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Isolation and culture of protoplasts from cotyledons of Pinus coulteri D. Don. (1984)

Patel, Kamlesh R. ; Shekhawat, Narpat S. ; Berlyn, Graeme P. ; [et al.]

Springer

Plant cell, tissue and organ culture 3 (1984), S. 85-90

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ISSN: 1573-5044

Keywords: Pinus coulteri ; cotyledons ; protoplast culture ; phenolics

Source: Springer Online Journal Archives 1860-2000

Topics: Biology

Notes: Abstract A protocol was developed for the isolation and culture of protoplasts from the cotyledons of seedlings of Pinus coulteri D. Don. Incubation of cotyledon pieces in a mixture consisting of cellulase Onozuka R10 2%, Pectolyase Y-23 0.1%, mannitol 10%, CaCl2 500 mg/l and other macro and micro-nutrients yielded viable protoplasts. After 24 hours of culture in a complex nutrient medium, the protoplasts regenerated new cell walls and the first divisions were observed within 7–10 days. Small cell colonies were formed within 15–20 days, but these started to accumulate phenolics and no further growth of the colonies was observed.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00033729

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