IN VITRO MUTAGENESIS IN GINGER (Zingiber officinale Rosc.) FOR THE SELECTION OF PLANTS RESISTANT TO Fusarium oxysporum f.sp. zingiber
Loading...
Date
2019-12
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
UHF,NAUNI
Abstract
ABSTRACT
In the present investigation, an attempt has been made to develop Fusarium yellows resistant plants of
ginger (Zingiber officinale Rosc.) var. using in vitro mutagenesis and shoot selection technique. In vitro
propagation was done using buds as explants. Maximum in vitro bud establishment was achieved in spring season
(62.85%). 0.2% HgCl2 for 5 minutes was found to be the best concentration of surface sterilant with 42.59 per cent
survival. 72.85% buds established on MS medium supplemented with 1 mg/l BA and 0.1 mg/l NAA. MS medium
fortified with 0.5 mg/l BA and 0.1 mg/l NAA was found to be best multiplication medium resulting in 5.2 average
number of shoots per explant with average shoot length of 4.5 cm. Rooting could be achieved on the same medium
after second subculture. With the increase in number of subcultures, shoot multiplication rate, average shoot length,
per cent rooting, thickness of roots and average number of roots per shoot showed an increase. In vitro cultures were
subjected to gamma irradiation (10-100 Gy) for in vitro mutagenesis. LD50 was calculated after every four weeks and
it was calculated to be 15 Gy after 16 weeks of irradiation. The non irradiated and irradiated plantlets were hardened
with 100 per cent survival rate in potting mixture comprising of soil: cocopeat: FYM (2:1:1). On biochemical
evaluation of rhizomes, it was observed that essential oil content was found to be same in all the samples (2%)
whereas, highest oleoresin content was found in tissue culture propagated rhizomes (3.16%) and lowest crude fibre
content (5.3%) was obtained in rhizomes collected from gamma irradiated plants. HPLC studies on gingerol content
showed its highest value in rhizomes produced by gamma irradiated plants. No difference in chromosome number was
observed in tissue culture propagated and gamma irradiated shoots. Gamma irradiated shoots were cultured on
selective medium containing different concentrations (0-20%) of fungal culture filtrate (FCF) for in vitro selection.
17.5% FCF was found to be the highest concentration on which 5% shoots survived after 1st selection cycle which
reduced to 1.15% after 3rd alternate, discontinuous selection cycle. Selected shoots from 15 and 17.5% FCF were
multiplied on multiplication medium. Rooted plantlets were successfully hardened with 100% survival. 46.4 and 52%
plants selected at 15 and 17.5% FCF were found to be highly resistant on in vivo evaluation. SCoT and SSR analysis
distinguished between FCF selected, tissue culture propagated and gamma irradiated plants. In SSR analysis, two
unique bands were obtained in plantlets selected on 17.5% FCF. On sequencing, the band obtained with GIN6 showed
98% homology with disease resistance protein-like gene CC-NBS-LRR from clone ZwP627 and band obtained with
primer GIN9 showed 97% homology with same gene from clone ZoP620 of Z. officinale.After in vivo bioassay, SSR
analysis of selected highly resistant plants again confirmed the unique bands with both the primers and the unique
DNAsequences obtained have been submitted to NCBI under accession number MN497252 and MN497253 and will
be published in GenBank after complete processing. Therefore, in the present study we have successfully developed
Fusarium yellows resistant plants of ginger (Zingiber officinale Rosc.) var. Himgiri using in vitro mutagenesis and
selection technique.
Description
Keywords
null