Srinivasan Samineni

Fast-track introgression of “QTL-hotspot” for root traits and other drought tolerance traits in JG 11, an elite and leading variety of chickpea

A “QTL‐hotspot” containing quantitative trait loci (QTL) for several root and drought tolerance traits was transferred through marker‐assisted backcrossing into JG 11, a leading variety of chickpea (Cicer arietinum L.) in India from the donor parent ICC 4958. Foreground selection with up to three simple sequence repeat markers, namely TAA170, ICCM0249, and STMS11, and background selection with up to 10 amplified fragment length polymorphism primer combinations was undertaken. After undertaking three backcrosses with foreground and background selection and selfing, 29 BC3F2 plants homozygous for two markers (ICCM0249 and TAA170) were selected and referred as introgression lines (ILs). Root trait phenotyping of these ILs showed higher rooting depth (RDp) (average 115.21 ± 2.24 cm) in all 29 ILs, better root length density (RLD) (average 0.41 ± 0.02 cm cm−3) in 26 ILs, and higher root dry weight (RDW) (average 1.25 ± 0.08 g per cylinder) as compared to the recurrent parent, JG 11 (111.70 cm for RDp, 0.39 cm cm−3 for RLD, and 1.10 g per cylinder for RDW), as well as the donor parent, ICC 4958 (114.20 cm for RDp, 0.45 cm cm−3 for RLD, and 1.25 g per cylinder for RDW). These ILs, developed in 3 yr, after multilocation field trials may be released as improved variety with enhanced drought tolerance.

Allelic relationships of flowering time genes in chickpea

Flowering time and crop duration are the most important traits for adaptation of chickpea (Cicer arietinum L.) to different agro-climatic conditions. Early flowering and early maturity enhance adaptation of chickpea to short season environments. This study was conducted to establish allelic relationships of the early flowering genes of ICC 16641, ICC 16644 and ICCV 96029 with three known early flowering genes, efl-1 (ICCV 2), ppd or efl-2 (ICC 5810), and efl-3 (BGD 132). In all cases, late flowering was dominant to early-flowering. The results indicated that the efl-1 gene identified from ICCV 2 was also present in ICCV 96029, which has ICCV 2 as one of the parents in its pedigree. ICC 16641 and ICC 16644 had a common early flowering gene which was not allelic to other reported early flowering genes. The new early flowering gene was designated efl-4. In most of the crosses, days to flowering was positively correlated with days to maturity, number of pods per plant, number of seeds per plant and seed yield per plant and negatively correlated or had no correlation with 100-seed weight. The double-pod trait improved grain yield per plant in the crosses where it delayed maturity. The information on allelic relationships of early flowering genes and their effects on yield and yield components will be useful in chickpea breeding for desired phenology.

Recent breeding programs enhanced genetic diversity in both desi and kabuli varieties of chickpea ( Cicer arietinum L.)

In order to understand the impact of breeding on genetic diversity and gain insights into temporal trends in diversity in chickpea, a set of 100 chickpea varieties released in 14 countries between 1948 and 2012 were re-sequenced. For analysis, the re-sequencing data for 29 varieties available from an earlier study was also included. Copy number variations and presence absence variations identified in the present study have potential to drive phenotypic variations for trait improvement. Re-sequencing of a large number of varieties has provided opportunities to inspect the genetic and genomic changes reflecting the history of breeding, which we consider as breeding signatures and the selected loci may provide targets for crop improvement. Our study also reports enhanced diversity in both desi and kabuli varieties as a result of recent chickpea breeding efforts. The current study will aid the explicit efforts to breed for local adaptation in the context of anticipated climate changes.

Plant growth-promotion and biofortification of chickpea and pigeonpea through inoculation of biocontrol potential bacteria, isolated from organic soils

Seven strains of bacteria [Pseudomonas plecoglossicida SRI-156, Brevibacterium antiquum SRI-158, Bacillus altitudinis SRI-178, Enterobacter ludwigii SRI-211, E. ludwigii SRI-229, Acinetobacter tandoii SRI-305 and Pseudomonas monteilii SRI-360; demonstrated previously for control of charcoal rot disease in sorghum and plant growth-promotion (PGP) in rice] were evaluated for their PGP and biofortification traits in chickpea and pigeonpea under field conditions. When treated on seed, the seven selected bacteria significantly enhanced the shoot height and root length of both chickpea and pigeonpea over the un-inoculated control. Under field conditions, in both chickpea and pigeonpea, the plots inoculated with test bacteria enhanced the nodule number, nodule weight, root and shoot weights, pod number, pod weight, leaf weight, leaf area and grain yield over the un-inoculated control plots. Among the seven bacteria, SRI-229 was found to significantly and consistently enhance all the studied PGP and yield traits including nodule number (24 and 36%), nodule weight (11 and 44%), shoot weight (22 and 20%), root weight (23 and 16%) and grain yield (19 and 26%) for both chickpea and pigeonpea, respectively. When the harvested grains were evaluated for their mineral contents, iron (up to 18 and 12%), zinc (up to 23 and 5%), copper (up to 19 and 8%), manganese (up to 2 and 39%) and calcium (up to 22 and 11%) contents in chickpea and pigeonpea, respectively, were found enhanced in test bacteria inoculated plots over the un-inoculated control plots. This study further confirms that the selected bacterial isolates not only have the potential for PGP in cereals and legumes but also have the potential for biofortification of mineral nutrients.

Advances in Chickpea Genomics

Chickpea (Cicer arietinum L.), the second largest consumed pulse crop of the world after common bean, is grown in over 50 countries and traded across 140 countries. After several decades of slow progress, the recent years have witnessed spectacular progress in development of genetic (mapping populations) and genomic resources (structural and functional molecular markers, integrated genetic map and mapping of genes/quantitative trait loci; QTLs) in this crop. QTLs associated with traits of interest including resistance against wilt, Ascochyta blight, Botrytis grey mould and rust; tolerance against salinity and drought and agronomic traits have been identified and validated. A more than 30 genetic linkage maps available in this crop are useful resources for genetic analysis and marker assisted breeding. Genomic tools like bacterial artificial chromosome (BAC) libraries, expressed sequence tags (ESTs) and targeting induced local lesions in genome (TILLING) mutants have been developed in chickpea to facilitate the genome sequencing efforts in this crop. A major landmark in chickpea genomics has been the publication of 738 Mb draft whole genome sequence assembly of a kabuli variety, CDC Frontier. Now, chickpea is one of the most advanced grain legumes in terms of availability of genomic resources. Efforts have already begun on application of these genomics resources in chickpea improvement. This book chapter provides an update on the development of genetic and genomic resources for chickpea and their current and potential uses in chickpea improvement.

Molecular Mapping of Flowering Time Major Genes and QTLs in Chickpea (Cicer arietinum L.)

Flowering time is an important trait for adaptation and productivity of chickpea in the arid and the semi-arid environments. This study was conducted for molecular mapping of genes/quantitative trait loci (QTLs) controlling flowering time in chickpea using F2 populations derived from four crosses (ICCV 96029 × CDC Frontier, ICC 5810 × CDC Frontier, BGD 132 × CDC Frontier and ICC 16641 × CDC Frontier). Genetic studies revealed monogenic control of flowering time in the crosses ICCV 96029 × CDC Frontier, BGD 132 × CDC Frontier and ICC 16641 × CDC Frontier, while digenic control with complementary gene action in ICC 5810 × CDC Frontier. The intraspecific genetic maps developed from these crosses consisted 75, 75, 68 and 67 markers spanning 248.8 cM, 331.4 cM, 311.1 cM and 385.1 cM, respectively. A consensus map spanning 363.8 cM with 109 loci was constructed by integrating four genetic maps. Major QTLs corresponding to flowering time genes efl-1 from ICCV 96029, efl-3 from BGD 132 and efl-4 from ICC 16641 were mapped on CaLG04, CaLG08 and CaLG06, respectively. The QTLs and linked markers identified in this study can be used in marker-assisted breeding for developing early maturing chickpea.

Estimation of genetic components of variation for salt tolerance in chickpea using the generation mean analysis

Chickpea (Cicer arietinum L.) is known to be salt-sensitive and in many regions of the world its yields are restricted by salinity. Recent identification of large variation in chickpea yield under salinity, if genetically controlled, offers an opportunity to develop cultivars with improved salt tolerance. Two chickpea land races, ICC 6263 (salt sensitive) and ICC 1431 (salt tolerant), were inter-crossed to study gene action involved in different agronomic traits under saline and control conditions. The generation mean analysis in six populations, viz. P1, P2, F1, F2, BC1P1 and BC1P2, revealed significant gene interactions for days to flowering, days to maturity, and stem Na and K concentrations in control and saline treatments, as well as for 100-seed weight under salinity. Seed yield, pods per plant, seeds per plant, and stem Cl concentration were controlled by additive effects under saline conditions. Broad-sense heritability values (>0.5) for most traits were generally higher in saline than in control conditions, whereas the narrow-sense heritability values for yield traits, and stem Na and K concentrations, were lower in saline than control conditions. The influence of the sensitive parent was higher on the expression of different traits; the additive and dominant genes acted in opposite directions which led to lower heritability estimates in early generations. These results indicate that selection for yield under salinity would be more effective in later filial generations after gene fixation.

Molecular Mapping of QTLs for Heat Tolerance in Chickpea

Chickpea (Cicer arietinum L.), a cool-season legume, is increasingly affected by heat-stress at reproductive stage due to changes in global climatic conditions and cropping systems. Identifying quantitative trait loci (QTLs) for heat tolerance may facilitate breeding for heat tolerant varieties. The present study was aimed at identifying QTLs associated with heat tolerance in chickpea using 292 F8-9 recombinant inbred lines (RILs) developed from the cross ICC 4567 (heat sensitive) × ICC 15614 (heat tolerant). Phenotyping of RILs was undertaken for two heat-stress (late sown) and one non-stress (normal sown) environments. A genetic map spanning 529.11 cM and comprising 271 genotyping by sequencing (GBS) based single nucleotide polymorphism (SNP) markers was constructed. Composite interval mapping (CIM) analysis revealed two consistent genomic regions harbouring four QTLs each on CaLG05 and CaLG06. Four major QTLs for number of filled pods per plot (FPod), total number of seeds per plot (TS), grain yield per plot (GY) and % pod setting (%PodSet), located in the CaLG05 genomic region, were found to have cumulative phenotypic variation of above 50%. Nineteen pairs of epistatic QTLs showed significant epistatic effect, and non-significant QTL × environment interaction effect, except for harvest index (HI) and biomass (BM). A total of 25 putative candidate genes for heat-stress were identified in the two major genomic regions. This is the first report on QTLs for heat-stress response in chickpea. The markers linked to the above mentioned four major QTLs can facilitate marker-assisted breeding for heat tolerance in chickpea.

Genetic studies for seed size and grain yield traits in kabuli chickpea

Seed size, determined by 100-seed weight, is an important yield component and trade value trait in kabuli chickpea. In the present investigation, the small seeded kabuli genotype ICC 16644 was crossed with four genotypes (JGK 2, KAK 2, KRIPA and ICC 17109) and F1, F2 and F3 populations were developed to study the gene action involved in seed size and other yield attributing traits. Scaling test and joint scaling test revealed the presence of epistasis for days to first flower, days to maturity, plant height, number of pods per plant, number of seeds per plant, number of seeds per pod, biological yield per plant, grain yield per plant and 100-seed weight. Additive, additive × additive and dominance × dominance effects were found to govern days to first flower. Days to maturity and plant height were under the control of both the main as well as interaction effects. Number of seeds per pod was predominantly under the control of additive and additive × additive effects. For grain yield per plant, additive and dominance × dominance effects were significant in the cross ICC 16644 × KAK 2, whereas, additive × additive effects were important in the cross ICC 16644 × JGK 2. Additive, dominance and epistatic effects influenced seed size. The study emphasized the existence of duplicate epistasis for most of the traits. To explore both additive and non-additive gene actions for phenological traits and yield traits, selection in later generations would be more effective.

Botany of Chickpea

Chickpea is one of the important food legumes cultivated in several countries. It originated in the Middle East (area between south-eastern Turkey and adjoining Syria) and spread to European countries in the west to Myanmar in the east. It has several vernacular names in respective countries where it is cultivated or consumed. Taxonomically, chickpea belongs to the monogeneric tribe Cicereae of the family Fabaceae. There are nine annuals and 34 perennial species in the genus Cicer. The cultivated chickpea, Cicer arietinum, is a short annual herb with several growth habits ranging from prostrate to erect. Except the petals of the flower, all the plant parts are covered with glandular and non-glandular hairs. These hairs secrete a characteristic acid mixture which defends the plant against sucking pests. The stem bears primary, secondary and tertiary branches. The latter two branch types have leaves and flowers on them. Though single leaf also exists, compound leaf with 5–7 pairs of leaflets is a regular feature. The typical papilionaceous flower, with one big standard, two wings and two keel petals (boat shaped), has 9 + 1 diadelphous stamens and a stigma with 1–4 ovules. Anthers dehisce a day before the flower opens leading to self-pollination. In four weeks after pollination, pod matures with one to three seeds per pod. There is no dormancy in chickpea seed. Based on the colour of chickpea seed, it is desi type (dark-coloured seed) or kabuli type (beige-coloured seed). Upon sowing, germination takes a week time depending on the soil and moisture conditions.