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Query: EC:4.2.2.10 (
PNL
)
341
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
A study was undertaken to develop a protoplast regeneration system for pinellia. A yield of 19 29 x 10(5) protoplasts/g F. W. could be obtained from cell suspension cultures incubated in a digestion enzyme solution with 2% cellulase Onzuka R-10, 10% pectinase (Sigma), 0.01%
pectolyase
Y23. K8P and modified MS media were used to culture protoplasts in: a) liquid, b) liquid-solid double layer, or c) agarose embedded protoplast culture. The former two were conducive to colony formation from protoplast-derived cells. The frequency of cell division was about 8% after 3 days in culture. Gradually adding fresh medium of lower osmotic pressure into the medium for protoplast culture favored cell division. Calli (1-2 mm in diameter) formed after 30-40 days in culture. The calli transferred onto medium supplemented with KT (0.5 mg 1(-1)) and
NAA
(0.2 mg 1)(-1)) could regenerate plants after 40-50 days. Of 47 plantlets transplanted into plots, 29 flowered and were fertile.
...
PMID:Protoplast culture and plant regeneration ofPinellia ternata. 2417 63
Protoplasts were isolated from hypocotyls of 7-d-old seedlings of three genotypes of Brassica carinata after enzymatic digestion in cellulase R-10 (0.5%) and
pectolyase
Y-23 (0.025%). The protoplasts were stabilized with 0.4 M mannitol used as osmoticum, and were cultured in darkness in Kao's liquid medium containing 0.4 M glucose and the growth regulators 2,4-D (1.0 mg/l),
NAA
(0.1 mg/l) and zeatin riboside (0.5 mg/l). Protoplasts were transferred to 16 h photoperiod conditions after 3 d of dark culture, and the medium was diluted to reduce the osmoticum on the seventh and tenth days of culture. Microcolonies were thus obtained which, upon transfer to MS agarose medium with 2,4-D (0.1 mg/l), BAP (1 mg/l) and 0.1 M sucrose, proliferated further to produce callus clumps. The plating efficiency of the three genotypes varied from 1 to 2%. Calli 2-3 mm in diameter were transferred to MS agarose plates with zeatin (2 mg/l) where they produced shoot buds and shoots with frequencies ranging from 22.5 to 74.2% for the three genotypes. The shoots were rooted in medium with IBA (1 mg/l) and were then established in soil. The time required for protoplast to plant development was 8 to 10 weeks.
...
PMID:Rapid and efficient plant regeneration from hypocotyl protoplasts of Brassica carinata. 2421 51