Studies on in vitro plant regeneration in Capsicum (Capsicum annuum L.)
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Date
2014
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Publisher
CCSHAU
Abstract
Development of an efficient plant regeneration system is a pre-requisite for any plant biotechnological approach for crop improvement. In this regard the in vitro regeneration from cotyledon and hypocotyl explants from aseptically raised seedlings of popular Capsicum F1 hybrids Bharat and Indra was achieved. The anther culture was also tried to produce double haploids. Seeds of both hybrids were exposed to different treatments for proper germination. The treated seeds were decontaminated and placed in a half-strength MS medium and then incubated in 16h photoperiod for germination. The seed soaked in distilled water along with GA3 at 2 mg/l for two days prior to sowing had more pronounced effect on both capsicum hybrids recording maximum germination (90.45 and 84.59 %, respectively) and that also in minimum number of days (9.67 days and 10.33 days, respectively). Tissue culture responses to morphogenesis varied with the genotypes, source of explants, type of explants and combinations of growth regulators used. Per cent regeneration (96.30%), number of shoots per explants (4.56) and per cent elongation (82.10%) was maximum in hybrid Indra from cotyledons having abaxial side in contact with growing medium supplemented with zeatin at 7.5 mg/l along with GA3 at 2.0 mg/l. Cent per cent rooting, optimum number of roots (27.56 and 23.65, respectively) and root length (4.94 cm and 7.71 cm, respectively) were observed when regenerated shoots cultured on MS media supplemented with IBA at 0.5 mg/l in both the hybrids. The survival percentage of regenerated plantlets was higher in Indra (92.31%) as compared to Bharat (85.71%) during hardening in pots containing sterile mixture of coco-peat and vermiculite (1:1) and thereafter in pots containing soil. Anthers of both hybrids were collected at three different stages of microspore development and cultured on medium supplemented with different ratios of growth regulators and additives. The effect of heat treatment at 35oC for seven days on androgenesis was also examined. Androgenic callus induction was best on heat treated anthers cultured on media supplemented with zeatin at 1.0 mg/l along with 2,4-D at 0.2 mg/l and AgNO3 at15 mg/l in both the hybrids (54.02% and 60.92%, respectively) and hybrid Indra responded better than Bharat. The effect of different cytokinins (BAP, kinetin, TDZ and zeatin) along with NAA on regeneration of androgenic callus was also studied, however, shoot induction could not be achieved. Hence, greening of callus in particular medium was recorded. The MS media supplemented with kinetin at 7.5 mg/l along with NAA at 0.2 mg/l recorded maximum per cent greening of callus i.e. 53.33% and 51.11% in hybrid Indra and Bharat, respectively.
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Keywords
regeneration, cytokinins, auxins, planting, biological phenomena, hybrids, pepper, vegetative propagation, developmental stages, organ culture