In vitro studies in acid lime (Citrus aurantifolia Swingle) cv. kagzi lime
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Date
2006
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Publisher
CCSHAU
Abstract
The experiment was conducted during 2004-2006 on in
vitro studies in acid lime (Citrus aurantifolia Swingle) var. Kagzi Lime in
the laboratory of Centre for Research and Application in Plant Tissue
Culture CRATPC CCS HAU, Hisar Haryana. The experiment consisted
of in vitro raising of seedlings and direct shoot regeneration in acid lime
var. Kagzi Lime. The experimental material comprised of seeds and
different explants (in vitro shoot tip and nodal segment, in vivo shoot tip
and nodal segment) with different combinations of growth regulators
(GA3, BAP, NAA, IBA) supplementing the MS basal medium. For axenic
production of seedling through in vitro seed germination, the best
medium was MS medium + GA3 0.5 mg/l and for seedling growth best
medium was MS medium + GA3 0.5 + BAP 0.5 mg/l. Direct
regeneration of shoots, roots and whole plant without intervention of
callus from both in vitro and in vivo shoot tip and nodal segment on MS
basal medium containing BAP along with NAA at different
concentrations was observed. The MS medium + BAP 0.25 mg/l wa
observed to be the best medium for shoot regeneration inducing 90
percent in both in vitro shoot tip and nodal segment and 77.77 and
78.59 percent in in vivo shoot tip and nodal segment respectively.
The half MS medium + IBA 1.0 mg/l was found to be best
for different aspects of root differentiation like number of days required
for root initiation (13.80) and completion of rooting (25.60), percent
rooting (100%), number of roots/plantlet (3.80), root length (3.88cm)
and percent survival of rooted plantlets (100%). The maximum survival
percentage of micro propagated plantlets under green house condition
and open condition after hardening period of four weeks was 100
percent in the potting mixture having soil, sand and FYM in an equal
proportion (1:1:1 v/v). Improved transformation frequencies were
obtained by co-cultivation the explants with Agrobacterium on feeder
plants by using kanamycin at 100 mg/l as selective agent. Attempts to
use geneticin as selection antibiotic were not successful. The presence
and expression of transferred genes was verified by -glucuronidase
histochemical expressed as blue spots in co-cultivated explants.