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A Simple Method for Isolation of Soybean Protoplasts and Application to Transient Gene Expression Analyses
Published on: January 25, 2018
Development of a highly efficient, repetitive system of organogenesis in soybean (Glycine max (L.) Merr)
Zhihui Shan1, Krit Raemakers, Emmanouil N Tzitzikas
1Laboratory of Plant Breeding, Wageningen University, 386, 6709 AJ Wageningen, The Netherlands.
Abstract:
A highly efficient, repetitive system of organogenesis was developed in soybean. Seeds of soybean cv. 'White hilum' pretreated with TDZ formed multiple bud tissue(s) (MBT) at the cotyledonary nodes. MBT initiation occurred only if the axillary buds were not removed from the cotyledonary node. The best MBT formation was achieved by pretreating the seeds for 1 week on medium supplemented with 0.1 mg/l TDZ, followed by culture of the cotyledonary node on medium supplemented with 0.5 mg/l BA for 4 weeks. Culture of the MBT on medium supplemented with 0.1 mg/l TDZ resulted in the proliferation of MBT. MBT was maintained in this way for 12 months. Three hundred thirty six shoots were obtained when 1 g of MBT was subcultured on medium supplemented with 0.5 mg/l BA. Plants were rooted on medium without growth regulators. The regenerated plants grew normally in the greenhouse. Unfortunately, they did not set seeds because of the long-day conditions during growth. This system was successfully applied in three other genotypes.
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