Plant regeneration through callus initiation from thin mature embryo fragments of wheat |
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Authors: | Delporte F. Mostade O. Jacquemin J.M. |
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Affiliation: | (1) Centre de Recherches agronomiques - Département de Biotechnologie, Chaussée de Charleroi 234, B5030 Gembloux, Belgium |
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Abstract: | A wheat regeneration system was developed using mature embryos. Embryos were removed from surface-sterilised mature caryopses (winter wheat Odeon cultivar and spring wheat Minaret cultivar) and ground to pieces through a sterile nylon mesh. The fragments were characterised by means of the image analysis technique. They were 500 M mean diameter and most of them were elongated. They were used as explants to initiate embryogenic calli on solid medium supplemented with 10 M 2,4-dichlorophenoxyacetic acid. The morphogenic pathway of the initiated calli was followed for a 40-day culture period. Active cellular division occurred within 24 hours of cultivation. Several hundred calli were produced from 100 fragmented embryos within 3 days. A 90% callus induction rate was achieved and proembryos appeared by the 8th day of culture. The highest embryogenic calli induction rate of 47% was obtained when 2,4-dichlorophenoxyacetic acid was suppressed after a 3–4 week induction period. Two regeneration methods were finally compared. A total of 513 plantlets were produced. The optimal protocol produced 25–30 plants per 100 embryos. This regeneration method may be suitable for transformation applications. |
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Keywords: | in vitro regeneration mature embryo somatic embryogenesis Triticum aestivum L |
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