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Title: IN VITRO CULTURE OF KENAF FOR TRANSFORMATION

Author
item SMITH, ROBERTA - TEXAS A&M
item SRIVATANAKUL, METINEE - TEXAS A&M
item SIJ, JOHN - TEXAS A&M - BEAUMONT, TX
item Webber Iii, Charles

Submitted to: Kenaf Association International Conference Proceedings
Publication Type: Proceedings
Publication Acceptance Date: 3/15/1997
Publication Date: N/A
Citation: N/A

Interpretive Summary: Weed control is a very important aspect in the commercial production of kenaf, especially during the early establishment of the crop. The development of herbicide resistant kenaf varieties through genetic engineering would be a tremendous benefit to the kenaf industry. Herbicide resistant genes are available, and there has been success in production of herbicide resistant cotton and other important crop plants. The first ste in the development of a herbicide resistant kenaf was to develop a cell culture system for kenaf which was rapid, genotype independent, and resulted in rooted plants at a high frequency. A gene vector was used in the transformation of kenaf. Seeds from kenaf varieties Everglades 71, Tainung 1, and Tainung 2 were used in the transformation process. Contamination was controlled and was minimal for all three cultivars. Shoots were isolated, elongated rapidly and formed roots. Some shoots rooted within seven days. All three cultivars develop into rooted plants in 1 to 3 months. In vitro derived plants appear to be normal. Shoots exposed to the gene vector survived 4 weeks of the selection process and are capable of establishment in soil. The next step is to analyze the plants at the molecular level to determine if the herbicide resistant gene is present.

Technical Abstract: A major restraint for commercial production of kenaf is weed competition during early establishment of the crop. An approach to solving this constraint is the development of herbicide resistant kenaf using the new technologies of genetic engineering. Herbicide resistant genes are available, and there has been success in production of herbicide resistant cotton and other important crop plants. The first priority in development of herbicide resistant kenaf is to develop a cell culture system for kenaf which is rapid, genotype independent, and results in rooted plants at a high frequency. This coupled with using Agrobacterium tumefaciens as the gene vector would give a practical system for transformation of kenaf. Seedlings from aseptically-germinated seeds from cultivars Everglades 71, Tainung 1, and Tainung 2 were established on a Murashige and Skoog inorganic salt medium. Contamination was minimal in all three cultivars. Shoot apices were isolated at three to five days and cultured on a Murashige and Skoog inorganic salt medium with vitamins, inositol, and 30% sucrose. Shoot apex cultures rapidly elongated and formed roots in vitro. Some shoots rooted within seven days in culture. All three cultivars developed into rooted plants in 1 to 3 months after shoot apex culture. In vitro derived plants appear to be normal in phenotype. Selection of non-treated shoot apices at 1.5 mg/L ppt for 3 weeks did not allow any plant survival. Shoot apices exposed to Agrobacterium containing the gene for herbicide resistance survived 4 weeks of ppt selection and plants can be established in soil.