Abstract
We developed an efficient method for inducing high-frequency in vitro flowering and fruiting in 10 cultivars (Cap-1 to -10) of three different species of Capsicum (C. annuum, C. chinense, and C. frutescens) collected from diverse geographical regions of India and Mexico. Shoot tips of in vitro germinated seedlings were cultured on MS media supplemented with different concentrations of 6-benzylaminopurine (BAP) alone or in combination with silver nitrate or silver thiosulphate. Low BAP concentration was a good inducer of flower buds in vitro. Synergism between silver ions and cytokinin improved flower induction. Flowering frequency improved significantly when silver nitrate or silver thiosulphate was supplemented with optimum BAP. Maximum responses, 7.5 ± 0.20 (in Cap-1) to 15.8 ± 0.27 (in Cap-5) flowers, were induced in the medium containing 0.75 mg l−1 BAP and 30 μM silver thiosulphate within 60 days of culture. Both in vitro hand-pollination and high sucrose concentration (5.0–6.0 %) in the medium improved fruit setting. In general, fruits were very small (mini-fruit) and seedless; however, three cultivars (Cap-3, -5, and -6) produced normal fruits with seeds. On an average, 144 days were required from seed implantation to fruit ripening. The frequency of meiotic abnormalities was higher in flowers produced in vitro than in those produced in vivo, and pollen viability was relatively lower in the in vitro flowers than in the in vivo flowers. The present protocol offers a repeatable system for studying the physiological mechanism of flowering and fruiting as well as providing the basis for further investigation for rapid in vitro cross-breeding programmes in Capsicum spp. for commercial hybrid production.
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Acknowledgments
SMH acknowledges the Ministry of Minority Affairs (MOMA) and the University Grant Commission (UGC) of India for providing Maulana Azad National Fellowship (MANF). BG thanks the Department of Science and Technology, West Bengal (DST-WB) for providing a research Grant. All authors acknowledge Dr. Paul W. Bosland, Professor of the Department of Agronomy and Horticulture, New Mexico State University, Mexico for gifting us the seeds of Mexican cultivars of Capsicum, and Swami Kamalasthananda, Principal, Ramakrishna Mission Vivekananda Centenary College, Rahara, Kolkata, India for the facilities provided during the present study. The DST-FIST programme for infrastructural facilities is also acknowledged.
Authors’ contribution
SMH carried out all the experimental works and statistical analysis under the guidance of BG. SP help to SMH during experimental work. The manuscript was initially drafted by SMH and critically revised by BG, then finally checked and approved by all authors. BG obtained a research grant [Grant Number 616(Sanc.)/ST/P/S&T/1G-6/2011] for conducting the work.
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Haque, S.M., Paul, S. & Ghosh, B. High-frequency in vitro flowering, hand-pollination and fruit setting in ten different cultivars of Capsicum spp. (C. annuum, C. chinense, and C. frutescens): an initial step towards in vitro hybrid production. Plant Cell Tiss Organ Cult 127, 161–173 (2016). https://doi.org/10.1007/s11240-016-1039-9
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DOI: https://doi.org/10.1007/s11240-016-1039-9