ЧУТЛИВІСТЬ РИЗОБАКТЕРІЙ РІЗНИХ ВИДІВ ДО ВПЛИВУ ҐРУНТОВИХ ГЕРБІЦИДІВ В УМОВАХ IN VITRO
Objective. To investigate the sensitivity of Bradyrhizobium japonicum, Bradyrhizobium elkanii, Azospirillum brasilense, Pseudomonas fluorescens, and Azotobacter chroococcum to the effects of soil-applied herbicides under in vitro conditions. Methods. Microbiological and statistical. The sensitivity...
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| author | Коць, С. Я. Хоменко, В. Д. Кукол, К. П. Пухтаєвич, П. П. |
| author_facet | Коць, С. Я. Хоменко, В. Д. Кукол, К. П. Пухтаєвич, П. П. |
| author_institution_txt_mv | [
{
"author": "С. Я. Коць",
"institution": "Інститут фізіології рослин і генетики НАН України"
},
{
"author": "В. Д. Хоменко",
"institution": "Інститут фізіології рослин і генетики НАН України"
},
{
"author": "К. П. Кукол",
"institution": "Інститут фізіології рослин і генетики НАН України"
},
{
"author": "П. П. Пухтаєвич",
"institution": "Інститут фізіології рослин і генетики НАН України"
}
] |
| author_sort | Коць, С. Я. |
| baseUrl_str | https://smic.in.ua/index.php/journal/oai |
| collection | OJS |
| datestamp_date | 2026-07-22T10:10:52Z |
| description | Objective. To investigate the sensitivity of Bradyrhizobium japonicum, Bradyrhizobium elkanii, Azospirillum brasilense, Pseudomonas fluorescens, and Azotobacter chroococcum to the effects of soil-applied herbicides under in vitro conditions. Methods. Microbiological and statistical. The sensitivity of rhizobacteria of different species to herbicidal preparations was studied using the well diffusion method on agarized nutrient medium plates. Results. As a result of the laboratory experiments series, it was established that the rhizobial strains Bradyrhizobium japonicum 634b, РС07, РС08, B20, T66, T21-2, B78 and Bradyrhizobium elkanii SAF18 were not sensitive to average recommended rate herbicide formulations containing 312.5 g/l S-metolachlor + 187 g/l terbuthylazine, 900 g/l acetochlor and 960 g/l S-metolachlor. A low level of sensitivity was observed in certain rhizobial strains to a herbicide formulation containing 500 g/l prometryn. Under the influence of the specified active ingredient, the growth inhibition zones on bacterial lawns of the analytically selected strains B. elkanii SAF18 and B. japonicum РС07 measured 5.0 mm and 11.0 mm, respectively. Exposure to prometryn at the tested concentration resulted in the formation of growth inhibition zones measuring 8.0 mm and 6.5 mm on bacterial lawns of Tn5 mutants of B. japonicum strains B20 and B78, respectively. As a result of examining the growth characteristics of A. brasilense 410, A. chroococcum T79, and P. fluorescens strains 33 and 267 under the influence of soil-applied herbicides, no growth inhibition zones were observed on the bacterial lawns of any of the tested strains. The formation and appearance of bacterial colonies were characteristic of each of the species studied. Conclusions. It was found that, under in vitro conditions, the chemical plant protection products used in the study did not exhibit bactericidal activity against rhizobacteria of various species under the conditions of applying the manufacturer’s recommended average rate. In vegetation and field experiments, it is advisable to determine the effect of herbicidal compounds on the realization of the symbiotic potential of rhizobia. It is also important to assess how these substances influence the plant growth-promoting properties of Pseudomonas, Azotobacter and Azospirillum strains. Such studies will contribute to resolving the issue of compatibility between the application of chemical plant protection products and complex bacterial inoculation. |
| doi_str_mv | 10.35868/1997-3004.42.19-29 |
| first_indexed | 2026-03-18T02:00:08Z |
| format | Article |
| fulltext |
19
Сільськогосподарська мікробіологія. 2025. Вип. 42. С. 19–29.
ISSN 1997-3004
https://doi.org/10.35868/1997-3004.42.19-29
UDC 631.847.211:579.64:632.9
SENSITIVITY OF RHIZOBACTERIA OF DIFFERENT
SPECIES TO THE INFLUENCE OF SOIL HERBICIDES
UNDER IN VITRO CONDITIONS
S. Ya. Kots, https://orcid.org/0000-0002-3477-793X,
V. D. Khomenko, https://orcid.org/0009-0000-7391-2391,
K. P. Kukol, https://orcid.org/0000-0002-2889-9957,
P. P. Pukhtaievych, https://orcid.org/0000-0002-6179-6239
Institute of Plant Physiology and Genetics, NAS of Ukraine
31/17 Vasylkivska str., Kyiv, 03022, Ukraine; e-mail: azotfixation@gmail.com
Objective. To investigate the sensitivity of Bradyrhizobium japonicum, Bradyrhizobium elkanii,
Azospirillum brasilense, Pseudomonas fluorescens, and Azotobacter chroococcum to the effects of
soil-applied herbicides under in vitro conditions. Methods. Microbiological and statistical. The
sensitivity of rhizobacteria of different species to herbicidal preparations was studied using the well
diffusion method on agarized nutrient medium plates. Results. As a result of the laboratory
experiments series, it was established that the rhizobial strains Bradyrhizobium japonicum 634b,
РС07, РС08, B20, T66, T21-2, B78 and Bradyrhizobium elkanii SAF18 were not sensitive to
average recommended rate herbicide formulations containing 312.5 g/l S-metolachlor + 187 g/l
terbuthylazine, 900 g/l acetochlor and 960 g/l S-metolachlor. A low level of sensitivity was obser-
ved in certain rhizobial strains to a herbicide formulation containing 500 g/l prometryn. Under the
influence of the specified active ingredient, the growth inhibition zones on bacterial lawns of
the analytically selected strains B. elkanii SAF18 and B. japonicum РС07 measured 5.0 mm and
11.0 mm, respectively. Exposure to prometryn at the tested concentration resulted in the formation
of growth inhibition zones measuring 8.0 mm and 6.5 mm on bacterial lawns of Tn5 mutants of
B. japonicum strains B20 and B78, respectively. As a result of examining the growth characteristics
of A. brasilense 410, A. chroococcum T79, and P. fluorescens strains 33 and 267 under the
influence of soil-applied herbicides, no growth inhibition zones were observed on the bacterial
lawns of any of the tested strains. The formation and appearance of bacterial colonies were
characteristic of each of the species studied. Conclusions. It was found that, under in vitro
conditions, the chemical plant protection products used in the study did not exhibit bactericidal
activity against rhizobacteria of various species under the conditions of applying the manufac-
turer’s recommended average rate. In vegetation and field experiments, it is advisable to determine
the effect of herbicidal compounds on the realization of the symbiotic potential of rhizobia. It is also
important to assess how these substances influence the plant growth-promoting properties of Pseu-
domonas, Azotobacter and Azospirillum strains. Such studies will contribute to resolving the issue
of compatibility between the application of chemical plant protection products and complex bacte-
rial inoculation.
Key words: Bradyrhizobium japonicum, Bradyrhizobium elkanii, Azospirillum brasilense,
Pseudomonas fluorescens, Azotobacter chroococcum, soil-applied herbicide, sensitivity, growth
inhibition zone.
Introduction. In soybean cultivation tech-
nologies, it is common practice to introduce
microorganisms into agroecosystem that are
capable of providing plants with nutrients and
© S. Ya. Kots, V. D. Khomenko, K. P. Kukol, P. P. Pukhtaievych, 2025
20
synthesizing various biologically active sub-
stances to stimulate the growth and develop-
ment of the cultivated crop. At the same time,
soybeans are very sensitive to weeds, especially
in the early stages of vegetation. This necessi-
tates the use of effective chemical plant protec-
tion products (PPPs) to reduce the number of
weeds in crops. Synthetic chemical compounds
may negatively affect the viability and biologi-
cal traits of microorganisms possessing agro-
nomically beneficial properties. Therefore, in
production conditions, it is important to study
their sensitivity to herbicides in vitro.
Analysis of recent studies and publica-
tions. Soybean (Glycine max (L.) Merr.) origi-
nates from China. Its cultivation history spans
over 5,000 years. Under modern agricultural
conditions, soybean holds the largest cultivated
area among leguminous crops worldwide and
serves as an important source of edible oil and
high-quality plant protein, as well as a raw ma-
terial for animal feed production [1].
Given the need to restore and preserve soil
fertility, as well as to produce environmentally
safe agricultural products, scientific interest in
the processes of biological nitrogen fixation is
steadily increasing. Particular attention is paid
to the study of symbiotic and associative inte-
ractions between diazotrophic microorganisms
and plants [2; 3].
Soybean plants, in symbiosis with root no-
dule bacteria, are capable of fixing atmospheric
nitrogen. This accounts for approximately 77 %
of the total amount of this element biologically
assimilated by all leguminous crops [4].
The intensity of biological nitrogen fixation
in soybean agroecosystems is influenced by nu-
merous factors, including soil temperature,
moisture, aeration, pH, salinity, the amount of
available nitrogen, the properties of the rhizobi-
al strain and plant genotype, as well as the ap-
plication of pesticides and other agrochemicals
[5; 6].
In Ukraine, the typical microsymbiont of
soybean is Bradyrhizobium japonicum. Nume-
rous strains of this species have been isolated
from soils in various climatic zones of the
country, and they show considerable differences
in their phenotypic and genotypic characteristics
[7; 8]. Ongoing research focuses on a more
detailed study of strains obtained both through
analytical selection and by means of genetic
engineering. The virulence of new strains, the
intensity of N2 assimilation by root nodules
formed with their participation, and the effects
of inoculation on plant growth, development,
and grain productivity are evaluated. Based on
these assessments, rhizobia with improved pro-
perties are selected for use as effective micro-
symbionts [9].
Among the bacteria commonly used for
seed inoculation of both grain and leguminous
crops are representatives of the genus Azospiril-
lum. The most extensively studied among them
are Azospirillum brasilense and Azospirillum li-
poferum. The benefits of introducing azospirilla
to plants are mainly attributed to their ability
to fix atmospheric nitrogen and to synthesize
phytohormones. In recent years, researchers
have also highlighted the important role of Azo-
spirillum in enhancing plant tolerance to abiotic
and biotic stresses [10]. Rhizobacteria are conti-
nuously screened for the development of bio-
preparations, as they are often characterized by
the synthesis of plant-beneficial metabolites and
can provide both growth-promoting and phyto-
protective functions by acting against pathogens
of various etiologies [11]. To preserve the via-
bility and biological activity of microbial
agents, various carrier materials, sterilization
methods, and storage temperatures for the final
products are developed [12].
There are reports in the literature on the
effectiveness of combined inoculation of soy-
bean seeds with root nodule bacteria B. japo-
nicum together with rhizobacteria, including
A. brasilense [13], P. fluorescens [14], and
A. chroococcum [15]. These studies confirmed
the synergistic effect resulting from the combi-
ned functions of the applied microorganisms,
which contributed to an increase in soybean
grain yield.
One of the factors affecting the viability
and realization of the nitrogen-fixing potential
of microorganisms introduced into agroecosys-
tems is the impact of chemical plant protection
products. In soybean weed control systems, soil-
applied herbicides play a key role by providing
prolonged control over weed germination, the-
reby creating more favorable conditions for the
growth and development of the cultivated crop
[16; 17].
It should be taken into account that herbi-
cides, as highly active chemical compounds,
may affect the viability of microorganisms with
agronomically beneficial properties even at
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2025. Вип. 42.
21
production-recommended application rates un-
der in vitro conditions. They can also alter the
abundance and activity of these microorganisms
in soil [18]. Researchers have analyzed a sub-
stantial body of scientific literature addressing
the effects of herbicidal compounds on rhizobia
and have pointed out the potential impact on the
host plant as well as on the establishment and
development of symbiosis in vivo [19].
The investigation of rhizobacterial sensiti-
vity to soil-applied herbicides under laboratory
conditions is relevant, particularly for forecas-
ting the potential impact of chemical prepara-
tions on the efficiency of plant-microbe interac-
tions when applied simultaneously within agro-
ecosystems.
Materials and methods. The following
strains were used in the study: soybean nodule
bacteria B. japonicum 634b, РС07, РС08, B20,
B78, T66, and T21-2; B. elkanii SAF18; rhizo-
bacteria A. brasilense 410; P. fluorescens 10
and 267; and A. chroococcum T79. All strains
are maintained in the collection of symbiotic
and associative nitrogen-fixing microorganisms
at the Institute of Plant Physiology and Genetics
of the National Academy of Sciences of Ukraine
(IPPG NASU).
The nodule bacteria B. japonicum and B. el-
kanii were grown on a mineral medium of yeast
mannitol agar (YMA), which contained:
K2HPO4 — 0.5 g/l; MgSO4·7H2O — 0.4 g/l;
NaCl — 0.1 g/l; mannitol — 10 g/l; yeast ext-
ract — 0.5 g/l; agar — 15 g/l, distilled water,
pH 6.4. Strains A. brasilense and P. fluorescens
were cultivated on LPG medium, which contai-
ned yeast extract — 3 g/l; bacillus peptone —
5 g/l; glucose — 5 g/l; agar — 16 g/l, distilled
water, pH 7.0. A. chroococcum bacteria were
grown on Ashby nutrient medium, which inc-
luded K2HPO4 — 0.2 g/l; sucrose — 20 g/l;
MgSO4·7H2O — 0.2 g/l; NaCl — 0.2 g/l;
K2SO4 — 0.1 g/l; CaCO3 — 5 g/l; microele-
ments (according to Fedorov) — 1 ml/l; agar —
16 g/l; distilled water, pH 7.2–7.4.
The sensitivity of the microorganisms used
in this study to soil-applied herbicides was
assessed using a modified well-diffusion me-
thod [20]. In Petri dishes containing nutrient
medium under aseptic conditions, wells with a
diameter of 10 mm were cut using a metal
cylinder. According to standard procedures, the
surface of the nutrient medium in the dishes was
inoculated with a uniform bacterial lawn using
a microbial cell suspension with a titer of
106 CFU/mL [21]. Each well was filled with
80 µL of the herbicide solution and incubated in
a thermostat for 5–7 days at a temperature of
28 °C. The control consisted of wells in the
center of the agar medium filled with 80 µL of
sterile tap water. All experimental variants were
performed in triplicate.
Table 1. List and characteristics of the pes-
ticides used in the study [22]
Trade Name Active
Ingredient
Recommended
Application
Rate
Gezagard
500 FW SC
Prometryn,
500 g/L 2.0‒5.0 L/ha
Primextra TZ
Gold 500 SC
S-metolachlor,
312.5 g/L +
terbuthylazine,
187.5 g/L
4.0–4.5 L/ha
Harness EC Acetochlor,
900 g/L 1.5–2.5 L/ha
Dual Gold
900 EC
S-metolachlor,
960 g/L 1.0‒1.6 L/ha
Working solutions of the herbicides were
prepared using the average concentration reco-
mmended by the manufacturer for application
in soybean cultivation. The study also took into
account and applied the recommended amount
of water (250 L/ha) for dissolving each of the
herbicides.The sensitivity of B. japonicum,
B. elkanii, A. brasilense, P. fluorescens, and
A. chroococcum strains to the herbicides in
laboratory experiments was assessed by measu-
ring the size of the growth inhibition zones of
the bacterial culture around wells containing the
chemical compounds. The intensity of bacterial
growth around the wells was evaluated visually:
«+ + +» — intensive growth, «+ + –» — slight
inhibition, «+ – –» — stronger inhibition,
«– – –» — complete absence of bacterial
growth. Intensive bacterial growth and the
absence of growth inhibition zones around the
wells with herbicides indicated the resistance of
the tested microorganisms to the applied formu-
lations.
Results and discussion. It was established
that all the strains of root nodule bacteria
involved in the study — B. japonicum 634b,
РС07, РС08, B20, T66, T21-2, B78, and
B. elkanii SAF18 — were not sensitive to the
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2025. Вип. 42.
22
action the manufacturer’s recommended avera-
ge rate of herbicide formulations containing
312.5 g/L S-metolachlor + 187.5 g/L terbuthy-
lazine, 900 g/L acetochlor, and 960 g/L S-meto-
lachlor.
The soil-applied herbicide based on 500 g/L
prometryn caused growth inhibition zones on
the bacterial lawn measuring 5.0 mm and
11.0 mm for the analytically selected strains
B. elkanii SAF18 and B. japonicum РС07, re-
spectively (Fig. 1). Under the influence of this
active ingredient, growth inhibition zones
measuring 8.0 mm and 6.5 mm were observed
on the bacterial lawns of B. japonicum B20 and
B78, respectively (Table 2).
The formation of growth inhibition zones
on the bacterial lawn with diameters of up to
15 mm in various strains indicates their low
sensitivity, in pure culture, to the action of the
herbicidal substances used in this study.
An examination of the growth characteris-
tics of A. brasilense 410, A. chroococcum T79,
and P. fluorescens strains 33 and 267 on agar
nutrient media with soil-applied herbicides
added to the wells showed that none of the
strains exhibited any growth inhibition zones on
Fig. 1. Growth inhibition zones on bacterial lawns of Bradyrhizobium japonicum РС07 under
the influence of soil-applied herbicides: A — 500 g/L prometryn; B — 312.5 g/L S-metolach-
lor + 187.5 g/L terbuthylazine; C — 900 g/L acetochlor.
Table 2. Sensitivity of soybean nodule bacteria to the production-rate application of soil-
applied herbicides
Bacterial
strain
Method
of production
Prometryn,
500 g/L
S-metolachlor,
312.5 g/L +
terbuthylazine,
187.5 g/L
Acetochlor,
900 g/L
S-metolachlor,
960 g/L
Size of growth inhibition zones on the bacterial lawn around
wells containing the herbicide, mm / bacterial growth rate
B. japonicum 634b
Analytical selection
0 / + + + 0 / + + + 0 / + + + 0 / + + +
B. japonicum РС07 11.0 / + + + 0 / + + + 0 / + + + 0 / + + +
B. japonicum РС08 0 / + + + 0 / + + + 0/ + + + 0 / + + +
B. elkanii SАF18 5.0 / + + − 0 / + + + 0 / + + + 0 / + + +
B. japonicum Т66 B. japonicum 646 +
Е. coli S17-1
(pSUP2021::Tn5)
0 / + + + 0 / + + + 0 / + + + 0 / + + +
B. japonicum Т21-2 0 / + + + 0 / + + + 0 / + + + 0 / + + +
B. japonicum В20 B. japonicum 646 +
Е. coli S17-1
(pSUP5011::Tn5 mob)
8.0 / + + + 0 / + + + 0 / + + + 0 / + + +
B. japonicum В78 6.5 / + + + 0 / + + + 0 / + + + 0 / + + +
Note. In tables 2 and 3: «+ + +» — intensive growth, «+ + –» — slight inhibition, «+ – –» — stronger
inhibition, «– – –» − complete absence of bacterial growth.
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2025. Вип. 42.
23
the bacterial lawn (Table 3). The formation of
bacterial colonies was typical for each of the
specified species.
As an example, Fig. 2 shows the growth of
the bacterial lawn of A. chroococcum T79 under
the influence of the soil-applied herbicides used
in this study.
There is a great diversity of microorganisms
used as the basis for biopreparations, as well
as a wide range of chemical compounds applied
to control weeds in agricultural crops, including
soybean. Therefore, the literature contains nu-
merous studies investigating the effects of vari-
ous herbicidal substances on different groups of
beneficial microbiota. It should also be noted
that there are considerable differences in the
research methodologies employed, particularly
under laboratory conditions.
The response of Bradyrhizobium and Azo-
tobacter strains to different concentrations of
herbicides in liquid nutrient media was studied
by Ubogu and Akponah. An inhibitory effect
of atrazine at concentrations above 1.5 % on the
viability of Azotobacter was observed. When
2.0 % glyphosate was added to the cultivation
medium of rhizobia, a decrease in the number of
viable cells was recorded after 24 hours of co-
cultivation, followed by recovery and stimu-
lation of growth. Under similar conditions,
growth activity of pure cultures of Azotobacter
and Bradyrhizobium was suppressed by para-
quat. The toxic effect of paraquat was evident at
all tested concentrations (0.5 %, 1.0 %, 1.5 %,
and 2.0 %). Root nodule bacteria are more sen-
sitive to the effects of herbicides than free-living
diazotrophs [23]. In our study, strains of Pseu-
domonas, Azospirillum, and Azotobacter also
demonstrated resistance to the action — to the
average recommended rate of soil herbicides
based on 500 g/L prometryn, 312.5 g/L S-meto-
lachlor + 187.5 g/L terbuthylazine, 900 g/L ace-
tochlor, and 960 g/L S-metolachlor. Prometryn
caused slight growth inhibition zones (ranging
from 5 to 11 mm) in rhizobial strains differing
in origin and activity. Despite the slight sensi-
tivity to this active ingredient observed in some
Table 3. Sensitivity of various rhizobacterial species to production-rate applications of soil-
applied herbicides
Bacterial
strain
Method
of production
Prometryn,
500 g/L
S-metolachlor,
312.5 g/L +
terbuthylazine,
187.5 g/L
Acetochlor,
900 g/L
S-metolachlor,
960 g/L
Size of growth inhibition zones on the bacterial lawn around
wells containing the herbicide, mm / bacterial growth rate
A. brasilense 410
Analytical
selection
0 / + + + 0 / + + + 0 / + + + 0 / + + +
A. chroococcum T79 0 / + + + 0 / + + − 0 / + + + 0 / + + +
P. fluorescens 33 0 / + + + 0 / + + + 0 / + + + 0 / + + +
P. fluorescens 267 0 / + + + 0 / + + + 0 / + + + 0 / + + +
Fig. 2. Growth of the bacterial lawn of Azotobacter chroococcum T79 under exposure to:
A — 500 g/L prometryn; B — 960 g/L S-metolachlor.
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2025. Вип. 42.
24
strains such as B. japonicum РС07, B20, B78,
and B. elkanii SAF18, the production of poly-
saccharide slime by these strains remained
intensive in all tested treatments. According to
the literature, extracellular polymers such as
exopolysaccharides provide microorganisms
with an adaptive advantage in the environment.
The functions of this heteropolymer include
protection against various stresses, biofilm for-
mation, and attachment to the roots of host
plants [24]. Rhizobia defective in polysaccha-
ride synthesis are unable to establish an effec-
tive symbiosis [25].
We previously evaluated the effect of fun-
gicides on soybean root nodule bacteria in pure
culture. Growth inhibition zones measuring 13.0
and 14.0 mm were observed in B. japonicum
strains B20 and B78, respectively, when ex-
posed to the recommended field dose of a for-
mulation containing 500 g/kg benomyl. Upon
doubling this dose, strain B. japonicum B20 was
classified as slightly sensitive, whereas the
growth of B. japonicum B78 was completely sup-
pressed, exhibiting an inhibition zone ≥ 25 mm
[26].
A difference was noted in the levels of her-
bicide tolerance between B. japonicum and
B. elkanii strains, which is attributed to dif-
ferences in the absorption characteristics of
these substances by the respective species [27].
It was found that the P. fluorescens strain
CMA55, isolated from water used for washing
chemical pesticide containers, exhibited changes
in the activity of antioxidant enzymes and pro-
duced biofilms when exposed to certain herbi-
cides. The authors therefore suggested the pre-
sence of a specific model of phenotypic plas-
ticity in these microorganisms for adaptation to
environments contaminated with xenobiotics.
This should be considered in studies on herbi-
cide bioremediation [28]. Some strains have
shown resistance to chemical plant protection
products of various types. For example, P. flu-
orescens 8 demonstrated high resistance to ten
different insecticides and herbicides (quizalofop
ethyl 5 % EC, sodium pyrithiobac 10 % EC,
oxyfluorfen 3.5 % EC, cyhalofop-butyl 10 % EC,
glyphosate + ammonium sulfate 71SG, pendi-
methalin 30 % EC, imazethapyr 10 % EC, atra-
zine 50 % WP, glyphosate 41 % SL) when app-
lied at concentrations of 100, 500, and 1000 mg/L
in agar nutrient medium. Among fungicidal
substances, azoxystrobin and a mixture of car-
boxin with thiram did not affect the viability of
this strain, regardless of the concentration used.
The authors consider it highly probable that
there is an additive effect when using biopre-
parations based on P. fluorescens 8 together
with the tested pesticides [29].
Researchers working with the pesticide-
resistant strain Azotobacter vinelandii AZ6 no-
ted that herbicides at elevated concentrations
were more toxic compared to fungicidal and
insecticidal formulations. The effects of kitazin,
hexaconazole, metalaxyl, glyphosate, quizalo-
fop, atrazine, fipronil, monocrotophos, and imi-
dacloprid on the synthesis by the bacteria of
indole-3-acetic, salicylic, and 2,3-dihydroxy-
benzoic acids were determined. With increasing
pesticide concentrations, a decrease in their so-
lubilization capacity was observed, which was
attributed to changes in the pH of the nutrient
medium. Microorganisms of this strain conti-
nued to synthesize biologically active compo-
unds even under the influence of high pesticide
concentrations, indicating their potential for use
as effective inoculants for agricultural crops
[30]. Therefore, it is advisable to determine the
effects of herbicides on the biological properties
of the studied rhizosphere microorganisms and
their production of plant growth-promoting and
antibiotic substances.
In some cases, the addition of herbicides to
the bacterial cultivation medium results in re-
duced growth intensity of bacteria with agro-
nomically beneficial properties. This is associ-
ated with a decreased efficiency in resource uti-
lization by the bacteria. Under stress conditions,
a portion of the energy available to the organism
is expended to maintain cellular and biochemi-
cal mechanisms of tolerance to the stressor [31].
Singh & Wright investigated the effects of
terbutryn with terbuthylazine, trietazine with
simazine, prometryn, and bentazon on the
growth of R. leguminosarum, using herbicide
concentrations significantly higher than those
recommended by the manufacturers. The reduc-
tion in microorganism viability observed under
in vitro conditions is considered unlikely to
occur under field conditions. They explain this
by noting that even in cases of non-compliance
with herbicide application regulations or gradual
accumulation of residues over the years, the
amounts present in soils decrease due to adso-
rption processes, degradation, leaching, and
other factors [32]. In general, a number of
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2025. Вип. 42.
25
scientific studies emphasize the high tolerance
of certain representatives of the rhizosphere
microbiota to xenobiotics.
It is scientifically justified to limit the use
of herbicides that have demonstrated an inhibi-
tory effect on beneficial microbiota or on the
physiological and biochemical processes in
plants. The use of compounds that are highly
persistent in the environment, capable of remai-
ning in the soil for years and affecting the
effectiveness of plant-microbe interactions over
a prolonged period, should be avoided [19].
Conclusions. It was found that, under in
vitro conditions, the chemical plant protection
products under the conditions of the average
manufacturer-recommended rate used in this
study containing 312.5 g/L S-metolachlor +
187.5 g/L terbuthylazine, 900 g/L acetochlor,
960 g/L S-metolachlor, and 500 g/L prometryn
did not exhibit bactericidal activity against
B. japonicum strains 634b, РС07, РС08, B20,
B78, T66, T21-2; B. elkanii SAF18; the rhi-
zobacteria A. brasilense 410, P. fluorescens 10,
267, and A. chroococcum T79. The growth inhi-
bition zones ranging from 5 to 11 mm caused by
the tested concentration of prometryn in certain
soybean root nodule bacterial strains indicate
their low sensitivity to this herbicide.
In future research, it should be taken into
account that, in soil, changes in the viability or
reproduction intensity of microorganisms de-
pend on the processes of pesticide detoxifica-
tion. Therefore, in vegetation and field experi-
ments, it is advisable to determine the effect of
herbicidal compounds on the realization of the
symbiotic potential of rhizobia and the growth-
promoting properties of Pseudomonas, Azoto-
bacter and Azospirillum. This will help address
the issue of compatibility between the applica-
tion of chemical plant protection products and
pre-sowing bacterial inoculation.
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Received: 21.08.2025
Accepted: 06.10.2025
Published online: 29.12.2025
https://doi.org/10.35868/1997-3004.42.19-29
УДК 631.847.211:579.64:632.9
ЧУТЛИВІСТЬ РИЗОБАКТЕРІЙ РІЗНИХ ВИДІВ ДО ВПЛИВУ
ҐРУНТОВИХ ГЕРБІЦИДІВ В УМОВАХ IN VITRO
С. Я. Коць, https://orcid.org/0000-0002-3477-793X,
В. Д. Хоменко, https://orcid.org/0009-0000-7391-2391,
К. П. Кукол, https://orcid.org/0000-0002-2889-9957,
П. П. Пухтаєвич, https://orcid.org/0000-0002-6179-6239
Інститут фізіології рослин і генетики НАН України, м. Київ
e-mail: azotfixation@gmail.com
Мета. Дослідити чутливість бактерій Bradyrhizobium japonicum, Bradyrhizobium elka-
nii, Azospirillum brasilense, Pseudomonas fluorescens та Azotobacter chroococcum до впливу
ґрунтових гербіцидів в умовах in vitro. Meтоди. Мікробіологічні, статистичні. Зокрема
чутливість ризобактерій різних видів до впливу препаратів з гербіцидною активністю вив-
чали методом лунок, висічених у пластинках агаризованого живильного середовища.
Результати. У серії лабораторних досліджень встановлено, що бульбочкові бактерії
B. japonicum 634б, РС07, РС08, В20, Т66, Т21-2, В78 і B. elkanii САФ18 є не чутливими до
дії середньої рекомендованої норми препаратів на основі 312,5 г/л S-метолахлору + 187 г/л
тербутилазину; 900 г/л ацетохлору та 960 г/л S-метолахлору. Водночас виявлено слабку
чутливість в окремих штамів ризобій до гербіциду, в складі якого міститься 500 г/л промет-
рину. Зокрема, за впливу вказаної діючої речовини зони затримки росту бактеріального га-
зону в аналітично селекціонованих штамів B. elkanii САФ18 та B. japonicum РС07 станови-
ли 5,0 і 11,0 мм відповідно. Окрім цього, за дії прометрину в досліджуваній концентрації від-
значали наявність зон затримки росту бактеріального газону розміром 8,0 та 6,5 мм у Tn5-
мутантів B. japonicum В20 і В78. Дослідивши особливості росту A. brasilense 410, A. chroo-
coccum T79 та P. fluorescens 33 і 267 за дії ґрунтових гербіцидів, відзначили відсутність зон
затримки росту бактеріального газону у всіх вивчених штамів. Формування бактеріальних
колоній було типовим для кожного із вказаних видів. Висновки. Таким чином з’ясовано, що
в умовах in vitro залучені у роботу хімічні ЗЗР за умов застосування середньої рекомендова-
ної виробником норми не чинять бактерицидної дії на ризобактерії різних видів. Водночас
у вегетаційних та польових дослідах доцільно з’ясувати вплив речовин із гербіцидною актив-
ністю на реалізацію симбіотичного потенціалу ризобій і ростостимулювальні властивості
псевдомонад, азотобактера й азоспірил, що сприятиме розв’язанню питання сумісності
застосування хімічних ЗЗР із комплексною бактеризацією.
Ключові слова: Bradyrhizobium japonicum, Bradyrhizobium elkanii, Azospirillum brasilense,
Pseudomonas fluorescens, Azotobacter chroococcum, ґрунтовий гербіцид, чутливість, зона за-
тримки росту.
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2025. Вип. 42.
28
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01117.x
Отримано: 21.08.2025
Прийнято до друку: 06.10.2025
Опубліковано онлайн: 29.12.2025
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2025. Вип. 42.
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| id | oai:ojs2.smic.in.ua:article-544 |
| institution | Agriciltural microbiology |
| keywords_txt_mv | keywords |
| language | English |
| last_indexed | 2026-07-23T01:16:25Z |
| publishDate | 2025 |
| publisher | Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine |
| record_format | ojs |
| resource_txt_mv | smicinua/22/b4048a26ca39d435a957f9a2ffec4b22.pdf |
| spelling | oai:ojs2.smic.in.ua:article-5442026-07-22T10:10:52Z SENSITIVITY OF RHIZOBACTERIA OF DIFFERENT SPECIES TO THE INFLUENCE OF SOIL HERBICIDES UNDER IN VITRO CONDITIONS ЧУТЛИВІСТЬ РИЗОБАКТЕРІЙ РІЗНИХ ВИДІВ ДО ВПЛИВУ ҐРУНТОВИХ ГЕРБІЦИДІВ В УМОВАХ IN VITRO Коць, С. Я. Хоменко, В. Д. Кукол, К. П. Пухтаєвич, П. П. Bradyrhizobium japonicum, Bradyrhizobium elkanii, Azospirillum brasilense, Pseudomonas fluorescens, Azotobacter chroococcum, soil-applied herbicide, sensitivity, growth inhibition zone Bradyrhizobium japonicum, Bradyrhizobium elkanii, Azospirillum brasilense, Pseudomonas fluorescens, Azotobacter chroococcum, ґрунтовий гербіцид, чутливість, зона затримки росту Objective. To investigate the sensitivity of Bradyrhizobium japonicum, Bradyrhizobium elkanii, Azospirillum brasilense, Pseudomonas fluorescens, and Azotobacter chroococcum to the effects of soil-applied herbicides under in vitro conditions. Methods. Microbiological and statistical. The sensitivity of rhizobacteria of different species to herbicidal preparations was studied using the well diffusion method on agarized nutrient medium plates. Results. As a result of the laboratory experiments series, it was established that the rhizobial strains Bradyrhizobium japonicum 634b, РС07, РС08, B20, T66, T21-2, B78 and Bradyrhizobium elkanii SAF18 were not sensitive to average recommended rate herbicide formulations containing 312.5 g/l S-metolachlor + 187 g/l terbuthylazine, 900 g/l acetochlor and 960 g/l S-metolachlor. A low level of sensitivity was observed in certain rhizobial strains to a herbicide formulation containing 500 g/l prometryn. Under the influence of the specified active ingredient, the growth inhibition zones on bacterial lawns of the analytically selected strains B. elkanii SAF18 and B. japonicum РС07 measured 5.0 mm and 11.0 mm, respectively. Exposure to prometryn at the tested concentration resulted in the formation of growth inhibition zones measuring 8.0 mm and 6.5 mm on bacterial lawns of Tn5 mutants of B. japonicum strains B20 and B78, respectively. As a result of examining the growth characteristics of A. brasilense 410, A. chroococcum T79, and P. fluorescens strains 33 and 267 under the influence of soil-applied herbicides, no growth inhibition zones were observed on the bacterial lawns of any of the tested strains. The formation and appearance of bacterial colonies were characteristic of each of the species studied. Conclusions. It was found that, under in vitro conditions, the chemical plant protection products used in the study did not exhibit bactericidal activity against rhizobacteria of various species under the conditions of applying the manufacturer’s recommended average rate. In vegetation and field experiments, it is advisable to determine the effect of herbicidal compounds on the realization of the symbiotic potential of rhizobia. It is also important to assess how these substances influence the plant growth-promoting properties of Pseudomonas, Azotobacter and Azospirillum strains. Such studies will contribute to resolving the issue of compatibility between the application of chemical plant protection products and complex bacterial inoculation. Мета. Дослідити чутливість бактерій Bradyrhizobium japonicum, Bradyrhizobium elkanii, Azospirillum brasilense, Pseudomonas fluorescens та Azotobacter chroococcum до впливу ґрунтових гербіцидів в умовах in vitro. Meтоди. Мікробіологічні, статистичні. Зокрема чутливість ризобактерій різних видів до впливу препаратів з гербіцидною активністю вивчали методом лунок, висічених у пластинках агаризованого живильного середовища. Результати. У серії лабораторних досліджень встановлено, що бульбочкові бактерії B. japonicum 634б, РС07, РС08, В20, Т66, Т21-2, В78 і B. elkanii САФ18 є не чутливими до дії середньої рекомендованої норми препаратів на основі 312,5 г/л S-метолахлору + 187 г/л тербутилазину; 900 г/л ацетохлору та 960 г/л S-метолахлору. Водночас виявлено слабку чутливість в окремих штамів ризобій до гербіциду, в складі якого міститься 500 г/л прометрину. Зокрема, за впливу вказаної діючої речовини зони затримки росту бактеріального газону в аналітично селекціонованих штамів B. elkanii САФ18 та B. japonicum РС07 становили 5,0 і 11,0 мм відповідно. Окрім цього, за дії прометрину в досліджуваній концентрації відзначали наявність зон затримки росту бактеріального газону розміром 8,0 та 6,5 мм у Tn5- мутантів B. japonicum В20 і В78. Дослідивши особливості росту A. brasilense 410, A. chroococcum T79 та P. fluorescens 33 і 267 за дії ґрунтових гербіцидів, відзначили відсутність зон затримки росту бактеріального газону у всіх вивчених штамів. Формування бактеріальних колоній було типовим для кожного із вказаних видів. Висновки. Таким чином з’ясовано, що в умовах in vitro залучені у роботу хімічні ЗЗР за умов застосування середньої рекомендованої виробником норми не чинять бактерицидної дії на ризобактерії різних видів. Водночас у вегетаційних та польових дослідах доцільно з’ясувати вплив речовин із гербіцидною активністю на реалізацію симбіотичного потенціалу ризобій і ростостимулювальні властивості псевдомонад, азотобактера й азоспірил, що сприятиме розв’язанню питання сумісності застосування хімічних ЗЗР із комплексною бактеризацією. Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine 2025-12-29 Article Article Рецензована Стаття application/pdf https://smic.in.ua/index.php/journal/article/view/544 10.35868/1997-3004.42.19-29 Agricultural microbiology; Vol. 42 (2025): Agriciltural microbiology; 19-29 Сільськогосподарська мікробіологія; Том 42 (2025): Сільськогосподарська мікробіологія; 19-29 1997-3004 10.35868/1997-3004.42 en https://smic.in.ua/index.php/journal/article/view/544/612 Авторське право (c) 2025 S. Ya. Kots, V. D. Khomenko, K. P. Kukol, P. P. Pukhtaievych https://creativecommons.org/licenses/by/4.0 |
| spellingShingle | Bradyrhizobium japonicum Bradyrhizobium elkanii Azospirillum brasilense Pseudomonas fluorescens Azotobacter chroococcum ґрунтовий гербіцид чутливість зона затримки росту Коць, С. Я. Хоменко, В. Д. Кукол, К. П. Пухтаєвич, П. П. ЧУТЛИВІСТЬ РИЗОБАКТЕРІЙ РІЗНИХ ВИДІВ ДО ВПЛИВУ ҐРУНТОВИХ ГЕРБІЦИДІВ В УМОВАХ IN VITRO |
| title | ЧУТЛИВІСТЬ РИЗОБАКТЕРІЙ РІЗНИХ ВИДІВ ДО ВПЛИВУ ҐРУНТОВИХ ГЕРБІЦИДІВ В УМОВАХ IN VITRO |
| title_alt | SENSITIVITY OF RHIZOBACTERIA OF DIFFERENT SPECIES TO THE INFLUENCE OF SOIL HERBICIDES UNDER IN VITRO CONDITIONS |
| title_full | ЧУТЛИВІСТЬ РИЗОБАКТЕРІЙ РІЗНИХ ВИДІВ ДО ВПЛИВУ ҐРУНТОВИХ ГЕРБІЦИДІВ В УМОВАХ IN VITRO |
| title_fullStr | ЧУТЛИВІСТЬ РИЗОБАКТЕРІЙ РІЗНИХ ВИДІВ ДО ВПЛИВУ ҐРУНТОВИХ ГЕРБІЦИДІВ В УМОВАХ IN VITRO |
| title_full_unstemmed | ЧУТЛИВІСТЬ РИЗОБАКТЕРІЙ РІЗНИХ ВИДІВ ДО ВПЛИВУ ҐРУНТОВИХ ГЕРБІЦИДІВ В УМОВАХ IN VITRO |
| title_short | ЧУТЛИВІСТЬ РИЗОБАКТЕРІЙ РІЗНИХ ВИДІВ ДО ВПЛИВУ ҐРУНТОВИХ ГЕРБІЦИДІВ В УМОВАХ IN VITRO |
| title_sort | чутливість ризобактерій різних видів до впливу ґрунтових гербіцидів в умовах in vitro |
| topic | Bradyrhizobium japonicum Bradyrhizobium elkanii Azospirillum brasilense Pseudomonas fluorescens Azotobacter chroococcum ґрунтовий гербіцид чутливість зона затримки росту |
| topic_facet | Bradyrhizobium japonicum Bradyrhizobium elkanii Azospirillum brasilense Pseudomonas fluorescens Azotobacter chroococcum soil-applied herbicide sensitivity growth inhibition zone Bradyrhizobium japonicum Bradyrhizobium elkanii Azospirillum brasilense Pseudomonas fluorescens Azotobacter chroococcum ґрунтовий гербіцид чутливість зона затримки росту |
| url | https://smic.in.ua/index.php/journal/article/view/544 |
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