Agrobacterium‐mediated transformation of sea urchin embryos

Abstract

Agrobacterium-mediated transformation of higher plants is a well-known and powerful tool for transgene delivery to plant cells. In the present work, we studied whether Agrobacterium can transfer genetic information to animal (sea urchin) embryos. Sea urchin embryos were co-cultivated with A. tumefaciens strains carrying binary vectors containing the nptII marker gene and agrobacterial rolC and rolB oncogenes. Bacterial plasmid T-DNA-sea urchin DNA junction sites were identified in the genome of these embryos, thus indicating successful transformation. The nptII and both rol genes were expressed in the transformed embryos. The processes of transgene integration and transgene expression were suppressed when Agrobacteria contained mutated virA, virB or virG genes, suggesting that Agrobacterium transforms sea urchin cells by a mechanism similar to that which mediates T-DNA transfer to plants. Some of the embryos co-cultivated with Agrobacterium developed teratoma-like structures. The ability of Agrobacterium strains to trigger formation of teratoma-like structures was diminished when they contained the mutated vir genes. In summary, our results demonstrate that Agrobacterium is able to transform animal (sea urchin) embryonic cells, thus indicating a potential of this natural system for gene delivery to animal hosts. We also discuss the possibility of horizontal gene transfer from Agrobacterium to marine invertebrates.

Natural plant genetic engineer Agrobacterium rhizogenes: role of T-DNA in plant secondary metabolism

“…Introducing Ri (root inducing) plasmids into the cured strain converts the bacterium into a rhizogenic strain (Lam et al 1984). Agrobacterium has a wide range of host infection including transfer of DNA to human cells (Kunik et al 2001), sea urchin embryos (Bulgakov et al 2006), angiosperms, gymnosperms, fungi, including yeasts, ascomycetes and basidiomycetes (Gelvin 2003). Several monocotyledonous plants like rice, corn, wheat, sugarcane, Linum flavum have been transformed successfully.…”

T-DNA Binary Vectors and Systems

“…Agrobacterium transfers T-DNA, which makes up a small (approximately 5%-10%) region of a resident Ti-plasmid or root-inducing plasmid (Ri-plasmid), to numerous species of plants (DeCleene and DeLey, 1976;Anderson and Moore, 1979), although the bacterium can be manipulated in the laboratory to transfer T-DNA to fungal (Bundock et al, 1995;Piers et al, 1996;de Groot et al, 1998;Abuodeh et al, 2000;Kelly and Kado, 2002;Li et al, 2007) and even animal cells (Kunik et al, 2001;Bulgakov et al, 2006). Transfer requires three major elements: (1) T-DNA border repeat sequences (25 bp) that flank the T-DNA in direct orientation and delineate the region that will be processed from the Ti/Ri-plasmid (Yadav et al, 1982); (2) vir genes located on the Ti/Ri-plasmid; and (3) various genes (chromosomal virulence [chv] and other genes) located on the bacterial chromosomes.…”

Agrobacterium tumefaciens-Induced Bacteraemia Does Not Lead to Reporter Gene Expression in Mouse Organs

“…This idea is consistent with our observations that A. tumefaciens can persist in the mouse bloodstream without detectible expression of its T-DNA in the host tissues as well as with previous findings that A. tumefaciens genetically transforms cultured mammalian cells inefficiently [20], probably at the levels undetectable in whole animal tissues. Interestingly, embryonic tissues may be more susceptible to transformation which has been recently reported for sea urchin embryos [21].…”