Surendra BARPETE, Muhammad AASIM, Khalid Mahmood KHAWAR, Sancar Fatih ÖZCAN, Sebahattin ÖZCAN

Preconditioning effect of cytokinins on in vitro multiplication of embryonic node of grass pea (Lathyrus sativus L.) cultivar Gürbüz

Lathyrus sativus L. (grass pea) is an important grain legume crop grown all over the world. It is very recalcitrant and difficult to regenerate under in vitro conditions. The present study determined the effect of preconditioning of cytokinins [10 and 20 mg L-1 thidiazuron (TDZ) and 2-isopentenyl adenine (2iP)] and postconditioning of dissimilar levels of TDZ-indole-3-butyric acid (IBA) and 2iP-IBA on in vitro shoot multiplication using embryonic node explants. The results showed that preconditioning followed by postconditioning on Murashige and Skoog (MS) medium with 0.25, 0.50, 0.75, and 1.0 mg L-1 2iP or TDZ combined with 0.10 mg L-1 IBA had positive effects on shoot multiplication. It seemed as if the TDZ-preconditioned and TDZ-IBA-postconditioned explants were more regenerative compared to 2iP-preconditioned or 2iP-IBA-postconditioned explants. Maximum shoot induction (100%) was achieved on MS medium containing 1 mg L-1 TDZ and 0.10 mg L-1 IBA, with a mean number of 31.10 shoots per explant. In comparison to the other cytokinin tested, 2iP-preconditioned and 2iP-IBA-postconditioned explants had longer shoots, with a mean shoot length of 8.10 cm. Root initiation was observed on all cultures 4 weeks after the transfer of shoots with the best rooting on MS medium containing 2 mg L-1 IBA. The rooting response was 76.56% with a mean number of 8.03 roots per shoot and root length of 8.2 cm. The healthy plants were transferred to a greenhouse for acclimatization and exhibited 90% survival. It was concluded that TDZ preconditioning was necessary for grass pea multiplication along with TDZ-IBA postconditioning. This regeneration protocol may facilitate future genetic transformation and breeding studies to reduce \Beta-N-oxalyl-L-\Alpha,\Beta-di-amino propionic acid content in grass pea.

Anahtar Kelimeler:

Embryonic node, grass pea, Lathyrus sativus L., regeneration

Preconditioning effect of cytokinins on in vitro multiplication of embryonic node of grass pea (Lathyrus sativus L.) cultivar Gürbüz

Keywords:

Embryonic node, grass pea, Lathyrus sativus L., regeneration,

PDF

___

Aasim M (2012). Micropropagation of lentil (Lens culinaris Medik.) using pulse treatment of immature plumular apices. Pak J Agric Sci 49: 149–154.
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, Rezai F, Hajyzadeh M, Mahmud ST, Ozcan S (2011). In vitro shoot regeneration from pre-conditioned explants of chickpea (Cicer arietinum L.) cv. Gokce. African J Biotech 10: 2020–2023.
Aasim M, Khawar KM, Özcan S (2009). In vitro micropropagation from plumular apices of Turkish cowpea (Vigna unguiculata L.) cultivar Akkiz. Sci Horti 122: 468–471.
Aasim M, Khawar KM, Özcan S (2010). Efficient in vitro propagation from pre-conditioned embryonic axes of Turkish cowpea (Vigna unguiculata L.) cultivar Akkiz. Arch Biol Sci 62: 1047–1052.
Anwar F, Sharmila P, Pardha Saradhi P (2008). An optimal protocol for in vitro regeneration, efficient rooting and stable transplantation of chickpea. Physiol Mol Biol Plants 14: 329–335.
Barik DP, Mohapatra U, Chand PK (2005). Transgenic grass pea (Lathyrus sativus L.): factors influencing Agrobacterium-mediated transformation and regeneration. Plant Cell Rep 24: 523–531.
Barpete S, Parmar D, Sharma NC (2010). Callus culture and plantlet formation using axillary shoot explant of grass pea (Lathyrus sativus L.). In: Pandey BN, Singh SP, Singh R, editors. Sustainable Management and Conservation of Biodiversity. New Delhi, India: Narendra Publishing House, pp. 55–68.
Barpete S, Parmar D, Sharma NC, Kumar S (2012). Karyotype analysis in grass pea (Lathyrus sativus L.). J Food Legumes 25: 14–17.
Barpete S, Sharma NC, Parmar D, Dhingra M (2008). In-vitro regeneration of Lathyrus sativus L. Nat J Life Sci 05: 207–210.
Belide S, Sajjalaguddam RR, Paladugu A (2010). Cytokinin preconditioning enhances multiple shoot regeneration in Pongamia pinnata (L.) Pierre - a potential, non-edible tree seed oil source for biodiesel. Electron J Biotechn 13: 1–9.
Chen J, Witham FH, Heuser CW (1995). Inhibition of NAA-induced adventitious roots in mung bean cuttings by kinetin, zeatin, ethidium bromide and other DNA intercalators. World Wide Web J Biol 1: 1–8.
Jana S, Iyyakkannu S, Jeong BR (2013). Effect of cytokinins on in vitro multiplication of Sophora tonkinensis. Asian Pac J Trop Biomed 3: 549–553.
Kumar S, Bejiga G, Ahmed S, Nakkoul H, Sarker A (2011). Genetic improvement of grass pea for low neurotoxin (ODAP) content. Food Chem Toxicol 49: 589–600.
Kumar S, Gupta P, Barpete S, Sarker A, Amri A, Mathur PN, Baum M (2013). Grass pea. In: Singh MH, Upadhyaya D, Bisht IS, editors. Genetic and Genomic Resources for Grass Pea Improvement. Amsterdam, the Netherlands: Elsevier, pp. 269–293.
Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15: 473–497.
Ochatt SJ, Conreux C, Jacas L (2013). Flow cytometry distinction between species and between landraces within Lathyrus species and assessment of true-to-typeness of in vitro regenerants. Plant Syst Evol 299: 75–85.
Ochatt SJ, Pontecaille C, Rancillac M (2000). The growth regulators used for bud regeneration and shoot rooting affect the competence for flowering and seed set in regenerated plants of protein pea. In Vitro Cell Dev-Pl 36: 188–193.,
Özcan S (1995). Assessment of the susceptibility of different pea (Pisum sativum L.) genotypes to Agrobacterium tumefaciens. Turk J Bot 19: 417–422.
Özcan S, Barghchi M, Firek S, Draper J (1993). Efficient adventitious shoot regeneration and somatic embryogenesis in pea. Plant Cell Tiss Org 34: 271–277.
Özcan S, Yıldız M, Sancak C, Özgen M (1996). Adventitious shoot regeneration in sainfoin (Onobrychis viciifolia Scop.). Turk J Bot 20: 497–501.
Snedecor GW, Cochran WG (1967). Statistical Methods. Ames, IA, USA: Iowa State University Press.
Sriskandarajah S, Frello S, Serek M (2001). Induction of adventitious shoots in vitro in Campanula carpatica. Plant Cell Tiss Org 67: 295–298.
Vaz Patto MC, Skiba B, Pang ECK, Ochatt SJ, Lambein F, Rubiales D (2006). Lathyrus improvement for resistance against biotic and abiotic stresses: from classical breeding to marker assisted selection. Euphytica 147: 133–147.
Zheng, Z, Wang HB, Chen GD, Yan GJ, Liu CJ (2013). A procedure allowing up to eight generations of wheat and nine generations of barley per annum. Euphytica 191: 311–316.