OPTIMIZATION OF CALLUS INDUCTION FROM POMEGRANATE (Punica granatum L.) LEAF EXPLANTS

Abstract

The traditional method of pomegranate plant propagation does not ensure disease-free development of plants and is time-consuming and labor-intensive process. A large numbers of cuttings do not survive during plantation. The present study was undertaken to optimize callus induction from Punica granatum leaf explants. Young leaf explants from mature pomegranate tree was used as explants. leaf segments was investigated on full-strength Murashige and Skoog (MS) medium supplemented with different combination 6-benzylaminopurine (BAP) with αnaphthaleneacetic acid(NAA).The result revealed that the survival percent differed significantly and ranged from 0.00 to 84% in different treatment. Treatment consisting of 70% ethanol (EtOH) for 0.75 min, 0.2% Bavistin for 15 min, and 0.5% sodium hypo chlorite (NAOCL) solution for 1 min was optimal with the least contamination (17.71%) of cultures and caused the least damage to explants. Subculturing (three or four times) of explants on fresh solidified MS basal medium without any additive at the interval of 24 h was found to be an effective control measure of browning. 5mM BAP (6-benzylaminopurine) in combination with 2.5mM NAA had the maximum callus fresh weight (1.34g), callus induction rate (69%), and callus diameter (13.10mm). At the maximum amount of BAP 6mM combination with 3mM NAA callus induction rate (58.00%) was lower than 5mM BAP concentrations. It can be concluded from the result of the present study that longer exposure is not recommended for obtaining the maximum percentage of servival cultures. The percentage of callus induction increases when BAP is added to the medium, but further increase in BAP levels was decreased the percentage of callus induction