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Callus formation from root protoplasts of Quercus rubra L. (red oak)

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Abstract

Root protoplasts of Quercus rubra L. were isolated from 12 day old seedlings with an enzyme mixture containing Cellulase R1O + Rhozyme HP150 + Macerozyme R1O, supplemented with cysteine and bovine serum albumin.

Protoplasts were purified by a Ficoll density gradient centrifugation and cultured at low density in a liquid medium. The modified woody plant medium, containing 2.2 μM benzyladenine + 1.8; μM zeatin + 5.3 μM α-naphthaleneacetic acid + 2.2 μM dichlorophenoxyacetic acid, allowed sustained divisions and formation of microcalluses.

Protoplast — derived microcallus developed into green and compact callus when transferred to an agarose solidified medium, supplemented with casein hydrolysate and indole 3-acetic acid (devoid of 2,4 dichlorophenoxyacetic acid) and placed under low illumination.

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Abbreviations

IAA:

indole-3-acetic acid

IBA:

indole-3-butyric acid

BA:

benzyladenine

BSA:

bovine serum albumin

2,4-D:

2,4-dichlorophenoxyacetic acid

FW:

fresh weight

GA3 :

gibberellic acid

MES:

2-(N-morpholino) ethanesulfonic acid

MS medium:

Murashige and Skoog medium (1962)

NAA ⪤:

α-naphthaleneacetic acid

WPM:

woody plant medium (Lloyd and Mc Cown, 1981)

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Communicated by A. M Boudet

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Brison, M., Lamant, A. Callus formation from root protoplasts of Quercus rubra L. (red oak). Plant Cell Reports 9, 139–142 (1990). https://doi.org/10.1007/BF00232090

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Keywords

  • Cellulase
  • Zeatin
  • Benzyladenine
  • Casein Hydrolysate
  • Woody Plant Medium