wboratory experiments were conducted at National Citrus Research Program, Dhankuta in
two consecutive years (200212003 to 200312004) to optimize tl1e protocol for in vitro plalll
regeneration'of Madame Vinous Cultivar of Sweet Orange (MVSO). Five different levels of
BAP and two levels of fAA were tested in a factorial Randomized Complete Block Design
(RCBD) with six replications. The best treatment, in terms of response frequency and mean
number of axillary shoots per explant was 011 MS medium ( f962) supplemented with 0.5 mg/1
BAP and 0.2 mg/1/AA (4.8 shoots per explant). Effective rooting was observed on MS medium
supplemented with 0.5 mg/1 fAA. The in vitro rooted plantlets were acclimatized in Cello
polyethylene containers for four weeks in room temperature followed by transferring them
i11to the glasshouse. The results reveled that a11 efficient system for in vitro plant regeneration
by multiple shoot formation from in vitro shoot tip explams of MVSO has been successfully
established, thus it is useful for the production of disease free plallling materials. The
pathogen free plants of MVSO could be used as indicator plallf for Biological Indexing of
Citrus Greening Disease caused by gram-negative bacterium (Liberibacter asiatic:um).

wboratory experiments were conducted at National Citrus Research Program, Dhankuta in
two consecutive years (200212003 to 200312004) to optimize tl1e protocol for in vitro plalll
regeneration'of Madame Vinous Cultivar of Sweet Orange (MVSO). Five different levels of
BAP and two levels of fAA were tested in a factorial Randomized Complete Block Design
(RCBD) with six replications. The best treatment, in terms of response frequency and mean
number of axillary shoots per explant was 011 MS medium ( f962) supplemented with 0.5 mg/1
BAP and 0.2 mg/1/AA (4.8 shoots per explant). Effective rooting was observed on MS medium
supplemented with 0.5 mg/1 fAA. The in vitro rooted plantlets were acclimatized in Cello
polyethylene containers for four weeks in room temperature followed by transferring them
i11to the glasshouse. The results reveled that a11 efficient system for in vitro plant regeneration
by multiple shoot formation from in vitro shoot tip explams of MVSO has been successfully
established, thus it is useful for the production of disease free plallling materials. The
pathogen free plants of MVSO could be used as indicator plallf for Biological Indexing of
Citrus Greening Disease caused by gram-negative bacterium (Liberibacter asiatic:um).