Heavy metals-resistant PGPR strains of Pseudomonas sp. stimulating the growth of alfalfa under cadmium stress

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Abstract

Three bacteria strains of Pseudomonas sp. resistant to heavy metals were isolated from the chemically contaminated soil. According to the results on the nucleotide sequences of the 16S rRNA and rpoD genes, strain Pseudomonas sp. 17 НМ was identified as Pseudomonas capeferrum, and the strains of Pseudomonas sp. 65 НМ и 67 НМ were most closely related to the type strain of Pseudomonas silesiensis и Pseudomonas umsongensis, respectively. It has been shown that strains of Pseudomonas sp. 17 НМ, 65 НМ, 67 НМ are characterized by different levels of resistance of heavy metals: maximum tolerance concentration (MTC) of zinc was 1 mМ for all strains, cadmium 1, 1.5, 1 mМ, lead 5, 5, 4 mМ, nickel 7, 9, 7 mМ, respectively. All pseudomonad strains can form biofilms and have the properties of PGPR bacteria. Treatment of alfalfa seeds (Medicago sativa L.) with strains Pseudomonas sp. 17 НМ, 65 НМ, 67 НМ under cadmium stress led to an increase in the dry weight of alfalfa seedling up to 40 % and the content of chlorophyll a and b in the leaves by 25-33% relative to the control.

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O. V. Chubukova

Institute of Biochemistry and Genetics—Subdivision of the Ufa Federal Research Centre of the Russian Academy of Sciences

Author for correspondence.
Email: chubukova@bk.ru
Russian Federation, st. Prospect Oktyabrya, 71, Ufa, 450054

L. R. Khakimova

Institute of Biochemistry and Genetics—Subdivision of the Ufa Federal Research Centre of the Russian Academy of Sciences

Email: chubukova@bk.ru
Russian Federation, st. Prospect Oktyabrya, 71, Ufa, 450054

R. T. Matnyazov

Institute of Biochemistry and Genetics—Subdivision of the Ufa Federal Research Centre of the Russian Academy of Sciences

Email: chubukova@bk.ru
Russian Federation, st. Prospect Oktyabrya, 71, Ufa, 450054

Z. R. Vershinina

Institute of Biochemistry and Genetics—Subdivision of the Ufa Federal Research Centre of the Russian Academy of Sciences; Federal State Budgetary Educational Institution of Higher Education “Ufa State Petroleum Technological University”

Email: chubukova@bk.ru
Russian Federation, st. Prospect Oktyabrya, 71, Ufa, 450054; st. Kosmonavtov, 1, Ufa, 450000

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2. Fig. 1. Phylogenetic tree of bacteria constructed based on comparative analysis of 16S rRNA gene sequences. The numbers show the statistical reliability of the branching order determined using bootstrap analysis (the values ​​of the bootstrap analysis indicator from 70% are shown). The horizontal axis shows the weight of this alignment, expressed as the number of nucleotide substitutions (×100). The nucleotide sequence of the 16S rRNA gene of E. coli ATCC 11775T was used as an outgroup.

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3. Fig. 2. Phylogenetic tree of bacteria constructed based on comparative analysis of rpoD gene sequences. The numbers show the statistical significance of the branching order determined using bootstrap analysis (the values ​​of the bootstrap analysis indicator from 70% are shown). The horizontal axis shows the weight of this alignment, expressed as the number of nucleotide substitutions (×100). The nucleotide sequence of the rpoD gene of E. coli ATCC 11775T was used as an outgroup.

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4. Fig. 3. Biofilms formed by Pseudomonas sp. strains on the plate surface after 7 days of cultivation: a – Pseudomonas sp. 17 HM, LB medium; b – Pseudomonas sp. 65 HM, YM medium; c – Pseudomonas sp. 67 HM, MH medium.

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5. Fig. 4. Formation of biofilms on the surface of alfalfa seedling roots after 24 hours of co-cultivation: a – Pseudomonas sp. 17 HM; b – Pseudomonas sp. 65 HM; c – Pseudomonas sp. 67 HM.

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6. Fig. 5. Dynamics of inorganic phosphate mobilization by Pseudomonas sp. strains: 1 – 17 HM; 2 – 67 HM; 3 – 65 HM. The abscissa axis shows the day on which the phosphate mobilization area was measured; the ordinate axis shows the area of ​​phosphate mobilization zones in mm².

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7. Fig. 6. Effect of Pseudomonas sp. strains on dry biomass of alfalfa plants (ordinate axis: change in dry biomass of plants in g under normal conditions (a) and under cadmium stress (b). Designations of Pseudomonas sp. strains (1–4): 1 – control; 2 – 17 HM; 3 – 65 HM; 4 – 67 HM.

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