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Transgenic technology serves to introduce gene sequences for expression of a desired trait. Production of transgenic plants is reported in many crops, but commercialization is limited to a few selected crops, such as cotton (Gossypium hirsutum L.), corn (Zea mays L.), soybean [Glycine max (L.) Merr.], and canola (Brassica napus L. and B. rapa L.). This paper presents the sequential processes of transgenic event design, event selection, and “cleaning up” genetic background for forward breeding programs. Expression of the foreign gene cannot be viewed in isolation and is more complex than has been assumed because of the interaction of transgene with native genes. Variations among clones and within the progeny are observed, and hence all the clones are taken into account for evaluation. Plant breeding must be involved to move transgenes from transformable but agronomically unacceptable genotypes into elite breeding lines with two backcrosses. Production of transgenic plants in large numbers is difficult and laborious and requires large investments. It is worthwhile investing in parallel efforts to incorporate the transgene into improved plant material to satisfy commercial interests.

Embryogenic, friable and small (ca. 3 mm) calli showed optimum gus expression and were best suited for selection during genetic transformation of rice through particle bombardment. Through prolonged culture of mature seeds on original callus induction medium, this type of calli could be produced in large numbers across several elite rice genotypes. To minimize the non-transformed escapes 50 mg dm -3 hygromycin and 8 mg dm -3 glufosinate ammonium were found to be critical during selection. Addition of selection marker during regeneration was essential. Regular and frequent (every 15 d) transfer of calli to fresh selection medium for three cycles was also important. A simple and economic procedure for screening large number of putative resistant plants was described.

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Transgenic sorghum plants were produced through particle bombardment and Agrobacterium methods in two elite, but recalcitrant genotypes of Sorghum bicolor L. Moench. Use of target cells from developing tissues (immature embryos and multiple shoot buds), pre-culture of target tissue, small size of target tissue (2-3 mm), and regular subculture improved the selection and regeneration efficiencies. Addition of amino acid l-cysteine during co-cultivation and blotting sheet interface was helpful for complete decontamination of Agrobacterium tumefaciens and regeneration of transgenic plants. We demonstrated production of transgenic sorghum plants expressing a Bacillus thuringiensis lepidopteran toxin, through tailored in vitro protocols. Our results showed that decontamination of agrobacteria employing subtle treatments aided recovery of transgenic plants in recalcitrant genotypes. We generated 14 independent transgenic lines carrying different classes of B. thuringiensis toxin genes, cry1Aa and cry1B. Many single copy events were generated in two elite parental lines, CS3541 and 296B. Accumulation of the B. thuringiensis protein in leaves during the susceptible period of plant growth ranged from 35 to 500 ng/g fresh leaf tissue. Comprehensive insect bioassays for tolerance to spotted stem borer (Chilo partellus) were conducted through leaf disk and whole plant assays. Transgenic progeny plants showed 20-30% of damage as compared to 70-80% in non-transformed controls. (Résumé d'auteur