Pigeonpea Hybrids Research Articles

Development of CGMS Systems in Pigeonpea with Special Reference to A2 Source of Cytoplasm

Background: Exploitation of hybrid vigour is quite possible in cross-pollinated crops. However, pigeonpea is a grain legume crop with a moderate level of cross-pollination (20-70%), with diploid (2n = 2×) chromosome number of 22 and genome size of 1C = 858 Mbp. Outcrossing mainly aided by insect pollinators. Commercialization of CMS based hybrid is constrained because of the labor intensiveness of seed production and concerns about seed purity. In pigeonpea, two dominant genes (Rf1 and Rf2) have been identified and reported by Saxena et al., (2011), which impart fertility restoration to the hybrid plants. The fertility restorer (Rf or Fr) genes in the nucleus suppress the male-sterile phenotype and allow the production of high yielding CGMS-based hybrids. Cytoplasmic male-sterile would effectively circumvent these constraints and revolutionize the hybrid seed industry. CGMS based hybrid breeding can be found more reliable to give productive hybrids in pigeonpea but need some expansion. Methods: In the present study, the objective was to develop cytoplasmic-genetic male-sterile (CGMS) lines in pigeonpea through wide hybridization involving conventional backcrossing. Five CGMS lines viz. BDN 2004-1A, BDN 2004-2A, BDN 2004-3A, BDN 2004-4A and BSMR 736A were developed from Cajanus scarabaeoides cytoplasmic background. This paper discusses development of A and B lines by using A2 CMS systems available for pigeonpea. Result: Our investigations on Development of CGMS systems in Pigeonpea with special reference to A2 source of cytoplasm have allowed us to diverse the male sterile lines based on Cajanus scarabaeoides (A2) cytoplasm in pigeonpea which can be used as marker for easy identification, however characterization of these mentioned five CGMS lines will also help in the predicting the performance of progenies in the different breeding programmes.

Revolutionizing plant breeding: a historical perspective from cross-pollination to gene editing

The history of plant breeding can be broadly divided into four eras: the Pre-Mendelian era (before 1900), the Mendelian era (1900-1920), the post-Mendelian era (1921-1950), and the Modern era (after 1950). Significant milestones include the release of the first hybrid maize varieties in 1961, the first sorghum hybrid (CSH-1) in 1964, the first bajra hybrid (HB-1) in 1965, and the first pigeon pea hybrid (ICPH-8) in 1991. Achieving a world with zero hunger requires a sustainable increase in food production and distribution and elimination of poverty. This goal demands scientific, logistical, and humanitarian approaches to ensure food security from farmers and breeders to policymakers and governments. Conventional breeding techniques alone are insufficient to meet the challenges posed by climate change, biotic and abiotic stress, and the growing global population projected to reach 10 billion by 2050. Novel plant breeding techniques, such as genome editing, speed breeding, and omics technology integration, offer precise, cost-effective, and less time-consuming solutions. These techniques enable the editing of agriculturally significant genes, promoting hybrid seed production, induced apomixis, and resistance to biotic and abiotic stress. Genome editing technologies have evolved, with CRISPR/Cas9 and its variants, base editing (BE), and prime editing (PE) playing pivotal roles in generating transgene-free plants. These advancements have significant implications for crop improvement, meeting food and nutrition security, and catering to regional preferences. Syngenta's initiative to open rights to CRISPR-based technologies through the Shoots by Syngenta platform exemplifies the collaborative efforts needed to drive agricultural innovation and sustainability. The review discusses the current regulatory regimes governing genome-edited crops, prospects of new tools such as DNA-free editing systems and nanotechnology, and their applicability in crop improvement. A multidisciplinary approach involving political, social, economic, and scientific stakeholders is essential for the successful adoption of genome editing technologies, which will ultimately make agriculture a lucrative profession and attract youth to the field.

Comparison of the Agronomic Performance between a Pigeonpea Hybrid and Five Breeding Lines Developed through Conventional Breeding

Pigeonpea [Cajanus cajan (L.) Mill.] may have cross- or self-pollination that allows the use of hybrids or improved genotypes by conventional breeding approaches in pigeonpea production. The objectives of this research were to: 1) compare the agronomic performance of a hybrid vs. breeding lines (BLs) developed by pedigree and bulk methods, and 2) determine if there were average and useful heterosis and heterobeltiosis for quantitative traits. The hybrid 13KLAF1 and UPE-1, UPE-2, UPE-3, UPE-4, UPE-5 BLs, their parents (13KPP-264-05 and ‘Lázaro’), and one early maturity check ‘ICPL 86012’ were evaluated in field trials in Isabela and Lajas, PR. Data of plant height, days to flowering and harvest, seed yield, and weight of 100 dry-seeds were noted for all genotypes. There were no significant differences between 13KLAF1 and UPE BLs for plant height in both locations. ‘Lázaro’ was late maturity in Isabela (157 days) and Lajas (124 days), as expected. The weight of 100 seeds for all genotypes did not reach values more than 18.5 and 16.3 g such as those observed for 13KPP-264-05 in Isabela and Lajas, respectively. The higher seed yields were observed for 13KLAF1 (3112 kg·ha−1), UPE-4 (2970 kg·ha−1), and ‘ICPL 86012’ (2739 kg·ha−1) in Isabela, whereas 13KLAF1 (1599 kg·ha−1) and ‘ICPL 86012’ (1450 kg·ha−1) produced the higher yields in Lajas. The 13KLAF1 showed an average heterosis and heterobeltiosis of 87% and 60%, and 78% and 30% for seed yield in Isabela and Lajas, respectively. The useful heterosis was >125% in both locations.

Growth Response and Economics Variability in Hybrid Pigeonpea to Nitrogen and Phosphorus Fertilization

Aims: To study the growth response and economic variability in hybrid pigeonpea to nitrogen and phosphorus fertilization. Study Design: Factorial randomized block design. Place and Duration of Study: A field experiment was conducted on deep black cotton soils of Regional Agricultural Research Station, Lam during kharif season, 2018-19 in hybrid pigeonpea (ICPH-2740). Methodology: The experiment was laid out in factorial randomized block design (FRBD) comprised of four levels of nitrogen in first factor (N1: 20 kg ha-1, N2: 40 kg ha-1, N3: 60 kg ha-1, N4: 80 kg ha-1) and three levels of phosphorus allotted to second factor (P2O5) (P1: 50 kg ha-1, P2: 75 kg ha-1, P3: 100 kg ha-1). Results: The higher values of grain yield (1900 kg ha-1 & 1833 kg ha-1), net returns (Rs.60522/- & Rs.57181/-) and B:C ratio (1.62 & 1.54) were found with application of nitrogen (60 kg ha-1) and phosphorus (75 kg ha-1), respectively. Conclusion: It was concluded that in Krishna agro-climatic zone of Andhra Pradesh, an application of 60 kg N ha-1 and 75 kg P2O5 ha-1 was applied for getting economically higher grain yield in hybrid pigeonpea.

Pigeonpea hybrid breeding in India

The constantly increasing gap in the demand to supply for pigeon pea has been a matter of concern to pigeon pea researchers in India to increase productivity and production. The entomophily abet cross-pollination behavior of pigeon pea is a desirable crop to develop and establish the hybrid system to exploit the commercial heterosis. Keeping this in view, pigeon pea research was directed towards a new initiative on hybrid pigeon pea breeding at ICRISAT, Hyderabad immediately after the identification of a male sterile line in 1974. Which in turn led to the development of a new GMS hybrid called ICPH 8 in 1991 for cultivation in the central zone. Then five GMS hybrids (PPH 4, CoPH 1, CoPH 2, AKPH 4101, and AKPH 2022) in the early maturing group were released by the state and central varietal release committee. Nevertheless, the GMS-based hybrids did not yield much success due to difficulty in the production of commercial F1 seed. Trifurcation of All India Coordinated Pulses Improvement (AICPIP) and further strengthening of AICPRP on pigeon pea led to the development of national level strategic and basic research vis-àvis provided testing platform for varieties and hybrids. The advertent GMS system led to the development of a stable and economically viable CGMS system in pigeon pea after 26 years of its first GMS initiative. The first CGMS line GT 288A and its maintainer B line was registered by Pulse Research Station, SDAU, GAU, SK Nagar, Gujarat in 2000. Consequently, in 2006 the first CMS hybrid GTH 1 was developed by SDAU, Gujarat, and released by CVRC for cultivation in the central zone. Thirty-nine CGMS lines have been registered with ICAR-NBPGR and four CMS-based hybrids (ICPH 2671, ICPH 2740, IPH 15-03, and IPH 09-5) are released for cultivation. It is to endorse that the joint efforts of the ICAR-NARS and ICRISAT led to the establishment of the hybrid system in pigeon pea by sharing the materials and technology. Nonetheless, supplying quality hybrid seed is a mammoth task to reap the true potential of hybrid technology in pigeon pea.

Hybrid Pigeonpea (ICPH 2740) Nodular Activity and Biological Nitrogen Fixation as Influenced by Agronomic Practices

Background: To explore the impact of agronomic practices on nodulation, nitrogenase activity and biological nitrogen fixation in hybrid pigeonpea, a field investigation was under taken at BW5 block ICRISAT development center (IDC), International Crops Research Institute for Semi-Arid Tropics (ICRISAT), Patancheru, Hyderabad, during kharif, 2021 and 2022. Methods: The seedlings were raised in portrays in advance to onset of monsoon during last week of May (28th May in 2021 kharif and 30th May in 2022 kharif) in both the years by filling portrays with cocopeat and vermicompost in 1:1 ratio. Transplanting of 24 days and 25 days old seedlings were done on 20th June in 2021 kharif and 23th June in 2022 kharif, respectively, on the same day of dibbling in the main field at 100 cm × 100 cm ,120 cm × 120 cm square geometry and 150 cm × 60 cm. Nutrient management treatments were imposed one week after sowing. Result: Among the planting methods higher nodule number and nitrogenase activity with transplanting compared to dibbling. 120 cm × 120 cm found higher nodule number plant-1 and nitrogenase activity but biological nitrogen fixation was higher with 100 cm × 100 cm ha-1 basis. With respect on nutrient management practices, control performed well in terms of higher nitrogenase activity and biological nitrogen fixation and on par with integrated approach of 100% soil test based NPK + vermicompost + PSB + seed treatment with Rhizobium. So, with transplanting method of establishment, 100 ´ 100 cm square plant geometry and 100% soil test based NPK + vermicompost + PSB + seed treatment with Rhizobium was recommended agronomic practices for hybrid pigeonpea.

Bio-inoculated Nutrient Management Influence on Soil Nutrient Availability Pattern and Growth of Hybrid Pigeonpea (ICPH 2740) under Establishment Methods and Crop Geometry

Background: A two-year field study was conducted at the ICRISAT research farm during the rainy seasons of 2021 and 2022 to investigate the impact of crop geometry, crop establishment method and sustainable nutrient management practices on nutrient availability pattern and growth of hybrid pigeon pea. Methods: The experiment followed a split-split plot design. The collected data was analysed using radar graph and heat maps for nutrient availability and dry matter respectively. Result: Data revealing that transplanted plots registered higher nutrient availability and proportionate root and total dry matter production at various growth stages. Among plant geometry the root and total dry matter production was higher with 100×100 cm. when considering planting methods, transplanting with a square system of 100×100 cm, combined with an integrated nutrient management approach consisting of 150% (or) 100% soil test based NPK, vermicompost at a rate of 5 t ha-1, phosphate-solubilizing bacteria (PSB) and seed treatment with Rhizobium, resulted in average of 31.5% higher dry matter production over alone inorganic nutrient management practices. Thus, a square geometry of 100×100 cm, along with sustainable integrated nutrient management (100% soil test based NPK, vermicompost at a rate of 5 t ha-1, PSB and seed treatment with Rhizobium), resulted in higher nutrient availability and dry matter production. These findings highlight the importance of careful selection of planting methods, crop geometry and nutrient management practices for maximizing the nutrient mining for production of high dry matter production of hybrid pigeonpea.

Identification of fertility restorers for A2 cytoplasm-based CGMS lines and development of short duration hybrids in pigeonpea (Cajanus cajan (L.) Millsp.)

Identification of fertility restorers for A2 cytoplasm-based CGMS lines and development of short duration hybrids in pigeonpea (Cajanus cajan (L.) Millsp.)

Genetics of fertility restoration in A2‐based cytoplasmic genetic male sterility system in pigeonpea (Cajanus cajan)

AbstractThe three short duration cytoplasmic genetic male sterility (CGMS) hybrids developed using A2 (Cajanus scarabeoides) cytoplasm‐based male sterile lines (CORG 990047A, CORG 990052A and CORG 7A) and the restorer inbred AK 261322 and their segregating populations (F2 and BC1F1) were subjected to the study of inheritance of fertility restoration in pigeonpea. The fertility restoration was studied based on three different criteria, namely, anther colour, pollen grain fertility and pollen grain morphology and staining. The F2 and BC1F1 populations of the three crosses, namely, CORG 990047A × AK 261322, CORG 990052A × AK 261322 and CORG 7A × AK 261322, segregated in the ratio of 3:1 and 1:1, for anther colour (yellow:pale yellow), pollen grain fertility (fertile:sterile) and for pollen grain morphology and staining. The above study confirmed that the trait fertility restoration was controlled by single dominant gene. This finding can be utilized for the identification of potential restorers, which can be further used in the development of CGMS‐based hybrids in pigeonpea.

Genomic Basis of Defining Heterotic Pools in Pigeonpea (Cajanus cajan L.): A Review

Hybrid breeding strategy leading to define heterotic pools expedited execution of hybrid technology for amelioration of yield, quality, wide adaptability and resistance to biotic and abiotic stresses. In case of pigeonpea conventional hybrid breeding programme has yielded good dividends, yet it can be further honed up by understanding the genome wide variations in the hybrid parental lines for defining heterotic pools. Also, combining ability basis of defining heterotic pools offer enormous opportunities for pragmatic exploitation of pigeonpea hybrid technology with high yield advantages realized in farmers’ fields.

Current Status and Way Forward for Scaling up Productivity of Pigeonpea in Madhya Pradesh: A Review

Madhya Pradesh ranks first both in terms of area (25%) and production (32%) of pulses in India. However, it ranks sixth after Uttar Pradesh, Bihar, Haryana, West Bengal and Gujarat in terms of productivity. To realize better yield; recommended production and protection technologies have to be followed. Most of the pigeonpea varieties have been developed by traditional breeding either by pedigree method or selection from landraces or by bi-parental mating. After getting stable male sterile and restorer lines, development of hybrid pigeonpea have been major breeding objectives. Availably of genomic tools by decoding the genome sequence, pigeonpea have been equipped with modern genetic and genomic tools for use in the research programmes. Different approached like mining superior alleles by re-sequencing of wild species, fine mapping for drought tolerance, fertility restoration and developing superior lines by genomic selection, identification of candidate genes which are associated with hybrid vigour by using epi-genomics and mitochondrial genome sequencing and identification of candidate genes and functional markers for water-logging tolerance are already in progress. Transgenic approach has also been initiated to develop resistance against pod borer. Collaborative efforts among ICRISAT, IARI and IIPR-Kanpur are on the way for use of different transgenes and promoter options for developing transgenic plants and their evaluations for effectiveness and bio-safety concerns.

Characterization of heterosis and genomic prediction-based establishment of heterotic patterns for developing better hybrids in pigeonpea.

Whole-genome resequencing (WGRS) of 396 lines, consisting of 104 hybrid parental lines and 292 germplasm lines, were used to study the molecular basis of mid-parent heterosis (MPH) and to identify complementary heterotic patterns in pigeonpea [Cajanus cajan (L.) Millsp.] hybrids. The lines and hybrids were assessed for yield and yield-related traits in multiple environments. Our analysis showed positive MPH values in 78.6% of hybrids, confirming the potential of hybrid breeding in pigeonpea. By using genome-wide prediction and association mapping approaches, we identified 129 single nucleotide polymorphisms and 52 copy number variations with significant heterotic effects and also established a high-yielding heterotic pattern in pigeonpea. In summary, our study highlights the role of WGRS data in the study and use of heterosis in crops where hybrid breeding is expected to boost selection gain in order to ensure global food security.

​Effect of Foliar Spray of Nutrients, Growth Regulators on Seed Yield and Quality in Hybrid Pigeonpea [Cajanus cajan (L.) Millsp

Background: The new generation special fertilizers have been introduced exclusively for foliar feeding and fertilization. The increased supply of nutrients and good response by plants resulted in enhanced translocation of nutrients to reproductive structures.Therefore, it is planned to study the effect of different nutrients as a foliar spray for growth, yield and seed quality parameters of hybrid pigeon pea. Methods: The experiment on effect of foliar spray of nutrients, growth regulators on seed yield, quality in hybrid pegionpea [Cajanus cajan (L.) Millsp] was conducted at Seed Farm, College of Agriculture, Vijayapura, Karnataka during kharif 2019-2020. The treatment consisting of foliar spray of control (T1), 1% Pulse magic (T2), 1% 19:19:19 (T3), 0.2% Borax (T4), 50 ppm NAA (T5), 1% Pulse magic+1%19:19:19 (T6), 1% Pulse magic+0.2% Borax (T7), 1% Pulse magic+50 ppm NAA (T8), 1% Pulse magic+1% 19:19:19+0.2% Borax (T9), 1% Pulse magic+ 1% 19:19:19+50 ppm NAA (T10), 1% Pulse magic+1% 19:19:19+50 ppm NAA+0.2% Borax (T11). Foliar spray was taken at flower initiation and peak flowering stage. Result: The results revealed that, foliar application of Pulse magic @1% +19:19:19@1%+ NAA@50 ppm + Borax@0.2% recorded higher number of branches (10.60), number of pods per plant (239.33), number of seeds per plant (5.75), test weight (12.18g), hybrid seed yield per plant (48.17g), hybrid seed yield per plot (1188.83g), hybrid seed yield per ha (1499kg /ha) and seed quality parameters viz: seed germination (93%), shoot length (15.08 cm), root length (18.67 cm), seedling vigour index(2649), lower electric conductivity (0.59 dSm-1) and protein percentage (22.18 %) respectively.

Enhancement of Rural Income and Nutrition by Cultivating Pigeonpea Hybrids

Pigeonpea [Cajanus cajan (L.) Millsp.] is a favourite crop of rain-fed farmers due to its high food value, drought tolerance and various soil improving properties. The productivity enhancement of this crop has been a long-term goal at the national level but with a little success. In this context, the advent of hybrid breeding technology with over 30% on-farm yield advantages has provided a much-needed breakthrough. The hybrids in pigeonpea were bred using a stable CMS system and natural out-crossing. It is believed that the adoption of locally adapted hybrids would contribute significantly towards both family income and nutrition.

High resolution mapping of restoration of fertility (Rf) by combining large population and high density genetic map in pigeonpea [Cajanus cajan (L.) Millsp

BackgroundRestoration of fertility (Rf) is an important trait for pigeonpea hybrid breeding. Few coarse quantitative trait locus (QTL) studies conducted in the past identified QTLs with large confidence intervals on the genetic map and could not provide any information on possible genes responsible for Rf in pigeonpea. Therefore, a larger population comprising of 369 F2s derived from ICPA 2039 × ICPL 87119 was genotyped with high density Axiom Cajanus SNP Array with 56 K single nucleotide polymorphism (SNPs) for high resolution mapping of Rf.ResultsA genetic map with 4867 markers was developed and a total of four QTLs for Rf were identified. While one major effect QTL (qRf8.1) was co-localized with the QTL identified in two previous studies and its size was refined from 1.2 Mb to 0.41 Mb. Further analysis of qRf8.1 QTL with genome sequence provided 20 genes including two genes namely flowering locus protein T and 2-oxoglutarate/Fe (II)-dependent dioxygenases (2-ODDs) superfamily protein with known function in the restoration of fertility.ConclusionThe qRf8.1 QTL and the potential candidate genes present in this QTL will be valuable for genomics-assisted breeding and identification of causal genes/nucleotides for the restoration of fertility in the hybrid breeding program of pigeonpea.

Genotype × Environment Interaction for Fertility Restoration and Yield Traits in Pigeonpea (Cajanus cajan) Hybrids

In cytoplasmic genetics male-sterility (CGMS)-based hybrid seed production technology, instability of fertility restoration is one of the prime causes behind instable yield performance of hybrids over the wide range of environments which makes better hybrids adaptive to only a specific environment. Therefore, the present study was conducted to investigate the stability of fertility restoration and yield traits of 10 promising CGMS-based pigeonpea hybrids during 2012–2013 at three different environments of India. The results revealed a significant variability among hybrids for fertility restoration and yield traits over the environments. All the hybrids except ICPH 3494 and ICPH 3491 exhibited high (> 80%) pollen fertility and fully male-fertile plants across the environments indicating complete fertility restoration in these hybrids. Hybrids ICPH 2671, ICPH 2740, ICPH 3933, and ICPH 3461 were stable for pollen fertility, fertility restoration, and for days to maturity except ICPH 3461. Hybrids ICPH 2671, ICPH 2751, and ICPH 3461 were stable for number of pods plant−1, whereas ICPH 2671, ICPH 2740, ICPH 3933, ICPH 3461, and ICPH 3762 were stable for grain yield plant−1 with high mean, unit regression coefficient, and non-significant deviation from the regression line. The varying fertility status of hybrids across the environments indicates that fertility restoration is sensitive to environmental conditions.

Application of heterotic grouping, standard heterosis and per se performance in breeding high-yielding inbred and hybrid cultivars of pigeonpea

A set of 102 hybrid combinations, developed by crossing three A4 CMS lines and 34 diverse testers, was evaluated to identify potential parental genotypes for breeding new high yielding cultivars. The hybrid combinations were assessed for their per se performance, standard heterosis and combining ability. Significant differences were recorded for parents versus hybrids for yield and its related traits, suggesting the presence of substantial heterotic responses. Highly significant GCA effects for yield were recorded within the female parents and testers. Relatively more number of hybrids involving ICPA 2092, as female parent, exhibited significant positive SCA values for yield, days to flower, plant height, primary branches, pods/plant, seed size and seeds/pod. A total of 45 hybrids recorded significant SCA effects for productivity. The yield superiority of hybrids over the inbred control cultivar ranged from -50.81 to 76.89%. Interestingly, 23 hybrids were also found superior to the first released pigeonpea hybrid ICPH 2671 by a margin of 5.48% to 64.31%. On the basis of their performance in hybrid combinations, the testers were classified into seven heterotic groups. The present study identified two CMS lines (ICPA 2092 and ICP 2043) and 13 testers including ICP 3525, ICPL 20196, BDN 2001-6 and BSMR 198 etc. as potential parental lines for use in breeding medium duration pigeonpea pure line and hybrid cultivars.

Can pigeonpea hybrids negotiate stresses better than inbred cultivars?

Pigeonpea [Cajanus cajan (L.) Millsp.] is an important rainfed pulse crop of tropics and sub-tropics, and during its long growth cycle of 6–9 months it encounters a number of biotic and abiotic stresses. The recently developed CMS-based pigeonpea hybrids have demonstrated large gains in yield and stability over the traditional inbred cultivars. In this review, the authors argue that the heterosis expressed in traits like seed germination, radicle growth, root biomass production and moisture retention during water stress confers advantages to hybrid plants in negotiating a few abiotic and biotic stresses in much better way than pure line cultivars.

Natural cross-pollination is both a boon and bane for pigeonpea breeders

Natural cross-pollination in pigeonpea, mediated by insects, is a universal phenomenon with the first report appearing in 1919. Considerable information is now available on various aspects of this biological phenomenon including degree of out-crossing and pollinating agents in different parts of the world. A large variation (0-60%) has been recorded in 30 different environments across 11 countries. The role of cross-pollination in rapid deterioration of the genetic purity of cultivars and elite genetic stocks is well understood and documented by breeders and seed producers. Pigeonpea breeders have converted this constraint into an opportunity by way of visually selecting natural hybrids from landraces in farmers’ fields and deriving high yielding cultivars. In a key development, the occurrence of natural cross-pollination in the wild relatives of pigeonpea has also been recorded; and in three wild species such natural hybrids have yielded valuable cytoplasmic male sterility systems. This has encouraged breeders to develop a hybrid breeding technology with natural out-crossing playing a key role in the large-scale seed production of commercial pigeonpea hybrids.

Molecular mapping and inheritance of restoration of fertility (Rf) in A4 hybrid system in pigeonpea (Cajanus cajan (L.) Millsp.)

Key messageWe report molecular mapping and inheritance of restoration of fertility (Rf) in A4 hybrid system in pigeonpea. We have also developed PCR-based markers amenable to low-cost genotyping to identify fertility restorer lines.Commercial hybrids in pigeonpea are based on A4 cytoplasmic male sterility (CMS) system, and their fertility restoration is one of the key prerequisites for breeding. In this context, an effort has been made to understand the genetics and identify quantitative trait loci (QTL) associated with restoration of fertility (Rf). One F2 population was developed by crossing CMS line (ICPA 2039) with fertility restorer line (ICPL 87119). Genetic analysis has shown involvement of two dominant genes in regulation of restoration of fertility. In parallel, the genotyping-by-sequencing (GBS) approach has generated ~ 33 Gb data on the F2 population. GBS data have provided 2457 single nucleotide polymorphism (SNPs) segregating across the mapping population. Based on these genotyping data, a genetic map has been developed with 306 SNPs covering a total length 981.9 cM. Further QTL analysis has provided the region flanked by S8_7664779 and S8_6474381 on CcLG08 harboured major QTL explained up to 28.5% phenotypic variation. Subsequently, sequence information within the major QTLs was compared between the maintainer and the restorer lines. From this sequence information, we have developed two PCR-based markers for identification of restorer lines from non-restorer lines and validated them on parental lines of hybrids as well as on another F2 mapping population. The results obtained in this study are expected to enhance the efficiency of selection for the identification of restorer lines in hybrid breeding and may reduce traditional time-consuming phenotyping activities.