The present study was conducted to find out an in vitro efficient method for multiple shoot regeneration of two local chickpea varieties. The mature embryos were excised of two chickpea varieties i.e. Bittle-98 and Dasht-2000 (with cotyledon and without cotyledon) used as explants. The explants were cultured on Murashige and Skoog (MS) medium supplemented with three concentrations of (2, 3, 4 mg/l) 6-benzylaminopurine (BAP) using explant with and without cotyledon. Further, 0.5 mg/l ?-naphthaleneacetic acid (NAA) along with varied concentrations of BAP (2, 3, 4 mg/ l) was also tested using explant with cotyledon. 3 mg/l BAP alone and 3 mg/l BAP with 0.5 mg/l NAA were found the most effective cytokinin in multiple shoot induction in both tested varieties. Bittle-98 and Dasht-2000 showed 82% and 76% elongation in shoots induction with 0.2 mg/l Indole-3-acetic acid (IAA). Root formation was recorded 80% and 60% in Bittle-98 and Dasht-2000 with 1.0 mg/l Indole-3-butyric acid (IBA). Whereas, recorded root formation was 40 and 20% in Bittle-98 and Dasht-2000 with 1.0 mg/l NAA. The best response for rooting observed in Bittle-98 as its roots were thick, long and strong. Plantlets of Bittle-98 were acclimatized in solid medium for 7-14 days. The successful invitro regeneration of Bittle-98 was observed, when excised embryo with cotyledon were used as explant, made it valuable for genetic transformation.
___
Aasim M, Day S, Rezaei F, Hajyzadeh M. 2013. Multiple shoots regeneration of plumular apices of chickpea. Turk. J. Agric. For., 37:33-39.
Aasim M, Day S, Rezael F, Hajyzadeh M, Mahmud ST. Ozcan, S. 2013b. In vitro shoot regeneration from preconditioned explants of chickpea (Cicer arietinum L.) cv. Gokce. . Afr. J. Biotechnol., 10: 2020-2023.
Barpete S, Khawar, KM, Ozcan S. 2014. Differential competence for in vitro adventitous rooting of grass pea (Lathyrus sativus L.). Plant Cell, Tissue and Organ Culture (PCTOC), 119: 39-50.
Chahal, GS, Gosal, SS. 2002. Principles and procedures of plant breeding: biotechnological and conventional approaches. Alpha Science Int'l Ltd.
Chakraborti D, Sarkar A, Das S. 2006. Efficient and rapid in vitro plant regeneration system for Indian cultivars of chickpea (CicerarietinumL.) Pl. Cell, Tissue and Organ Culture, 86: 117- 123.
Daryanto S, Wang L, Jacinthe PA. 2015. Global synthesis of drought effects on food legume production. PloS one, 10(6). Fatima Z, Aslam M, Bano A. 2008. Chickpea nitrogen fixation increases production of subsequent wheat in rainfed system. Pak. J. Bot., 40: 369-376.
Gaur PM, Tripathi S, Gowda CLL, Ranga Rao GV, Sharma HC, Pande S, Sharma M. 2010. Chickpea Seed Production Manual. Patancheru 502 324, Andhra Pradesh, India: International Crops Research Institute for the Semi-Arid Tropics. 28 pp.
Geetha N, Venkatachalam P, Prakash V, Sita LG. 1998. High frequency induction of multiple shoots and plant regeneration from seedling explants of pigeonpea (Cajanus cajan L.). Curr. Sci., 75: 1036-1041.
Ghanti SK, Sujata KG, Rao MS, Kishor PK. 2010. Direct somatic embryogenesis and plant regeneration from immature explants of chickpea. Biologia Plantarum, 54: 121-125.
Ignacimuthu S, Prakash S. 2006. Agrobacterium mediated transformation of chickpea with ?-amylase inhibitor gene for insect resistance. J. Biol. Sci., 31: 339-345.
Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Pl., 15: 473- 497.
Paul V, Polisetty R, Chandra , Suresh K. 2008. Age of seedling explant and regeneration potential in chickpea (Cicerarietinum L.).Phytomorphol., 58: 41-48.
Pawar BD, Jadhav AS, Kale AA, Chimote VP, Pawar SV. 2012. Thidiazuron improves in vitro multiple shoot induction in chickpea (Cicer arietinum L.) cotyledon with embryonic axis. Journal of Food Legumes, 25: 9-13.
Reinert J., Bajaj, YPS. 1977. Applied and Fundamental Aspects of Plant Cell, Tissue and Organ Cultures, Springer-Verlag, Berlin (West).
Sarker RH, Tarannum F, Hoque MI. 2005. In vitro direct regeneration of three indigenous chickpea (CicerarietinumL.) varieties of Bangladesh. Pl. Tissue Cult.Biotech., 15: 135-144.
Singh R, Jat RS. Sahoo PD. 2002. Thidiazuron induced multiple shoot formation in chickpea (Cicerarietinum L). J. Pl. Biochem. Biotech., 11: 129-131.
Somers DA, Samac DA, Olhoft PM. 2003. Recent advances in legume transformation. Plant Physiol., 131: 892-899.
Stat Soft INC. 1995.Statistica for Windows (Computer program manual). Stat soft Inc., Tulsa, OK
Sujatha G, Jayabalan N, Kumari BDR. 2007. Rapid in vitro micropropagation of Cicerarietinum L. Hort. Sci., 34: 1-5.
Tripathi L, Singh AK, Singh S, Singh R, Chaudhary S, Sanyal I, Amla DV. 2013. Optimization of regeneration and Agrobacterium-mediated transformation of immature cotyledons of chickpea (Cicer arietinum L.). Plant Cell, Tissue and Organ Culture (PCTOC), 113(3), 513-527.
Yousefiara M, Bagheri A, Moshtaghi N. 2008. Optimizing regeneration condition in chickpea (Cicerarietinum L.). Pak. J. Biol. Sci., 11: 1009-1014.