Issue |
Agron. Sustain. Dev.
Volume 28, Number 4, October-December 2008
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Page(s) | 481 - 488 | |
DOI | https://doi.org/10.1051/agro:2008025 | |
Published online | 21 June 2008 |
DOI: 10.1051/agro:2008025
Transgenic resistance of Bulgarian potato cultivars to the Colorado potato beetle based on Bt technology
Ivanka Kamenova1, Rossitza Batchvarova1, Stanislaw Flasinski2, Lidia Dimitrova3, Petya Christova1, Slavcho Slavov1, Atanas Atanassov1, Plamen Kalushkov4 and Wojciech Kaniewski51 AgroBioInstitute, 8 Dragan Tzankov Blvd., 1164 Sofia, Bulgaria
2 Monsanto, GG3220, 700 Chesterfield Parkway West, St. Louis, MO 63017, USA
3 Potato Laboratory, 2000 Samokov, Bulgaria
4 Institute of Zoology, Bulgarian Academy of Sciences, 1 Zar Osvoboditel, 1000 Sofia, Bulgaria
5 Adam Mickiewicz University, 85 Umultowska ul. 61-614 Poznan, Poland
Accepted 30 April 2008; published online 21 June 2008
Abstract - Colorado potato beetle, Leptinotarsa decemlineata Say, is the most destructive
insect pest of potatoes. When the population of beetles is high, plants can
be completely defoliated and commercial potato production is nearly
impossible without control of the beetle. The beetles have shown a
tremendous ability to develop resistance against insecticides. Previously, a
biotechnology approach to control Colarado potato beetle based on the use of
the synthetic Bt gene was developed. In this article, a
transformation procedure for three commercial Bulgarian potato
cultivars was developed and potentially commercial transgenic lines have
been selected based on field resistance to Colorado potato beetles and
yield. Plants were transformed with the Bacillus thuringiensis (Bt) cry3A gene using
Agrobacterium-mediated transformation. 110 plants from the three cultivars were
regenerated and tested by double antibody sandwich enzyme-linked
immunosorbent assay (DAS-ELISA). The Cry3A protein accumulation varied
across the transgenic lines, rating from very low to 71.5 g/g fresh
weight. 21 transgenic lines expressing the Cry3A protein at levels above 10
g/g fresh weight were tested in two successive years in filed
conditions at two different locations of the country. All
transgenic lines compared with the controls, nontransgenic potatoes from
the respective cultivar, were consistently protected from foliar damages from
all developmental stages of the beetle. The comparison of all properties of
the tested transgenic lines, including variety phenotypes and tuber yield,
allowed the selection of the most promising 2–3 lines per cultivar. Selected
lines produced tuber amounts 80–100% higher compared with the control,
non-transgenic plants. Those lines were grown for mass propagation during
the third year of field experiments. The presence of the transgene in these
lines was confirmed with the use of primers specific to the transgene by
polymerase chain reaction (PCR). Additionally, the results from the
insect bioassay showed that these lines were highly resistant to insect
feeding, leading to 100% of mortality of larval populations. In summary,
we generated potentially commercial potato lines highly resistant to
Colorado potato beetle using Bt technology that may have a profound impact
on development of sustainable agriculture in Bulgaria. This is one of the
several agriculture biotechnology products entirely developed and tested in
Bulgaria.
Key words: potato / Bt Cry3A / Colorado potato beetle resistance / field tests / genetic transformation
Corresponding author: vanjakamenova@yahoo.com
© INRA, EDP Sciences 2008