Adventitious Shoot Regeneration from in Vitro Subcultured Callus of Rhododendron Exbury Hybrids
James F. Harbage, Dennis P. Stimart
Abstract
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James F. Harbage, Dennis P. Stimart
Abstract
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Abstract Callus culture establishment and adventitious shoot development from callus of Rhododendron × ‘Gibraltar’ (G) and R. × ‘Old Gold’ (OG) after serial subculture were determined. Callus grew on Anderson's Rhododendron medium containing 2,4- D at 0.0045, 0.009, or 0.018 m m , but was optimum at 0.018 m m , since shoot organogenesis was suppressed at that level. After two callus subcultures, adventitious shoot development from callus on media containing 0, 0.017, 0.034, 0.068, or 0.136 m m zeatin (mixed isomers) was optimum at 0.034 and 0.068 m m zeatin for G and OG, respectively. After 17 weeks (five subcultures), 70- to 75-mg pieces of G and OG callus regenerated ≈20 shoots each. Adventitious shoot regeneration from callus declined with prolonged callus subculture. Regenerated plants are currently being evaluated for somaclonal variation. Chemical names used: (2,4-dichlorophenoxy)acetic acid (2,4-D); (E)-2-methyl-4-(1 H -purin-6-ylamino)-2-buren-1-ol (zeatin).
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Abstract Callus culture establishment and adventitious shoot development from callus of Rhododendron × ‘Gibraltar’ (G) and R. × ‘Old Gold’ (OG) after serial subculture were determined. Callus grew on Anderson's Rhododendron medium containing 2,4- D at 0.0045, 0.009, or 0.018 m m , but was optimum at 0.018 m m , since shoot organogenesis was suppressed at that level. After two callus subcultures, adventitious shoot development from callus on media containing 0, 0.017, 0.034, 0.068, or 0.136 m m zeatin (mixed isomers) was optimum at 0.034 and 0.068 m m zeatin for G and OG, respectively. After 17 weeks (five subcultures), 70- to 75-mg pieces of G and OG callus regenerated ≈20 shoots each. Adventitious shoot regeneration from callus declined with prolonged callus subculture. Regenerated plants are currently being evaluated for somaclonal variation. Chemical names used: (2,4-dichlorophenoxy)acetic acid (2,4-D); (E)-2-methyl-4-(1 H -purin-6-ylamino)-2-buren-1-ol (zeatin).
Key concepts: Callus, Subculture (biology), Zeatin, Shoot, Botany, Somaclonal variation, Biology, Organogenesis