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High frequency organogenesis in hypocotyl, cotyledon, leaf and petiole explants of broccoli (Brassica oleracea L. var. italica), an important vegetable crop

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Abstract

Broccoli (Brassica oleracea L. var. italica) is an important, nutritionally rich vegetable crop, but severely affected by environmental stresses, pests and diseases which cause massive yield and quality losses. Genetic manipulation is becoming an important method for broccoli improvement. In the present study, a reproducible and highly efficient protocol for obtaining organogenesis from hypocotyl, cotyledon, leaf and petiole explants of broccoli (Brassica oleracea L. var. italica cv. Solan green head) has been developed. Hypocotyl and cotyledon explants were used from 10 to 12 days old aseptically grown seedlings whereas leaf and petiole explants were excised from 18 to 20 days old green house grown seedlings and surface sterilized. These explants were cultured on shoot induction medium containing different concentration and combination of BAP and NAA. High efficiency shoot regeneration has been achieved in hypocotyl (83.33 %), cotyledon (90.11 %), leaf (62.96 %) and petiole (91.10 %) explants on MS medium supplemented with 3.5 mg/l BAP + 0.019 mg/l NAA 2.5 mg/l BAP + 0.5 mg/l NAA, 4.0 mg/l BAP + 0.5 mg/l NAA and 4.5 mg/l BAP + 0.019 mg/l NAA respectively. Petiole explants showed maximum shoot regeneration response as compared to other explants. MS medium supplemented with 0.10 mg/l NAA was found best for root regeneration (100 %) from in vitro developed shoots. The regenerated complete plantlets were transferred to the pots containing cocopeat and successfully acclimatized. This optimized regeneration protocol can be efficiently used for genetic transformation in broccoli. This is the first comparative report on multiple shoot induction using four different types of explants viz. hypocotyl, cotyledon, leaf and petiole.

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Abbreviations

MS :

Murashige & Skoog

NAA :

Naphthalene acetic acid

BAP- 6 :

Benzyl aminopurine

IAA :

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

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Acknowledgments

We are grateful to the Professor and Head, Department of Vegetable Science, Dr. Y. S. Parmar University of Horticulture and Forestry- Nauni, Solan for providing us the germplasm of broccoli cv. Solan green head. The senior author (P.K.) thankfully acknowledge the award of DST INSPIRE fellowship New Delhi.

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Authors and Affiliations

  1. Department of Biotechnology, Dr. Y S Parmar University of Horticulture and Forestry, Nauni, Solan, 173230, Himachal Pradesh, India

    Pankaj Kumar & D. K. Srivastava

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  1. Pankaj Kumar
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  2. D. K. Srivastava
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Correspondence to D. K. Srivastava.

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Kumar, P., Srivastava, D.K. High frequency organogenesis in hypocotyl, cotyledon, leaf and petiole explants of broccoli (Brassica oleracea L. var. italica), an important vegetable crop. Physiol Mol Biol Plants 21, 279–285 (2015). https://doi.org/10.1007/s12298-015-0282-6

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