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Use of Buckwheat Seed Protease Inhibitor Gene for Improvement of Tobacco and Potato Plant Resistance to Biotic Stress

N. V. Khadeeva1, E. Z. Kochieva2,3, M. Yu. Tcherednitchenko3, E. Yu. Yakovleva1, K. V. Sydoruk4, V. G. Bogush4, Y. E. Dunaevsky5, and M. A. Belozersky5*

1Vavilov Institute of General Genetics, Russian Academy of Sciences, ul. Gubkina 3, 119991 Moscow, Russia

2Bioengineering Center, Russian Academy of Sciences, pr. 60-letiya Oktyabrya 7/1, 117312 Moscow, Russia

3Russian State Agricultural University, Timiryazev Moscow Agricultural Academy, ul. Timiryazevskaya 49, 127550 Moscow, Russia

4FSUE State Scientific Center of Russian Federation “State Scientific Research Institute of Genetics”, 1-yi Dorozhnyi Proezd 1, 117545 Moscow, Russia

5Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119992 Moscow, Russia; fax: (495) 939-3181; E-mail: mbeloz@belozersky.msu.ru

* To whom correspondence should be addressed.

Received July 4, 2008; Revision received September 29, 2008
The possibility to use agrobacterial transformation of leaf discs to produce resistance to bacterial infections in tobacco and potato plants by introduction of a single gene encoding the serine proteinase inhibitor BWI-1a (ISP) from buckwheat seeds is shown. All studied PCR-positive transgenic plants exhibited antibacterial activity in biotests. It was shown that the presence of just a single gene of serine proteinase inhibitor provides sufficient protection at least against two bacterial phytopathogens, Pseudomonas syringae pv. tomato and Clavibacter michiganensis sbsp. michiganensis. The biotest including tobacco plant infection by the white wings butterfly in the green house has also demonstrated the existence of protective effect in transgenic tobacco plants. Significant genotypic variations in the protection efficiency were found between members of different genera of the same family (potato and tobacco) as well as between different lines of the same species. Northern blot analysis of four transgenic potato lines and three tobacco lines transformed by a vector plasmid containing the ISP gene of serine proteinases BWI-1a from buckwheat seeds has shown the presence of the expected size mRNA transcript.
KEY WORDS: agrobacterial transformation, serine proteinase inhibitor gene, tobacco, potato, protection against phytopathogens

DOI: 10.1134/S0006297909030031