Isolation of protoplasts from cardamom (Elettaria cardamomum Maton.) and ginger (Zingiber officinale Rose.)
2000
Ravindran, K V Peter, S P Geetha, K Nirmal Babu, J Rema, P N
Protoplasts were isolated from leaf mesophyll tissue, collected from in vitro grown plantlets and cell suspension cultures of cardamom (Elettaria cardamomum) and ginger (Zingiber oJficinale). In cardamom, a protoplast yield of 3.5 x 105/ g of leaf tissue was obtained when incubated in an enzyme solution containing 0.5% macerozyme RIO, 2% cellulase Onozuka RIO and 9% mannitol for 18-20 h at 25°C in dark. The yield of protoplasts from cell suspension culture was 1.5 x 105 / g tissue, when incubated in I % macerozyme RIO and 2% cellulase Onozuka RIO for 24 h at 25°C with gentle shaking at 53 rpm in dark. The viability of leaf mesophyll protoplast was 75% and that of cell suspension was 40% on Evan's blue staining. In ginger, a protoplast yield of 2.5 x lOS / g of leaf tissue was obtained on digestion in an enzyme solution containing 0.5% macerozyme RIO, 3% hemicellulase and 5% cellulase Onozuka RIO, when incubated for 10 h at 15°C followed by 6 h at 30°C. The protoplast viability was 55%. Protoplast yield from cell suspension culture was 1 x lOS /g of callus when digested with an enzyme solution of I % macerozyme RIO, 3% hemicellulase and 6% cellulase Onozuka RIO and incubated for 10h at 15°C and later at 30°C for 8 h. Seventy two per cent of the protoplasts were viable. The protoplasts from both the species could be cultured and made to develop up to microcalli stage.
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