A bacterial consortium attenuates the low-dose gamma-irradiation effect in kalanchoe plantlets

1Burlak, OP, 2Lar, ЕV, 3Rogutskyy, IS, 3Danilchenko, BA, 4Mikheev, OM, 2Zaets, IYe., 5de Vera, J-PP, 6Foing, BH, 2Kozyrovska, NA
1Institute of Molecular Biology & Genetics of NASU, Kyiv, Ukraine
2Institute of Molecular Biology and Genetics of the National Academy of Sciences of Ukraine, Kyiv, Ukraine
3Institute of Physics of the National Academy of Sciences of Ukraine, Kyiv, Ukraine
4Institute of Cell Biology & Genetic Engineering of the National Academy of Sciences of Ukraine, Kyiv, Ukraine
5German Aerospace Center (DLR) Berlin, Institute of Planetary Research, FRG, Berlin,Germany
6ESA/ESTEC/SRE-S, postbus 299 NL-2200 AG, Noordwijk, The Netherlands
Kosm. nauka tehnol. 2010, 16 ;(2):75-80
https://doi.org/10.15407/knit2010.02.075
Publication Language: English
Abstract: 
The ability of plants to protect themselves against ionizing radiation is limited. One of the ways how to alleviate irradiation consequences in plants is to use plant-associated bacteria for inoculation. Two defined plant growth promoting bacterial strains were used for inoculation Kalanchoe daigremontiana plantlets before acute irradiation with γ-quanta (60 Co). The lethal γ-rays doses were 3.0 kGy for Klebsiella oxytoca IMBG26, and 500 Gy for Paenibacillus sp. IMBG156. The bacteria expressed the increase of the pelX promoter activity after sublethal dose irradiation. The pelX promoter activity that was measured as activity of β-galactosidase of the pelX::lacZ fusion in K. oxytoca (pGalP) was 0,88 mkM/ml·min after exposure to 2.0 kGy, e.a. 80 % of the control (untreated) bacterial activity, although the irradiated bacterial population comprised 1.25 % of control one. Integrated index of plantlets development which was relied on both root number and root length reflected fluctuations in metabolic processes in irradiated plantlets without treatment with bacteria.
          Stabilizing stress-reactions occurred during 10 days after irradiation at different doses (30, 50, 70 Gy), however, index of growth (IG) remained at the level of 30‒60 % to control plantlets. The effect of irradiation on kalanchoe plantlets was relieved by bacteria at doses of 30 and 50 Gy, moreover, IG was observed at levels of 500 and 200 %, respectively. A 30 Gy dose was obviously stimulating for K. daigremontiana plantlets. Intense root elongation, instead of development of new coronal roots, led to fast adaptation to stressful conditions and normalization of metabolic processes in kalanchoe plantlets. However, integrated index showed inhibition of both inoculated and non-inoculated plantlets development after getting a 70 Gy dose.
Keywords: bacteria, gamma-irradiation, inoculation
References: 
1. Arkhipov N. P., Kuchma N. D., Askbrant S., et al. Acute and long-term effects of irradiation on pine (Pinus sylvestris) stands post-Chernobyl. Sci. Tot. Environ., 157, 383—386 (1994).
https://doi.org/10.1016/0048-9697(94)90601-7
2. Daly M. J. Engineering radiation-resistant bacteria for environmental biotechnology. Curr. Opin. Biotechnol., 11, 280—285 (2000).
https://doi.org/10.1016/S0958-1669(00)00096-3
3. Daly M. J., Gaidamakova E. K., Matrosova V. Y., et al. Accumulation of Mn(II) in Deinococcus radiodurans facilitates gamma-radiation resistance. Science, 306, 1025—1028 (2004).
https://doi.org/10.1126/science.1103185
4. Daly M. J., Gaidamakova E. K., Matrosova V. Y., et al. Protein oxidation implicated as the primary determinant of bacterial radioresistance. PLoS Biol., 5, 769—779 (2007).
https://doi.org/10.1371/journal.pbio.0050092
5. Danchenko M., Skultety L., Rashydov N. M., et al. Proteomic analysis of mature soybean seeds from the Chernobyl area suggests plant adaptation to the contaminated environment. J. Proteome Res., 8 (6), 2915—2922 (2009).
https://doi.org/10.1021/pr900034u
6. de Groot A., Dulermo R., Ortet P., et al. Alliance of proteomics and genomics to unravel the specificities of Sahara bacterium Deinococcus deserti. PLoS Genet., 5 (3), e1000434 (2009).
https://doi.org/10.1371/journal.pgen.1000434
7. Dittami S., Scornet D., Petit J.-L., et al. Global expression analysis of the brown alga Ectocarpus siliculosus (Phaeo-phyceae) reveals large-scale reprogramming of the tran-scriptome in response to biotic stress. Genome Biology, 10, R66 (2009).
https://doi.org/10.1186/gb-2009-10-6-r66
8. Kovalchuk I., Abramov V., Pogribny I., et al. Molecular aspects of plant adaptation to life in the Chernobyl zone. Plant Physiol., 135, 357—363 (2004).
https://doi.org/10.1104/pp.104.040477
9. Kovalchuk O., Burke P., Arkhipov A., et al. Genome hypermethylation in Pinus silvestris of Chernobyl — a mechanism for radiation adaptation. Mutat. Res., 529, 13—20 (2003).
https://doi.org/10.1016/S0027-5107(03)00103-9
10. Kozyrovska N., Negrutska V., Kovalchuk M., et al. Paenibacillus sp., a promising candidate for development of a novel technology of plant inoculant production. Biopolymers and Cell., 21 (4), 312—319 (2005).
https://doi.org/10.7124/bc.0006F7
11. Lar O. V., Kovtunovych G. L., Kozyrovska N. O. Cloning and analysis of the gene encoding pectate lyase, the Klebsiella oxytoca VN13 pelX. Biopolymers and Cell., 18 (5), 417—422 (2002).
https://doi.org/10.7124/bc.000620
12. Lar O. V., Kovtunovych G. L., Kozyrovska N. O. A study of Klebsiella oxytoca exopectate lyase the pelX gene. Biopolymers and Cell., 21 (3), 264—270 (2005).
https://doi.org/10.7124/bc.0006F0
13. Le-Tien C., Lafortune R., Shareck F., et al. DNA analysis of a radiotolerant bacterium Pantoea agglomerans by FT-IR spectroscopy. Talanta, 71 (5), 1969—1975 (2007).
https://doi.org/10.1016/j.talanta.2006.09.003
14. Makarova K. S., Omelchenko M. V., Gaidamakova E. K., et al. Deinococcus geothermalis: The pool of extreme radiation resistance genes shrinks. PLoS ONE, 2 (9), 955 (2007).
https://doi.org/10.1371/journal.pone.0000955
15. Melki M., Sallami D. Studies the effects of low dose of gamma rays on the behaviour of chickpea under various conditions in Pakistan. J. Biol. Sci., 11 (19), 2326—2330 (2008).
16. Miller J. H. Experiments in Molecular Genetics, 432 p. (Cold Spring Harbor Laboratory Press, New York, 1972).
17. Mittler R., Vanderauwera S., Gollery M., et al. Reactive oxygen gene network of plants. Trends Plant Sci., 9, 490—498 (2004).
https://doi.org/10.1016/j.tplants.2004.08.009
18. Nishimura A., Morita M., Sugino Y. A. Rapid and highly efficient method for preparation of competent Esherichia coli cells. Nucl. Acid Res., 18, 6169 (1990).
https://doi.org/10.1093/nar/18.20.6169
19. Sambrook J., Fritsch E. F., Maniatis T. Molecular Cloning: a laboratory manual. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N. Y., 1989).
20. Sorochinskiy B., Kozyrovska N. Biotechnological aspects of problem associated with phytoremediation of the environment from radionuclide pollution. Agrobiotechnologia, 2, 123—130 (1998).
21. Sorochinskiy B., Prokhnevskiy O., Ruchko M. Some mechanisms of somatic effects of irradiation indicated in plants from the 10 km zone of Chernobyl APS. Cytologyia i Genetika, 30 (4), 15—19 (1996).
22. Yang J., Kloepper J. W., Rye C. M. Rhizosphere bacteria help plants tolerate abiotic stress. Trends Plant Sci., 14, 1—4 (2009).
https://doi.org/10.1016/j.tplants.2008.10.004
23. Zaetz I. E., Kozyrovska N. O. Effect of a bacterial consortium on oxidative stress in soybean plants in cadmium-contaminated soil. Biopolymers and Cell., 24, 246—253 (2008).
https://doi.org/10.7124/bc.0007A7
24. Zaka R., Vandecasteele C. M., Misset M. T. Effects of low chronic doses of ionizing radiation on antioxidant enzymes and G6PDH activities in Stipa capillata (Poaceae). J. Exp. Bot., 53 (376), 1979—1987 (2002).
https://doi.org/10.1093/jxb/erf041