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Diversity of soybean bacterial blight pathogen Pseudomonas savastanoi pv. glycinea in the Russian Federation
- Authors: Tarakanov R.I.1, Evseev P.V.2, Troshin K.S.1, Ignatov A.N.3, Dzhalilov F.S.1
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Affiliations:
- Russian State Agrarian University - Moscow Timiryazev Agricultural Academy
- Shemyakin - Ovchinnikov Institute of Bioorganic Chemistry
- RUDN University
- Issue: Vol 19, No 1 (2024): Factors of sustainable animal productivity: from genomics to therapy
- Pages: 139-154
- Section: Plant protection
- URL: https://agrojournal.rudn.ru/agronomy/article/view/19997
- DOI: https://doi.org/10.22363/2312-797X-2024-19-1-139-154
- EDN: https://elibrary.ru/AIEVBJ
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Abstract
One of the most harmful bacterial diseases of soybean is bacterial blight caused by bacterium Pseudomonas savastanoi pv. glycinea. The pathogen can reduce soybean yield (up to 40 %), oil content and seed germination. To manage the pathogen damage, protection measures should be comprehensive, the most cardinal of which is breeding for resistance. To obtain resistant varieties, it is necessary to understand the diversity of the pathogen in the area and to breed against the most common and harmful forms of the pathogen. In this regard, the aim of the study was to characterize Pseudomonas savastanoi pv. glycinea strains circulating in the Russian Federation as casual agents of bacterial blight of soybean. 12 strains of the soybean bacterial blight pathogen were isolated from soybean plant parts and seeds grown in different regions of the Russian Federation. The isolated strains were identical to the reference strain Psg CFBP 2214 in fluorescence, colony morphology on King B medium and LOPAT test results (+, –, –, –, –, +) and formed an amplicon in specific PCR analysis of cfl (coronafacate ligase) gene. The strains had different virulence to Kasatka soybean plants, and the width of symptomatic zone when leaves were artificially inoculated ranged from 3.23 mm (in strain G7) to 6.53 mm in strain G4. Comparison of the obtained gltA and ITS 16S-23S rRNA gene sequences showed a high (95.8…98.8 %) identity to the bacterial blight pathogen strains deposited to NCBI Genbank, and phylogenetic analysis showed a low intra-strain genetic polymorphism. Analysis of the race composition of the strains showed that the race 4 was predominant in the Russian Federation.
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Table 1. Pseudomonas savastanoi pv. glycinea strains used in the research
Strain | Host | Part of plant | Source | Year of isolation |
Strains of Pseudomonas savastanoi pv. glycinea | ||||
G1 | Soybean (Glycine max) | seeds | Voronezh region | 2019 |
G2 | Amur region | 2019 | ||
G3 | Amur region | 2019 | ||
G4 | Khabarovsk region | 2020 | ||
G5 | Voronezh region | 2020 | ||
G6 | Voronezh region | 2021 | ||
G7 | Voronezh region | 2021 | ||
G8 | leaves | Primorsky region | 2021 | |
G9 | seeds | Khabarovsk region | 2021 | |
G10 | Amur Region | 2021 | ||
G11 | Khabarovsk region | 2021 | ||
G17 | Khabarovsk region | 2021 | ||
CFBP 2214 | leaves | New Zealand | 1968 | |
Pseudomonas sp. | ||||
P. fluorescens | — | soil | Krasnodar region | 2021 |
P. putida | — | soil | 2021 | |
Fig. 1. Virulence of Pseudomonas savastanoi pv. glycinea strains: A – difference in chlorosis zone width between strains and control; Б – Values of leaf lesion zone width 12 days after inoculation with different strains. Different letters near bars indicate statistical difference by Duncan test (P < 0.05)
Source: created by the authors
Fig. 2. PCR detection of cfl gene in Pseudomonas savastanoi pv. glycinea strains: M – 100+bp molecular weight marker (Eurogen) (#NL002); K– – negative control (reaction without DNA); G1-G17 — analyzed strains
Source: created by the authors
Table 2. Pseudomonas savastanoi pv. glycinea strains identified by gltA and ITS 16S-23S rRNA genes sequences
Strain | Closest species when comparing sequences with Genbank database by | Sequence identification number in Genbank | ||
citrate synthase gene (gltA) | ITS rRNA 16S-23S gene | by citrate synthase gene (gltA) | by ITS rRNA 16S-23S gene | |
G1 | P. syringae; P. savastanoi | P. amygdali; P. syringae | OQ743493 | OR750531 |
G2 | P. savastanoi; P. syringae | P. amygdali; P. syringae | OQ743494 | OR750532 |
G3 | P. savastanoi | P. syringae; P. savastanoi | OQ743495 | OR750533 |
G4 | P. savastanoi; P. syringae | P. syringae | OQ743496 | OR750534 |
G5 | P. savastanoi | P. syringae; P. amygdali | OQ743497 | OR750535 |
G6 | P. savastanoi; P. syringae | P. syringae | OQ743498 | OR750536 |
G7 | P. syringae; P. amygdali | P. amygdali; P. syringae | OQ743499 | OR750537 |
G8 | P. savastanoi | P. syringae; P. savastanoi | OQ743500 | OR750538 |
G9 | P. savastanoi; P. syringae | P. syringae | OQ743501 | OR750539 |
G10 | P. savastanoi; P. syringae | P. syringae; P. amygdali | OQ743502 | OR750540 |
G11 | P. savastanoi; P. syringae | P. amygdali; P. syringae | OQ743503 | OR750541 |
G17 | P. savastanoi | P. syringae; P. amygdali | OQ743504 | OR750542 |
Fig. 3. Phylogenetic trees of nucleotide sequences of citrate synthase (gltA) (A) and ITS 16S‑23S rRNA (Б) genes of Pseudomonas spp. Orange boxes indicate the strains studied in the research
Source: created by the authors
Fig. 4. Heat map of ITS 16S‑23S rRNA gene sequences showing pairwise genetic distances of P. savastanoi pv. glycinea strains and other P. syringae pathovars. Different colors correspond to the percent of genetic difference in pairwise comparison of DNA from different strains
Source: created by the authors
Table 3. Results of inoculation of differentiator varieties with Pseudomonas savastanoi pv. glycinea strains used in this work (S = susceptible, R = resistant)
Сорт-дифференциатор /Differentiator variety | Штамм / Strain | ||||||||||||
G1 | G2 | G3 | G4 | G5 | G6 | G7 | G8 | G9 | G10 | G11 | G17 | CFBP 2214 | |
Acme | S | S | S | S | S | S | S | S | S | S | S | S | S |
Chippewa | S | S | S | S | S | S | S | S | S | S | S | S | S |
Flambeau | S | S | S | S | S | S | S | S | S | S | S | S | S |
Harosoy | S | S | S | S | S | S | S | S | S | S | S | S | S |
Lindarin | S | S | S | S | S | S | S | S | S | S | S | S | S |
Merit | S | S | S | S | S | S | S | S | S | S | S | S | S |
Norchief | S | S | S | S | S | S | S | S | S | S | S | S | S |
Hardee | S | S | S | S | S | S | S | S | S | S | S | S | S |
Peking | S | S | S | S | S | S | S | S | S | S | S | S | S |
Centennial | S | S | S | S | S | S | S | S | S | S | S | S | S |
Номер расы / Number of race | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 | 4 |
About the authors
Rashit I. Tarakanov
Russian State Agrarian University - Moscow Timiryazev Agricultural Academy
Author for correspondence.
Email: tarakanov.rashit@mail.ru
ORCID iD: 0000-0002-3235-8467
SPIN-code: 9049-7157
assistant, PhD student, Department of Plant Protection
49 Timiryazevskaya st., Moscow, 127434, Russian FederationPeter V. Evseev
Shemyakin - Ovchinnikov Institute of Bioorganic Chemistry
Email: petevseev@gmail.com
ORCID iD: 0000-0002-1646-9802
SPIN-code: 4275-9187
Candidate of Biological Sciences, Researcher
16/10 Miklouho-Maklaya st., Moscow, 117997, Russian FederationKonstantin S. Troshin
Russian State Agrarian University - Moscow Timiryazev Agricultural Academy
Email: konstantinetr@gmail.com
ORCID iD: 0009-0004-5018-1265
SPIN-code: 6032-4313
Master student, Junior researcher, Department of Plant Protection
49 Timiryazevskaya st., Moscow, 127434, Russian FederationAleksandr N. Ignatov
RUDN University
Email: an.ignatov@gmail.com
ORCID iD: 0000-0003-2948-753X
SPIN-code: 3324-4985
Doctor of Biological Sciences, Professor, Department of Agrobiotechnology
6 Miklouho-Maklaya st., Moscow, 117198Fevzi S.U. Dzhalilov
Russian State Agrarian University - Moscow Timiryazev Agricultural Academy
Email: labzara@mail.ru
ORCID iD: 0000-0002-5014-8375
SPIN-code: 3033-3991
Doctor of Biological Sciences, Professor, Head of the Department of Plant Protection
49 Timiryazevskaya st., Moscow, 127434, Russian FederationReferences
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