STUDIES ON MICROPROPAGATION AND CALLUS INDUCTION IN POMEGRANATE (Punica granatum L.) CV. MRIDULA
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Date
2002-06-29
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MAHATMA PHULE KRISHI VIDYAPEETH, RAHURI-413 722, DIST. AHMEDNAGAR MAHARASHTRA STATE (INDIA)
Abstract
In the present study an attempt was made to develop an in vitro procedure for large scale multiplication of pomegranate (Punica granatum L.) cv. Mridula. The main objectives of the study was to standardize the prolocol for micropropagation, callus induction and regeneration. The explains like cotyledon, leaf segment, shoot tip and nodal segment were used for culturing in the basal MS medium fortified with growth regulators like BAP, NAA and IBA in different concentrations and combinations. During the culturing of explant the in culture medium often browning of the explant/media was noticed due to the exudation of polyphenols. Inoculation of explants was followed by sub-culturing twice, firstly one day after inoculation and secondly at third day after inoculation which effectively controlled browning. Besides this, the cotyledon explant was noticed to escape browning as compared to other explants. Contd... The culture media containing MS + NAA 0.4 mg L-1 + BAP 1.0 mg L-1 was revealed as an effective medium for shoot differentiation. This culture media induced shooting in 77.77 per cent and 81.25 per cent of shoot tip and nodal segment explant after inoculation with 6.8 and 6.4 days, respectively. An average of 1.75 shoots/shoot tip explants and 4.25 shoots/nodal segment explant were also obtained in the same media. 66.4 per cent of shoots successfully rooted when transferred to1/2 MS + NAA 0.5 mg L-1. Rooted plantlets of 5-6 cm height were transferred to plastic cups containing vermiculture under mist house conditions before they were finally exposed to external environment. Of these 49 per cent plantlets were survived. Additionally, an attempt was made to propagate plantlets of pomegranate by intervention of callus. The culture media containing MS + NAA 0.4 mg L-1 + BAP 1.0 mg L-1 was identified as the most effective medium for getting maximum callus proliferation and growth. The proliferated callus when cultured for differentiation, it continued to proliferate as callus instead of differentiating into shoot and root.
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