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Improved transformation of alfalfa somatic embryos using a superbinary vector

100%

Ninkovic S., Miljus-Dukic J., Vinterhalter B., Neskovic M.

Acta Biologica Cracoviensia. Series Botanica

2004

tom 46

139-143

Increased transformation efficiency of alfalfa (Medicago sativa L. cv. Zaječarska 83), was achieved using Agrobacterium tumefaciens LBA4404 carrying the superbinary vector pTOK233 and the hpt gene for hygromycin resistance. Evidence for transformation was based on GUS activity and the presence of a fragment of the uidA gene in transformed embryos and regenerated plants, as demonstrated by the polymerase chain reaction. Efficiency was 1% (LBA4404/pBI121) to 14% (LBA4404/pTOK233) higher than that of some other vectors used in previous work with the same cultivar. The higher efficiency may be due to the presence of an extra set of vir genes in pTOK233. Moreover, the presence of the hpt gene, enabling the use of hygromycin instead of kanamycin for selection, was better for the development of secondary somatic embryos, thus contributing to a higher final number of transgenic plants.

Reproducible transformation in two grain legumes - soybean and azuki bean - using different systems

100%

El-Shemy H. A., Khalafalla M., Wakasa K., Ishimoto M.

Cellular and Molecular Biology Letters

2002

tom 07

nr 2B

Two plasmid vectors were introduced into soybean (Glycine max (L.) Merr.) and azuki bean (Vigna angularis Willd. Ohwi & Ohashi) using different transformation systems. Azuki bean epicotyl explants were prepared from etiolated seedlings and co-cultivated with Agrobacterium tumefaciens for 2 days. Adventitious shoots were developed from the callus of the explants on a regeneration medium containing hygromycin, and the shoots were excised and transferred to a rooting medium containing hygromycin at the same concentration. Rooting shoots were transferred to soil and grown in a glass-house to produce viable seeds. PCR analysis confirmed clearly the presence of the hpt gene in most of the azuki beans regenerated under hygromycin selection. A soybean embryogenic suspension culture was generated from immature cotyledons, and used for the introduction of plasmids by particle bombardment. Hygromycin-resistant embryogenic clones were isolated after 8 weeks of hygromycin selection, and then the green clones were matured on the differentiation medium. After desiccation, the embryos were germinated on the rooting medium, and the plants were transferred to soil in a glass-house. More than 50% of the regenerated soybean plants tolerant to hygromycin yielded the hpt fragment on PCR analysis. The azuki bean transformants were obtained more rapidly and with higher efficiency than the soybean transformants.

Ograniczanie wyników

1 Acta Biologica Cracoviensia. Series Botanica

1 Cellular and Molecular Biology Letters

1 El-Shemy H. A.

1 Ishimoto M.

1 Khalafalla M.

1 Miljus-Dukic J.

1 Neskovic M.

1 Ninkovic S.

1 Vinterhalter B.

1 Wakasa K.

1 2004

1 2002

Wyniki wyszukiwania

Improved transformation of alfalfa somatic embryos using a superbinary vector

Reproducible transformation in two grain legumes - soybean and azuki bean - using different systems