To improve Agrobacterium-mediated transformation of tobacco, factors influencing gene delivery, including genotype of the plant, bacterial strain, and Agrobacterium transformation procedure, were tested via direct somatic embryogenesis. Leaf tissue of three different tobacco genotypes (Nicotiana tabacum L. cvs. Samsun, and Xanthi, and N. benthamiana) we...

Transformed Arabidopsis thaliana plants have been produced by a modified leaf disk transformation-regeneration method. Leaf pieces from sterilely grown plants were precultured for 2 days and inoculated with an Agrobacterium tumefaciens strain containing an avirulent Ti (tumor-inducing) plasmid with a chimeric gene encoding hygromycin resistance. After cocultivation for 2 days, the leaf pieces w...

In vitro and in vivo screening methods were established for the discovery of new active substances against Treponema hyodysenteriae. During the screening methods, hygromycin A produced by Streptomyces hygroscopicus KA-355 was found to be active against T. hyodysenteriae. Hygromycin A did not show high antitreponemal activity in in vitro test using the paper disc method on the agar plate inocula...

A protocol for the generation of high yields of viable protoplasts has been developed for several highly branched (colonial) strains of the Quorn2 myco-protein fungus, Fusarium graminearum A3}5. Driselase was found to produce higher protoplast yields (ca 10* g−" wet weight) than the other lytic enzymes tested (Glucanex, Novozyme, β-glucuronidase, Sigma lytic enzyme, or ICN lytic enzyme), althou...

An efficient Agrobacterium tumefaciens-mediated genetic transformation method was successfully established for a newly isolated Taxol-producing fungus, Ozonium sp EFY21. A specific hygromycin B resistance expression vector, pCAMBIA1304'AN7-1, was constructed for fungal transformation. Key factors affecting transformation efficiency were thoroughly investigated and optimized. PCR amplification a...

Of the three dominant marine microalgal groups, dinoflagellates and diatoms can undergo genetic transformation; however, no transformation method has been established for haptophytes to date. Here, we report the first stable genetic transformation of a coccolithophore, Pleurochrysis carterae, by means of polyethylene glycol (PEG)-mediated transfer of a bacterial hygromycin B-resistance gene. To...

With the binary vectors pBIG and pBIB it is possible to: i) exchange the plant selectable marker/promoter fusion and the reporter gene, ii) use one vector and Agrobacterium strain system, thus reducing variables associated with using different vectors and strains, iii) allow for divergent transcription of the selectable marker gene and of promoter/reporter gene fusions. The two plasmids are der...

Hygromycin A, an antibiotic produced by Streptomyces hygroscopicus, is an inhibitor of bacterial ribosomal peptidyl transferase. The antibiotic binds to the ribosome in a distinct but overlapping manner with other antibiotics and offers a different template for generation of new agents effective against multidrug-resistant pathogens. Reported herein are the results from a series of stable-isoto...