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An efficient in vitro regeneration system of fieldpea (Pisum sativum L.) via. shoot organogenesis

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Abstract

In-vitro regeneration in fieldpea was achieved from immature embryonic axes and cotyledonary node explants of six genotypes on modified MS media supplemented with different concentration of plant growth regulators, 6-Benzylamino purine (BAP) and Naphthalene acetic acid (NAA). The best regeneration response, leading to multiple shoot formation efficiency (22.34 shoots/explant) was observed in the medium supplemented with 1.0 mg/L BAP and 0.2 mg/L NAA and best frequency (67.55 ± 4.74) was achieved on medium containing 2.0 mg/L BAP and 0.4 mg/L NAA. The shoots were subcultured on a medium supplemented with a combination of 1.0 mg/L GA3, 2.0 mg/L BAP and 0.4 mg/L NAA, which resulted in elongation of 85 % of shoots. Rooting attempted from the elongated shoots, on half strength MS medium and supplemented with three different auxins IBA, IAA and NAA separately, exhibited similar results. Alternatively, micro-grafting of in vitro regenerated shoots onto pre-germinated root stocks raised in green house facility was attempted with high success rate (75 %). The grafted plants could be successfully hardened, fertigated with Hoagland solution and distilled water in a ratio of (1:10) for acclimatization and further development. All the genotypes tested, produced multiple shoots that could be established to mature fertile plant, hence, the medium combinations used were found to be genotype neutral.

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Abbreviations

BAP:

6-benzylaminopurine

B5 :

Gamborg medium

GA3 :

Gibberellic acid

IAA:

Indole-3-acetic acid

MS:

Murashige and Skoog medium (1962)

NAA:

α-naphthalene acetic acid

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Acknowledgments

The authors are thankful to National Agricultural Innovation Project (NAIP), Indian Council of Agricultural Research (4-767-04) for financial support. Thanks are due to Dr Sripad Bhat, IIPR, for his help with statistical analysis and Dr G P Dixit, IIPR for providing fieldpea genotypes.

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Authors and Affiliations

  1. Division of Crop Improvement, Indian Institute of Pulses Research, Kanpur, 208 024, India

    Alok Das, Sumit Kumar, P. Nandeesha, Indu Singh Yadav, Jyoti Saini, S. K. Chaturvedi & Subhojit Datta

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  1. Alok Das
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  2. Sumit Kumar
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  3. P. Nandeesha
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  4. Indu Singh Yadav
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  5. Jyoti Saini
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  6. S. K. Chaturvedi
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  7. Subhojit Datta
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Correspondence to Alok Das.

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Alok Das and Sumit Kumar contributed equally

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Das, A., Kumar, S., Nandeesha, P. et al. An efficient in vitro regeneration system of fieldpea (Pisum sativum L.) via. shoot organogenesis. J. Plant Biochem. Biotechnol. 23, 184–189 (2014). https://doi.org/10.1007/s13562-013-0200-3

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  • DOI: https://doi.org/10.1007/s13562-013-0200-3

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