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GrainGenes Reference Report: PCT-44-71

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Reference	PCT-44-71
Title	Plant regeneration from embryogenic cell suspensions and protoplasts in sugarcane (Saccharum spp. hybrid cv. CoL-54)
Journal	Plant Cell Tissue and Organ Culture
Year	1996
Volume	44
Pages	71-78
Author	Aftab F Zafar Y Malik K Iqbal J
Abstract	Embryogenic callus was developed from young leaves of sugarcane (Saccharum spp hybrid, cv CoL-54) A good embryogenic callus response was achieved using MS basal medium containing 2.0 micromole (0.5 mg l(-1) picloram under dark conditions at 27 +/- 1 degrees C. Initiation of fast growing homogeneous cell suspension cultures was achieved in MS and AA media, both supplemented with g micromole (2 mg l(-1)) 2,4-D and 500 mg l(-1) CH. Embryogenic callus was reinitiated from embryogenic cell suspension cultures using MS medium containing 30 g 1(-1) sucrose, 500 mg l(-1) CH and 2.26 micromole (0.5 mg l(-1)) 2,4-D after 4-6 weeks of culture under 16-h photoperiod conditions. Plant regeneration was achieved after about 4 weeks in MS medium lacking growth regulators but containing CH (500 mg l(-1)) and sucrose (60 g l(-1)). Rooting was enhanced by transferring regenerated plantlets to half strength MS basal medium. Totipotent protoplasts with an average yield of 2.0 X 10(7) to 1.0 X 10(8) ml-1 were obtained from embryogenic cell suspension cultures at log phase, i.e., 4-5 days after transfer to fresh media. The best growth response was achieved when protoplasts were cultured in a modified KM8P medium at the density of 2.0 X 10(5) m l(-1). Protoplasts were mainly embedded in 0.8% sea plaque agarose. Division efficiency of 22.2% was achieved after 20 days of culture and 0.26% of microcolonies continued growth and formed microcalluses after 30 days of culture under dark conditions. Microcalluses were proliferated in MS medium having 2,4-D (2 mg l(-1)) under 16-h photoperiod. Transferring these embryogenic calluses in MS medium + 9.29 micromole kinetin (2 mg l(-1) + 5.37. micromole NAA (1.0 mg l(-1) + activated charcoal (200 mg l(-1)) for 5 weeks favoured plant regeneration. Shoots and roots were further proliferated in half strength MS basal medium for 2-4 weeks. Regenerated plants were transferred to autoclaved sand for 2 weeks under 16-h photoperiod in growth room and transferred to soil in a greenhouse to raise to maturity.
Keyword	[ Hide all but 1 of 31 ] 2,4-d callus casein hydrolysate cell division cell growth cell suspensions charcoal culture media dark greenhouse culture growth stage kinetin leaves methodology micropropagation muller and grafe medium murashige and skoog medium naa photoperiod picloram plant embryos plant morphology protoplast regenerative ability root rooting capacity saccharum sand shoots soil sucrose

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