MICROSHOOT TIP CULTURE THERAPY FOR DISEASE ELIMINATION IN DIFFERENT VARIETIES OF GRAPES CVs. PRINCESS SEEDLESS AND GOLDEN ITALIAN MUSACT IN PAKISTAN
S. Ilyas1, M. Khalid1 and S. Naz1*
1Dept. of Biotechnology, Lahore College for Women University, Lahore.
*Corresponding Author E-mail: drsnaz31@hotmail.com
ABSTRACT
Grape is considered as a vital commercial crop in Pakistan especially in Baluchistan. An effort has been made to establish a rapid protocol for virus free production of grapes varieties i.e. Princess seedless and Golden Italian musact through microshoot tip culture technology and callogenesis. The meristem (0.5mm) of grapes was cultured in Murashige and Skoogbasal medium provided with different levels of phytohormones (BAP, TDZ and NAA). The best results for shoot initiation were noted in MS+1.0mg/l BAP. In Princess seedless the shoot length was 1.9a±0.06 cm in 12c±0.43 days and in Golden Italian muscat the shoot length was 1.8a±0.36 cm in 13c±0.48 days. The extreme shoot multiplication was noticed inMS+BAP 1.0 mg/l+TDZ 6 µM for both varieties. The in vitro grown plantlets were then acclimatized in greenhouse in four different potting mixtures (redwood bark + coir + sand, sand+soil+ cocopeat, soil + cocopeat + leaf manure, sand + soil + leaf manure,).The frequency of plant survival was found best (95%) and (90%) in redwood bark-coir-sand for Princess seedless and Golden Italian muscat, respectively. For callogenesis, fresh leaves were inoculated on MS media supplemented with NAA, IAA, 2, 4-D and BAP. The highest percentages of callus formation i.e. 97% and 95% were obtained in MS+2,4-D 0.1mg/l+BAP 1 mg/l in Princess seedless and Golden Italian musact, respectively.In future, microshoot tip tissue culture therapy would be highly beneficial to produce virus free planting material and control spread of diseases in food crops at larger scale.
Keywords: Microshooting, virus eradication, disease free plants, Grape varieties and Acclimatization
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