AN EFFICIENT METHOD FOR THE ESTABLISHMENT OF CELL SUSPENSION CULTURES IN POTATO (SOLANUM TUBEROSUM L.)

摘要

Cell suspension cultures offers an In vitro system that can be used as a tool for various studies involving mutantselection, mass propagation, protoplast isolation, gene transfer and selection of cell-lines which are resistant to various bioticor abiotic stresses. Research work on the development of cell suspension cultures was carried out to establish the mostefficient method in Potato (cv. Desiree). Healthy, well-proliferating tissues from different types of callus cultures (compact,friable, embryogenic or non-embryogenic) were inoculated on various media combinations, i.e., MS, MS2 or AA liquidmedium containing 18.09 μM 2, 4-D. A fixed quantity (0.5-1.0 g) of callus tissue from 60-day-old callus cultures wastransferred to 10-25 ml of liquid medium in 100 ml Erlenmeyer flask. Cultures were placed on an orbital shaker and agitatedat different speeds (75, 100 or 125 rpm) under 16-h photoperiod at 25 ± 2°C. Medium was changed after every 3 days andfractionated tissue was filtered after every 6 days through sterile mesh (100-800 μm) to develop a cell-line by transferringresulting suspension to fresh medium under the same conditions. Results indicated that eight-week-old translucent, friable,off-white callus cultures were an excellent starting material for the initiation of homogeneous cell suspension cultures ascompared to other tested sources. Of the three tested media (MS, MS2 or AA medium containing 18.09 μM 2, 4-D), MS2was found to be a better medium for the initiation of cell suspension cultures. Cell suspension cultures, placed in 16-hphotoperiod at 25 ± 2°C and agitated at 120 rpm using a gyratory shaker showed excellent results. Several other factorsinfluencing quick establishment of cell suspension cultures in this cultivar are also discussed in this communication.