In Vitro Regeneration of Pigeonpea [Cajanus cajan (L.) Millsp.] Genotype GT 101 using Cotyledonary Node (original) (raw)
Objectives: A protocol was developed for plant regeneration using cotyledonary node of high-yielding indigenous pigeonpea [Cajanus cajan (L.) Millsp.] genotype GT 101. Methods: Induction of multiple shoots directly was completed from cotyledonary node as an explants on Murashige and Skoog's (MS) medium supplemented with various concentrations and combination of 6-benzylaminopurine (BAP), kinetin and α-naphthalene acetic acid (NAA). Elongation of multiplied shoots was performed on MS medium supplemented with different combination of BAP and GA 3. These well elongated plantlets were further transferred on MS medium supplemented with various concentrations of indole-3-butyric acid (IBA) for root induction. Regenerated plants were transferred to cocopeat:soil:vermiculite (2:1:2) for acclimation. Findings: The frequency of multiplication and number of multiple shoots from cotyledonary node explant were influenced on various types and concentrations of cytokinin. For multiple shoots induction, 3.0 mg/L BAP with 0.5 mg/L NAA was superior as compared to other combinations. The elongation of multiplied shoots was carried out on MS medium supplemented with 0.5 mg/L BAP and 0.5 mg/L GA 3. The developed shoots were advanced to rooting on the medium supplemented with 0.5 mg/L IBA. They were subsequently grown in pots with 80% survival rate and these plants produced viable seeds. Improvement: The protocol for the production of in vitro multiple shoots with high frequency and their successive conversion to whole plants agreements potential for use in the improvement of protocol for development of transgenic in pigeonpea.
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Plant Cell Reports, 1998
A method for regenerating pigeonpea [Cajanus cajan (L.) Millsp.] plants has been developed using distal cotyledonary segments of mature seeds as explants. A large number of shoot buds were induced directly from explants of genotypes T-15-15 and GAUT-82-90 when cultured on six different basal media fortified with 22.2 μm N6-benzylaminopurine, 2.3 μm kinetin, and 271 μm adenine sulfate. The shoot buds developed into shoots when they were subcultured on the same medium but with one-tenth concentrations of cytokinins and adenine sulfate. The shoots elongated by subculturing first two to three times on Murashige and Skoog (MS) basal medium supplemented with 2.22 μm N6-benzylaminopurine and 0.54 μm α-naphthaleneacetic acid or on half-strength MS medium containing 2.89 μm gibberellic acid, and then once on the same medium without growth regulators. Elongated shoots were rooted with 80–85% efficiency on MS medium with 4.92 μm indole-3-butyric acid and the plantlets were transferred for hardening. Plants survival in pots was 70–75%. This method may be useful for improving the crop through genetic manipulations.
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Pigeon pea is legume crop play a crucial role as source of dietary protein in diet, growing extensively in the rainfed and dryland spots of India and worldwide. Plant tissue regenerate through in-vitro system attempting organogenesis as well as embryogenesis pathway, which are in support of unfamiliar genes assimilation targeted for development of transgenic plants. Present study was undertaken to investigate the most appropriate explant type in Pigeon pea regeneration by virtue of in vitro culture system. Genotype Durga (NTL-30) was breed and used as principal material for regeneration studies. Explants isolation from in vitro elevated germinating 6-8 days old seedlings were used for embryonic axis and cotyledonary node, whereas isolation of Scutellum (IZE) explants from overnight imbibed seed. Isolated explants were cultured on Murashige and Skoog medium supplemented with zeatine (0.572, 1.35, 1.47, 2.32 μM) and kinetin (0.46, 0.93, 1.39 & 1.86 μM) as plant growth regulator for re...
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