The transcriptome of cultivated chickpea (Cicer arietinum L.), an important crop legume, has recently been sequenced. Here, we report sequencing of the transcriptome of wild chickpea, C. reticulatum (PI489777), the progenitor of cultivated chickpea, by GS-FLX 454 technology. The optimized assembly of C. reticulatum transcriptome generated 37 265 transcripts in total with an average length of 946 bp. A total of 4072 simple sequence repeats (SSRs) could be identified in these transcript sequences, of which at least 561 SSRs were polymorphic between C. arietinum and C. reticulatum. In addition, a total of 36 446 single-nucleotide polymorphisms (SNPs) were identified after optimization of probability score, quality score, read depth and consensus base ratio. Several of these SSRs and SNPs could be associated with tissue-specific and transcription factor encoding transcripts. A high proportion (92-94%) of polymorphic SSRs and SNPs identified between the two chickpea species were validated successfully. Further, the estimation of synonymous substitution rates of orthologous transcript pairs suggested that the speciation event for divergence of C. arietinum and C. reticulatum may have happened approximately 0.53 million years ago. The results of our study provide a rich resource for exploiting genetic variations in chickpea for breeding programmes.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1467-7652.2012.00712.x

Transcriptome sequencing of wild chickpea as a rich resource for marker development.

Chickpea is an important grain legume of the semi arid tropics and warm temperate zones, and forms one of the major components of human diet. However, a narrow genetic base of cultivated chickpea (Cicer arietinum L.) has hindered the progress in realizing high yield gains in breeding programs. Furthermore, various abiotic and biotic stresses are the major bottlenecks for increasing chickpea productivity. Systematic collection and evaluation of wild species for useful traits has revealed presence of a diverse gene pool for tolerance to the biotic and abiotic stresses. Relationships among the species of genus Cicer are presented based on crossability, karyotype and molecular markers. The reproductive barriers encountered during interspecific hybridization are also examined. Recent information on genetic linkage maps, comparison of isozymes and different DNA marker systems used for diversity analysis in chickpea germplasm, tagging of genes/QTLs for qualitative and quantitative traits and progress in application of marker assisted selection and genomics in chickpea are presented.

https://www.tandfonline.com/doi/pdf/10.5661/bger-25-267

Chickpea Improvement: Role of Wild Species and Genetic Markers

Random amplified polymorphic DNA markers were used to distinguish between nine different Cicer taxa representing the cultivated chickpea and eight other related annual wild species. Of the 75 random10-mer primers tested, only 8 amplified genomic DNA across all the species. A total of 115 reproducibly scorable RAPD markers were generated, all except 1 polymorphic, and these were utilized to deduce genetic relationships among the annual Cicer species. Four distinct clusters were observed and represented C. arietinum, C. reticulatum and C. echinospermum in first cluster followed by C. chorassanicum and C. yamashitae in the second cluster, while C. pinnatifidum, C. judaicum and C. bijugum formed the third cluster. Cicer cuneatum did not cluster with any of the species and was most distantly placed from the cultivated species. Except for the placement of C. chorassanicum and C. yamashitae, deduced species’ relationships agreed with previous studies. In addition, species-diagnostic amplification products specific to all the nine species were identified. The results clearly demonstrate a methodology based on random-primed DNA amplification that can be used for studying Cicer phylogeny and chickpea improvement.

Random amplified polymorphic DNA (RAPD) analysis reveals genetic relationships among the annual Cicer species

Efforts to improve the yield and quality of cultivated chickpea (Cicer arietinum L.) are constrained by a low level of intraspecific genetic diversity. Increased genetic diversity can be achieved via the hybridisation of the cultivated species with the unimproved 'wild' relatives from within the 43 species of the Cicer genus. To date, the 8 species sharing an annual growth habit and chromosome number with C. arietinum have been the primary focus of screening and introgression efforts. Screening of these species has uncovered morphological characteristics and resistance to a number of abiotic and biotic stresses that are of potential value to chickpea improvement programs. Detailed analysis of protein and DNA, karyotyping, and crossability studies have begun to elucidate the relationships between the annual Cicer species. In comparison, perennial species have received little attention due to difficulties in collection, propagation, and evaluation. This review discusses the progress towards an understanding of genetic relationships between the Cicer species, and the introgression of genes from the wild Cicer species into the cultivated species.

Utilisation of wild Cicer in chickpea improvement — progress, constraints, and prospects

Three molecular markers, including start codon targeted (SCoT) polymorphism, directed amplification of minisatellite-region DNA polymerase chain reaction (DAMD-PCR), and inter simple sequence repeat (ISSR) markers, were compared in terms of their informativeness and efficiency for analysis of genetic relationships among 38 accessions of eight annual Cicer species The results were as follows: (1) the highest level of detected polymorphism was observed for all three marker types; (2) the rate of diversity for the three marker techniques was approximately equal, and the correlation coefficients of similarity were statistically significant for all three marker systems; (3) the three molecular markers showed relatively similar phylogenetic grouping for examined species Diversity analysis showed that Cicer reticulatum is the closest wild species to the cultivated chickpea, and this finding supports the hypothesis that C reticulatum is the most probable progenitor of the cultivated species C bijugum, C judaicum, and C pinnatifidum were clustered together, and in other clusters C yamashitae and C cuneatum were grouped close together To our knowledge, this is the first detailed comparison of performance among two targeted DNA region molecular markers (SCoT and DAMD-PCR) and the ISSR technique on a set of samples of Cicer The results provide guidance for future efficient use of these molecular methods in genetic analysis of Cicer

Comparison of genetic variation and differentiation among annual Cicer species using start codon targeted (SCoT) polymorphism, DAMD-PCR, and ISSR markers

Somatic karyotypes of the nine annual species of Cicer (2n = 16) were analyzed using C-banding. Highly significant differences in haploid genome length and C-band positive heterochromatin content were observed. The haploid genome length ranged from 20.0 μm in the wild species C. judaicum to 28.7 μm in the cultivated species C. arietinum, and significant differences for this character were observed between accessions within several species. Based on their heterochromatin content, the species were divided into two groups: low heterochromatin content (average of 41.7%), which included C. arietinum, C. chorassanicum, C. echinospermum, C. judaicum, C. pinnatifidum, C. reticulatum, and C. yamashitae, and high heterochromatin content (average of 59.5%), which included C. bijugum and C. cuneatum. Within-group variation for heterochromatin content was insignificant, while differences between groups were highly significant. There seemed to be a trend for reduction in C-heterochromatin content in the course of evolution in Cicer. In all species studied, C-bands were located proximally around the centromere with occasional bands in intercalary and distal positions. C-banding patterns allowed for chromosome identification and matching pairs of homologues in all species analyzed.

A. J. Lukaszewski

Papers

Chromosome banding patterns in the annual species of Cicer.