Cajanus cajanifolius, the putative progenitor species of pigeonpea (C. cajan), has many agro-economic traits that deserve attention for introgression into pigeonpea cultivars. However, unidirectional compatibility creates barriers for interspecific hybridization and alien gene transfer between these two species. In the present study, very low percentage of pod setting and F1 seed germination were observed. Thus, attempts were made for the propagation of interspecific F1 hybrids in vitro, and an efficient in vitro propagation protocol has been optimized. Excised cotyledonary explants of F1 hybrids were cultured on both MS and Blaydes’ medium supplemented with NAA, BAP and GA3 at various combinations and concentrations. MS medium with BAP (25 μM) was most suitable for multiple shoot induction. MS medium was found to be superior for both multiple shoot and root induction from cotyledonary explants. Cyto-morphological and SSR marker based analysis confirmed the hybridity of the micro-propagated F1 plantlets. Meiocytes of the regenerated F1 hybrids experienced chromosome heteromorphism during diakinesis leading to laggard and bridge formation at anaphase-I. The present study evidenced the scope of interspecific hybridization between C. cajan and C. cajanifolius aiming at transmission of stress tolerant genes from C. cajanifolius to the cultivated gene pool of pigeonpea for broadening its genetic base.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Blaydes DF. Interaction of kinetin and various inhibitors in the growth of soybean callus. Physiol Plant. 1966;19:748–53.
Bohra A, Dubey A, Saxena RK, Penmetsa RV, Poornima KN, Kumar N, et al. Analysis of BAC-end sequences (BESs) and development of BES-SSR markers for genetic mapping and hybrid purity assessment in pigeonpea (Cajanus spp.). BMC Plant Biol. 2011;11:56.
Dongre AB, Raut MP, Bhandarkar MR, Meshram KJ. Identification and genetic purity testing of cotton F1 hybrids using molecular markers. Indian J Biotechnol. 2011;10:301–6.
Geetha N, Venkatachalam P, Prakash V, Lakhmisita G. High frequency induction of multiple shoots and plant regeneration from seedling explants of pigeonpea (Cajanus cajan L.). Curr Sci. 1998;75:1036–41.
George L, Eapen S. Organogenesis and embryogenesis from diverse explants in pigeonpea (Cajanus cajan L.). Plant Cell Rep. 1994;13:417–20.
Gupta PK, Varshney RK. The development and use of microsatellite markers for genetic analysis and plant breeding with emphasis on bread wheat. Euphytica. 2000;113:163–5.
Gutierrez OA, Basu S, Saha S, Jenkins JN, Shoemaker DB, Cheatham CL, et al. Genetic distances among selected cotton genotypes and its relationship with F2 performance. Crop Sci. 2002;42:1841–7.
Hashemi SH, Mirmohammadi-Maibody MSA, Nematzadeh GA, Arzani A. Identification of rice hybrids using microsatellite and RAPD markers. Afr J Biotechnol. 2009;8:2094–101.
Hipi A, Surahman M, Ilyas S. Seed genetic purity assessment of maize hybrid using microsatellite markers (SSR). Int J Appl Sci Tech. 2013;3:66–71.
Huchett BI, Botha FC. Stability and potential use of RAPD markers in sugarcane genealogy. Euphytica. 1995;86:117–25.
Jha SS, Ohri D. Phylogenetic relationships of Cajanus cajan (L.) Millsp. (Pigeonpea) and its wild relatives based on seed protein profiles. Genet Resour Crop Evol. 1996;43:275–81.
Kassa MT, Penmetsa RV, Carrasquilla-Garcia N, Sarma BK, Datta S, Upadhyaya HD, et al. Genetic patterns of domestication in pigeonpea (Cajanus cajan (L.) Millsp.) and wild Cajanus relatives. PLoS ONE. 2012;7:e39563.
Krishna G, Reddy PS, Ramteke PW, Rambab UP, Tawar KB, Bhattacharya P. Agrobacterium mediated genetic transformation of pigeonpea (Cajanus cajan (L.) Millsp.) for resistance to legume pod borer Helicoverpa armigera. J Crop Sci Biotechnol. 2011;14:197–204.
Kumar AS, Ready TP, Ready GM. Multiple shoots from the cultured explants of pigeonpea and Atylosia species. SABRAO J. 1984;16:101–5.
Kumar MRC, Vishwanath K, Shivakumar N, Prasad SR, Radha BN. Ramegowda. Utilization of SSR markers for seed purity testing in popular rice hybrids (Oryza sativa L.). Anals. Plant Sci. 2012;1:1–5.
Liu S, Cantrell RG, McCartyJr JC, McStewart JD. Simple sequence repeat based assessment of genetic diversity in cotton race stock accessions. Crop Sci. 2000;40:1459–69.
Majumdar S, Banerjee S, De KK. Efficient shoot regeneration in pigeon pea, Cajanus cajan (L) Millisp. using seedling petioles. Curr Sci. 2004;86:30–2.
Mallikarjuna N, Saxena K, Lakshmi J, Varshney R, Srikanth S, Jadhav D. Differences between Cajanus cajan (L.) Millspaugh and C. cajanifolius (Haines) van der Maesen, the progenitor species of pigeonpea. Genet Resour Crop Evol. 2012;59:411–7.
Mehta U, Mohanram HY. Regeneration of plantlets from the cotyledons of Cajanus cajan. Indian J Exp Biol. 1980;18:800–2.
Mishra RR, Sahu A, Rath SC, Mishra SP, Panigrahi J. Cyto-morphological and molecular characterization of Cajanus cajan × C. scarabaeoides F1 hybrid. Nucleus. 2012;55:27–35.
Mishra SK, Panigrahi J. Identification of inter-specific hybrid between Cajanus cajan (L.) and C. cajanifolius (H.) using cyto-morphological and DNA markers. Electron J Plant Breed. 2017;8:63–71.
Mohan ML, Krishnamurthy KV. Plant regeneration in pigeonpea (Cajanus cajan (L.) Millsp.) by organogenesis. Plant Cell Rep. 1998;17:705–10.
Mohanty M, Patnaik SN. Cytomorphological analysis of F1 Hybrids between Cajanus cajan (L.) Millsp. and Atylosia cajanifolia (Haines). Cytologia. 1989;54:121–8.
Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant. 1962;15:473–97.
Nadimpalli RJ, Jarret RL, Pathak SC, Kochert G. Phylogenetic relationships of the pigeonpea (C. Cajan) based on nuclear restriction fragment length polymorphisms. Genome. 1993;36:216–23.
Nayak A, Mohanty M, Patnaik SN. Multiple shoot regeneration in Cajanus cajan and Atylosia cajanifolia. Proc Conf Cyto Genet. 1992;3:127–32.
Odeny DA, Jayashree B, Ferguson M, Hoisington D, Crouch J, Gebhardt C. Development, characterization and utilization of microsatellite markers in pigeonpea. Plant Breed. 2007;126:130–6.
Panigrahi J, Patnaik SN, Kole C. Identification of putative hybrids between pigeon pea (Cajanus cajan) and C. cajanifolius using seed protein markers. Indian. J Genet. 2001;61:367–8.
Parani M, Singh KN, Rangasamy S, Ramalingam RS. Identification of Sesamum alatum × Sesamum indicum hybrid using protein, isozyme and RAPD markers. Indian J Genet. 1997;57:381–8.
Pundir RPS, Singh RB. Cytogenetics of F1 hybrids between Cajanus and Atylosia species and its phylogenetic implications. Theor Appl Genet. 1985;71:216–20.
Rangasmy P, Krishnakumar JC, Vanniarajan C, Ramalingam J. Heterotic vigour in intergenetic hybrids of pigeonpea for protein and methionine content. Crop Improv. 1991;18:152–3.
Shivaprakash N, Pental D, Bhallasarin N. Regeneration of pigeonpea (C. cajan) from cotyledonary node via multiple shoot formation. Plant Cell Rep. 1994;13:623–7.
Singh U, Jambunathan R. Protease inhibitors and in vitro protein digestibility of pigeonpea (Cajanus cajan L.) and its wild relatives. J Food Sci Technol. 1981;18:83–5.
Sivaramakrishnan S, Seetha K, Rao AN, Singh L. RFLP analysis of cytoplasmic male sterile line of pigeonpea (Cajanus cajan L. Millsp.) developed by interspecific crosses. Euphytica. 1997;93:307–12.
Smith JF, Burke CC, Wangner WL. Interspecific hybridization in natural population of Cyrtandra (Gesneriaceae) on the Hawaian islands: evidence from RAPD markers. Plant Syst Evol. 1996;200:61–7.
Subrahmanyam NC, Kumar PS, Faris DG. Genome relationships in pigeonpea and its implication in crop improvement. In: Gupta PK, Bhal JR, editors. Genetics and crop improvement. Meerut: Rastogi and Co; 1986. p. 309–20.
Thiruvengadam V, Muthiah A. Interspecific hybridization between Cajanus cajan and C. cajanifolius. Crop Breed Appl Biotechnol. 2007;7:204–11.
Tripathi SN, Patil BD, Shukla GP. Phylogenic and hybridization potentials in Atylosia and Cajanus species. Forage Res. 1984;10:5–9.
van der Maesen LJG. Pigeonpea: origin, history, evolution and taxonomy. In: Nene YL, Hall SD, Sheila VK, editors. Wallingford, Oxen. UK: CAB International; 1990. p. 15–46.
Yadav PBS, Padmaja V. Shoot organogenesis and plantlet regeneration from leaf segments of pigeonpea. Plant Cell, Tissue Organ Cult. 2003;73:197–200.
The authors are grateful to the International Foundation for Science, Sweden for the financial assistance through the Young Scientist scheme (C-4584-1) to the author (JP), and to the Department of Science and Technology, New Delhi for providing INSPIRE Fellowship (DST/INSPIRE Fellowship/2013/383) to the author (SKM).
About this article
Cite this article
Panda, S., Mishra, S.K., Naik, A. et al. In vitro propagation of Cajanus cajan × C. cajanifolius hybrid and its characterization using cyto-morphological and SSR markers. Nucleus 61, 83–93 (2018). https://doi.org/10.1007/s13237-018-0232-0
- Interspecific hybrid
- In vitro culture
- SSR marker