AGROBACTERIAL TRANSFORMATION OF ST. JOHN'S WORT HIPERICUM PERFORATUM L.

Issue: 
10
Year: 
2016

V.N. Ovchinnikova
Ph.D. (Biol.), Leading Research Scientist, Russian Federal Research Institute of Agricultural Biotechnology (Moscow)
E-mail: vera.ovchinnikova.1957@mail.ru
N.V. Nikiforova
Junior Research Scientist, Russian Federal Research Institute of Agricultural Biotechnology (Moscow)
E-mail: nikif_natali@mail.ru
P.N. Kharchenko
Academician RANS, Russian Federal Research Institute of Agricultural Biotechnology (Moscow)
Е-mail: kharchenko@iab.ac.ru; iab@iab.ac.ru

We attempted to hold agrobacterial transformation of explants of Hypericum perforatum L. green plant parts (stem callus) by using A. tumefacience. Object of research are test-tube plant of St. John's wort (Hypericum perforatum L.) (wild ecotype). As explants for ATMT used stem callus of Hypericum. For agrobacterial transformation was used a strain AGL0 Agrobacterium tumefaciens. Vector design contains a marker gene GUS. As an selective factor appeared a resistance gene of regenerants to an antibiotic hygromycin. Micro shoots of Hypericum perforatum L. tested by PCR analysis for the presence of the insert hpt in the plant DNA (resistance to the antibiotic Hg), as well as for contamination of agrobacterium (virE- gene abstnce). Thus, it can be concluded about the possibility of genetic transformation of Hypericum perforatum L. by using Agrobacterium tumefaciens. Result we have obtained allows us to continue works on getting of the expression construction containing a target gene Hyp1.1, which is believed to affect the synthesis of hypericin Hypericum perforatum L. for further transformation of the culture to increase the synthesis of hypericin.

Keywords: 
Key words: St. John’s wort (Hypericum perforatum L.)
Agrobacterium tumefaciens-mediated transformation (АТМТ)
hygromycin (Hg)
in vitro shoot regeneration
PCR analysis.