БАКТЕРІАЛЬНА МІКРОБІОТА НАСІННЯ НАСІННЯ ЦУКРОВИХ БУРЯКІВ
The paper shows the results of bacterial microflora of sugar beet seeds study, with the analysis of morphological, culture-biochemical and virulent characteristics of isolated bacteria. The presence of pathogenic bacteria was observed on seeds pretreated with fungicide. The antagonistic activity of...
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| Date: | 2013 |
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| Language: | English Ukrainian |
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Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine
2013
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| Journal Title: | Agriciltural microbiology |
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Agriciltural microbiology| _version_ | 1871465717955559424 |
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| author | Дворак , К.П. Буценко , Л.М. |
| author_facet | Дворак , К.П. Буценко , Л.М. |
| author_institution_txt_mv | [
{
"author": "К.П. Дворак ",
"institution": "Інститут біоенергетичних культур і цукрових буряків НААН"
},
{
"author": "Л.М. Буценко ",
"institution": "Інститут мікробіології і вірусології ім.Заболотного НАН України"
}
] |
| author_sort | Дворак , К.П. |
| baseUrl_str | https://smic.in.ua/index.php/journal/oai |
| collection | OJS |
| datestamp_date | 2026-07-22T10:10:44Z |
| description | The paper shows the results of bacterial microflora of sugar beet seeds study, with the analysis of morphological, culture-biochemical and virulent characteristics of isolated bacteria. The presence of pathogenic bacteria was observed on seeds pretreated with fungicide. The antagonistic activity of bacterial microflora of sugar beet seeds in relation to known bacterial diseases agents of this culture was investigated. |
| doi_str_mv | 10.35868/1997-3004.17.48-57 |
| first_indexed | 2025-07-17T12:24:25Z |
| format | Article |
| fulltext |
BACTERIAL MICROFLORA OF SUGAR BEET SEEDS
1Dvorak K. P., 2Butsenko L. M.
1Institute of Bioenergy Crops and Sugar Beet of NAAS of Ukraine, Kyiv
25, Klinichna str., Kyiv, 03141
2Zabolotny Institute of Microbiology and Virology NAS of Ukraine, Kyiv
154, Zabolotnogo str., Kyiv, D03680
e-mail: ekaterina-dvorak@rambler.ru
The paper shows the results of bacterial microflora of sugar beet seeds study,
with the analysis of morphological, culture-biochemical and virulent characteristics
of isolated bacteria. The presence of pathogenic bacteria was observed on seeds
pretreated with fungicide. The antagonistic activity of bacterial microflora of sugar
beet seeds in relation to known bacterial diseases agents of this culture was
investigated.
Keywords: sugar beet, seeds, bacterial microflora, antagonism, artificial
infection.
Crop seeds is a substrate for various microorganisms: micromycetes, bacteria,
mycoplasma and viruses [10]. Seeds microbiota is divided into the several groups.
Epiphytic microbiota – microorganisms that colonize the seed surface and consume
plant cells waste products. Under the normal conditions, these organisms do not
invade the internal tissues and do not harm the plant [4]. Under the favorable
conditions, epiphytes proliferate on the surface of plants and can cause a biological
barrier that prevents parasites penetration into the plant tissue. Endophytic bacteria
colonize internal tissues, but do not cause disease symptoms and do not have adverse
effects on plants [11]. However, the composition of epiphyte and endophyte
microbiota can include microorganisms that may be able not only to penetrate into
the inner parts of plants and develop there, but also to cause diseases of seeds and
plants [4].
Use of pesticides is required nowadays as the basis of plant protection.
However, the use of compounds toxic to the certain types of pathogens, can cause the
appearance of the resistant forms of pathogens and development of pathogenic
organisms of other taxonomic groups. It was shown [3] that on the surface of sugar
beet seeds treated with highly toxic insecticides Carbofuran and Gaucho, fungicide
Tachygaren and moderately toxic fungicide TMTD, contained even more bacterial
cells than on non treated seeds (100 – 160 thousand and 30 – 40 thousand,
respectively).
Therefore, the aim of our work was to analyze the composition of biological
properties and microbiota composition of sugar beet seeds, intact and treated with
pesticides.
Materials and methods. Seeds of sugar beet hybrids Olexandria and Umansky
CS-97, selected on the breeding station of the Institute of Bioenergy Crops and Sugar
Beet of NAAS of Ukraine were used in the studies.
Hybrid Olexandria – single seed triploid on sterile base, with high yield
potential and sugar content. It was entered into the Registry of Plant Varieties of
Ukraine in 1997 [7]. The studied seeds of Olexandria hybrid were treated with
fungicide Maxim XL 035 FS.
Umansky CS-97 – is single seed diploid MS hybrid oriented on high sugar
content yield. It is resistant to premature seeding and tolerant to swift moth and
Cercosporella spot. It was entered into the Registry of Plant Varieties of Ukraine in
2003 [7]. The intact seeds of Umansky CS-97 hybrid were used.
Germinating energy and germination of studied hybrids were determined
according to DSTU [5].
For bacteria isolation the seeds were rinsed for 15 minutes with running and
sterile water and grounded in a mortar with 2 ml of saline solution. Than the small
amount of received suspension was plated on potato agar (PA) with thick strokes
from one edge of the Petri dish to another and cultivated at a 28 °C temperature.
For comparative studies the collection strains of sugar beet bacterial diseases
were used: Pseudomonas wieringae (Elliot) Savulesku 1947 strains 7923, 7921;
Pseudomonas syringae pv. aptata (Brown & Jamieson 1913) Yong, Dye & Wilkie
1978 strains 8544, 8545; Hanthomonas ahonopodis Starr & Garces 1950 strains 6,
10, 22, 7325, 8715, Rhizobium vitis (Smith & Towsend 1907) Conn 1942 strains
9052, 9054, 8628, stored in the collection of cultures of pathogenic bacteria in the
Zabolotny Institute of Microbiology and Virology NAS of Ukraine
Cultural, physiological, biochemical and morphological properties of the
isolated bacteria were determined using the classical methods [1, 9]. The formation of
pigments was determined visually under ultraviolet light for bacteria cultivated on
King B. medium. Bacteria were identified with the Bergey identification guide [6].
To determine the presence of pectinase enzymes the ability of isolates to
macerate potato pieces was studied. Selected isolates were also tested for their ability
to induce hypersensitivity reactions and antagonistic activity against the bacterial
diseases strains of sugar beet [1].
Pathogenic characteristics of isolates was determined by artificial infection of
sugar beet (hybrids Baccara, Olzhych) and beans (Mavka variety). For this the
suspension of bacteria (at concentration of 109 cells / ml of sterile tap water) was
applied to the surface of plant leaves, followed by their injury with needle [1].
Experiments repetition – 5–7 – fold.
Results and discussion. The observation survey of sugar beet crops in
Uladovo-Lyulenetskiy experimental breeding station (Vinnytsia region) conducted in
2012 had showed that during the growing season plants has symptoms of bacterial
leaf spot, bacterial gummosis and root canker. To determine the presence of
pathogenic bacteria on seeds and influence of seed protectants on microbiota
composition the bacteriological analysis of the samples was performed.
Seeds of both hybrids, taken for analysis, was visually healthy and were
characterized by the parameters given in Table 1. Seedlings from seed samples were
also healthy, without visible signs of damage.
18 different types of bacterial isolates were isolated from seeds samples, 12 of
them – from hybrid Umansky CS-97 and 6 – from hybrid Olexandria (Table 2). The
larger number of bacteria isolated from the untreated seeds of hybrid Umansky CS-97
indicate the greater diversity of its microbiota.
TABLE 1. Germinating rate and laboratory germination of sugar beet seeds
Hybrid Germinating energy
(day 4), %
Laboratory germination
(day 10), %
Umansky CS-97 68 93
Olexandria 70 90
TABLE 2. Results of bacteriological analysis of sugar beet seeds
Samples Number of bacterial isolates
Hybrid Treatment
with chemicals Total
With colonies
Crème/ orange
color
Grey/white
color
Umansky CS-
97
Without
treatment 12 11 1
Olexandria Maxim XL
035 FS 6 3 3
In total, the single isolate with grayish white colony and 11 isolates with
colonies of yellow color (from cream to orange) from untreated seeds of hybrid
Umansky CS-97 were selected. Three isolates of yellow and 3 isolates of grayish-
white colored colonies were selected from the seeds of hybrid Olexandria. The color
intensity of the grayish- white yellow isolates had depended a lot on the batch of
potato agar. All selected isolates were analyzed by their morphological and
biochemical properties (Table 3).
All identified isolates were Gram-negative, oxidase negative, and unable to
form spores. 61% of them were mobile. 83% of isolated bacteria had bacilli shape of
the cells, while 17% – small cocci. 44% of selected bacteria were facultative
anaerobes (B-27, B-28, B-29, B-31, B-35, B-46, B-47, B-39). The presence of
fluorescent pigment was observed in isolates B-30, B-36 and B-40.
As a carbon source isolates B-27, B-28, B-29, B-30, B-31, B-33, B-47, B-36,
B-38, B-39, B-40 in different extent can utilize rhamnose, sucrose, xylose, maltose,
arabinose, mannitol and sorbitol. Isolates B-41, B-45 and B -37 were able to utilize
only some of the sugars: B-41 and B-45 – arabinose, B-37 – xylose, maltose and
arabinose.
Seventeen percent of isolates were identified as the members of the
Pseudomonas genus (B-30, B-36, B-40) – aerobic mobile bacilli, capable to produce
a fluorescent pigment and cannot form spores.
Isolates B-27, B-28, B-35, B-46 and B-39 were identified as Pantoea
agglomerans species.
Isolates B-41 and B-45 were related to the methylotrophic bacteria. These
bacteria constantly inhabit the philosphere of agricultural, medicinal, ornamental and
wild plants [8].
The remaining eight isolates had represented saprophyte microflora, which
cannot be determined in our tests.
TABLE 3. Morphological and cultural-biochemical properties of bacteria isolated from the sugar beet seeds.
Tests
Bacteria, isolated from sugar beet seeds
Umansky CS-97 Olexandria
B-27 B-28 B-29 B-30 B-31 B-32 B-33 B-34 B-35 B-41 B-45 B-47 B-36 B-37 B-38 B-39 B-40 B-46
Gram staining − − − − − − − − − − − − − − − − − −
Mobility + + + + − − − − + + + − − + − + + +
Shape of cell B B C B B B B B B C B B B B C B B B
Spores – – – – – – – – – – – – – – – – – –
Fluorescent
pigment
− − − + − − − − − − − − + − − − + −
Oxidase − − − − − − − − − − − − – − − − − −
Carbon source:
Glucose
(anaerobic)
К К К – К − − − К − − К – − − К − К
Glucose
(aerobic)
К К К К К К − − К − − К К К − К К К
Rhamnosa К К К К К К К± − n/d − − К К± − К± К К n/d
Sucrose К К К К К К К − n/d − − К К± − К± К К± n/d
Xylose К К К К К К К − n/d − − К К К К К К n/d
Maltose К К К К К К К − n/d − − К К К± К К К n/d
Arabinose К К К К К К К − n/d К± К± К К К К К К n/d
Mannitol К К К К К К К − n/d − − К К − К± К К± n/d
Sorbitol К К К К К К К − n/d − − К К − К± − К± n/d
Colony color
ye
llo
w
ye
llo
w
gr
ay
ye
llo
w
ye
llo
w
ye
llo
w
ye
llo
w
ye
llo
w
ye
llo
w
pi
nk
pi
nk
ye
llo
w
gr
ay
gr
ay
gr
ay
ye
llo
w
cr
èm
e
ye
llo
w
Notes: +/- – presence or absence of the sign; K – acid production (change of medium color); K± – less intensive acid production; ± –
partial presence of sign;
B – bacilli; C – cocci; n/d – not determined
Virulence and antagonistic properties of the selected isolates are depicted in
Table 4.
Anticipating the presence of pectinase bacteria among the selected isolates that
cause soft rot, the ability of isolates to macerate potato pieces was studied. It was
established that none of isolated isolates were able to cause maceration of potato, thus
indicating the absence of rot pathogens of sugar beet among them (Table 4).
TABLE 4. Virulent and antagonistic properties of bacteria isolated from sugar beet
seeds
Isolates
Test
Hypersensitive
reaction
Potato
maceration Antagonism
Artificial infection
Sugar
beet Beans
B-27 n/d − − − −
B-28 n/d − − − −
B-29 n/d − − − −
B-30 + − − + +
B-31 n/d − − − −
B-32 n/d − +* − −
B-33 n/d − − − −
B-34 n/d − − − −
B-35 n/d − − − −
B-41 n/d − − − −
B-45 n/d − − − −
B-47 n/d − − − −
B-36 − − − − +
B-37 n/d − − − −
B-38 n/d − − − −
B-39 n/d − − − −
B-40 + − +** + +
B-46 n/d − − − −
Notes: +/- – presence or absence of the sign; +* – antagonism to Хanthomonas
aхоnopodis 6; +** – antagonisms Pseudomonas syringae pv. аptata 8545; n/d
– not determined
The ability of isolates to induce hypersensitivity reactions was examined in
bacteria identified as Pseudomonas sp. Isolates B-30 and B-40 initiated the formation
of brown necrosis on leaves of tobacco. The ability to induce hypersensitivity
reactions is associated with virulent bacterial properties. Isolate B-36 was not able to
induce the hypersensitivity reactions and obviously represents the saprophytes
microbiota.
At artificial infection of sugar beet plants, it was revealed that isolate B-30
causes appearance of the yellow stripes on the leaves and weak deformation of veins
in the injection site. Isolate B-40 had caused leaves spotting – aqueous during first 3
– 4 days, and later increasing in size and darken. The remaining isolates had not
reveal any pathogenic properties at artificial infection of sugar beet plants.
Artificially infected bean plants with isolates B-30, B-40 and B-36 had yellow stripes
on the leaves.
Thus, pathogenic for sugar beet bacteria were found on intact seeds of hybrid
Umansky CS-97, and on the seeds of Olexandria hybrid treated with fungicide
Maxim XL 035. That had confirmed our previous investigations which have shown
that Maxim fungicide has no effect on phytopathogenic bacteria – pathogens of sugar
beet [2].
The antagonistic activity of bacteria isolated from the seeds to known
pathogens of sugar beet was tested. The weak antagonistic activity was determined in
isolate B-40, in relation to Hanthomonas ahonopodis 6 (halo of growth inhibition –
18 mm), and isolate B-32, in relation to Pseudomonas syringae pv. aptata 8545 (halo
of growth inhibition – 30 mm). The remaining isolates had no antagonistic activity.
It was established that microbiota of untreated seeds of sugar beet is more
diverse than on seeds, treated with fungicide. However, sugar beet pathogenic
bacteria (isolates B-30 and B-40) were isolated from the both samples. This indicates
that the fungicidal disinfectant Maxim XL 035 FS used for pre-sowing seeds
treatment was unable to protect sugar beet seeds from bacterial pathogens.
|
| id | oai:ojs2.smic.in.ua:article-198 |
| institution | Agriciltural microbiology |
| keywords_txt_mv | keywords |
| language | English Ukrainian |
| last_indexed | 2026-07-23T01:06:29Z |
| publishDate | 2013 |
| publisher | Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine |
| record_format | ojs |
| resource_txt_mv | smicinua/a1/a1bf820b56f29b2d7e8bdbc3d6c4f2a1.pdf |
| spelling | oai:ojs2.smic.in.ua:article-1982026-07-22T10:10:44Z BACTERIAL MICROFLORA OF SUGAR BEET SEEDS БАКТЕРІАЛЬНА МІКРОБІОТА НАСІННЯ НАСІННЯ ЦУКРОВИХ БУРЯКІВ Дворак , К.П. Буценко , Л.М. sugar beet, seeds, bacterial microflora, antagonism, artificial infection цукрові буряки, насіння, мікробіота, антагонізм, штучне зараження The paper shows the results of bacterial microflora of sugar beet seeds study, with the analysis of morphological, culture-biochemical and virulent characteristics of isolated bacteria. The presence of pathogenic bacteria was observed on seeds pretreated with fungicide. The antagonistic activity of bacterial microflora of sugar beet seeds in relation to known bacterial diseases agents of this culture was investigated. Проаналізовано бактеріальну мікробіоту насіння цукрових буряків, вивчено морфологічні, культурально–біохімічні властивості виділених ізолятів. Показано, що на протруєному фунгіцидом насінні зберігаються патогенні бактерії. Вивчено антагоністичну активність мікробіоти насіння цукрових буряків щодо відомих збудників бактеріальних хвороб цієї культури. Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine 2013-10-01 Article Article Рецензована Стаття application/pdf application/pdf https://smic.in.ua/index.php/journal/article/view/198 10.35868/1997-3004.17.48-57 Agricultural microbiology; Vol. 17 (2013): Agriciltural microbiology; 48-57 Сільськогосподарська мікробіологія; Том 17 (2013): Сільськогосподарська мікробіологія; 48-57 1997-3004 10.35868/1997-3004.17 en uk https://smic.in.ua/index.php/journal/article/view/198/277 https://smic.in.ua/index.php/journal/article/view/198/278 Авторське право (c) 2013 K. P. Dvorak , L. M. Butsenko https://creativecommons.org/licenses/by/4.0 |
| spellingShingle | цукрові буряки насіння мікробіота антагонізм штучне зараження Дворак , К.П. Буценко , Л.М. БАКТЕРІАЛЬНА МІКРОБІОТА НАСІННЯ НАСІННЯ ЦУКРОВИХ БУРЯКІВ |
| title | БАКТЕРІАЛЬНА МІКРОБІОТА НАСІННЯ НАСІННЯ ЦУКРОВИХ БУРЯКІВ |
| title_alt | BACTERIAL MICROFLORA OF SUGAR BEET SEEDS |
| title_full | БАКТЕРІАЛЬНА МІКРОБІОТА НАСІННЯ НАСІННЯ ЦУКРОВИХ БУРЯКІВ |
| title_fullStr | БАКТЕРІАЛЬНА МІКРОБІОТА НАСІННЯ НАСІННЯ ЦУКРОВИХ БУРЯКІВ |
| title_full_unstemmed | БАКТЕРІАЛЬНА МІКРОБІОТА НАСІННЯ НАСІННЯ ЦУКРОВИХ БУРЯКІВ |
| title_short | БАКТЕРІАЛЬНА МІКРОБІОТА НАСІННЯ НАСІННЯ ЦУКРОВИХ БУРЯКІВ |
| title_sort | бактеріальна мікробіота насіння насіння цукрових буряків |
| topic | цукрові буряки насіння мікробіота антагонізм штучне зараження |
| topic_facet | sugar beet seeds bacterial microflora antagonism artificial infection цукрові буряки насіння мікробіота антагонізм штучне зараження |
| url | https://smic.in.ua/index.php/journal/article/view/198 |
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