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PROTOCOL OPTIMIZATION FOR EFFICIENT CALLUS INDUCTION AND REGENERATION IN THREE PAKISTANI RICE CULTIVARS

ZAHID HUSSAIN¹, MOHAMMAD HAROON KHAN¹, RAISA BANO¹, HAMID RASHID^2* AND ZUBEDA CHAUDHRY³

Abstract: This study was undertaken to standardize an efficient and effective protocol for callus induction, subsequent growth and regeneration in three Pakistani rice (Oryza sativa L.) varieties viz., GNY-53, Basmati-370 and JP-5. MS and N6 media were used for callus induction. Overall, MS medium was found better for callus induction as compared to N6 medium. The growth regulator 2,4-D with varying concentrations (1-3mg/l) were tested for their callus induction and subsequent growth. GNY-53 (83%) and JP-5 (96.66%) showed maximum callus induction frequency on MS medium supplemented with 3mg/l 2,4-D while Basmati-370 (99%) showed higher callus induction frequency on the same medium supplemented with 1mg/l 2,4-D. The callus growth frequencies for varieties GNY-53 (60%) and Basmati-370 (94.73%) were best achieved on MS medium supplemented with 3mg/l 2,4D while JP-5 (73.68%) showed maximum callus growth frequency on the same medium with 1mg/l 2,4-D concentration. Mean square value for variety, treatment and their interactions were recorded for callus induction and growth on both MS and N6 media at probability level of p≤0.05. The regeneration efficiency of these varieties were tested alone on MS medium fortified with two different combinations of NAA and BAP (1mg/l NAA: 2mg/l BAP and 1mg/l NAA:4mg/l BAP). The two varieties GNY-53 (70.27%) and JP-5 (41.81%) showed maximum plantlets formation frequency on 1:2mg/l combination of NAA and BAP whereas Basmati-370 (43.33%) showed higher plantlets formation frequency on 1:4mg/l combination of NAA and BAP. Mean square value for variety, treatment and their interactions were also recorded for regeneration efficiency at probability level of p≤0.05 which showed that these entire factors significantly affect plantlets formation frequency.

¹Department of Biotechnology, University of Malakand, NWFP, Pakistan,

²Department of Bioinformatics, Mohammad Ali Jinnah University, Islamabad, Pakistan,

³Department of Botany, Hazara University Mansehra, NWFP, Pakistan.