ЛЕКТИНОВА АКТИВНІСТь у РІЗНИх ОРгАНАх ЛюЦЕРНИ ПРИ ФОРМуВАННІ СИМбІОЗу ІЗ Sinorhizobium meliloti ЗА РІЗНОгО ВОДОЗАбЕЗПЕчЕННЯ
The paper covers study of insufficient water supply effect on lectin activity in different organs of alfalfa plants inoculated with Sinorhizobium meliloti 441 (active strain) and SHM1-48 (inactive strain) during the early stages of symbiosis formation and its active functioning. The correlations bet...
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| Datum: | 2013 |
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Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine
2013
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Agriciltural microbiology| _version_ | 1871465702663127040 |
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| author | Михалків , Л.М. |
| author_facet | Михалків , Л.М. |
| author_institution_txt_mv | [
{
"author": "Л.М. Михалків ",
"institution": "Інститут фізіології рослин і генетики НАН України"
}
] |
| author_sort | Михалків , Л.М. |
| baseUrl_str | https://smic.in.ua/index.php/journal/oai |
| collection | OJS |
| datestamp_date | 2026-07-22T10:10:44Z |
| description | The paper covers study of insufficient water supply effect on lectin activity in different organs of alfalfa plants inoculated with Sinorhizobium meliloti 441 (active strain) and SHM1-48 (inactive strain) during the early stages of symbiosis formation and its active functioning. The correlations between the changes of lectin activity in alfalfa plants influenced by different water supply and strain characteristics, extent of plant development and stage of symbiosis formation. |
| doi_str_mv | 10.35868/1997-3004.17.21-30 |
| first_indexed | 2025-07-17T12:24:24Z |
| format | Article |
| fulltext |
LECTIN ACTIVITY IN PLANT ORGANS AT SYMBIOSIS FORMATION of
ALFALFA WITH SINORHIZOBIUM MELILOTI UNDER DIFFERENT
WATER CONDITIONS
Mykhalkiv L.M.
Institute of Plant Physiology and Genetics NAS of Ukraine,
31/17, Vasylkivska Str., Kyiv, 03022
E-mail: mykhalkiv@mail.ru
The paper covers study of insufficient water supply effect on lectin activity in
different organs of alfalfa plants inoculated with Sinorhizobium meliloti 441 (active
strain) and SHM1-48 (inactive strain) during the early stages of symbiosis formation
and its active functioning. The correlations between the changes of lectin activity in
alfalfa plants influenced by different water supply and strain characteristics, extent of
plant development and stage of symbiosis formation.
Keywords: lectin activity, alfalfaa, Sinorhizobium meliloti, water supply.
Application of scientific research findings on biological nitrogen fixation into
the modern agricultural practices can reduce the cost of energy and raw materials for
the production of mineral nitrogen fertilizers and provide an ecologically clean
production. At introduction of nitrogen-fixing plant-microbial systems into the
agrocoenoses the impact of environmental factors on their formation, functioning and
productivity should be taken into the account. Therefore, studies of resistance
mechanisms of symbiotic systems to external stress as well as search for the
adaptation means to harsh environmental conditions remain important. In this view,
the role of plant lectins is little-studied, but is promising.
External factors of abiotic nature (drought, salinity, temperature fluctuations,
heavy metals, etc.) can cause greater genes expression that are responsible for lectins
synthesis, increasing the level of these proteins in plants and changing their properties
[1-4]. Obviously, the accumulation of lectins is a general plant response to the
different types of stress, however, the ways of their involvement to the adaptation
processes remain unclear. O. Timofeeva et al. [5] have proved the involvement of cell
wall lectins into the non-specific defense mechanisms of plants. It was shown that
during the initial stage of formation of non-specific defense response in plants a rapid
changes of lectins activity and composition in the cell wall take place, which depends
on the functioning of calcium signaling system and may be associated with the role of
lectins in signal transduction into the cell or in apoplast space. A direct relationship
between the activity of cell wall lectin and the extend of winter wheat resistance to
biotic and abiotic stressors was established, together with genetically determined and
organ-specific correlation between the level of cell lectins activity and structural
condition of cytoskeleton. Pattern for Ca2+ signaling pathway and cytoskeleton
structures role in the regulating mechanism of lectins activity in the formation of
adaptive responses of plants was suggested. Findings on possible cryoprotection
effect of galactospesific lectins on isolated thylakoid membranes of chloroplasts are
also of a big interest [6]. Due to the relative hydrophobicity, these lectins can bind to
membrane glycolipids, strengthen thylakoid membrane structures and reduce their
fluidity caused by damage during freezing. It was also shown [7] that induction of
nonspecific resistance of legumes may be ensured by regulation of protective
inhibition stage in general adaptation syndrome due to the activation of protein
proteinase inhibitors and lectin-type proteins, which leads to a decrease of stressors
toxic effect on nitrogen fixing system and plant productivity. Soluble lectins can
function as elicitors [8], be involved in the regulation of gene expression in plants
under stress by modulating O-GlcNAc-dependent transduction signals through
specific protein-carbohydrate interactions with regulatory cytoplasmic and nuclear
glycoproteins [9].
Taking into the account lectins role as an important factor of efficient
symbiosis [10], as well as component of nonspecific plant response to stressors [11]
the determination of lectins activity in alfalfa plants influenced by rhizobia
inoculation of various activity under the optimal and insufficient water supply
conditions was performed.
Materials and methods. The studies were conducted in greenhouse conditions
at the Institute of Plant Physiology and Genetics, NAS of Ukraine. Alfalfa plants of
Nadezhda cultivar were grown in sand culture (10 plants / pot) in a 4 kg Wagner
containers (washed river sand, moisture content – 60% TMC) at natural lighting,
temperature and humidity. Mineral nutrition supply was prepared according to
Hellriegel with 0.25 dose of nitrogen and addition of trace elements B, Mo and Fe.
Prior to sowing seeds were sterilized with concentrated sulfuric acid, washed in
running tap water and inoculated with two strains of Sinorhizobium meliloti nodule
bacteria – 441 (active) and СХМ1-48 (inactive). Bacterial cultures were grown on
solid mannitol-yeast medium at a temperature of 28 °C for 4 days. Duration of seeds
inoculation – 1 hour, titer of inoculation suspension – 109 cells / ml.
On the 7th (experiment #1, trifoliate leaf emergence) or 20th day (experiment
#2, shooting stage) the appropriate plants were exposed to drought conditions (30%
TMC) by controlled irrigation. Analysis samples were taken: in experiment #1 – after
3 and 10 days after exposure to drought conditions, while in experiment #2 – on 3, 7
and 11 day of drought.
Nitrogen fixation activity of root nodules was determined with acetylene
reduction method [12]on gas chromatograph “Agilent 6850”. Biological repetition –
5 times. Vegetative plants mass (g dry matter / plant) was measured with 10 – 15
biological repetition.
Phytohemagglutins (lectins) were isolated from roots, stems and leaves of
alfalfa plants using ethanol elution method [13], with minor modification. The plant
material was grinded in a porcelain mortar with the addition of 0.5 M phosphate
buffer (pH 5.0), used as extraction solution. Extraction was carried out for 20 hours in
a cool dark place. After that, mixture was centrifuged at 3000 rev / min for 10 min.
Supernatant was decanted and mixed with 96%-s' ethyl alcohol so that the output
alcohol content in the mixture was 76%. Received solution was centrifuged at 6000
rev / min for 15 min. Supernatant was poured out while the precipitate of protein was
dissolved in saline. Determination of lectin activity in the received protein solution
was performed by hemagglutination reactions in immunological plates with U-shaped
holes [14]. Lectin activity was determined after samples incubation with human red
blood cells a solution during 2 hours at room temperature corresponding to the
biggest dilution, where hemagglutination was observed.
The results of experiments were statistically analyzed using standard method
[15]. Figure presents the arithmetic means and their standard errors.
Results and discussion. On the third day of plants cultivation under different
water conditions (10-day plant) neither nodules on the roots of alfalfa plants or
molecular nitrogen fixation activity were observed, although, as is well known [16],
that this period of plant growth is characterized with an active internal changes in the
structure and functioning of root caused by bacterial infection. Study of lectin activity
in alfalfa plants inoculated with active strain 441 have shown the inhibition of
erythrocyte sedimentation in the variants with insufficient water supply, while no
changes of lectin activity in the plants inoculated with inactive strains were observed.
No differences were revealed in plants inoculated with different strains (441 and
CXM1-48) under optimal water supply.
After 10 days of plants growing under different water conditions the nodules
formation was observed on the roots of alfalfa plants in variants with 60% TMC. At
this, inoculation with S. meliloti 441 in some plants resulted in formation of
botryoidal nodules. It should be noted that S. meliloti strain CXM1-48 have formed
about 2 – 2.5 times more nodules on the plant roots than the active strain 441.
Evaluation of nitrogen fixation activity revealed minor ethylene peaks in the
variant inoculated with S. meliloti 441 on optimal water supply background. As a
result, of drought the significant reduction of dry plants matter was observed during
this period, while no significant differences were noticed between the plants
inoculated with different strains, under the same water conditions (Figure).
FIGURE. Above-ground mass (g dry matter / plant) of 17-day old alfalfa plants
inoculated with different strains of Sinorhizobium meliloti on the background of
optimal and low (10-day drought) water conditions (experiment #1): 1 – inoculation
with S. meliloti CXM1-48, 30% TMC; 2 – S. meliloti inoculation with CXM1-48,
60% TMC; 3 – inoculation with S. meliloti 441, 30% TMC; 4 – inoculation with S.
meliloti 441, 60% TMC.
Estimation results of lectin activity in the roots and vegetative parts of the
plants during this period are presented in Table 1. It was established that neither
different levels of moisture nor nitrogen fixation activity of symbiosis have affected
this index in plant roots. However, in vegetative mass the increase of lectin activity in
variants with SHM1-48 strain application on both levels of water supply compared to
the inoculation with 441 strain was observed.
TABLE 1. Lectin activity (OA/50 ml) in 17-day old alfalfa plants at different levels
of water supply (10-day drought) and inoculation with Sinorhizobium meliloti
(experiment 1)
Variants Root Aboveground parts
S.meliloti СХМ1-48, 30%
TMC 2 4
S.meliloti СХМ1-48, 60%
TMC 2 4
S.meliloti 441, 30% TMC 2 2
S.meliloti 441, 60% TMC 2 2
Taking into the account that lectin activity is directly related to the nitrogen
fixation activity [17] the opposite effect of inoculation with studied strains would be
expected. Obviously, in this case, the influence of another factor, like, for example,
the nodulation activity of the strain, which in S. meliloti SHM1-48 is higher than that
of S. meliloti 441, or the culture characteristics or physiological peculiarities of the
early stages of symbiosis formation have played the determining role.
Studying the impact of moisture deficit on the lectin activity in alfalfa plants at
later stages of ontogenesis (early drought – shooting stage, 20-day old plants) the
decrease of lectin activity in leaves of plants at 3rd, 7th and 11th days of drought were
observed on the background of inoculation with inactive strain SHM1-48 (Table 2).
AS for the inoculation with 441 strain the lectin activity in leaves of plants with low
water supply have increased after 3 days of drought, if compared to 60% TMC, while
after 11 days, on the contrary – have decreased.
Regardless of inoculant strain, the increase of lectin activity in plant roots on
water deficit background was observed.
In plant stems the influence of 3-day water deficit was noticed in variants with
both strains – lectin activity was higher in 2 times. At 7th day of drought it was
established that in plants inoculated with inactive strain the lectin activity in stems
was higher, regardless of the water conditions. But, after 11th days the differences
between all studied variants were leveled. Thus, it was established that the lectin
activity in leaves and plant stems at drought conditions varies differently depending
on the activity of inoculant strain.
Hence, performed studies have revealed that different water conditions affect
lectin activity in different organs of alfalfa, which depend on the properties of
inoculant strain, level of plant development and symbiosis formation.
TABLE 2. Lectin activity (OA/50 ml) in different organs of alfalfa under the
different water supply conditions and inoculation with Sinorhizobium meliloti
(experiment 2)
Variants
Plant development stages
Shooting,
3rd day of
draught
Shooting,
7th day of
draught
Beginning of
budding,
11th day of
draught
Leaves
S.meliloti СХМ1-48, 30%
TMC 8 2 2
S.meliloti СХМ1-48, 60%
TMC 16 4 4
S.meliloti 441, 30% TMC 16 2 2
S.meliloti 441, 60% TMC 8 2 4
Roots
S.meliloti СХМ1-48, 30%
TMC 2 4 2
S.meliloti СХМ1-48, 60%
TMC 2 2 0
S.meliloti 441, 30% TMC 2 4 2
S.meliloti 441, 60% TMC 2 2 0
Stems
S.meliloti СХМ1-48, 30%
TMC 2 2 2
S.meliloti СХМ1-48, 60%
TMC 0 2 2
S.meliloti 441, 30% TMC 2 0 2
S.meliloti 441, 60% TMC 0 0 2
In the initial period of symbiosis formation (10-day old plant), the increase of
lectin activity due to the insufficient water supply in alfalfa plants inoculated with
active strain 441 was observed, whereas at plant inoculation with inactive strain
SHM1-48 at remained unchanged. With start of biological nitrogen fixation (17-day
old plants), the lectin activity in vegetative parts of alfalfa plants inoculated with
inactive strain was higher than in variants with active rhizobia inoculation, remaining
unchanged in plant roots. At this, no differences in variants with different water
supply were noticed.
As the result of influence of water deficit on formed symbiosis (shooting –
beginning of budding stages), the changes of lectin activity in leaves and stems of
alfalfa plants were determined by the activity of the inoculant strain, while no
differences were observed in the roots under the use of both strains.
|
| id | oai:ojs2.smic.in.ua:article-195 |
| institution | Agriciltural microbiology |
| keywords_txt_mv | keywords |
| language | English Ukrainian |
| last_indexed | 2026-07-23T01:06:14Z |
| publishDate | 2013 |
| publisher | Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine |
| record_format | ojs |
| resource_txt_mv | smicinua/08/04bed2dea75b8a9fe32cf346b3752208.pdf |
| spelling | oai:ojs2.smic.in.ua:article-1952026-07-22T10:10:44Z LECTIN ACTIVITY IN PLANT ORGANS AT SYMBIOSIS FORMATION of ALFALFA WITH SINORHIZOBIUM MELILOTI UNDER DIFFERENT WATER CONDITIONS ЛЕКТИНОВА АКТИВНІСТь у РІЗНИх ОРгАНАх ЛюЦЕРНИ ПРИ ФОРМуВАННІ СИМбІОЗу ІЗ Sinorhizobium meliloti ЗА РІЗНОгО ВОДОЗАбЕЗПЕчЕННЯ Михалків , Л.М. lectin activity, alfalfaa, Sinorhizobium meliloti, water supply лектинова активність, люцерна, Sinorhizobium meliloti, водозабезпечення The paper covers study of insufficient water supply effect on lectin activity in different organs of alfalfa plants inoculated with Sinorhizobium meliloti 441 (active strain) and SHM1-48 (inactive strain) during the early stages of symbiosis formation and its active functioning. The correlations between the changes of lectin activity in alfalfa plants influenced by different water supply and strain characteristics, extent of plant development and stage of symbiosis formation. Досліджено вплив недостатнього водозабезпечення на лектинову активність у різних органах люцерни, інокульованої штамами Sinorhizobium meliloti 441 (активний) та СХМ1-48 (неактивний), на початкових етапах формування симбіозу та у період його активного функціонування. Показано залежність змін лектинової активності у рослинах люцерни, які відбуваються під впливом різного водозабезпечення, від властивостей штамуінокулянта, ступеня розвитку рослин та сформованості симбіозу. 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/195 10.35868/1997-3004.17.21-30 Agricultural microbiology; Vol. 17 (2013): Agriciltural microbiology; 21-30 Сільськогосподарська мікробіологія; Том 17 (2013): Сільськогосподарська мікробіологія; 21-30 1997-3004 10.35868/1997-3004.17 en uk https://smic.in.ua/index.php/journal/article/view/195/271 https://smic.in.ua/index.php/journal/article/view/195/272 Авторське право (c) 2013 L.M. Mykhalkiv https://creativecommons.org/licenses/by/4.0 |
| spellingShingle | лектинова активність люцерна Sinorhizobium meliloti водозабезпечення Михалків , Л.М. ЛЕКТИНОВА АКТИВНІСТь у РІЗНИх ОРгАНАх ЛюЦЕРНИ ПРИ ФОРМуВАННІ СИМбІОЗу ІЗ Sinorhizobium meliloti ЗА РІЗНОгО ВОДОЗАбЕЗПЕчЕННЯ |
| title | ЛЕКТИНОВА АКТИВНІСТь у РІЗНИх ОРгАНАх ЛюЦЕРНИ ПРИ ФОРМуВАННІ СИМбІОЗу ІЗ Sinorhizobium meliloti ЗА РІЗНОгО ВОДОЗАбЕЗПЕчЕННЯ |
| title_alt | LECTIN ACTIVITY IN PLANT ORGANS AT SYMBIOSIS FORMATION of ALFALFA WITH SINORHIZOBIUM MELILOTI UNDER DIFFERENT WATER CONDITIONS |
| title_full | ЛЕКТИНОВА АКТИВНІСТь у РІЗНИх ОРгАНАх ЛюЦЕРНИ ПРИ ФОРМуВАННІ СИМбІОЗу ІЗ Sinorhizobium meliloti ЗА РІЗНОгО ВОДОЗАбЕЗПЕчЕННЯ |
| title_fullStr | ЛЕКТИНОВА АКТИВНІСТь у РІЗНИх ОРгАНАх ЛюЦЕРНИ ПРИ ФОРМуВАННІ СИМбІОЗу ІЗ Sinorhizobium meliloti ЗА РІЗНОгО ВОДОЗАбЕЗПЕчЕННЯ |
| title_full_unstemmed | ЛЕКТИНОВА АКТИВНІСТь у РІЗНИх ОРгАНАх ЛюЦЕРНИ ПРИ ФОРМуВАННІ СИМбІОЗу ІЗ Sinorhizobium meliloti ЗА РІЗНОгО ВОДОЗАбЕЗПЕчЕННЯ |
| title_short | ЛЕКТИНОВА АКТИВНІСТь у РІЗНИх ОРгАНАх ЛюЦЕРНИ ПРИ ФОРМуВАННІ СИМбІОЗу ІЗ Sinorhizobium meliloti ЗА РІЗНОгО ВОДОЗАбЕЗПЕчЕННЯ |
| title_sort | лектинова активність у різних органах люцерни при формуванні симбіозу із sinorhizobium meliloti за різного водозабезпечення |
| topic | лектинова активність люцерна Sinorhizobium meliloti водозабезпечення |
| topic_facet | lectin activity alfalfaa Sinorhizobium meliloti water supply лектинова активність люцерна Sinorhizobium meliloti водозабезпечення |
| url | https://smic.in.ua/index.php/journal/article/view/195 |
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