Transformation was performed as described previously (38). In brief, friable, embryogenic calli (cv. 60444) were transformed with Agrobacterium (strain LBA4404) harboring the expression constructs described above. Following cocultivation, somatic embryogenic tissue was regenerated on MS-based media (58) containing hygromycin B for plant selection and the synthetic auxin NAA. Differentiating somatic embryos were transferred to MS-based media containing the synthetic cytokinin BAP to induce shoot development. Emerging juvenile shoots were propagated on standard media (lacking auxins or cytokinins) to establish in vitro plantlets. Following normal growth, the apical shoot tip was transferred to MS-based media containing hygromycin B to select further for transgenic material. Only plantlets positive for the plant selection marker (hptII) would form roots and were used for molecular analyses and propagated for glasshouse cultivation.

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