Abstract
Objectives: A reliable in vitro regeneration protocol by direct organogenesis was developed in ICP 8863 variety of pigeon pea using leaf petiole and cotyledonary node explants. Methods: For direct shoot bud induction, leaf petiole explants from seven-day-old in vitro grown seedlings and cotyledonary node explants from twelve-day-old were cultured on MS medium supplemented with various combinations and concentrations of BAP, NAA and Kinetin. Induced shoot buds of both the explants were elongated on MS medium fortified with different concentrations of BAP, NAA and GA3. The wellelongated shoots of both the explants were transferred to MS medium supplemented with various concentrations of IBA. Finally, the regenerated plants were transferred to soil and vermiculate mixture in 1:1 ratio for acclimatization. Further, molecular characterization of the in vitro regenerated plants was carried out using eight OPP and OPAZ RAPD primer series. Findings: High frequency of shoot bud induction (92 %) was observed in leaf petiole explants with 2.0 mg/L6-BAP concentration compared to cotyledonary node explants. The induced shoots were kept for elongation and maximum percentage of elongation (93 %) was noticed in leaf petiole explants with 1.0 BAP + 0.1 NAA + 2.0 GA3 mg/L concentrations compared to cotyledonary node explants. The well-developed shoots of both the explants showed profuse rooting, where high percentage of rooting (95 %) was observed in leaf petiole explants with 0.5 mg/L IBA concentration. The pattern of amplification resulted through RAPD analysis confirmed the genetic stability of in vitro regenerated plants. Improvement: The regeneration protocol standardized in this study is suitable and reliable to develop transgenic pigeon pea plants by agrobacterium mediated genetic transformation. Keywords: Auxins, Cytokinins, Gibberellins, Organogenesis, Pigeonpea
Highlights
Pigeon pea [Cajanus cajan (L.) Millsp.] is one of the most important perennial food legume crops of rain-fed agriculture in the semi-arid tropics, which occupies fifth position in area, compared to the other legume crops like soybean, common bean, peanut and chickpea[1]
Leaf petiole and cotyledonary node explants collected from seven day old and twelve day old in vitro germinated seedlings showed high percentage of shoot bud induction and no shoot bud induction was noticed on plain MS medium without any plant growth regulators
In pigeon pea, in vitro regeneration was reported from various explants like, cotyledonary node[11,13,25,26,27,28], leaves[10,11,13,15,29], epicotyl[14], shoot apices[12], leaf petiole[10,30,31], leaf and apical meristem[32] and auxiliary buds[33]
Summary
Pigeon pea [Cajanus cajan (L.) Millsp.] is one of the most important perennial food legume crops of rain-fed agriculture in the semi-arid tropics, which occupies fifth position in area, compared to the other legume crops like soybean, common bean, peanut and chickpea[1]. Because of its high protein content, it is used as an important source of protein in Indians vegetarian diet It contains 3-5 % of soluble sugar, 1-2 % fats, 3-4 % crude fiber, 45-55 % starch and 3-4 % ash[5]. Pigeon pea is cultivated as an intercrop with other crops like sorghum (Sorghum bicolor), pearl millet (Pennisetum glaucum), maize (Zea mays) or with other legumes like peanuts (Arachis hypogaea) to maintain soil
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