest in C. pinnatifidum followed by C. reticulatum and lowest in C. macracanthum. Three main species groups were identified by UPGMA clustering using Nei's pair‐wise distance calculations. Group I included the cultivated species C. arietinum, C. reticulatum and C. echinospermum. Within this group, C. reticulatum accessions were clustered closest to the C. arietinum cultivars ‘Lasseter’, ‘Kaniva’ and ‘Bumper’, supporting the hypothesis that C. reticulatum is the most probable progenitor of the cultivated species. Cicer bijugum, C. judaicum and C. pinnatifidum were clustered together creating group II. Group III contained all nine perennial species assessed and two annual species C. yamashitae and C. cuneatum. The genetic distance detected between group I and group III (0.13) was equivalent to the genetic distance detected between group I and group II (the primary and annual tertiary species, respectively; 0.14). This indicated that the perennial tertiary species may be as valuable for increasing variation to incorporate novel germplasm in the cultigen as the annual tertiary species.     

Evaluation of Early Generation Testing in Chickpea1)

Inter-generation correlations between F₂-F₃, F₂-F₄ and F₃-F₄ in six crosses of chickpea were calculated by using individual plant/progeny means. In general, correlation values in case of plant height, seeds per pod and 100-seed weight were higher between F₃ and F₄ than those between F₂ and F₃ and F₂ and F₄. However, inter-generation correlations were mostly non-significant in case of pods per plant and grain yield. These results clearly show that selection in F₃ can be fruitful for seeds per pod and 100-seed weight. However, selection for pods per plant and grain yield in early generations may not show any relationship with later generation performance for these traits.     

Transferability of Chickpea Selection Indices from Normal to Drought-prone Growing Conditions in a Mediterranean Environment*

Chickpea (Cicer arietinum L.) is normally grown as a spring-sown rainfed crop in the Mediterranean region in areas receiving more than 400 mm of rainfall annually. Because of large fluctuations in the total amount and distribution of the rainfall, the crop occasionally suffers from extreme terminal drought. Breeders face a dilemma in selecting materials for both normal moisture regimes and drought conditions because it is often difficult to conduct two parallel breeding programmes. This study examines the transferability of the selection indices (developed as a multiple regression of yield on most influential traits) from one environment to another with the objective of selecting an appropriate environment for the development of cultivars adapted to both environments. Experiments were conducted for 3 years (1989-1991) during the spring at two locations with normal moisture regime (hereafter referred to as drought-free) in Syria and Lebanon and one drought-prone location in Syria. Each year, 192-240 newly bred lines were evaluated in replicated trials for seed yield, days to flowering and maturity, plant height and 100-seed weight. Correlation analyses showed that increased seed size, early maturity and reduced plant height at the drought-prone location and early maturity at the drought-free location were of prime importance in increasing seed yield. Regression equations developed to predict seed yield showed that days to flower and maturity accounted from 67-80 % variation in seed yield at the dry location, whereas at the drought-free locations the contribution of days to maturity was smaller except in 1991 in Lebanon. The percent success in the transferability of the selection indices from the drought-free environments to the drought-prone environment was higher than that from the drought-prone environment to the drought-free environment. These results indicate that chickpea breeding material developed under drought-free environments could be useful under drought-prone conditions. Early-maturing lines selected under normal environments would also perform well under drier conditions.     

Pollen grain germination and pollen tube growth following in vivo and in vitro self and interspecific pollinations in annual Cicer species

Summary Germination of pollen grains and growth of pollen tubes were studied to determine the cause of barreness in crosses among annual Cicer species. In vivo and in vitro time-course studies and fluorescent microscopy revealed no pollination incompatibility among the selfs, crosses and reciprocals of C. arietinum L., C. reticulatum Lad. and C. cuneatum Rich. In general, Cicer pollen grains germinated and grew on styles of Cicer species. Pollen tube growth was characterized by irregularly spaced and intermittent callose deposits. Failure of seed formation in interspecific pollinations may be attributed to the slowness of pollen tube growth or collapse of fertilized ovules. In addition to these causes, shortness of stamens and sparsity of pollen grains were responsible for flower drop in natural selfs. Although the number of pollen tubes entering the micropyle in interspecific pollinations was low, it may be possible to grow the fertilized ovules on an artificial medium to obtain F₁ plants.     

Inheritance of resistance to fusarium wilt in chickpea

This preliminary study indicated that the resistance to race 2 of fusarium wilt is controlled by two genes, the first of which must be present in the homozygous recessive form, and the other in the dominant form, whether homozygous or heterozygous for complete resistance. Early wilting results if the other gene is homozygous recessive. Late wilting occurs if both loci are dominant. The existence of differences among chickpea cultivars in the time taken to express the initial symptoms of fusarium wilt were observed.     

Genetic control of seed weight and calcium concentration in chickpea seed

Chickpea, Cicer arietinum L., is a staple protein source in many Asian and Middle Eastern countries. Hence, the mineral content of its seed, especially that of calcium, is of nutritional importance. Calcium is transported through plants and to legume pods almost exclusively via the xylem stream, with Ca accretion in developing seeds resulting primarily from diffusion of Ca from the adjoining pod wall. Thus, for seeds of differing surface‐to‐mass ratios, Ca concentration is expected to correlate inversely with seed weight. The relationship between seed weight and Ca concentration in chickpea seeds was studied using a range of germplasm and derivatives from crosses between types differing in seed Ca concentration. Among the cultivars tested, low seed mass was associated with high Ca concentration. However, the study of hybrid progeny indicated that seed Ca content was mainly determined by genetic factors other than grain weight genes. This finding may assist in future breeding of high nutritional quality chickpea cultivars.     

Inheritance of resistance to Fusarium wilt (race 2) in chickpea

The inheritance of resistance to Fusarium wilt (race 2) of chickpea was studied in a set of three crosses, i.e. ‘WR315’בC104’ (resistant × susceptible), ‘WR315’בK850’ (resistant × tolerant) and ‘K850’בGW5/7’ (tolerant × tolerant) in order to investigate the number of genes involved, their complementation and to find out whether resistant segregants are possible in a cross between two tolerant cultivars. Tests of F₁, F₂ and F₃ generations of these crosses under controlled conditions at ICRISAT, Patancheru, India, indicated involvement of three loci (two recessive and one dominant alleles). The homozygous recessive form at the first two loci conferred resistance whereas susceptibility occurred when the first two loci were in the dominant form. A dominant allele at the third locus can complement the dominant alleles at the other two loci to confer tolerance. Occurrence of resistant segregants in a cross between two tolerant cultivars was observed.     

Inheritance of growth vigour and its association with other characters in chickpea

Growth vigour plays an important role in the establishment of a normal crop. The F₂ population of a cross between high‐ and low‐growth vigour varieties of chickpea segregated into 15 high : 1 low growth vigour. The results for recombinant inbred lines and BC1P₂ showed a good fit to the expected 3 : 1 ratio. The results indicated that growth vigour is controlled by two genes with duplicate dominant epistasis. No gene has so far been identified for growth vigour in chickpea. Correlation between growth vigour and other characters showed that high growth vigour had significant negative correlation with days to first flower, days to 50% flowering, days to first pod and days to maturity.     

Genotypic Variation in Root Systems of Chickpea (Cicer arietinum L.) across Environments

Root systems of various chickpea genotypes were studied over time and in diverse environments, – varying in soil bulk density, phosphorus (P) levels and moisture regimes. In a pot study comparing a range of chickpea genotypes, ICC 4958 and ICCV 94916‐4 produced higher root length density (RLD) and root dry weight (RDW), which were better expressed under P stress conditions. In two field experiments in soils of intermediate and high soil bulk densities, ICC 4958 also had greater RLD and RDW, particularly under soil moisture stress conditions. The expression of greater rooting ability of ICC 4958 under a wide range of environmental conditions confirms its suitability as a parent for genetically enhancing drought resistance and P acquisition ability. The superiority of ICC 4958 over other genotypes was for root proliferation expressed through RLD. Thus, the variation in RLD can be the most relevant root trait that reflects chickpea's potential for soil moisture or P acquisition.     

Genetic Variations and Adaptations in Chickpea

A total of 329 chickpea lines, comprising 130 kabiili and 199 desi types, originating from six regions, viz., Indian sub-continent, Middle East, North Africa, America, Europe and USSR, were sown in augmented design. The data recorded on agronomic characters on all entries from each region were evaluated in terms of group (region) means. Among the six regions, greater plant height and lateness in flowering were specific to USSR entries. Group means of USSR accessions, both desi and kabuli, differed significantly from those of Indian varieties for a majority of the characters. These observations indicate that the degree of expression of certain characters could be ascribed to specific areas, which led to area-specific adaptations. Furthermore, type-specific differentiating characters, differing in degree of expression, were observed in desi and kabuli types. For example, high mean values of characters, like branches, pods, grain yield and harvest index, were associated with desi types, and 100-seed weight with kabulis. History of cultivation and selection for specific purpose has been cited as cause for area-specific adaptations and type-specific differentiating characters.     

Independent selection for seed free tryptophan content and vernalization response in chickpea domestication

Chickpea shows a distinct domestication trajectory vis‐a‐vis pod dehiscence and growth cycle mediated by vernalization insensitivity compared with its companion Near Eastern legumes. Our objectives were: (i) to map the quantitative trait loci (QTLs) associated with vernalization response and seed free tryptophan in domesticated × wild chickpea progeny and (ii) estimate the genetic correlation between vernalization response and free tryptophan content. A domesticated × wild chickpea cross was used to document phenotypic segregation in both traits and to construct a skeletal genetic map for QTL detection. A number of vernalization response and seed free tryptophan content QTLs were documented in both F₂ and F₃ generations. No significant genetic correlation between these two traits was observed. Epistatic relationship between two free tryptophan loci was documented. It is evident that selection for high seed tryptophan is easier to accomplish relative to selection for vernalization insensitivity. This suggests that the two traits were selected independently in antiquity, thereby corroborating earlier claims for conscious selection processes associated with chickpea domestication.     

On improving crossing success in chickpea

Artificial hybridization in chickpea (Cicer arietinum L.) is a tedious operation with known low rate of success. A study conducted at the University of Saskatchewan showed that crossing success was influenced by parental identity in this crop. Percent pod set to the total flowers pollinated in chickpea crosses was significantly increased when the smaller‐seeded parent was used as the female. Also, success rate increased with the size of the leaf subtending female parent's flowers at the time of crossing.     

A gene producing one to nine flowers per flowering node in chickpea

Chickpea (Cicer arietinum L.) has a racemose type of inflorescence and at each axis of the raceme usually one or two and rarely three flowers are borne. Plants producing 3 to 9 flowers, arranged in acymose inflorescence, at many axis of the raceme, were identified in F₂ of an interspecific cross ICC 5783 (C. arietinum) × ICCW 9 (C. reticulatum)in which both the parents involved were single-flowered. A spontaneous mutation in one of the two parents or in the F₁was suspected. However, the possibility for establishment of a rare recombination of two interacting recessive genes could not be ruled out. The number of pods set varied from 0 to 5 in each cyme. Inheritance studies indicated that a single recessive gene, designated cym, is responsible for cymose inflorescence. The allelic relationship of cym with sfl, a gene for double-flowered trait, was studied from a cross involving multi flowered plants and the double-flowered line ICC 4929. Thecym gene was not allelic to sfl, suggesting that two loci control the number of flowers per peduncle in chickpea. The cym locus segregated independently of the locus sfl, ifc (inhibitor of flower color) and blv (bronze leave). This revised version was published online in August 2006 with corrections to the Cover Date.     

Screening of Chickpeas for Adaptation to Autumn Sowing

Dry matter accumulation was determined in 27 chickpea (Cicer arietinum) lines in time‐of‐sowing field trials and in controlled‐environment chambers at day/night temperatures of 13/5, 18/8 and 23/13 °C to assess tolerance to growth‐inhibiting temperatures. Field trials were based at Narrabri, NSW, Australia, in a region of summer‐dominant rainfall where winter crops are grown on stored soil moisture. Percentage emergence was lower than expected in some field trials and in the coolest controlled environment. Subsequent dry matter accumulation showed the effects of poor crop establishment until the onset of flowering. Kabuli types were more susceptible to poor emergence than desi types. Different lines yielded the greatest dry matter production at different stages of growth. In the seedling phase, to 30 days after emergence, kabuli accessions SP1.563 and Garnet showed significantly greater dry matter accumulation than all other accessions in all controlled environments, suggesting broad adaptation. One desi accession, Gully, was almost as productive as these two kabuli accessions in the intermediate environment but was much poorer in the other environments, indicating very narrow adaptation. In the vegetative phase, the greatest relative growth rates were found in the desi accessions. Line 940‐26 was identified as highly productive in both field and controlled‐environment experiments. Dry matter accumulation was not significantly affected by temperature, although it was slightly greater in the coolest controlled environment than in the other two. The accession by temperature interaction was not significant, showing that the breadth of adaptation was similar in all accessions during this growth phase. The optimum time of sowing for dry matter accumulation was late May, 4–6 weeks before the winter solstice. The results showed that chickpeas are well adapted to germination and seedling establishment in moderate conditions, followed by vegetative growth in cooler conditions. These conditions are typical following autumn sowing in a Mediterranean or temperate environment. Kabuli types appear to have stronger growth during the seedling phase and desi types during the vegetative phase. Recombination of these traits could lead to more productive cultivars.     

Two genes and linked RAPD markers involved in resistance to Fusarium oxysporum f. sp. Ciceris race 0 in chickpea

The inheritance of resistance to fusarium wilt race 0 of chickpea and linked random amplified polymorphic DNA (RAPD) markers were studied in two F₆:₇ recombinant inbred line (RIL) populations. These RILs were developed from the crosses CA2156 × JG62 (susceptible × resistant) and CA2139 × JG62 (resistant × resistant), and were sown in a field infected with fusarium wilt race 0 in Beja (Tunisia) over 2 years. A1:1 resistant to susceptible ratio was found in the RIL population from the CA2156 × JG62 cross, indicating that a single gene with two alleles controlled resistance. In the second RIL population (CA2139 × JG62) a 3:1 resistant to susceptible ratio indicated that two genes were present and that either gene was sufficient to confer resistance. Linkage analysis showed a RAPD marker, OPJ20600, linked to resistance in both RIL populations, which is present in the resistant parent JG62.     

Inheritance of Dry Root Rot (Rhizoctonia bataticola) Resistance in Chickpea (Cicer arietinum)

The inheritance of resistance to dry root rot of chickpea caused by Rhizoctonia bataticola was studied. Parental F₁ and F₂ populations of two resistant and two susceptible parents, along with 49 F₁ progenies of one of the resistant × susceptible crosses were rested for their reaction to dry root rot using the blotting-paper technique. All F, plants of the resistant × susceptible crosses were resistant; the F₂ generation fitted a 3 resistant: 1 susceptible ratio indicating monogenic inheritance, with resistance dominant over susceptibility. F³ family segregation data confirmed the results. No segregation occurred among the progeny of resistant × resistant and susceptible × susceptible crosses.