ВЗАЄМОДІЯ РОСЛИН ЛЮПИНУ ЗІ ШТАМАМИ БУЛЬБОЧКОВИХ БАКТЕРІЙ РОДУ BRADYRHIZOBIUM
Objective. Study the peculiarities of the interaction between three species of lupin and nodule bacteria of the genus Bradyrhizobium, isolated from various leguminous plants: lupin, seradella, soybean and cowpea. Methods. Microbiological (isolation of rhizobia from nodules and their cultivation), se...
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| Date: | 2024 |
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| Language: | English |
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Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine
2024
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Agriciltural microbiology| _version_ | 1871466321336598528 |
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| author | Крутило, Д. В. |
| author_facet | Крутило, Д. В. |
| author_institution_txt_mv | [
{
"author": "Д. В. Крутило",
"institution": "Інститут сільськогосподарської мікробіології та агропромислового виробництва НААН"
}
] |
| author_sort | Крутило, Д. В. |
| baseUrl_str | https://smic.in.ua/index.php/journal/oai |
| collection | OJS |
| datestamp_date | 2026-07-22T10:10:52Z |
| description | Objective. Study the peculiarities of the interaction between three species of lupin and nodule bacteria of the genus Bradyrhizobium, isolated from various leguminous plants: lupin, seradella, soybean and cowpea. Methods. Microbiological (isolation of rhizobia from nodules and their cultivation), serological (defining serological affiliation of strains), vegetation experiment (study of host specificity of bradyrhizobia, study of the formation and functioning of the symbiosis between lupin and nodule bacteria), gas chromatographic (determination of the activity of symbiotic nitrogen fixation), mathematical and statistical. Results. Under the conditions of vegetation experiments on sod-podzolic soil and sterile vermiculite, the peculiarities of the interaction between different species of lupin and specific and non-specific strains of nodule bacteria of the genus Bradyrhizobium were studied. It was established that rhizobia isolated from lupin nodules (B. lupini 367a, 30l, LD4 and Bradyrhizobium sp. LD8), seradella (Bradyrhizobium sp. СР1) and soybean (B. lupini 631) are able to form an active symbiosis with white and yellow lupin plants. Microsymbionts of soybean (B. japonicum КВ11) and cowpea (B. diazoefficiens В22) did not infect these two species of lupin. In the experiment with narrow-leaved lupin, all strains of B. lupini (367а, 30l, LD4, 631) and LD8 Bradyrhizobium sp. ensured a reliable increase in the above-ground mass of plants by 14.3–22.6 % compared to the control. The maximum values of this indicator were obtained in variants inoculated with B. lupini LD4 and B. lupine 631 strains (1.03 and 1.02 g/plant versus 0.84 g/plant in the control). With the use of non-specific lupin strains B. japonicum КВ11 and B. diazoefficiens В22, a significant increase in the nitrogenase activity of nodules was registered (1.5 and 1.9 times compared to the control), however, the increase in the above-ground mass of plants was insignificant — 7.1 % and 10.7 %, respectively. Conclusion. It has been established that lupin-specific bradyrhizobia of various origins (B. lupini and Bradyrhizobium sp.) have a positive effect on the formation of the symbiotic apparatus, the growth and development of white, yellow and narrowleaved lupin plants. Rhizobia of soybean and cowpea (B. japonicum КВ11 and B. diazoefficiens В22) non-specific for lupin, contribute to the activation of the interaction between narrowleaved lupin and representatives of the soil population of B. lupini and act as PGPR microorganisms. B. lupini LD4 and B. lupini 631 strains are the best in terms of most symbiotic indicators and efficiency of symbiosis with narrow-leaved lupin. |
| doi_str_mv | 10.35868/1997-3004.39.22-36 |
| first_indexed | 2025-07-17T12:26:50Z |
| format | Article |
| fulltext |
22
Сільськогосподарська мікробіологія. 2024. Вип. 39. С. 22–36.
ISSN 1997-3004
https://doi.org/10.35868/1997-3004.39.22-36
UDC 582.73:582.23:579.8:579.2
INTERACTION BETWEEN LUPIN PLANTS AND STRAINS
OF THE BRADYRHIZOBIUM GENUS
D. V. Krutylo
Institute of Agricultural Microbiology and Agroindustrial Manufacture, NAAS
97 Shevchenka Str., Chernihiv, 14030, Ukraine; e-mail: krutylodv@gmail.com
Objective. Study the peculiarities of the interaction between three species of lupin and nodule
bacteria of the genus Bradyrhizobium, isolated from various leguminous plants: lupin, seradella,
soybean and cowpea. Methods. Microbiological (isolation of rhizobia from nodules and their culti-
vation), serological (defining serological affiliation of strains), vegetation experiment (study of host
specificity of bradyrhizobia, study of the formation and functioning of the symbiosis between lupin
and nodule bacteria), gas chromatographic (determination of the activity of symbiotic nitrogen
fixation), mathematical and statistical. Results. Under the conditions of vegetation experiments on
sod-podzolic soil and sterile vermiculite, the peculiarities of the interaction between different spe-
cies of lupin and specific and non-specific strains of nodule bacteria of the genus Bradyrhizobium
were studied. It was established that rhizobia isolated from lupin nodules (B. lupini 367a, 30l, LD4
and Bradyrhizobium sp. LD8), seradella (Bradyrhizobium sp. СР1) and soybean (B. lupini 631) are
able to form an active symbiosis with white and yellow lupin plants. Microsymbionts of soybean
(B. japonicum КВ11) and cowpea (B. diazoefficiens В22) did not infect these two species of lupin.
In the experiment with narrow-leaved lupin, all strains of B. lupini (367а, 30l, LD4, 631) and LD8
Bradyrhizobium sp. ensured a reliable increase in the above-ground mass of plants by 14.3–22.6 %
compared to the control. The maximum values of this indicator were obtained in variants inocu-
lated with B. lupini LD4 and B. lupine 631 strains (1.03 and 1.02 g/plant versus 0.84 g/plant in
the control). With the use of non-specific lupin strains B. japonicum КВ11 and B. diazoefficiens
В22, a significant increase in the nitrogenase activity of nodules was registered (1.5 and 1.9 times
compared to the control), however, the increase in the above-ground mass of plants was insignifi-
cant — 7.1 % and 10.7 %, respectively. Conclusion. It has been established that lupin-specific
bradyrhizobia of various origins (B. lupini and Bradyrhizobium sp.) have a positive effect on the
formation of the symbiotic apparatus, the growth and development of white, yellow and narrow-
leaved lupin plants. Rhizobia of soybean and cowpea (B. japonicum КВ11 and B. diazoeffici-
ens В22) non-specific for lupin, contribute to the activation of the interaction between narrow-
leaved lupin and representatives of the soil population of B. lupini and act as PGPR microorgan-
isms. B. lupini LD4 and B. lupini 631 strains are the best in terms of most symbiotic indicators and
efficiency of symbiosis with narrow-leaved lupin.
Key words: nodule bacteria, Bradyrhizobium lupini, B. japonicum, B. diazoefficiens, lupin,
symbiotic interactions.
Introduction. Lupin (Lupinus L.) it is a ge-
nus of the legume family Fabaceae (Legumi-
nose), which includes more than 200 species of
annual and perennial plants (grasses, subshrubs,
shrubs) [1; 2]. Due to its ability to symbiosis
with nodule bacteria and fixation of molecular
nitrogen, lupin is an important agricultural crop
with significant biological and economic poten-
tial, which is widely used as a source of protein
in human and animal nutrition and is a good
green manure [3; 4]. In world agricultural prac-
tice, only a few annual species of lupin have be-
come the most widespread: L. albus (white lu-
pin), L. luteus (yellow lupin), L. angustifolius
(narrow-leaved or blue lupin) and L. mutabilis
(pearl lupin) [3]. Perennial species are mainly
© D. V. Krutylo, 2024
23
used as ornamental plants. In Ukraine, high-
yielding varieties of white, yellow and narrow-
leaved lupin are grown to solve the problem of
fodder protein [5]. There are 7 species of lupin
in the wild.
Analysis of recent studies and publica-
tions. Considering that in Ukraine, lupin has
been grown as a sidereal and fodder crop since
the beginning of the 20th century [5; 6], stable
local populations of nodule bacteria capable of
forming symbiosis with this plant have been
formed in the soil. Lupin microsymbionts of on-
ly one species — B. lupini [7–9] are described
in domestic literature. Previously, we isolated
several strains from peanut nodules that, in ad-
dition to the host plant, infect white and yellow
lupin, as well as a strain that additionally forms
nodules on soybean roots [9]. These strains are
assigned to the genus Bradyrhizobium, but their
species needs clarification. In general, the diver-
sity of lupin rhizobia in the soils of Ukraine re-
mains poorly studied.
The analysis of literary sources devoted to
the study of nodule bacteria of lupin in different
countries shows that this plant is characterized
by wide specificity and is able to form a symbi-
osis with many species of nitrogen-fixing bacte-
ria [10]. The great diversity of lupin species re-
flects the great diversity of their microsymbi-
onts. Currently, both slow-growing and fast-
growing nodule bacteria capable of nodulating
lupin have been described. It has been estab-
lished that slow-growing representatives of the
genus Bradyrhizobium predominate in the no-
dules of various lupin species: B. lupini [11–13],
B. japonicum [11; 13; 14], B. diazoefficiens [13],
B. elkanii [15], B. canariense [16], B. valenti-
num [17], B. cytisi [13] and B. hipponense [18].
It was also found that the roots of European lu-
pin species are usually infected by B. canari-
ense and B. japonicum [19], and American ones
by B. japonicum and B. elkanii [17].
Despite that the predominant microsymbi-
onts of lupin are bradyrhizobia, fast-growing
nodule bacteria were also found in their nodule
populations. For example, in Morocco and Po-
land, rhizobia belonging to the genera Allorhi-
zobium, Sinorhizobium and Rhizobium were iso-
lated from nodules on the roots of lupin plants
[20; 21]. In the Tunisian soils, white lupin was
infected by bacteria of the genera Rhizobium
and Neorhizobium [22]. Representatives of ge-
nera are also described among fast-growing
microorganisms, namely Mesorhizobium (M. lo-
ti) [23], Agrobacterium [22], Phyllobacterium
(P. trifolii) [11; 24], Ochrobactrum (O. lupini)
[25], Microvirga (M. tunisiensis, M. lupini) [11;
26; 27] and Devosia (Devosia sp.) [28].
It should be noted that the above microor-
ganisms are able to infect not only lupin, but
also other species of legumes, forming different
groups of cross-inoculation. For example, no-
dule bacteria B. japonicum, B. diazoefficiens,
B. elkanii, in addition to lupin, form nitrogen-
fixing nodules on the roots of soybean, cowpea
and mung bean plants [10; 29–31]. Among the
bacteria of the species B. lupini (formerly Bra-
dyrhizobium sp. (Lupinus)), microsymbionts of
soybean [32], seradella [33] and peanut [9] have
also been described. Representatives of other
genera of nodule bacteria can also migrate bet-
ween legumes and form different ranges of host
plants. Under such conditions, the specificity of
bacteria in relation to leguminous plants will be
different, and the efficiency of symbiotic sys-
tems may differ significantly.
In the domestic literature, no information
on the formation of an effective symbiosis be-
tween lupin and various species of specific and
non-specific nodule bacteria is available. Con-
sidering the above, the objective of our work
was to study the specifics of the interaction
between three species of lupin and nodule bacte-
ria of the genus Bradyrhizobium isolated from
various leguminous plants: lupin, seradella, soy-
bean and cowpea.
Materials and methods. The objects of re-
search were strains of lupin nodule bacteria
(B. lupini 367a (standard), B. lupini 30l, B. lupi-
ni LD4, B. lupini 631, Bradyrhizobium sp.
LD8); strains isolated from nodules of seradella
(Bradyrhizobium sp. CP1), cowpea (B. diazoef-
ficiens B22) and soybean (B. japonicum 46,
B. japonicum КВ11); plants of narrow-leaved
lupin (Lupinus angustifolius L.), white lupin
(Lupinus albus L.), yellow lupin (Lupinus luteus
L.), truncated lupin (Lupinus truncatus Hook. &
Arn.), fleshy lupin (Lupinus subcarnosus
Hook.) and seradella (Ornithopus sativus Brot.).
Lupin and seradella seeds were provided by the
Department of Scientific Support for Agroin-
dustrial Manufacture of the Institute of Agri-
cultural Microbiology and Agroindustrial Ma-
nufacture of the National Academy of Agra-
rian Sciences (IAMAM NAAS), Polissia Insti-
tute of Agriculture of the National Academy of
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 39.
24
Agrarian Sciences and Ternopil National Peda-
gogical University named after Volodymyr
Hnatiuk. Bradyrhizobia strains are stored in the
collection of the Laboratory of Plant-Microbial
Interactions and in the Collection of Beneficial
Soil Microorganisms of the IAMAM NAAS.
Isolation of nodule bacteria from lupin and
seradella nodules and study of their morpholo-
gical and cultural properties was carried out ac-
cording to methodical recommendations [34].
The serological affiliation of the strains iso-
lated from different leguminous plants was de-
termined in the agglutination reaction according
to the Gruber-Widal method [35]. Bradyrhizo-
bia were grown on solid legume medium at
28 °С. In the logarithmic phase of growth, the
bacterial mass was washed off the agar slopes,
precipitated by centrifugation and washed twice
with normal saline solution. 5 mL of saline solu-
tion and 5 mL of 2.5 % glutaraldehyde solution
(to remove non-specific flagellar H-antigens)
were added to the cell sediment and left in a re-
frigerator for a day. A day later, the bacterial
cells (antigen) were washed three times from
glutaraldehyde, the sediment was resuspended
with saline solution and the antigen titre was
brought up to 2·109 cells/mL. Specific immune
anti-nodule bacteria obtained with lupin (367a,
631), soybean (46, M8, КВ11, 634b, OR, HR,
NR) and cowpea (B1 and B2) strains were used.
The ability of bradyrhizobia strains to enter
into symbiosis with white lupin (Lybid variety)
and yellow lupin (Chernihivets variety) was
studied in a vegetation experiment. Plants were
grown in 2 litre vessels on a nitrogen-free sub-
strate (sterile vermiculite), which was moistened
with a 0.2 % solution of КН2РО4. Before sow-
ing, surface sterilized seeds were treated with
a suspension of nodule bacteria (titre
2·109 cells/mL). The inoculation load was 200–
300 thousand cells per 1 seed. The repetition
of the experiment was fourfold. Humidity was
maintained at 60 % maximum water-holding
capacity.
The interaction between bradyrhizobia and
narrow-leaved lupin was studied under growing
conditions on sod-podzolic soil (soil samples
were taken in the fields of the IAMAM NAAS).
Nodule bacteria were cultivated for 72 hours in
750 mL flasks on a rocker (220 rpm) at 26–
28 °C in a liquid bean medium [34]. Lupin
seeds of the Lokomotyv narrow-leaved variety
were treated with the studied strains. The inocu-
lation load was 200–300 thousand cells per
1 seed. In the control variant, the seeds were
moistened with tap water. The repetition of the
experiment was fivefold. Humidity was main-
tained at 60 % maximum water-holding capa-
city [36].
The activity and efficiency of legume-rhi-
zobial symbiosis was evaluated in the flowering
phase according to the following parameters:
plant height, weight of dry matter of the aerial
part of plants, number and weight of nodules,
activity of symbiotic nitrogen fixation.
Vegetation experiment on the stimulating
effect of nodule bacteria of lupin and soybean
on plants of the white lupin of Lybid variety
was carried out on a nitrogen-free substrate
(vermiculite) moistened with a 0.2 % solution of
КН2РО4. Before sowing, the seeds were treated
with strains of B. lupini 367a, B. japonicum 46
and B. japonicum КВ11. The inoculation load
was 200–300 thousand cells per 1 seed. In the
control variant, the seeds were moistened with
tap water. The repetition of the experiment was
fivefold. Humidity was maintained at 60 % ma-
ximum water-holding capacity.
The nitrogenase activity of nodules was de-
termined by the acetylene-ethylene method [37]
on a Chrom-4 gas chromatograph with a flame
ionization detector (column with β-β΄-oxydipro-
pionitrile).
Processing of experimental data was carried
out using methods of mathematical statistics
[38], Microsoft Office Excel 2016 and Statisti-
ca 8.0.
Results and discussion. To study the inte-
raction between nodule bacteria and narrow-
leaved lupin plants, we have selected strains of
various species, representatives of which, ac-
cording to literature data, are or may be poten-
tial microsymbionts of lupin. In the work,
8 strains of bradyrhizobia were studied, isolated
from nodules of lupin, seradella, soybean and
cowpea (Table 1). Both new and collection
strains were used (B. lupini 367а, 30l, 631,
B. japonicum КВ11).
It should be noted that the four studied
strains of lupin nodule bacteria are microsymbi-
onts of different host plants. For example,
strains B. lupini 367a and B. lupini 30l were iso-
lated from nodules of narrow-leaved lupin and
yellow lupin, respectively. It was established
that despite their different origin, both strains
belong to the same serogroup 367a (Table 1).
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 39.
25
Table 1. Characteristics of nodule bacteria of the genus Bradyrhizobium isolated from no-
dules of various leguminous plans
Species of
microorganisms Host plant Serogroup
Formation of active
nodules on roots:
Lupinus
albus
Lupinus
luteus
B. lupini 367а Lupinus angustifolius 367а + +
B. lupini 30l Lupinus luteus 367а + +
B. lupini LD4 Lupinus truncatus × + +
Bradyrhizobium sp. LD8 Lupinus subcarnosus × + +
Bradyrhizobium sp. СР1 Ornithopus sativus 367а + +
B. lupini 631 Glycine max 631 + +
B. japonicum КВ11 Glycine max КВ11 – –
B. diazoefficiens В22 Vigna unguiculata В2 – –
Notes: × — unspecified serogroup; + — formation of nitrogen-fixing nodules; – — no nodules.
We have isolated two new strains, LD4 and
LD8, from truncated and fleshy lupin nodules,
respectively. They form colonies 2–3 mm in di-
ameter, which appear on solid bean medium at
Day 5 to 6 of cultivation. According to the mor-
phology, at Day 7 of growth, bacterial cells are
mobile, slightly bent gram-negative rods, which
do not form spores. Cultures acidify the milk
with litmus, but do not form a zone of serum on
the surface. No growth on MPA was registered.
According to the morphological and cultural
properties, the strains are classified as Bradyrhi-
zobium. In previous studies, the LD4 strain was
identified as B. lupini based on the results of se-
quencing of the ITS region [39]. Strains of
B. lupini LD4 and Bradyrhizobium sp. LD8 did
not react with any of the 11 antisera used, their
serological affiliation remains uncertain.
According to our research, strain CP1, iso-
lated from nodules of seradella, also had charac-
teristic signs of bradyrhizobia. In the agglutina-
tion reaction, it reacted positively with antise-
rum 367a, obtained against the standard strain
B. lupini 367a. Additional research is needed to
determine its species affiliation.
It should be noted that B. lupini 367a, B. lu-
pini 30l and Bradyrhizobium sp. CP1 showed
a weak positive reaction with КВ11 antiserum,
which suggests the presence of antigenic deter-
minants shared with the soybean rhizobia strain
B. japonicum КВ11.
We have also used the well-known strain of
nodule bacteria Bradyrhizobium sp. 631 isolated
from soybean nodules. It is interesting because
it can form nodules both on soybean and lupin
roots [32]. According to the results of sequen-
cing of the intergenic ITS region, strain 631 was
assigned to the species B. lupini [39]. According
to antigenic properties, this strain belongs to
serogroup 631.
Since it is known from the literature that
nodule bacteria B. japonicum and B. diazoeffi-
ciens are able to nodulate lupin [11; 13; 14], we
chose two strains that belong to these species
for the study. They were isolated from soybean
nodules (B. japonicum КВ11) and cowpea
(B. diazoefficiens B22) and belong to serogro-
ups КВ11 and B2, respectively.
Under the conditions of the vegetation ex-
periment on vermiculite, it was established that
all bradyrhizobia strains isolated from lupin
nodules (B. lupini 367а, 30l, LD4 and Bradyrhi-
zobium sp. LD8) and seradella (Bradyrhizobi-
um sp. CP1), as well as B. lupini 631, formed
active red nodules on the roots of white and yel-
low lupin (Table 1). Microsymbionts of soybean
(B. japonicum КВ11) and cowpea (B. diazoeffi-
ciens В22) did not infect these two species of
lupin.
The next stage of our work was to investi-
gate the peculiarities of the interaction between
microsymbionts of various leguminous crops
and narrow-leaved lupin. The data obtained in
the vegetation experiment are shown in the
Table 2.
It was established that a significant number
of nodules were formed on the roots of non-ino-
culated plants of narrow-leaved lupin, namely
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 39.
26
Table 2. Influence of inoculation with Bradyrhizobium strains on symbiotic indicators of
narrow-leaved lupin of Lokomotyv variety (vegetation experiment, sod-podzolic soil)
Variants of the experi-
ment Host plant
Number of nodules Weight of nodules
units/plant % versus
control g/plant % versus
control
Without inoculation
(control) – 54.73 ± 1.36 – 0.23 ± 0.01 –
Inoculation with
B. lupini 367а
lupin
57.00 ± 1.41 4.1 0.29 ± 0.01* 26.1
Inoculation with
B. lupini 30l 59.73 ± 1.26* 9.1 0.28 ± 0.01* 21.7
Inoculation with
B. lupini LD4 57.60 ± 1.09 5.2 0.31 ± 0.02* 34.8
Inoculation with
Bradyrhizobium sp. LD8 57.27 ± 1.14 4.6 0.29 ± 0.01* 26.1
Inoculation with
Bradyrhizobium sp. СР1 seradella 61.00 ± 1.10* 11.5 0.28 ± 0.01* 21.7
Inoculation with
B. lupini 631 soybean 59.60 ± 1.26* 8.9 0.31 ± 0.01* 34.8
Inoculation with
B. japonicum КВ11 soybean 56.87 ± 1.53 3.8 0.29 ± 0.01* 26.1
Inoculation with
B. diazoefficiens В22 cowpea 57.20 ± 0.63 4.5 0.31 ± 0.01* 34.8
НІР05 3.52 0.04
Note. * — significant increase versus control.
55 units/plant. This fact indicates the presence
of a population of nodule bacteria capable of
nodulating lupin in the soil.
Against the background of spontaneous in-
fection of the roots of narrow-leaved lupin, all
studied strains, regardless of their origin, con-
tributed to the formation of a greater number of
nodules compared to the control. A significant
increase in this indicator by 9.1–11.5 % was re-
gistered after inoculation with B. lupini 30l and
B. lupini 631, as well as with the strain of
serogroup 367a isolated from the nodules of se-
radella — Bradyrhizobium sp. СР1. The fact
that microsymbionts of lupin and seradella can
cross-infect these leguminous plants is men-
tioned in the works of a number of researchers
[16; 33].
Bradyrhizobia strains isolated from lupin,
seradella, cowpea and soybean contributed to a
significant increase in the weight of narrow-
leaved lupin nodules by 21.7–34.8 % versus the
control (Table 2). It is important to note that this
indicator increased not only with the use of spe-
cific bacteria of B. lupini species, but also
strains of other species: B. japonicum КВ11 (by
26.1 %) and B. diazoefficiens В22 (by 34.8 %).
In addition to the positive effect on the
number and weight of nodules, the inoculation
of lupin seeds contributed to a significant inten-
sification of the process of symbiotic nitrogen
fixation (Fig. 1). A significant increase in nitro-
gen-fixing activity versus the control was regis-
tered when using strains of Bradyrhizobium sp.
LD8 (by 58.2 %) and B. lupini 631 (by 85.8 %),
capable of infecting lupins, as well as strains of
soybean nodule bacteria B. japonicum КВ11 (by
48.6 %) and B. diazoefficiens B22 (by 87.6 %).
Other strains — typical nodule bacteria of lupin:
B. lupini 367a, B. lupini 30l, B. lupini LD4 in-
creased this indicator only by 11.3–23.8 %.
The studied bradyrhizobia contributed to
the improvement of the growth and deve-
lopment of lupin plants. Thus, the greatest reli-
able increase in plant height was registered
when treating seeds with strains isolated from
lupin nodules (B. lupini 367a, B. lupini 301,
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 39.
27
Host plant → [ lupin ] [seradella] [soybean] [soybean] [cowpea]
Figure 1. Nitrogenase activity of lupin nodules of the Lokomotyv narrow-leaved variety after
inoculation with bradyrhizobia strains of different species (vegetation experiment, sod-podzolic
soil). НІР05 = 0.84. * — significant increase versus control.
B. lupini LD4, Bradyrhizobium sp. LD8) —
10.4–13.7 % and soybean nodules (B. lupi-
ni 631) — 12.9 % (Table 3). A slight increase in
this indicator (by 5.8–6.8 %) was reported after
inoculation with another strain from soybean
nodules, but of a different species (B. japoni-
cum КВ11), as well as with microsymbionts of
seradella (Bradyrhizobium sp. CP1) and cowpea
(B. diazoefficiens B22).
The use of bradyrhizobia strains isolated
from various leguminous plants had a positive
effect on the content of dry matter in the above-
ground mass of narrow-leaved lupin. Against
the background of a large local population of
lupin nodule bacteria, all specific strains of the
species B. lupini (367a, 30l, LD4, 631) and
Bradyrhizobium sp. LD8 provided a reliable in-
crease in the above-ground mass of plants ver-
sus the control by 14.3–22.6 %. The maximum
values of this indicator were obtained in variants
inoculated with B. lupini LD4 and B. lupini 631
(1.03 and 1.02 g/plant vs. 0.84 g/plant in the
control), which proved to be better for most
symbiotic traits. Despite the high nitrogenase
activity of lupin nodules with the strains B. ja-
ponicum КВ11 and B. diazoefficiens В22, the
increase in the above-ground mass of plants ver-
sus the control was lower and amounted to
7.1 % and 10.7 %, respectively.
We have also studied the influence of non-
specific nodule bacteria of the species B. japo-
nicum on white lupin plants. Lupin seeds were
inoculated with two highly active strains of soy-
bean rhizobia — B. japonicum КВ11 and B. ja-
ponicum 46. The use of a sterile substrate (ver-
miculite) in the vegetation experiment made it
possible to neutralize the effect of representa-
tives of local populations of lupin microsymbi-
onts present in the soil on plants.
The obtained data proved that the studied
strains of soybean rhizobia are not capable of
infecting white lupin (Table 4). Nitrogen-fixing
nodules on the roots (22 units/plant) were
formed only when seeds were inoculated with
a specific strain of nodule bacteria — B. lupi-
ni 367a, while the above-ground mass of lupin
plants increased by 60.0 % versus the control
(without inoculation). In variants with soybean
2,82 3,14 3,17 3,49
4,46
3,65
5,24
4,19
5,29
0
1
2
3
4
5
6
W
ith
ou
t i
no
cu
la
tio
n
(c
on
tro
l)
B.
lu
pi
ni
3
67
а
B
. l
up
in
i 3
0l
B
. l
up
in
i L
D
4
B
ra
dy
rh
iz
ob
iu
m
sp
. L
D
8
Br
ad
yr
hi
zo
bi
um
sp
. С
Р1
B
. l
up
in
i 6
31
B.
ja
po
ni
cu
m
К
В1
1
B.
di
az
oe
ffi
ci
en
s В
22
μg
N
/p
la
nt
p
er
h
ou
r
*
*
*
*
B.
lu
pi
ni
B.
lu
pi
ni
B.
lu
pi
ni
B.
lu
pi
ni
Br
ad
yr
hi
zo
bi
um
B.
ja
po
ni
cu
m
Br
ad
yr
hi
zo
bi
um
B.
d
ia
zo
ef
fic
ie
ns
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 39.
28
Table 3. Influence of inoculation with Bradyrhizobium strains on growth and development
of lupin plants of narrow-leaved Lokomotyv variety (vegetation experiment, sod-podzolic soil)
Variants
of the experiment Host plant
Plant height Content of dry matter in the
above-ground mass of plants
cm % versus
control g/plant % versus
control
Without inoculation
(control) – 30.97 ± 1.17 – 0.84 ± 0.05 –
Inoculation with
B. lupini 367а
lupin
35.15 ± 0.87* 13.5 0.96 ± 0.03* 14.3
Inoculation with
B. lupini 30l 34.71 ± 1.14* 12.1 0.99 ± 0.02* 17.9
Inoculation with
B. lupini LD4 35.20 ± 0.70* 13.7 1.03 ± 0.22* 22.6
Inoculation with
Bradyrhizobium sp. LD8 34.18 ± 0.71* 10.4 0.97 ± 0.02* 15.5
Inoculation with
Bradyrhizobium sp. СР1 seradella 32.97 ± 1.11 6.5 0.91 ± 0.04 8.3
Inoculation with
B. lupini 631 soybean 34.99 ± 0.78* 12.7 1.02 ± 0.05* 21.4
Inoculation with
B. japonicum КВ11 soybean 33.09 ± 0.56 6.8 0.90 ± 0.02 7.1
Inoculation with
B. diazoefficiens В22 cowpea 32.76 ± 0.40 5.8 0.93 ± 0.03 10.7
НІР05 2.50 0.10
Note. * — significant increases versus control.
Table 4. Influence of soybean and lupin nodule bacteria on symbiotic indicators of white
lupin (vegetation experiment, vermiculite)
Variants
of the experiment
Number
of nodules,
units/plant
Weight
of nodules,
g/plant
Nitrogen
fixation
activity,
μ N2/plant
per hour
Content of dry
matter in the
above-ground
mass of plants,
g/plant
Gain versus
control, %
Without inoculation
(control) 0 0 0 0.70 ± 0.02 –
Inoculation with
B. lupini 367а 21.83 ± 0.64 0.13 ± 0.01 6.39 ± 0.47 1.12 ± 0.01 60.0
Inoculation with
B. japonicum КВ11 0 0 0 0.81 ± 0.02 15.7
Inoculation with
B. japonicum 46 0 0 0 0.98 ± 0.01 40.0
rhizobia strains B. japonicum КВ11 and 46, de-
spite the absence of nodules, a significant in-
crease of this indicator by 15.7 % and 40.0 %,
respectively, was also registered.
The analysis of data from two vegetation
experiments shows that strains of nodule bacte-
ria species B. japonicum and B. diazoefficiens,
not specific for lupin, are able to positively
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 39.
29
influence both symbiotic indicators and the
growth and development of lupin plants. We be-
lieve this may be related to the production of
biologically active substances by bradyrhizobia,
in particular, phytohormones and exopolysac-
charides (EPS) [40]. When interacting with lu-
pin, the role of non-specific nodule bacteria can
be similar to the role played by growth-
promoting bacteria (PGPR) in the activation of
symbiotic relationships under simultaneous
inoculation with rhizobia [41–43]. There is evi-
dence that symbiotic nitrogen fixers are also
PGPR microorganisms and can be used as free-
living rhizobacteria capable of forming associa-
tive relationships with non-leguminous plants
[44; 45]. In his work Peňa-Cabrales J. et al.
demonstrated that bacteria of the Bradyrhizobi-
um genus develop well in the germinating zone
of soybean, bean, clover, cowpea, oat, wheat
and corn, stimulating root development in a si-
milar way to free-living rhizobacteria [46].
Other researchers found a connection between
the ability of nodule bacteria B. japonicum to
stimulate the growth of non-leguminous plants
and their production of indolyl-3-acetic acid
(IAC) [47]. It is generally known that rhizobia,
as PGP rhizobacteria, are able to produce a wide
range of phytohormones: auxins, cytokinins,
gibberellins, abscisic acid [48; 49]. Currently,
the participation of all groups of phytohormones
in the initiation, development and functioning of
nitrogen-fixing nodules has been established.
Strains of B. japonicum КВ11 and 46 that
we have used are also active producers of phy-
tohormones of auxin and cytokinin nature,
which was registered in previous studies [40]. In
addition, it was shown that not only soybean
nodule bacteria, but also the products of their
metabolism are able to change the ratio of cer-
tain strains of rhizobia in nodules and positively
influence the formation and functioning of sym-
biotic soybean systems. It should be also noted
that B. japonicum KB11 produces a significant
amount of exopolysaccharides [40], which can
exhibit hormone-like effects. It is known from
the literature that EPS of rhizobia are able to
change the virulence of other homologous
strains, enhance the fixation of molecular nitro-
gen and increase the efficiency of symbiotic
systems [50; 51]. Thus, possessing a complex of
features characteristic of PGP microorganisms,
the studied nodule bacteria B. japonicum and
B. diazoefficiens, which are not specific for lu-
pin, could directly or indirectly contribute to the
activation of the symbiosis between this plant
and representatives of the local population of
B. lupini. Further study of the interaction be-
tween bradyrhizobia of various species and lu-
pin will allow a better understanding of the
principles of formation of effective symbiotic
systems of this culture.
Conclusion. It was established that nodule
bacteria isolated from lupin (B. lupini 367a, 30l,
LD4 and Bradyrhizobium sp. LD8), seradella
(Bradyrhizobium sp. CP1) and soybean (B. lu-
pini 631) nodules are able to form an active
symbiosis with white and yellow lupin. Micro-
symbionts of soybean (B. japonicum КВ11) and
cowpea (B. diazoefficiens В22) did not infect
these two species of lupin.
In the experiment with narrow-leaved lupin,
all strains of B. lupini (367a, 30l, LD4, 631) and
Bradyrhizobium sp. LD8 provided a reliable in-
crease versus the control (without inoculation)
of the above-ground mass of plants by 14.3–
22.6 %. The maximum values of this indicator
were obtained in variants inoculated with B. lu-
pini LD4 and B. lupini 631 (1.03 and
1.02 g/plant vs. 0.84 g/plant in the control),
which proved to be better for most symbiotic
traits.
With the use of non-specific strains of
B. japonicum КВ11 and B. diazoefficiens В22,
despite the large mass of nodules and their high
nitrogenase activity, the increase in the abo-
ve-ground mass of narrow-leaved lupin plants
was not significant — 7.1 % and 10.7 %, res-
pectively.
REFERENCES
1. Sprent, J. I., Ardley, J. K., & James, E. K.
(2017). Biogeography of nodulated legumes and
their nitrogen-fixing symbionts. New Phytol., 215,
40–56. https://doi.org/10.1111/nph.14474
2. Drummond, C. S., Eastwood, R. J., Miot-
to, S. T. S., & Hughes, C. E. (2012). Multiple con-
tinental radiations and correlates of diversification
in Lupinus (Leguminosae): Testing for key in-
novation within complete taxon sampling. Syst.
Biol., 61, 443–460. https://doi.org/10.1093/sysbio/
syr126
3. Lucas, M. M., Stoddard, F. L., Annicchiari-
co, P., Frías, J., Martínez-Villaluenga, C., Suss-
mann, D. … Pueyo, J. J. (2015). The future of lupin
as a protein crop in Europe. Front. Plant Sci.,
6 (705), 1–6. https://doi.org/10.3389/fpls.2015.00705
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 39.
30
4. Mazur V. A., & Pantsyreva H. V. (2020). Rid
Lupinus L. v Ukraini: henofond, introduktsiia, nap-
riamy doslidzhen ta perspektyvy vykorystannia [Lu-
pinus L. genus in Ukraine: gene pool, introduction,
directions of research and prospects of use]. Vinny-
tsia: VNAU [in Ukrainian].
5. Holodna, A. V. (2018). Tekhnolohichni as-
pekty vyroshchuvannia kormovykh liupyniv u zoni
Lisostepu Ukrainy [Technological aspects of grow-
ing fodder lupins in the forest-steppe zone of Uk-
raine].Vinnytsia: TVORY [in Ukrainian].
6. Ratoshniuk, V. I., & Havryliuk, M. M.
(2020). Liupyn vuzkolystyi — kultura universalnoho
vykorystannia u zoni Polissia Ukrainy [Narrow-lea-
ved lupine — the culture of universal use in the
Polissia area of Ukraine]. Visnyk ahrarnoi nauky —
Bulletin of Agricultural Science, 8 (809), 26–38 [in Uk-
rainian]. https://doi.org/10.31073/agrovisnyk202008
7. Kovalevska, T. M., Hubanova, L. S., & Bar-
dakov, A. H. (2000). Rol bulbochkovykh bakterii
liupynu ta roslyny-khaziaina v formuvanni efektyv-
nykh symbiotychnykh vidnosyn [The role of lupine
nodule bacteria and the host plant in the formation of
effective symbiotic relationships], Biuleten Instytutu
silskohospodarskoi mikrobiolohii UAAN — Bulletin
of the Institute of Agricultural Microbiology UAAS,
8, 24–27 [in Ukrainian].
8. Vasyliuk, V. M., Melnykova, N. M., My-
khalkiv, L. M., Omelchuk, S. V., & Kots, S. Ya.
(2007). Formuvannia symbiotychnykh vzaiemovid-
nosyn roslyn liupynu z transpozonovymy mu-
tantamy Bradyrhizobium sp. (Lupinus) [Formation
of the symbiotic relationships between lupine plants
and transposon mutants Bradyrhizobium sp. (Lupi-
nus)]. Fiziologiya i biohimiya kulturnyh rastenij —
Physiology and biochemistry of cultivated plants,
39 (3), 233–241 [in Ukrainian].
9. Krutylo, D. V. (2021). Biological features of
groundnut microsymbionts widespread in the soils
of Ukraine. Silskohospodarska mikrobiolohiia —
Agricultural Microbiology, 34, 3–14. https://doi.org/
10.35868/1997-3004.34.3-14
10. Stępkowski, T., Banasiewicz, J., Grana-
da, C. E., Andrews, M., & Passaglia, L. M. P.
(2018). Phylogeny and phylogeography of rhizobial
symbionts nodulating legumes of the tribe Genis-
teae. Genes., 9 (3), 163. https://doi.org/10.3390/
genes9030163
11. Msaddak, A., Rejili, M., Durán, D., Rey, L.,
Imperial, J., Palacios, J. M. … Mars, M. (2017).
Members of Microvirga and Bradyrhizobium genera
are native endosymbiotic bacteria nodulating Lupi-
nus luteus in Northern Tunisian soils. FEMS Mic-
robiol. Ecol., 93 (6), fix068. https://doi.org/10.1093/
femsec/fix068
12. Msaddak, A., Mars, M., Quiñones, M. A.,
Lucas, M. M., & Pueyo, J. J. (2023). Lupin, a unique
legume that is nodulated by multiple microsym-
bionts: The role of horizontal gene transfer. Int.
J. Mol. Sci., 24 (7), 6496. https://doi.org/10.3390/
ijms24076496
13. Mellal, H., Yacine, B., Boukaous, L., Khou-
ni, S., Benguedouar, A., Castellano-Hinojosa, A., &
Bedmar, E. J. (2019). Phylogenetic diversity of
Bradyrhizobium strains isolated from root nodules
of Lupinus angustifolius grown wild in the North
East of Algeria. Syst. Appl. Microbiol., 42, 397–402.
https://doi.org/10.1016/j.syapm.2019.01.003
14. Stępkowski, T., Hughes, C. E., Law, I. J.,
Markiewicz, Ł., Gurda, D., Chlebicka, A., & Mou-
lin, L. (2007). Diversification of lupine Bradyrhizo-
bium strains: Evidence from nodulation gene trees.
Appl. Environ. Microbiol., 73 (10), 3254–3264.
https://doi.org/10.1128/AEM.02125-06
15. Granada, C. E., Beneduzi, A., Lisboa, B. B.,
Turchetto-Zolet, A. C., Vargas, L. K., & Passag-
lia, L. M. P. (2015). Multilocus sequence analysis
reveals taxonomic differences among Bradyrhizobi-
um sp. symbionts of Lupinus albescens Plants grow-
ing in arenized and nonarenized areas. Syst. Appl.
Microbiol., 38 (5), 323–329. https://doi.org/10.1016/
j.syapm.2015.03.009
16. Vinuesa, P., León-Barrios, M., Silva, C.,
Willems, A., Jarabo-Lorenzo, A., Pérez-Galdona, R.
… Martínez-Romero, E. (2005). Bradyrhizobium ca-
nariense sp. nov., an acid-tolerant endosymbiont that
nodulates endemic genistoid legumes (Papiliono-
ideae: Genisteae) from the Canary Islands, along
with Bradyrhizobium japonicum bv. genistearum,
Bradyrhizobium genospecies alpha and Bradyrhi-
zobium genospecies beta. Int. J. Syst. Evol. Micro-
biol., 55 (Pt 2), 569–575. https://doi.org/10.1099/
ijs.0.63292-0.
17. Durán, D., Rey, L., Sánchez-Canizares, C.,
Navarro, A., Imperial, J., & Ruiz-Argüeso, T. (2013).
Genetic diversity of indigenous rhizobial symbionts
of the Lupinus mariaejosephae endemism from alka-
line-limed soils within its area of distribution in
Eastern Spain. Syst. Appl. Microbiol., 36, 128–136.
https://doi.org/10.1016/j.syapm.2012.10.008
18. Rejili, M., Off, K., Brachmann, A., & Ma-
rín, M. (2020). Bradyrhizobium hipponense sp. nov.,
isolated from Lupinus angustifolius growing in the
Northern region of Tunisia. Int. J. Syst. Evol. Micro-
biol., 70 (10), 5539–5550. https://doi.org/10.1099/
ijsem.0.004445
19. Velázquez, E., Valverde, A., Rivas, R., Go-
mis, V., Peix, Á., Gantois, I. … Martínez-Molina, E.
(2010). Strains nodulating Lupinus albus on diffe-
rent continents belong to several new chromosomal
and symbiotic lineages within Bradyrhizobium.
Antonie van Leeuwenhoek, 97, 363–376. https://doi.
org/10.1007/s10482-010-9415-7
20. El Hilali, I., El Jamali, J., Thami Alami, I.,
& Filali Maltouf, A. (2016). Characterization and
biodiversity of a fast-growing rhizobacterial popula-
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 39.
31
tion nodulating lupine in Morocco. Int. J. New Tech.
Res., 2 (12), 27–37.
21. Pudełko, K. (2010). Diversity among field
populations of bacterial strains nodulating lupins in
Poland. Fragm Agron., 27 (1), 107–116.
22. Tounsi-Hammami, S., Le Roux, C., Dhane-
Fitouri, S., De Lajudie, P., Duponnois, R., &
Ben Jeddi, F. (2019). Genetic diversity of rhizobia
associated with root nodules of white lupin (Lupinus
albus L.) in Tunisian calcareous soils. Syst. Appl.
Microbiol., 42, 448–456. https://doi.org/10.1016/
j.syapm.2019.04.002
23. González-Sama, A., Lucas, M. M., de Fe-
lipe, M. R., & Pueyo, J. J. (2004). An unusual infec-
tion mechanism and nodule morphogenesis in white
lupin (Lupinus albus). New Phytol., 163, 371–380.
https://doi.org/10.1111/j.1469-8137.2004.01121.x
24. Valverde, A., Velázquez, E., Fernández-
Santos, F., Vizcaíno, N., Mateos, P. F., Martínez-
Molina, E. … Willems, A. (2005). Phyllobacterium
trifolii sp. nov., nodulating Trifolium and Lupinus in
Spanish soils. Int. J. Syst. Evol. Microbiol., 55 (Pt 5),
1985–1989. https://doi.org/10.1099/ijs.0.63551-0
25. Trujillo, M. E., Willems, A., Abril, A.,
Planchuelo, A. M., Rivas, R., Ludeňa, D. … Veláz-
quez, E. (2005). Nodulation of Lupinus albus by
strains of Ochrobactrum lupini sp. nov. Appl. Envi-
ron. Microbiol., 71 (3), 1318–1327. https://doi.org/
10.1128/AEM.71.3.1318-1327.2005
26. Msaddak, A., Rejili, M., Durán, D., Rey, L.,
Palacios, J. M., Imperial, J. … Mars, M. (2018).
Definition of two new symbiovars, sv. lupini and sv.
mediterranense, within the genera Bradyrhizobium
and Phyllobacterium efficiently nodulating Lupinus
micranthus in Tunisia. Syst. Appl. Microbiol., 41,
487–493. https://doi.org/10.1016/j.syapm.2018.04.004
27. Ardley, J. K., Parker, M. A., De Meyer, S. E.,
Trengove, R. D., O’Hara, G. W., Reeve, W. G. …
Howieson, J. G. (2012). Microvirga lupini sp. nov.,
Microvirga lotononidis sp. nov. and Microvirga
zambiensis sp. nov. are alphaproteobacterial root-
nodule bacteria that specifically nodulate and fix ni-
trogen with geographically and taxonomically sepa-
rate legume hosts. Int. J. Syst. Evol. Microbiol.,
62 (Pt 11), 2579–2588. https://doi.org/10.1099/ijs.0.
035097-0
28. Msaddak, A., Rey, L., Imperial, J., Palaci-
os, J. M., Mars, M., & Pueyo, J. J. (2021). Phyloge-
netic analyses of rhizobia isolated from nodules of
Lupinus angustifolius in Northern Tunisia reveal
Devosia sp. as a new microsymbiont of lupin spe-
cies. Agronomy., 11 (8), 1510. https://doi.org/10.
3390/agronomy11081510
29. Kuykendall, L., Saxena, B., Devine, T., &
Udell, S. (1992). Genetic diversity in Bradyrhizobi-
um japonicum Jordan 1982 and a proposal for Bra-
dyrhizobium elkanii sp. nov. Can. J. Microbiol., 38,
501–505. https://doi.org/10.1139/m92-082
30. Delamuta, J. R. M., Ribeiro, R. A., Ormeño-
Orrillo, E., Melo, I. S., Martınez-Romero, E., &
Hungria M. (2013). Polyphasic evidence supporting
the reclassification of Bradyrhizobium japonicum
group Ia strains as Bradyrhizobium diazoefficiens
sp. nov. Int. J. Syst. Evol. Microbiol., 63, 3342–
3351. https://doi.org/10.1099/ijs.0.049130-0
31. Silva, F. V., Simões-Araújo, J. L., Silva Jú-
nior, J. P., Xavier, G. R., & Rumjanek, N. G. (2012).
Genetic diversity of Rhizobia isolates from Amazon
soils using cowpea (Vigna unguiculata) as trap plant.
Brazil J Microbiol., 43, 682–691. https://doi.org/
10.1590/S1517-83822012000200033
32. Koc, S. Ya., Morgun, V. V., Patyka, V. F.,
Malichenko, S. M., Mamenko, P. M., Kirizij, D. A. …
Melnikova, N. M. (2011). Biologicheskaya fiksaciya
azota: v 4-h t. T. 2: Bobovo-rizobialnyj simbioz.
[Biological fixation of nitrogen: monograph]. Kiev:
Logos [in Russian].
33. Martyniuk, S., Oroń, J., & Martyniuk, M.
(2005). Diversity and numbers of root-nodule bacte-
ria (rhizobia) in Polish soils. Acta Societatis Botani-
corum Poloniae, 74 (1), 83–86. https://doi.org/10.
5586/asbp.2005.012
34. Kovalevska, T. M., Kozar, S. F., Kruty-
lo, D. V., Horban, V. P., Romanova, I. M., & Us-
manova, T. O. (2015). Metody kultyvuvannia ta try-
valoho zberihannia bulbochkovykh bakterii v kolek-
tsiiakh: metodychni rekomendatsii [The method of
cultivation and long-term storage of nodule bacteria
in collections: methodical recommendations]. Cher-
nihiv: ISMAV NAAN [in Ukrainian].
35. Kebot, E., Meyer, B. (1968). Eksperimen-
tal’naya immunologiya [Experimental immunology].
Moskva: Medicina [in Russian].
36. Volkohon, V. V. (Ed.). (2010). Ekspery-
mentalna gruntova mikrobiolohiia [Experimental
soil microbiology]. Kyiv: Ahrarna nauka [in Ukrain-
ian].
37. Hardy, R. W. F., Holsten, R. D., Jack-
son, E. K., & Burns, R. C. (1968). The Acetylene-
Ethylene Assay for N2 Fixation: Laboratory and
Field Evaluation. Plant Physiology, 43 (8), 1185–
1207. https://doi.org/10.1104/pp.43.8.1185
38. Rozhkov, A. O. et al. (2016). Doslidna
sprava v ahronomii: navch. posibnyk: u 2 kn. Kn. 1.
Teoretychni aspekty doslidnoi spravy [Research in
agronomy: science manual: in 2 books. Book 1.
Theoretical aspects of research]. A. O. Rozhkov
(Ed.). Kharkiv: Maydan [in Ukrainian].
39. Krutylo, D. V. (2020). Phenotypic and ge-
notypic properties of bradyrhizobia nodulating le-
guminous plants of the Glycine, Vigna and Lupinus
genera. Mikrobiolohichnyi zhurnal — Microbiologi-
cal journal, 82 (2), 38–50. https://doi.org/10.15407/
microbiolj82.02.038
40. Krutylo, D. V., & Leonova, N. O. (2016).
Symbiotic potential of Bradyrhizobium japonicum
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 39.
32
strains with different growth rates. Mikrobiolohich-
nyi zhurnal — Microbiological journal, 78 (5), 42–
52.
41. Cassán, F., Perrig, D., Sgroy, V., Masciarel-
li, O., Penna, C., & Luna, V. (2009). Azospirillum bra-
silense Az39 and Bradyrhizobium japonicum E109,
inoculated singly or in combination, promote seed
germination and early seedling growth in corn (Zea
mays L.) and soybean (Glycine max L.). European
Journal of Soil Biology., 45, 28–35. https://doi.org/
10.1016/J.EJSOBI.2008.08.005
42. Masciarelli, O., Llanes, A., & Luna, V.
(2014). A new PGPR co-inoculated with Bradyrhi-
zobium japonicum enhances soybean nodulation.
Microbiol. Res., 169 (7–8), 609–615. https://doi.org/
10.1016/j.micres.2013.10.001
43. Korir, H., Mungai, N. W., Thuita, M., Ham-
ba, Y., & Masso, C. (2017). Co-inoculation effect of
rhizobia and plant growth promoting rhizobacteria
on common bean growth in a low phosphorus soil.
Front. Plant Sci., 8 (141), 1–10. https://doi.org/10.
3389/fpls.2017.00141
44. García-Fraile, P., Carro, L., Robledo, M.,
Ramírez-Bahena, M. H., Flores-Félix, J. D., Fernán-
dez, M. T. … Velázquez, E. (2012). Rhizobium
promotes non-legumes growth and quality in several
production steps: towards a biofertilization of edible
raw vegetables healthy for humans. PLoS ONE,
7 (5), e38122. https://doi.org/10.1371/journal.pone.
0038122
45. Fahde, S., Boughribil, S., Sijilmassi, B., &
Amri, A. (2023). Rhizobia: A promising source of
plant growth-promoting molecules and their non-
legume interactions: examining applications and
mechanisms. Agriculture, 13 (7), 1279. https://doi.
org/10.3390/agriculture13071279
46. Peňa-Cabrales, J., & Alexander, M. (1983).
Growth of Rhizobium in unamended soil. Soil Sci.
Am. J., 47, 81–84. https://doi.org/10.2136/sssaj1983.
03615995004700010016x
47. Antoun, H., Beauchamp, C., Goussard, N.,
Chabot, R., & Lalande, R. (1998). Potential of Rhizo-
bium and Bradyrhizobium species as planta growth
promoting rhizobacteria on non-legumes: effect on
radishes (Raphanus sativus L.). Plant and Soil,
204 (1), 57–67. https://doi.org/10.1023/A:10043269
10584
48. Boiero, L., Perrig, D., Masciarelli, O., Pen-
na, C., Cassan, F., & Luna, V. (2007). Phytohor-
mone production by three strains of Bradyrhizobium
japonicum and possible physiological and techno-
logical implications. Applied Microbiology and Bio-
technology, 74 (4), 874–880. https://doi.org/10.1007/
s00253-006-0731-9
49. Kots, S. Ya., & Hryshchuk, O. O. (2015).
Fitohormony u formuvanni ta funktsionuvanni sym-
biotychnykh vzaiemovidnosyn bobovykh roslyn
i bulbochkovykh bakterii [Phytohormones in the
formation and functioning of symbiotic relationships
of leguminous plants and nodule bacteria]. Fyzyolo-
hyia rastenyi y henetyka — Plant physiology and
genetics, 47 (3), 187–206.
50. Kosenko, L. V., Hajlova, G. F., & Kore-
lov, V. E. (2001). Vliyanie ekzopolisaharidov Rhizo-
bium leguminosarum bv. vicea na nodulyaciyu i ri-
zogenez rastenij goroha [Effect exopolysaccharides
of Rhizobium leguminosarum bv. viciae on nodu-
lation and rhizogenesis of pea plants]. Fiziologiya
i biohimiya kulturnyh rastenij — Physiology and
biochemistry of cultivated plants, 33 (4), 347–354
[in Russian].
51. Melnykova, N. M. (2019). Vplyv polisakha-
rydiv ryzobii na prorostannia nasinnia soi ta bul-
bochkoutvorennia pry formuvanni soievo-ryzobial-
noho symbiozu [Effect of rhizobial exopolysaccha-
rides on soybean seed germination and nodule de-
velopment in the soybean-rhizobia symbiosis]. Fyzy-
olohyia rastenyi y henetyka — Plant physiology and
genetics, 51 (5), 436–446. http://jnas.nbuv.gov.ua/
article/UJRN-0001093049 [in Ukrainian].
Received 12.04.2024
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 39.
33
https://doi.org/10.35868/1997-3004.39.22-36
УДК 582.73:582.23:579.8:579.2
ВЗАЄМОДІЯ РОСЛИН ЛЮПИНУ ЗІ ШТАМАМИ
БУЛЬБОЧКОВИХ БАКТЕРІЙ РОДУ BRADYRHIZOBIUM
Д. В. Крутило
Інститут сільськогосподарської мікробіології та агропромислового виробництва НААН, м. Чернігів
e-mail: krutylodv@gmail.com
Мета. Вивчити особливості взаємодії трьох видів люпину з бульбочковими бактеріями
роду Bradyrhizobium, виділеними з різних бобових рослин: люпину, серадели, сої та вигни.
Методи. Мікробіологічні (виділення ризобій із бульбочок та їх культивування), серологічні
(визначення серологічної належності штамів), вегетаційного досліду (дослідження хазяйсь-
кої специфічності брадіризобій, вивчення формування та функціонування симбіозу люпину
з бульбочковими бактеріями), газохроматографічний (визначення активності симбіотичної
азотфіксації), математично-статистичні. Результати. За умов вегетаційних дослідів на
дерново-підзолистому ґрунті та стерильному вермикуліті досліджено особливості взаємо-
дії різних видів люпину зі специфічними та неспецифічними штамами бульбочкових бактерій
роду Bradyrhizobium. Встановлено, що ризобії, виділені із бульбочок люпину (B. lupini 367а,
30л, ЛД4 і Bradyrhizobium sp. ЛД8), серадели (Bradyrhizobium sp. СР1) та сої (B. lupini 631),
здатні формувати активний симбіоз із рослинами люпину білого та жовтого. Мікросимбі-
онти сої (B. japonicum КВ11) та вигни (B. diazoefficiens В22) не інфікували ці два види люпи-
ну. У досліді з люпином вузьколистим усі штами B. lupini (367а, 30л, ЛД4, 631) і штам
Bradyrhizobium sp. ЛД8 забезпечили достовірне збільшення до контролю надземної маси рос-
лин на 14,3–22,6 %. Максимальні значення цього показника отримані у варіантах з інокуля-
цією штамами B. lupini ЛД4 і B. lupini 631 (1,03 і 1,02 г/рослину проти 0,84 г/рослину у конт-
ролі). За використання неспецифічних для люпину штамів B. japonicum КВ11 і B. diazo-
efficiens В22 спостерігали суттєве підвищення нітрогеназної активності бульбочок (в 1,5
і 1,9 раза проти контролю), проте приріст надземної маси рослин був незначним — 7,1 %
і 10,7 % відповідно. Висновки. Встановлено, що специфічні для люпину брадіризобії різного
походження (B. lupini та Bradyrhizobium sp.) позитивно впливають на формування симбіо-
тичного апарату, ріст і розвиток рослин люпину білого, жовтого і вузьколистого. Неспе-
цифічні для люпину ризобії сої та вигни (B. japonicum КВ11 і B. diazoefficiens В22) сприяють
активізації взаємодії люпину вузьколистого з представниками ґрунтової популяції B. lupini
та діють як PGPR-мікроорганізми. Кращими за більшістю симбіотичних показників та
ефективністю симбіозу з люпином вузьколистим є штами B. lupini ЛД4 і B. lupini 631.
Ключові слова: бульбочкові бактерії, Bradyrhizobium lupini, B. japonicum, B. diazoeffi-
ciens, люпин, симбіотична взаємодія.
ЦИТОВАНА ЛІТЕРАТУРА
1. Sprent J. I., Ardley J. K., James E. K. Bio-
geography of nodulated legumes and their nitrogen-
fixing symbionts. New Phytol. 2017. Vol. 215.
P. 40–56. https://doi.org/10.1111/nph.14474
2. Drummond C. S., Eastwood R. J., Miot-
to S. T. S., Hughes C. E. Multiple continental radi-
ations and correlates of diversification in Lupinus
(Leguminosae): Testing for key innovation within
complete taxon sampling. Syst. Biol. 2012. Vol. 61.
P. 443–460. https://doi.org/10.1093/sysbio/syr126
3. Lucas M. M., Stoddard F. L., Annicchiari-
co P., Frías J., Martínez-Villaluenga C., Suss-
mann D. … Pueyo J. J. The future of lupin as
a protein crop in Europe. Front. Plant Sci. 2015.
Vol. 6, № 705. P. 1–6.
4. Мазур В. А., Панцирева Г. В. Рід Lupi-
nus L. в Україні: генофонд, інтродукція, напрями
досліджень та перспективи використання. Він-
ниця: ВНАУ, 2020. 200 с.
5. Голодна А. В. Технологічні аспекти ви-
рощування кормових люпинів у зоні Лісостепу
України. Вінниця: ТВОРИ, 2018. 380 с.
6. Ратошнюк В. І., Гаврилюк М. М. Люпин
вузьколистий — культура універсального вико-
ристання у зоні Полісся України. Вісник аграрної
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 39.
34
науки. 2020. № 8 (809). С. 26–38. https://doi.org/
10.31073/agrovisnyk202008
7. Ковалевська Т. М., Губанова Л. С., Барда-
ков А. Г. Роль бульбочкових бактерій люпину та
рослини-хазяїна в формуванні ефективних сим-
біотичних відносин. Бюл. Інституту с.-г. мікро-
біології УААН. 2000. № 8. С. 24–27.
8. Василюк В. М., Мельникова Н. М., Ми-
халків Л. М., Омельчук С. В., Коць С. Я. Форму-
вання симбіотичних взаємовідносин рослин лю-
пину з транспозоновими мутантами Bradyrhizo-
bium sp. (Lupinus). Физиология и биохимия культ.
растений. 2007. Т. 39, № 3. С. 233–241.
9. Krutylo D. V. Biological features of ground-
nut microsymbionts widespread in the soils of Uk-
raine. Сільськогосподарська мікробіологія. 2021.
Вип. 34. С. 3–14. https://doi.org/10.35868/1997-
3004.34.3-14
10. Stępkowski T., Banasiewicz J., Grana-
da C. E., Andrews M., Passaglia L. M. P. Phyloge-
ny and phylogeography of rhizobial symbionts
nodulating legumes of the tribe Genisteae. Genes.
2018. Vol. 9, № 3. 163. https://doi.org/10.3390/
genes9030163
11. Msaddak A., Rejili M., Durán D., Rey L.,
Imperial J., Palacios J. M. … Mars M. Members of
Microvirga and Bradyrhizobium genera are native
endosymbiotic bacteria nodulating Lupinus luteus in
Northern Tunisian soils. FEMS Microbiol. Ecol.
2017. Vol. 93, № 6. fix068. https://doi.org/10.1093/
femsec/fix068
12. Msaddak A., Mars M., Quiñones M. A.,
Lucas M. M., Pueyo J. J. Lupin, a unique legume
that is nodulated by multiple microsymbionts: The
role of horizontal gene transfer. Int. J. Mol. Sci.
2023. Vol. 24, № 7. 6496. https://doi.org/10.3390/
ijms24076496
13. Mellal H., Yacine B., Boukaous L., Khou-
ni S., Benguedouar A., Castellano-Hinojosa A., Bed-
mar E. J. Phylogenetic diversity of Bradyrhizobium
strains isolated from root nodules of Lupinus angus-
tifolius grown wild in the North East of Algeria.
Syst. Appl. Microbiol. 2019. Vol. 42. P. 397–402.
https://doi.org/10.1016/j.syapm.2019.01.003
14. Stępkowski T., Hughes C. E., Law I. J.,
Markiewicz Ł., Gurda D., Chlebicka A., Moulin L.
Diversification of lupine Bradyrhizobium strains:
Evidence from nodulation gene trees. Appl. Environ.
Microbiol. 2007. Vol. 73, № 10. P. 3254–3264.
https://doi.org/10.1128/AEM.02125-06
15. Granada C. E., Beneduzi A., Lisboa B. B.,
Turchetto-Zolet A. C., Vargas L. K., Passag-
lia L. M. P. Multilocus sequence analysis reveals
taxonomic differences among Bradyrhizobium sp.
symbionts of Lupinus albescens Plants growing in
arenized and nonarenized areas. Syst. Appl. Micro-
biol. 2015. Vol. 38, № 5. P. 323–329. https://doi.org/
10.1016/j.syapm.2015.03.009
16. Vinuesa P., León-Barrios M., Silva C., Wil-
lems A., Jarabo-Lorenzo A., Pérez-Galdona R. …
Martínez-Romero E. Bradyrhizobium canariense sp.
nov., an acid-tolerant endosymbiont that nodulates
endemic genistoid legumes (Papilionoideae: Genis-
teae) from the Canary Islands, along with Brady-
rhizobium japonicum bv. genistearum, Bradyrhizo-
bium genospecies alpha and Bradyrhizobium geno-
species beta. Int. J. Syst. Evol. Microbiol. 2005.
Vol. 55 (Pt 2). P. 569–575. https://doi.org/10.1099/
ijs.0.63292-0
17. Durán D., Rey L., Sánchez-Canizares C.,
Navarro A., Imperial J., Ruiz-Argüeso T. Genetic
diversity of indigenous rhizobial symbionts of the
Lupinus mariaejosephae endemism from alkaline-
limed soils within its area of distribution in Eastern
Spain. Syst. Appl. Microbiol. 2013. Vol. 36. P. 128–
136. https://doi.org/10.1016/j.syapm.2012.10.008
18. Rejili M., Off K., Brachmann A., Marín M.
Bradyrhizobium hipponense sp. nov., isolated from
Lupinus angustifolius growing in the Northern
region of Tunisia. Int. J. Syst. Evol. Microbiol. 2020.
Vol. 70, № 10. P. 5539–5550. https://doi.org/10.
1099/ijsem.0.004445
19. Velázquez E., Valverde A., Rivas R., Go-
mis V., Peix Á., Gantois I. … Martínez-Molina E.
Strains nodulating Lupinus albus on different conti-
nents belong to several new chromosomal and sym-
biotic lineages within Bradyrhizobium. Antonie van
Leeuwenhoek. 2010. Vol. 97. P. 363–376. https://
doi.org/10.1007/s10482-010-9415-7
20. El Hilali I., El Jamali J., Thami Alami I.,
Filali Maltouf A. Characterization and biodiversity
of a fast-growing rhizobacterial population nodula-
ting lupine in Morocco. Int. J. New Tech. Res. 2016.
Vol. 2, № 12. P. 27–37.
21. Pudełko K. Diversity among field populati-
ons of bacterial strains nodulating lupins in Poland.
Fragm Agron. 2010. Vol. 27, № 1. P. 107–116.
22. Tounsi-Hammami S., Le Roux C., Dhane-
Fitouri S., De Lajudie P., Duponnois R., Ben Jed-
di F. Genetic diversity of rhizobia associated with
root nodules of white lupin (Lupinus albus L.) in
Tunisian calcareous soils. Syst. Appl. Microbiol.
2019. Vol. 42. P. 448–456. https://doi.org/10.1016/
j.syapm.2019.04.002
23. González-Sama A., Lucas M. M., de Feli-
pe M. R., Pueyo J. J. An unusual infection mecha-
nism and nodule morphogenesis in white lupin (Lu-
pinus albus). New Phytol. 2004. Vol. 163. P. 371–
380. https://doi.org/10.1111/j.1469-8137.2004.01121.x
24. Valverde A., Velázquez E., Fernández-San-
tos F., Vizcaíno N., Mateos P. F., Martínez-Moli-
na E., Igual J. M., Willems A. Phyllobacterium
trifolii sp. nov., nodulating Trifolium and Lupinus in
Spanish soils. Int. J. Syst. Evol. Microbiol. 2005.
Vol. 55 (Pt 5). P. 1985–1989. https://doi.org/10.
1099/ijs.0.63551-0
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 39.
35
25. Trujillo M. E., Willems A., Abril A., Plan-
chuelo A. M., Rivas R., Ludeňa D. … Velázquez E.
Nodulation of Lupinus albus by strains of Ochro-
bactrum lupini sp. nov. Appl. Environ. Microbiol.
2005. Vol. 71, № 3. P. 1318–1327. https://doi.org/
10.1128/AEM.71.3.1318-1327.2005
26. Msaddak A., Rejili M., Durán D., Rey L.,
Palacios J. M., Imperial J. … Mars M. Definition of
two new symbiovars, sv. lupini and sv. mediterra-
nense, within the genera Bradyrhizobium and Phyl-
lobacterium efficiently nodulating Lupinus micran-
thus in Tunisia. Syst. Appl. Microbiol. 2018. Vol. 41.
P. 487–493. https://doi.org/10.1016/j.syapm.2018.
04.004
27. Ardley J. K., Parker M. A., De Meyer S. E.,
Trengove R. D., O’Hara G. W., Reeve W. G. …
Howieson J. G. Microvirga lupini sp. nov., Micro-
virga lotononidis sp. nov. and Microvirga zambi-
ensis sp. nov. are alphaproteobacterial root-nodule
bacteria that specifically nodulate and fix nitrogen
with geographically and taxonomically separate
legume hosts. Int. J. Syst. Evol. Microbiol. 2012.
Vol. 62 (Pt 11). P. 2579–2588. https://doi.org/10.
1099/ijs.0.035097-0
28. Msaddak A., Rey L., Imperial J., Palaci-
os J. M., Mars M., Pueyo J. J. Phylogenetic analyses
of rhizobia isolated from nodules of Lupinus
angustifolius in Northern Tunisia reveal Devosia sp.
as a new microsymbiont of lupin species. Agronomy.
2021. Vol. 11, № 8. 1510. https://doi.org/10.3390/
agronomy11081510
29. Kuykendall L., Saxena B., Devine T., Udell S.
Genetic diversity in Bradyrhizobium japonicum Jor-
dan 1982 and a proposal for Bradyrhizobium elkanii
sp. nov. Can. J. Microbiol. 1992. Vol. 38. P. 501–
505. https://doi.org/10.1139/m92-082
30. Delamuta J. R. M., Ribeiro R. A., Ormeño-
Orrillo E, Melo I. S., Martınez-Romero E., Hung-
ria M. Polyphasic evidence supporting the reclassi-
fication of Bradyrhizobium japonicum group Ia
strains as Bradyrhizobium diazoefficiens sp. nov.
Int. J. Syst. Evol. Microbiol. 2013. Vol. 63. P. 3342–
3351. https://doi.org/10.1099/ijs.0.049130-0
31. Silva F. V., Simões-Araújo J. L., Silva Júni-
or J. P., Xavier G. R., Rumjanek N. G. Genetic
diversity of Rhizobia isolates from Amazon soils
using cowpea (Vigna unguiculata) as trap plant.
Brazil J Microbiol. 2012. Vol. 43. P. 682–691.
https://doi.org/10.1590/S1517-83822012000200033
32. Коць С. Я., Моргун В. В., Патыка В. Ф.,
Маличенко С. М., Маменко П. М., Киризий Д. А.
… Мельникова Н. М. Биологическая фиксация
азота: бобово-ризобиальный симбиоз: в 4-х т.
Т. 2. Киев : Логос, 2011. 523 с.
33. Martyniuk S., Oroń J., Martyniuk M. Diver-
sity and numbers of root-nodule bacteria (rhizobia)
in Polish soils. Acta Societatis Botanicorum Polo-
niae. 2005. Vol. 74. № 1. P. 83–86. https://doi.org/
10.5586/asbp.2005.012
34. Ковалевська Т. М., Козар С. Ф., Крути-
ло Д. В., Горбань В. П., Романова І. М., Усмано-
ва Т. О. Методи культивування та тривалого збе-
рігання бульбочкових бактерій у колекціях. Ме-
тодичні рекомендації. Чернігів : ІСМАВ НААН,
2015. 36 с.
35. Кэбот Э., Мейер Б. Экспериментальная
иммунология. Москва : Медицина, 1968. 677 с.
36. Експериментальна ґрунтова мікробіоло-
гія / за ред. В. В. Волкогона. Київ : Аграрна нау-
ка, 2010. 464 с.
37. Hardy R. W. F., Holsten R. D., Jackson E. K.,
Burns R. C. The acetylene-ethylene assay for N2
fixation: laboratory and field evaluation. Plant Phy-
siol. 1968. № 43. P. 1185–1207. https://doi.org/
10.1104/pp.43.8.1185
38. Дослідна справа в агрономії: навч. посіб-
ник: у 2 кн. — Кн. 1. Теоретичні аспекти дослід-
ної справи / А. О. Рожков, В. К. Пузік, С. М. Ка-
ленська та ін.; за ред. А. О. Рожкова. Харків :
Майдан, 2016. 316 с.
39. Krutylo D. V. Phenotypic and genotypic
properties of bradyrhizobia nodulating leguminous
plants of the Glycine, Vigna and Lupinus genera.
Мікробіол. журн. 2020. Т. 82. № 2. С. 38–50.
https://doi.org/10.15407/microbiolj82.02.038
40. Krutylo D. V., Leonova N. O. Symbiotic
potential of Bradyrhizobium japonicum strains with
different growth rates. Мікробіол. журн. 2016.
Т. 78, № 5. С. 42–52.
41. Cassán F., Perrig D., Sgroy V., Masciarel-
li O., Penna C., Luna V. Azospirillum brasilense
Az39 and Bradyrhizobium japonicum E109, inocu-
lated singly or in combination, promote seed ger-
mination and early seedling growth in corn (Zea
mays L.) and soybean (Glycine max L.). European
Journal of Soil Biology. 2009. Vol. 45. P. 28–35.
https://doi.org/10.1016/J.EJSOBI.2008.08.005
42. Masciarelli O., Llanes A., Luna V. A new
PGPR co-inoculated with Bradyrhizobium japoni-
cum enhances soybean nodulation. Microbiol. Res.
2014. Vol. 169, № 7–8. P. 609–615. https://doi.org/
10.1016/j.micres.2013.10.001
43. Korir H., Mungai N. W., Thuita M., Ham-
ba Y., Masso C. Co-inoculation effect of rhizobia
and plant growth promoting rhizobacteria on com-
mon bean growth in a low phosphorus soil. Front.
Plant Sci. 2017. Vol. 8, № 141. P. 1–10. https://doi.
org/10.3389/fpls.2017.00141
44. García-Fraile P., Carro L., Robledo M., Ra-
mírez-Bahena M. H., Flores-Félix J. D., Fernán-
dez M. T. … Velázquez E. Rhizobium promotes
non-legumes growth and quality in several produc-
tion steps: towards a biofertilization of edible raw
vegetables healthy for humans. PLoS ONE. 2012.
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 39.
36
Vol. 7, № 5. e38122. https://doi.org/10.1371/journal.
pone.0038122
45. Fahde S., Boughribil S., Sijilmassi B., Am-
ri A. Rhizobia: A promising source of plant growth-
promoting molecules and their non-legume inte-
ractions: examining applications and mechanisms.
Agriculture. 2023. Vol. 13, № 7. 1279. https://doi.
org/10.3390/agriculture13071279
46. Peňa-Cabrales J., Alexander M. Growth of
Rhizobium in unamended soil. Soil Sci. Am. J. 1983.
Vol. 47. P. 81–84. https://doi.org/10.2136/sssaj1983.
03615995004700010016x
47. Antoun H., Beauchamp C., Goussard N.,
Chabot R., Lalande R. Potential of Rhizobium and
Bradyrhizobium species as planta growth promoting
rhizobacteria on non-legumes: effect on radishes
(Raphanus sativus L.). Plant and Soil. 1998.
Vol. 204, № 1. P. 57–67. https://doi.org/10.1023/
A:1004326910584
48. Boiero L., Perrig D., Masciarelli O., Pen-
na C., Cassan F., Luna V. Phytohormone production
by three strains of Bradyrhizobium japonicum and
possible physiological and technological implica-
tions. Applied Microbiology and Biotechnology.
2007. Vol. 74, № 4. P. 874–880. https://doi.org/10.
1007/s00253-006-0731-9
49. Коць С. Я., Грищук О. О. Фітогормони
у формуванні та функціонуванні симбіотичних
взаємовідносин бобових рослин і бульбочкових
бактерій. Физиология растений и генетика. 2015.
Т. 47, № 3. С. 187–206.
50. Косенко Л. В., Хайлова Г. Ф., Коре-
лов В. Е. Влияние экзополисахаридов Rhizobium
leguminosarum bv. vicea на нодуляцию и ризоге-
нез растений гороха. Физиология и биохимия
культ. растений. 2001. Т. 33, № 4. С 347–354.
51. Мельникова Н. М. Вплив полісахаридів
ризобій на проростання насіння сої та бульбоч-
коутворення при формуванні соєво-ризобіаль-
ного симбіозу. Физиология растений и генетика.
2019. Т. 51, № 5. С. 436–446. http://jnas.nbuv.
gov.ua/article/UJRN-0001093049
Отримано 12.04.2024
ISSN 1997-3004 Сільськогосподарська мікробіологія. 2024. Вип. 39.
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| id | oai:ojs2.smic.in.ua:article-523 |
| institution | Agriciltural microbiology |
| keywords_txt_mv | keywords |
| language | English |
| last_indexed | 2026-07-23T01:16:04Z |
| publishDate | 2024 |
| publisher | Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine |
| record_format | ojs |
| resource_txt_mv | smicinua/eb/ddd87a6a5b73ba2db4a2987006fc28eb.pdf |
| spelling | oai:ojs2.smic.in.ua:article-5232026-07-22T10:10:52Z INTERACTION BETWEEN LUPIN PLANTS AND STRAINS OF THE BRADYRHIZOBIUM GENUS ВЗАЄМОДІЯ РОСЛИН ЛЮПИНУ ЗІ ШТАМАМИ БУЛЬБОЧКОВИХ БАКТЕРІЙ РОДУ BRADYRHIZOBIUM Крутило, Д. В. nodule bacteria, Bradyrhizobium lupini, B. japonicum, B. diazoefficiens, lupin, symbiotic interactions бульбочкові бактерії, Bradyrhizobium lupini, B. japonicum, B. diazoefficiens, люпин, симбіотична взаємодія Objective. Study the peculiarities of the interaction between three species of lupin and nodule bacteria of the genus Bradyrhizobium, isolated from various leguminous plants: lupin, seradella, soybean and cowpea. Methods. Microbiological (isolation of rhizobia from nodules and their cultivation), serological (defining serological affiliation of strains), vegetation experiment (study of host specificity of bradyrhizobia, study of the formation and functioning of the symbiosis between lupin and nodule bacteria), gas chromatographic (determination of the activity of symbiotic nitrogen fixation), mathematical and statistical. Results. Under the conditions of vegetation experiments on sod-podzolic soil and sterile vermiculite, the peculiarities of the interaction between different species of lupin and specific and non-specific strains of nodule bacteria of the genus Bradyrhizobium were studied. It was established that rhizobia isolated from lupin nodules (B. lupini 367a, 30l, LD4 and Bradyrhizobium sp. LD8), seradella (Bradyrhizobium sp. СР1) and soybean (B. lupini 631) are able to form an active symbiosis with white and yellow lupin plants. Microsymbionts of soybean (B. japonicum КВ11) and cowpea (B. diazoefficiens В22) did not infect these two species of lupin. In the experiment with narrow-leaved lupin, all strains of B. lupini (367а, 30l, LD4, 631) and LD8 Bradyrhizobium sp. ensured a reliable increase in the above-ground mass of plants by 14.3–22.6 % compared to the control. The maximum values of this indicator were obtained in variants inoculated with B. lupini LD4 and B. lupine 631 strains (1.03 and 1.02 g/plant versus 0.84 g/plant in the control). With the use of non-specific lupin strains B. japonicum КВ11 and B. diazoefficiens В22, a significant increase in the nitrogenase activity of nodules was registered (1.5 and 1.9 times compared to the control), however, the increase in the above-ground mass of plants was insignificant — 7.1 % and 10.7 %, respectively. Conclusion. It has been established that lupin-specific bradyrhizobia of various origins (B. lupini and Bradyrhizobium sp.) have a positive effect on the formation of the symbiotic apparatus, the growth and development of white, yellow and narrowleaved lupin plants. Rhizobia of soybean and cowpea (B. japonicum КВ11 and B. diazoefficiens В22) non-specific for lupin, contribute to the activation of the interaction between narrowleaved lupin and representatives of the soil population of B. lupini and act as PGPR microorganisms. B. lupini LD4 and B. lupini 631 strains are the best in terms of most symbiotic indicators and efficiency of symbiosis with narrow-leaved lupin. Мета. Вивчити особливості взаємодії трьох видів люпину з бульбочковими бактеріями роду Bradyrhizobium, виділеними з різних бобових рослин: люпину, серадели, сої та вигни. Методи. Мікробіологічні (виділення ризобій із бульбочок та їх культивування), серологічні (визначення серологічної належності штамів), вегетаційного досліду (дослідження хазяйської специфічності брадіризобій, вивчення формування та функціонування симбіозу люпину з бульбочковими бактеріями), газохроматографічний (визначення активності симбіотичної азотфіксації), математично-статистичні. Результати. За умов вегетаційних дослідів на дерново-підзолистому ґрунті та стерильному вермикуліті досліджено особливості взаємодії різних видів люпину зі специфічними та неспецифічними штамами бульбочкових бактерій роду Bradyrhizobium. Встановлено, що ризобії, виділені із бульбочок люпину (B. lupini 367а, 30л, ЛД4 і Bradyrhizobium sp. ЛД8), серадели (Bradyrhizobium sp. СР1) та сої (B. lupini 631), здатні формувати активний симбіоз із рослинами люпину білого та жовтого. Мікросимбіонти сої (B. japonicum КВ11) та вигни (B. diazoefficiens В22) не інфікували ці два види люпину. У досліді з люпином вузьколистим усі штами B. lupini (367а, 30л, ЛД4, 631) і штам Bradyrhizobium sp. ЛД8 забезпечили достовірне збільшення до контролю надземної маси рослин на 14,3–22,6 %. Максимальні значення цього показника отримані у варіантах з інокуляцією штамами B. lupini ЛД4 і B. lupini 631 (1,03 і 1,02 г/рослину проти 0,84 г/рослину у контролі). За використання неспецифічних для люпину штамів B. japonicum КВ11 і B. diazoefficiens В22 спостерігали суттєве підвищення нітрогеназної активності бульбочок (в 1,5 і 1,9 раза проти контролю), проте приріст надземної маси рослин був незначним — 7,1 % і 10,7 % відповідно. Висновки. Встановлено, що специфічні для люпину брадіризобії різного походження (B. lupini та Bradyrhizobium sp.) позитивно впливають на формування симбіотичного апарату, ріст і розвиток рослин люпину білого, жовтого і вузьколистого. Неспецифічні для люпину ризобії сої та вигни (B. japonicum КВ11 і B. diazoefficiens В22) сприяють активізації взаємодії люпину вузьколистого з представниками ґрунтової популяції B. lupini та діють як PGPR-мікроорганізми. Кращими за більшістю симбіотичних показників та ефективністю симбіозу з люпином вузьколистим є штами B. lupini ЛД4 і B. lupini 631. Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine 2024-05-17 Article Article Рецензована Стаття application/pdf https://smic.in.ua/index.php/journal/article/view/523 10.35868/1997-3004.39.22-36 Agricultural microbiology; Vol. 39 (2024): Agriciltural microbiology; 22-36 Сільськогосподарська мікробіологія; Том 39 (2024): Сільськогосподарська мікробіологія; 22-36 1997-3004 10.35868/1997-3004.39 en https://smic.in.ua/index.php/journal/article/view/523/592 Авторське право (c) 2024 D. V. Krutylo https://creativecommons.org/licenses/by/4.0 |
| spellingShingle | бульбочкові бактерії Bradyrhizobium lupini B. japonicum B. diazoefficiens люпин симбіотична взаємодія Крутило, Д. В. ВЗАЄМОДІЯ РОСЛИН ЛЮПИНУ ЗІ ШТАМАМИ БУЛЬБОЧКОВИХ БАКТЕРІЙ РОДУ BRADYRHIZOBIUM |
| title | ВЗАЄМОДІЯ РОСЛИН ЛЮПИНУ ЗІ ШТАМАМИ БУЛЬБОЧКОВИХ БАКТЕРІЙ РОДУ BRADYRHIZOBIUM |
| title_alt | INTERACTION BETWEEN LUPIN PLANTS AND STRAINS OF THE BRADYRHIZOBIUM GENUS |
| title_full | ВЗАЄМОДІЯ РОСЛИН ЛЮПИНУ ЗІ ШТАМАМИ БУЛЬБОЧКОВИХ БАКТЕРІЙ РОДУ BRADYRHIZOBIUM |
| title_fullStr | ВЗАЄМОДІЯ РОСЛИН ЛЮПИНУ ЗІ ШТАМАМИ БУЛЬБОЧКОВИХ БАКТЕРІЙ РОДУ BRADYRHIZOBIUM |
| title_full_unstemmed | ВЗАЄМОДІЯ РОСЛИН ЛЮПИНУ ЗІ ШТАМАМИ БУЛЬБОЧКОВИХ БАКТЕРІЙ РОДУ BRADYRHIZOBIUM |
| title_short | ВЗАЄМОДІЯ РОСЛИН ЛЮПИНУ ЗІ ШТАМАМИ БУЛЬБОЧКОВИХ БАКТЕРІЙ РОДУ BRADYRHIZOBIUM |
| title_sort | взаємодія рослин люпину зі штамами бульбочкових бактерій роду bradyrhizobium |
| topic | бульбочкові бактерії Bradyrhizobium lupini B. japonicum B. diazoefficiens люпин симбіотична взаємодія |
| topic_facet | nodule bacteria Bradyrhizobium lupini B. japonicum B. diazoefficiens lupin symbiotic interactions бульбочкові бактерії Bradyrhizobium lupini B. japonicum B. diazoefficiens люпин симбіотична взаємодія |
| url | https://smic.in.ua/index.php/journal/article/view/523 |
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