Screening and peculiarity of the biological action of synthetic plant growth regulators
As a result of biological screening among synthetic compounds that have growth regulatory activities in culture tissues in vitro and in vivo in plants, 5 perspective substances were chosen as the substitutes of phytohormones. Thus, it was found that Ivin (N-oxide 2,6-dimethylpyridine) revealed predo...
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Tsygankova, V.A. Blume, Y.B. 2019-06-17T10:24:23Z 2019-06-17T10:24:23Z 1997 Screening and peculiarity of the biological action of synthetic plant growth regulators / V.A. Tsygankova, Y.B. Blume // Биополимеры и клетка. — 1997. — Т. 13, № 6. — С. 484-492. — Бібліогр.: 18 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.0004AB https://nasplib.isofts.kiev.ua/handle/123456789/155686 As a result of biological screening among synthetic compounds that have growth regulatory activities in culture tissues in vitro and in vivo in plants, 5 perspective substances were chosen as the substitutes of phytohormones. Thus, it was found that Ivin (N-oxide 2,6-dimethylpyridine) revealed predominant cytokinin activity in vitro and a cytokinin-gibberelin activity in vivo; Methyur (Na-derivative 6-methylthyouracyl) showed an expressive cytokinin-gibberellin activity in vivo; Triamelon (Iodide tris (2,2-trimethylammoniummethyl phosphate) revealed a cytokinin-anxin activity in vitro; preparation D-107 (l-acetyl-amino-1-acetylthyo-2-oxo-2-phenylethan) revealed an auxine-like effect and the preparation N 2622 (derivative tetrahydrothyophendioxide) showed an expressive auxin activity in vivo. The model of mediated action of plant growth regulators through phytohormones is proposed. У процесі біологічного скринінгу синтетичних препаратів, які виявляють ріст-регулюючу активність у культурах тканин in vitro і на рослинах in vivo, відібрано п'ять препаратів як перспективних замінників фітогормонів: Івін (N-oxide-2,6- dimethy[pyridine) з переважною цитокініновою активністю in vitro і цитокінін-гібереліновою активністю in vivo; Метіур (Na-derivative 6-methylthyouracyl) з вираженою цитокінін-гібереліновою активністю in vivo; Тріамелон (iodide tris (2,2-trimethylammonium,methyl phosphate)) – з цитокінін-ауксиновою активністю in vitro; D-107 (1 -acetylamino-1-acetylthyo-2-oxo-2-phenylethan) з ауксиноподібним ефектом та № 2622 (derivative tetrahydrothyophendoixide) з яскраво вираженою ауксиновою активністю in vivo. Запропоновано модель опосередкованої дії регуляторів росту рослин через фітогормони. В процессе биологического скрининга синтетических препаратов с рост-регуліірующей активностью в культурах тканей in vitro и на растениях in vivo были отобраны пять препаратов как перспективные заменители фитогормонов: Ивин (N-oxide-2,6-dimethylpyridine) с преобладающей цитокининовой активностью in vitro и цитокинин-гиббереллиновой in vivo; Метиур (Na-derivative 6-methylthyouracyl) с выраженной цитокинин-гиббереллиновой активностью in vivo; Триамелон (iodide tris (2,2-trimethylammoniummethyl phospate) – с ицтокинин-ауксиновой активностью in vitro; D-107 (1-acetylamina- 1-acetylthyo-2-oxo-2-phenylethan) с ауксиноподобным еффектом и № 2622 (derivative tetrahydrothyophendioxide) с ярко выраженной ауксиновой активностью in vivo. Предложена модель опосредованного действия регуляторов роста растений через фитогормоны. en Інститут молекулярної біології і генетики НАН України Биополимеры и клетка Клеточная биология Screening and peculiarity of the biological action of synthetic plant growth regulators Скринінг та особливості біологічної дії синтетичних регуляторів росту рослин Скрининг и особенности биологического действия синтетических регуляторов роста растений Article published earlier |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine |
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DSpace DC |
| title |
Screening and peculiarity of the biological action of synthetic plant growth regulators |
| spellingShingle |
Screening and peculiarity of the biological action of synthetic plant growth regulators Tsygankova, V.A. Blume, Y.B. Клеточная биология |
| title_short |
Screening and peculiarity of the biological action of synthetic plant growth regulators |
| title_full |
Screening and peculiarity of the biological action of synthetic plant growth regulators |
| title_fullStr |
Screening and peculiarity of the biological action of synthetic plant growth regulators |
| title_full_unstemmed |
Screening and peculiarity of the biological action of synthetic plant growth regulators |
| title_sort |
screening and peculiarity of the biological action of synthetic plant growth regulators |
| author |
Tsygankova, V.A. Blume, Y.B. |
| author_facet |
Tsygankova, V.A. Blume, Y.B. |
| topic |
Клеточная биология |
| topic_facet |
Клеточная биология |
| publishDate |
1997 |
| language |
English |
| container_title |
Биополимеры и клетка |
| publisher |
Інститут молекулярної біології і генетики НАН України |
| format |
Article |
| title_alt |
Скринінг та особливості біологічної дії синтетичних регуляторів росту рослин Скрининг и особенности биологического действия синтетических регуляторов роста растений |
| description |
As a result of biological screening among synthetic compounds that have growth regulatory activities in culture tissues in vitro and in vivo in plants, 5 perspective substances were chosen as the substitutes of phytohormones. Thus, it was found that Ivin (N-oxide 2,6-dimethylpyridine) revealed predominant cytokinin activity in vitro and a cytokinin-gibberelin activity in vivo; Methyur (Na-derivative 6-methylthyouracyl) showed an expressive cytokinin-gibberellin activity in vivo; Triamelon (Iodide tris (2,2-trimethylammoniummethyl phosphate) revealed a cytokinin-anxin activity in vitro; preparation D-107 (l-acetyl-amino-1-acetylthyo-2-oxo-2-phenylethan) revealed an auxine-like effect and the preparation N 2622 (derivative tetrahydrothyophendioxide) showed an expressive auxin activity in vivo. The model of mediated action of plant growth regulators through phytohormones is proposed.
У процесі біологічного скринінгу синтетичних препаратів, які виявляють ріст-регулюючу активність у культурах тканин in vitro і на рослинах in vivo, відібрано п'ять препаратів як перспективних замінників фітогормонів: Івін (N-oxide-2,6- dimethy[pyridine) з переважною цитокініновою активністю in vitro і цитокінін-гібереліновою активністю in vivo; Метіур (Na-derivative 6-methylthyouracyl) з вираженою цитокінін-гібереліновою активністю in vivo; Тріамелон (iodide tris (2,2-trimethylammonium,methyl phosphate)) – з цитокінін-ауксиновою активністю in vitro; D-107 (1 -acetylamino-1-acetylthyo-2-oxo-2-phenylethan) з ауксиноподібним ефектом та № 2622 (derivative tetrahydrothyophendoixide) з яскраво вираженою ауксиновою активністю in vivo. Запропоновано модель опосередкованої дії регуляторів росту рослин через фітогормони.
В процессе биологического скрининга синтетических препаратов с рост-регуліірующей активностью в культурах тканей in vitro и на растениях in vivo были отобраны пять препаратов как перспективные заменители фитогормонов: Ивин (N-oxide-2,6-dimethylpyridine) с преобладающей цитокининовой активностью in vitro и цитокинин-гиббереллиновой in vivo; Метиур (Na-derivative 6-methylthyouracyl) с выраженной цитокинин-гиббереллиновой активностью in vivo; Триамелон (iodide tris (2,2-trimethylammoniummethyl phospate) – с ицтокинин-ауксиновой активностью in vitro; D-107 (1-acetylamina- 1-acetylthyo-2-oxo-2-phenylethan) с ауксиноподобным еффектом и № 2622 (derivative tetrahydrothyophendioxide) с ярко выраженной ауксиновой активностью in vivo. Предложена модель опосредованного действия регуляторов роста растений через фитогормоны.
|
| issn |
0233-7657 |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/155686 |
| citation_txt |
Screening and peculiarity of the biological action of synthetic plant growth regulators / V.A. Tsygankova, Y.B. Blume // Биополимеры и клетка. — 1997. — Т. 13, № 6. — С. 484-492. — Бібліогр.: 18 назв. — англ. |
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I S S N 0233-7657. Биополимеры и клетка. 1997. Т. 13. № 6
Screening and peculiarity of the biological action
of synthetic plant growth regulators
V. A- Tsygankova, Ya. B. Blume
Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine
148 Zabolotnogo str., 252143 , Kiev
As a result of biological screening among synthetic compounds that have growth regulatory activities in
culture tissues in vitro and in vivo in plants, 5 perspective substances were chosen as the substitutes of
phytohormones. Thus, it was found that Ivin (N-oxide 2,6-diniethylpyridine) revealed predominant
cytokinin activity in vitro and a cytokinin-gibberelin activity in vivo; Methyur (Na-derivative 6-
methylthyouracyl) showed an expressive cytokinin-gibberellin activity in vivo; Triamelon (Iodide tris
(2,2-trimethylammoniummethyl phosphate) revealed a cytokinin-auxin activity in vitro; preparation D-107
(I-acetylamino-1-acetylthyo-2-oxo-2-phenylethan) revealed an auxine-like effect and the preparation N
2622 (derivative tetrahydrothyophendioxide) showed an expressive auxin activity in vivo. The model of
mediated action of plant growth regulators through phytohormones is proposed.
Introduction. The most important problem of cell
biology and biotechnology is the substitution of ex
pensive natural plant growth regulators (phytohor
mones) by synthetic compounds [1—6]. The world
wide experiments in substitution of natural phyto
hormones with artificial preparations (2,4-D, NAA,
NOAA, BAP) confirm a perspective this approach.
At the long time in the Institute of Organic
Chemistry and Institute of Bioorganic and Petroleum
Chemistry of National Academy of Sciences of Ukra
ine the synthesis and screening of plant growth
regulators are the significant direction of scientific
investigations.
For the biological screening of substances with
growth-regulative activity synthezing in these Ins
titutes we proposed to use the following test-systems:
1) cultures of isolated plant tissues of family Sola-
naceae (tobacco, tomato and potato) for which con
ditions of induction callus tissue formation in vitro
from differentiated cells, organogenesis and rege
neration of plants from cells of callus tissues were
determinated in detail (i. e. compositions of nutrient
media and which from phytohormones or its com
bination produce each of these processes) [7—9 ]; 2)
embryo seeds and leaf grafts of vegetating plants of
© V. A. TSYGANK.0 VA, YA. В BLUME, 1997
haricot that were used for a determination of phy
tohormones activities in vivo tests [10, 11 ].
Using above mentioned models at least 5 com
pounds Ivin (N-oxide 2,6-dimethylpyridine), Me
thyur (Na-derivative 6-methylthyouracyl), substance
Triamelon (N 2863) (Iodide tris (2,2-trimethylam-
moniurnmethyI phosphate)), D-107 (1 -Acetylamino-
l - a c e t y l t h y o - 2 - o x o - 2 - p h e n y l e t h a n ) , substance
N 2622 (derivative tetrahydrothyophendioxide) is un
der patent procedure, have been chosen as perspective
substitutions of some phytohormones for the practical
purposes.
Materials and Methods. Chemical structures of
chosen compounds are presented in the scheme.
Plant material. There are tobacco Nicotiana ta-
bacum (varieties R- l ; SR-2, visconsinia), potato So-
lanum tuberosum (varieties Zarevo and Nevskiy) and
2 types of tomato: cultural species (Lycopersicon
esculentum) of mutant line MO 393 and wild Peruvian
tomato (L. peruvianum var. dentatum 3767) were
used in test in vitro.
Before introducing in the culture the plant tissues
(leaf or stem tissues) were sterilized with diocid (or
hypochloride) and 70 % ethanol and then washed
with sterile distilled water.
For in vivo experiments seeds of asparagus ha
ricot of Belozernaya (Phaseolus vulgaris L.) were
used.
Seeds were germinated in humid atmosphere in
484
SCREENING OF T H E BIOLOGICAL A C T I O N G R O W T H REGULATORS
special trays in the thermostat at 25—26 °С during
2—3 days, and then seedlings were placed in the light
box.
The vegetation lasted a week at 4000—6000
luxes, 22—24 °С and 16 hours' light day. An auxin
activity was determined by Turetskaya method [10].
Leaf grafts of 12—14 day plants of haricot were
placed in aqueous solutions of investigated substances.
Every time 2 parallel experiments each with 5 grafts
were made.
Cultural media for plant tissues. Modified nut
rient media (RMKU; RMOP; NP; RMB; RMO), on
the basis of elaborated by Murashige and Skoog
(MS); Gamborg et al. (B5) and medium of Linsmaier,
Skoog (LS) containing similar to MS macro- and
microelements compositions were used. Names and
compositions of the media were taken from ref. [9].
To prepare the nutrient media the initial concentrated
solutions were used: 1) 10 times solution of macro-
salts; 2) 100 times solution of microsalts (besides salt
solutions of CoCl 2 x 6 H 2 0 and CuS0 4 x 5 H 2 0 were
prepared separately by a consistent dissolving of
25 mg of each salt in 10 ml of water. Then they were
united with solution of other microsalts. General
volume was increased to 100 ml); 3) an initial solution
of Fe-helat was prepared by a consistent dissolving
7.45 g of Na 2 EDTA and 5.57 g of FeSO, x 7H 2 0 in
water (the final volume was led to 1 1 and heated to
100 °С). To prepare 1 1 of an agar nutrient medium
we mixed 100 ml of the initial solution of macrosalts,
10 ml of microsalts, 5 ml of Fe-helat and cor
responding quantities of agar, vitamins, hormones,
hydrocarbons, etc.
Vitamin solutions were prepared just before expe
riments. Auxins were dissolved initially in a small
quantity of ethanol and brought to the necessary
volume with water under heating; cytokinin solutions
were prepared by a dissolving of a corresponding
cytokinin in a small volume of 0.5 N HC1 under
heating with the following mixing with necessary
amount of water.
Agar nutrient media (6—8 g of agar/1) brought
to certain pH were autoclaved. Isolation and cul
tivation of mesophyll protoplasts from leaf discs of
tobacco were realised by methods in detail described
in protocol part of the book [9].
Fig. 1. Undifferentiated growing tobacco tissue (callus) received on the RMKV medium containing 0.1 — 0.5 mg/1 of Triamelon
485
TSYGANKOVA V. A , BLUME Ya. B.
Results and Discussion. It is known that for
many kinds of family Solanaceae the formation of
callus tissue can be induced with the medium con
taining 1 mg/1 of auxin 2,4-D (2,4-dichlorphenoxy-
acetic acid) as well as organogenesis can be stimu
lated with cytokinin media — 1 mg/1 of BAP (5-
benzylaminopurine, 6-benzyladenin). However, for
some species the combination of auxins and cyto
kines is necessary. If the ratio of auxin: cytokinin is
more than one the callus initiation prevails over stem
organogenesis. The contrary situation takes place
when the ratio is less than 1.
With the aim of more detailed investigation of
chemical compounds we used both media with (NP,
BK) and without (RMNO, RMKU, RMP, RMO,
RMOP, RMB) above mentioned regularities.
The possibility to substitute cytokinin by Tria
melon was shown on 3 different media, that usually
used for the formation and support of tobacco callus
in regime of long term passing:
1) RMKU, where 0.1 mg/1 of Triamelon mixed
with 1 mg/1 of NAA;
2) RMP, where 0.1 mg/1 of Triamelon mixed
with 0.1 mg/1 of 2,4-D;
3) RMNO, where 0.05 mg/1 of Triamelon mixed
with 3.0 mg/1 of NAA and with 0.1 mg/1 2,4-D.
So Triamelon can be combined both with natural
auxins (IAA) and with synthetic analogues (2,4-D;
NAA). At all combinations the growth and support of
callus were observed (fig. 1). There has not been
callus growth inhibition.
The next stage of our experiment was to check
the action of synthetic compounds with the media of
organogenesis induction. The root formation in cul
tured tobacco callus and at following-passing the
formation of stem buds and then leaflets were obser
ved on RMO medium containing a mixture of Tria
melon (1 mg/1) and IAA (2 mg/I) (fig. 2, a).
Fig. 2. The stem bud formation and the appearance of leaves on the
RMO medium containing 1 mg/1 of Triamelon and 2 mg/1 of NAA
(a); the bud formation of tobacco tissue on the RMOP medium
containing 1.5 mg/1 of Triamelon and 0.1 mg/1 of NAA (b);
organogenesis in callus culture on the RMKU medium containing
2—5 mg/1 of Triamelon and 1 mg/1 of NAA (c)
486
S C R E E N I N G O F T H E BIOLOGICAL A C T I O N G R O W T H REGULATORS
Fig. 3. The embryogenic callus on the RMKU medium containing Fig. 4. The microscopic analysis of mesophyll protoplasts of tobacco
10—20 mg/1 of Triamelon and і mg/1 of NAA (magnifying * І 8 0 times)
There was only a formation of buds on the
RMOP medium (Triamelon 1.5 mg/1 and 0.1 mg/1
NAA) (fig. 2, b). When we used RMKU (2.5 mg/1
Triamelon and 1 mg/1 NAA) plant-regenerants were
noticed (fig. 2 c).
So we have found optimum concentrations of
auxins and substance N 2863 (Triamelon) that like
cytokinins can cause root formation and support
conditions for meristematization of tissue and for
mation of stem buds. Also it should be noticed that
under the influence of light callus became green
because indicated concentration of Triamelon induced
the synthesis of chlorophyll and formation of chloro-
plasts.
Previous works [9] as well as our investigations
show the essential role of auxins presented in nutrient
media for the formation of embryogenic structures.
Decrease of concentrations of 2,4-D, NAA, IAA,
CPAA or their absence in presence of cytokinins
promoted the maturity of embryogenic callus with
formation of solid globular structures embryoids and
then of shoots.
On the contrary, prevention of embryoids ma
turity was observed on media, where proembryoids
were passed in the presence of auxins. We have
induced the embryogenesis on RMKU medium at
higher concentrations of Triamelon (10—20 mg/1)
and on RMP medium at certain combination Tria
melon with artificial auxin (NAA) (fig. 3).
Also we have investigated the influence of Tria
melon concentrations (0.1—20 mg/1) on the induction
of cell division. Literature sources point out that this
test can be considered as specific one for cytokinin
activity. With this aim instead of kinetin indicated
quantities of the preparation were added to Linsmaier
and Skoog medium and callus culture of core pa
renchyma of tobacco stem was cultivated. The same
media without kinetin and minimum enriched with
kinetin served as control samples. Efficiencies of
preparations were estimated by increase of biomass
counted for dry and humid mass. So we suggest that
Triamelon evinces the cytokinin activity at con
centration 2 mg/1 in the best way and under this
conditions it is more effective than kinetin on 23 %.
In test experiments with mezophyll protoplasts of
tobacco (fig. 4) the possibility to use Triamelon
instead of kinetin on both W-5 medium (ussually
applied for formation of cells' colonies) and MS
medium (for induction of callus tissues formation and
producing of plant-regenerants) was also demon
strated.
It is known that conditions for active growth of
embryoids and connecting with it further elongation
of stem (i. e. transition from meristematic tissue to
phase of active growth) require essential changes in
cultivation conditions with new nutrient media [8, 9;
12—13]. We have received a good result using 2—4
consistent passages of embryogenic callus on the same
RMOP medium. The only difference was that we took
high concentrations of Triamelon (2.0—5.0 mg/1)
(fig. 5), and further placed callus tissue in medium P
without hormones (fig. 6, a) .
Also in the literature one can find that the
regeneration of tobacco stem can be observed on
487
TSYGANK.0VA V. A. , BLUME Ya. В.
Fig. 5. Plant growth on the RMOP medium containing 5 mg/I of
Triamelon and 0.1 mg/l of NAA
media containing no auxins, but enriched with cyto-
kinins [9, 14]. There was the appearants of plant-
regenerant in our experiment on RMB medium con
taining Triamelon (5 mg/1) instead of cytokinin BAP
(0.1—1.5 mg/1) (fig. 6, b). Then this test-tube plant
with the aim of root formation was placed in RM r a i Q
medium (without phytohormones) with 0.01 mg/1
Triamelon (fig. 7, a). Results of the experiment are
demonstrated on fig. 7, b, where plant-regenerants
grown with Triamelon are shown before planting in
ground.
Screening of induction of callus formation and
callus passing of wild species of tomato have been
made on BK medium. In this medium to two auxins
(NAA and 2,4-D) we added Triamelon (0.5 mg/l)
instead of kinetin. The same procedure was in
vestigated on RMOP medium. Concentration of Tria
melon was 1 mg/l. To support the callus of cultural
species of tomato we used RMKU medium with
Triamelon (0.2 mg/l) .
Thus, we have shown the first the possibility to
use Triamelon for cultural works with tomato and
potato tissues in media that are used for induction
and long-term passing of tobacco and potato cultures
as well as the possibility to combine Triamelon with
phytohormones of auxin type (NAA, 2,4-D) in this
Fig. 6. Formation of the plant from the embryoid on P medium
without phytohormones (a); the regeneration of whole tobacco plant
on the RMB medium containing 5 mg/l of Triamelon (h)
media. Screening the growth regulative activity of Ivin
and its derivatives (Ivin-s, Ivin-h) were made on
callus cultures of potato.
It has been shown that in NP medium used for
induction of growth and support in passage of potato
callus Ivin (0.05—10 mg/l) taken instead kinetin
stimulated growth of callus biomass comparing with
control (NP medium with 1 mg/l of kinetin). The best
results were received with concentration of Ivin
5 mg/l. Besides, Ivin delayed cells' ageing of callus
488
SCREENING OF T H E B I O L O G I C A L A C T I O N G R O W T H REGULATORS
Fig. 8. Plant regenerants of potato derived from embryoids on the
NP medium containing kinetin (a) and 5 — 1 0 mg/l of Ivin-s instead
of kinetin (b)
found that there was growth of callus mass at above
mentioned concentrations. Moreover, much more in
tensive process was observed on RMKU medium for
potato callus, which under light turned to dark green
after 2 months period.
Ivin-h (0.05—3 mg/l) on P medium evinced the
similar action. So, derivatives of Ivin increase the
growth of callus biomass, delay the cell' ageing and
activate photosynthesis in cells. Comparative studies
of Ivin-s (0.1—5 mg/l) and Triamelon (5 mg/l) with
potato medium NP showed that both preparations can
be used instead of kinetin for callus growth and
maturity. In the both cases callus turned to green
after 40 days period under the influence of light, i.
е., there was the activation of photosynthesis.
Ivin-s (concentration 5—10 mg/l) was used on
ST-2 medium for the induction of morphogenesis and
the creation of potato plant-regenerants (fig. 8).
Test-tube plants grown from the embryoids that were
received on the medium containing Ivin-s showed a
more powerful development (b), than plants grown
489
TSYGANKOVA V. А. , В Ы J ME Ya. B.
Comparative data of the stimulation of root formation by synthetic substitutes of phytohormones
+ m . — main root, !. — lateral root.
with kinetin medium (я). The root formation on ST-3
medium without hormones was made.
Also the screening of Ivin and Ivin-s were made
on callus culture of tobacco. Induction of callu-
sogenesis was found on RMKU medium, containing
1 mg/l of Ivin or 2 mg/1 of Ivin-s and on RMP
medium (0.2 mg/1 of Ivin). There was the callus
meristematization on RMOP medium, containing 5
mg/1 of Ivin-s. All experiments were carried out on
nutrient media in which kinetin was substituted by
Ivin or Ivin-s, combined with auxins (NAA; 2,4-D).
Action the chemical substance N 2622 was in
vestigated with the same model and the same me
dium, although instead of auxin-NAA (1 mg/1) we
used 1 mg/1 of substance N 2622. So we combining it
with cytokinin-kinetin. Appearance of leaf was obser
ved after one month callus cultivation in darkness.
Thus we can conclude that synthetic chemical
compounds (Triamelon (N 2863), Ivin, Ivin-s, N
2622) evince high biological activity. They can be
used for works with cell' cultural and plant organs of
family Solanaceae.
Experiments for screening of auxin activity with
leaf cuttings of haricot in vivo determined three
compounds possessing the auxin-like effect: D-107,
substance N 2622 and Triamelon (see table). Among
the preparations the most significant activity (that
was even more than IAA's one) was detected for
substance N 2622 at minimal concentration (1 mg/1).
Substance D-107 had similar effect with 10 mg/1 and
Triamelon shown the lowest activity in root formation
tests.
Studying the influence of synthetic plant growth
regulators on the germination of embryo axes of
haricot seeds we found that Ivin and Methyur have
essential cytokinin-gibberellin activities [11], i. e.
they increase significantly the germination energy of
seeds of haricot reducing germination terms almost in
2 times. In these experiments Triamelon didn't show
a phytohormone activity in the contrast of its above
mentioned biological activity in vitro on Solanaceae
cultures and preliminary data of its growth-promoting
influences on melons' plants in vivo.
Our and derived from the literature data [ 1 , 7 ,
12, 13, 15] witness that on the contrary to phyto
hormones synthetic plant growth regulators act upon
both cells of undifferented and differented tissues and
cells of different plant species not always in the same
way. In the one case the synthetic preparation works
as analogue of one phytohormone but under other
conditions the same preparation can evince properties
of another phytohormone. Thus some preparations
have multifunctional properties. All of it complicates
the interpretation of possible synthetic substances
mechanisms of action in particularly when synthetic
plant growth regulators have great differences in
structure from the natural phytohormones.
For example, it has been suggested that synthetic
auxins 1-NAA, 2-NOAA, 2,4-D demonstrate similar
to IAA biological effects (though varied in quantitative
values) despite of their structural differences, that
accounts by common steric configuration (at least for
some structural features). It allows the above men
tioned regulators interact with a certain receptor [8 ].
High activities of 2,4-D and 2,4,5-T (even more than
IAA) is explained so that of IAA's oxydase can't
hydrolyse these artificial compounds [8 ]. Another
hypothesis affirms that synthetic compounds prevent
an oxidation of endogenic phytohormones that allows
last ones demonstrate a maximum of their activity
[7]. Different levels of activity for natural and
synthetic compounds also could be connected with
distinctive membrane permeabilities and transport
mechanisms inside the cells and in intracellular space
490
for such substances. Another question about non
specifical action of synthetic auxins through increase
of N-containing amino acid — tryptophan biosyn
thesis (the predecessor of IAA) or growth of free IAA
concentration due to induction of hydrolysis of auxin-
protein complex by synthetic compounds are con
sidering [2, 7] .
Thus a great number of facts confirm non spesi-
fical influence of synthetic plant growth regulators on
plant cells, and especially those of them that differ in
structure from natural compounds essentially.
Indeed if the specific receptors for recognition
sites of structure and specific fitting points of hor
mones inside cells really exist in the case of natural
compounds (phytohormones) then one can suppose
that the action of synthetic regulators with «strange»
structures is indirect and unified. Most probably they
act unificationly by changing the active pool of
endogenic phytohormones (either by transfer of their
inactive forms to active ones or by non specifical
activation of genes encoding the synthesis of phy
tohormones similar to the action of stress factors or
by two mechanisms at the same time), that increase
a growth and a maturity of plants through the natural
metabolic pathways.
The data concerning the influence of chemical
preparations on cell division, biomass growth of yeast
and fungi in cultural media (submarsed fungus Pleu-
rotus sp. and strains yeast: N 563 for alcoholic
fermentation and bakers yeast U-1037). obtained in
our experiments from numerous investigations can be
indirect confirmation of this thesis.
It was found that no one of great number
compounds (including and used those in this work) in
physiological concentrations did not have marked
influence on the indicated exponents.
We suppose that this phenomenon is connected
with absence at above mentioned organisms adequate
hormonal regulatory system of growth and develop
ment, which contrary exists in plants.
In cells of undifferentiated and differentiated
tissues and cells of various plant species different
«targets» (or their absence at all) for synthetic
regulators with non specific configurations can be
realised and so there are various biological effects (or
their absence).
Their appearances connected with a multiplicity
of cell regulation systems. It is known that un
differentiated cells have more simple regulation sys
tem comparing with differentiated ones and specia-
lizated cells of various plant species have different
homeostaze (i. е., the constancy of intracellular
medium) and so different types of regulation for its
support. In as much as in cells of higher organisms
SCREENING OF T H E BIOLOGICAL A C T I O N G R O W T H REGULATORS
there are polycomponent principle of regulation and
not one but several compounds act simultaneously in
the regulatory process [16—18], we can suppose that
synthetic growth regulators increase the formation of
active pool of all compounds participating in such
process either everyone of separately, or one after
another on cascade mechanism.
Analysing differences of regulators' action in vivo
and in vitro also it should be noticed that multi-
elemental ompositions of nutrient media are selected
by their authors with the aim to provide maximum
and specific activities of exogenic phytohormones or
their close synthetic substitutes. Therefore synthetic
compounds with other chemical reactivities can inte
ract with chemical components of medium (sometimes
it contains over 20 elements) and as result of such
interactions change their structural and biological
properties.
Differences in the influence of growth regulators
on differentiated and undifferentiated cells can be
explained only with further experimental investi
gations. But despite of the differences the use of
growth regulators as substitutes of phytohormones are
very perspective not only for studies of morpho-
genetical processes in vitro, but for practical purposes
too (even if for solution of limited tasks).
Concerning the perspectives of use selected syn
thetic growth regulators in present work, we conclude
that their characteristics on the all criterions entirely
suit for the practical application.
Ivin and its derivatives (complexes of Ivin with
succinic — Ivin-s, hydrochloric — Ivin-h, phospho
ric — Ivin-p or boric acids — Ivin-b) are widely used
in a plant-growing to increase a productivity of some
agricultural plants [5]. Using these substances in
agriculture is permitted by State Chemical Committee
of Ukraine.
Also use of Methyur and Triamelon enlarges
rapidly. Toxities of chosen preparates and their
derivatives are lower than for known growth regu
lators.
That fact can be explained because chemical
structures of these compounds are close to natural
pyrimidines (for example, Methyur, substance D-107,
N 2622). On the other hand, Ivin and Triamelon have
no toxic groups. Manufacturing and commercial costs
of investigated compounds are 10—100 time lower
then for foreign ones. An important advantage of
suggested compounds is a solubility in water. Al
though it should be noticed that some of them can
disintegrate in aqueous solutions at a long-term
storage.
The data of this work also confirm the possibility
to use selected synthetic growth regulators not only
491
TSYGANKOVA V. A . , BLUME Ya. B.
for purposes of plant-growing, but for biotechnology
in vitro.
В. А. Циганкова, Я. Б. Блюм
Скринінг та особливості біологічної дії синтетичних регуляторів
росту рослин
Резюме
У процесі біологічного скринінгу синтетичних препаратів, які
виявляють ріст-регулюючу активність у культурах тканин in
vitro і на рослинах in vivo, відібрано п'ять препаратів як
перспективних замінників фітогормонів: J він (N-oxide-2,6-
dimethy[pyridine) з переважною цитокініновою активністю in
vitro і цитокінін-гібереліновою активністю in vivo; Метіур
(Na-derivative 6-methylthyouracyl) з вираженою цитокінін-гібе-
реліновою активністю in vivo; Тріамелон (iodide tris (2,2-
trimethylammonium,methyl phosphate)) — з цитокінін-ауксино-
вою активністю in vitro; D-107 (I -acetylamino-l-acetylthyo-2-
oxo-2-phenylethan) з ауксиноподібним ефектом та № 2622
(derivative tetrahydrothyophendoixide) з яскраво вираженою аук-
синовою активністю in vivo. Запропоновано модель опосеред
кованої дії регуляторів росту рослин через фітогормони.
В. А. Цыганкова, Я. Б. Блюм
Скрининг и особенности биологического действия
синтетических регуляторов роста растений
Резюме
В процессе биологического скрининга синтетических препара
тов с рост-регуліірующей активностью в культурах тканей in
vitro и на растениях in vivo были отобраны пять препаратов
как перспективные заменители фитогормонов: И вин (N-oxide-
2,6-dimethy[pyridine) с преобладающей цитокининовой актив
ностью in vitro и цитокинин-гиббереллиновой in vivo; Метиур
(Na-derivative 6-methylthyouracyl) с выраженной цитокинин-гиб
береллиновой активностью in vivo; Триаме:юн (iodide tris
(2,2-trimethylammoniummethyl phospate) — с ицтокинин-аукси-
новой активностью in vitro; D-107 (l-acetylamina- 1-acetylthyo-
2-0X0-2-phenylethan) с ауксиноподобным еффектом и № 2622
(derivative tetrahydrothyophendioxide) с ярко выраженной аукси-
новой активностью in vivo. Предложена модель опосредованно
го действия регуляторов роста растений через фитогормоны.
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