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Silver nitrate and 2-(N-morpholine) ethane sulphonic acid in culture medium promotes rapid shoot regeneration from the proximal zone of the leaf of Capsicum frutescens Mill.

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Abstract

The genetic manipulation of Capsicum has been unsuccessful, and a large bottleneck to transferring the desired genes is due to the difficulty in regenerating whole plants through tissue culture because of its highly recalcitrant and high genotype specificity. This study aimed to investigate and establish rapid shoot regeneration from the proximal ends of the leaves of Capsicum frutescens KT-OC and BOX-RUB varieties. A maximum of 8–10 shoot buds were obtained from the margins of the proximal portion of a cotyledonary leaf explant of C. frutescens variety KT-OC on medium I containing 44.44 µM 6-benzylaminopurine (BA), 5.71 µM indole-3-acetic acid (IAA), 10 µM silver nitrate (AgNO3) and 1.98 mg L−1 2-(N-morpholine) ethane sulphonic acid within 4 weeks of incubation, of which 60% of explants responded in terms of shoot buds. Petiole explants (40%) cultured on the same medium produced 2–4 shoots per explant from the distal portion. The cut portions of the cotyledonary leaf proximal portions responded well to shoot bud formation in the presence of 22.20 µM BA and 14.68 µM phenyl acetic acid (PAA), wherein 100% of explants responded in terms of shoot bud formation, with an average of 10 ± 1.7 and 8 ± 1.9 shoot buds per explant in KT-OC and BOX-RUB varieties, respectively. The differentiated shoots grew well and proliferated in the presence of 14.68 µM PAA + 22.20 µM BA and 10 µM AgNO3. Shoot elongation was obtained in presence of 1.44 µM gibberellic acid (GA3) and 10 µM AgNO3. These shoots were rooted on plant growth regulator-free half-strength MS medium and upon hardening; field survival rate was 70%. This reproducible regeneration method for C. frutescens, especially the Indian high pungent variety, from proximal portion of cotyledonary leaf and petiole explants, can be used for biotechnological improvement.

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Acknowledgments

The author AS acknowledge Council of Scientific and Industrial Research (CSIR) New Delhi for research fellowships and to Defence Research Development Organization (DRDO Pithoragarh) for providing the seeds of KT-OC and BOX-RUB. This research was financially supported by the Department of Biotechnology (BT/PR9091/PBD/17/504/2007), New Delhi, India.

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  1. Plant Cell Biotechnology Department, CSIR-Central Food Technological Research Institute, Mysore, Karnataka, 570 020, India

    Sharma Ashwani, Gokare Aswathanarayana Ravishankar & Parvatam Giridhar

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  1. Sharma Ashwani
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  2. Gokare Aswathanarayana Ravishankar
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  3. Parvatam Giridhar
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Correspondence to Parvatam Giridhar.

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Ashwani, S., Ravishankar, G.A. & Giridhar, P. Silver nitrate and 2-(N-morpholine) ethane sulphonic acid in culture medium promotes rapid shoot regeneration from the proximal zone of the leaf of Capsicum frutescens Mill.. Plant Cell Tiss Organ Cult 129, 175–180 (2017). https://doi.org/10.1007/s11240-016-1157-4

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