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The Proceedings of III conference

The Ufa Reaserch Centre of Russian Academy of Sciences Academy of Sciences of Republic Bashkortostan

The Proceedings of III conference
IMMUNOASSAY of PLANT GROWTH REGULATORS in PLANT PHYSIOLOGY, PLANT GROWING and BIOTECHNOLOGY
3 - 6 of October, 2000.

Ufa, 2000. 224 p.

 
 
 
 
Content

An.H. Bajmiyev, R.T. Matniyazov, F.R. Gimalov. Dynamics of the content of abscisic acid in seedlings of cabbage during cold acclimation.
The endogenous ABA levels were estimated in seedlings of Brassica oleracea during the cold acclimation.

V.A. Velikov, A.G. Laman, A.O. Shepelyakovskaya, N.Yu. Rakova, F.A. Brovko. Obtaining of combinatory library of single chain antibodies, selection and characteristics of mini antibodies to zeatin riboside.

S.Yu. Veselov, R. Valcke. Differential histochemical determination of glycosilated forms of cytokinins and their free bases in tobacco plants.

A.O. Shepelyakovskaya, V.S. Vasil'yeva, H.M. Boziyev, A.G. Laman, E.A. Burhanova, F.A. Brovko, O.N. Kulayeva. Application of sandwich variant of immunoenzyme assay for the study of the dynamics of distribution of cytokinin binding protein (CBP-70) in etiolated seelings of maize.

F.M. Shakirova, A.M. Aval'bayev, M.V. Bezrukova, F.R. Gimalov. Levels of hormonal regulation of lectin synthesis in roots of wheat seedligs.
Synthesis of wheat germ agglutinin (WGA) is controlled alongside with ABA by GA, IAA and epibrassinolide (EP). GA and EP are revealed to induce the expression of WGA gene independently from ABA, while IAA acts inderectly through IAA-induced accumulation of ABA.

M.V. Bezrukova, R.A. Fathutdinova, A.M. Aval'bayev, F.M. Shakirova. The effect of lectin on hormonal balance in regard to its action on root growth of wheat seedlings. Wheat germ agglutinin (1 mg/l) essentially stimulated the growth of wheat roots, which is likely to be related to induction under its influence of hormonal balance in favour of IAA and cytokinins.

I.F. Golovatskaya, R.A. Karnachuk, S.Yu. Tishchenko. Green light influence on plant growth and hormonal balance.
This work is devoted to study the green light influence on the endogenic hormones level and growth of the Arabidopsis thaliana (L.) Heynh seedlings. We used 7-days-old wild type (Ler) and hy4 mutant, deficient in cryptohrome 1 (NRY1) photoreseptor and observed hypocotyl elongation of the etiolated seedlings in green light. The inhibition of the hypocotyl lengthening and stimulation of the cotyledon growth by green light was connected with changes in level of some phytohormones (citokinins, GAs). Findings indicate the connection between light and hormonal signals in regulation of the seedling growth.

N.O. Zolotuhina, Ye.L. Rudashevskaya, V.V. Emel'yanov. Dynamics of phytohormone content in developing and germinating seeds of orchids.
The most important reason of the lack of germination is seed dormancy. Developing seeds and protocorms hormonal dynamics of hard-cultivated boreal orchids were studied. Dormant seeds of Epipactis atrorubens have a very high level of ABA and low level cytokinins. Endogenous physiological type of dormancy is discussed here.

A.Yu. Kalinina, N.V. Golubeva, S.S. Medvedev. Determination of IAA in plants and callus of Arabidopsis thaliana (L.) Heynh. With the help of method of ELISA.
Peculiarity of auxin status of Arabidopsis thaliana (L.) wild type (wt) plants and abruptus (abr) mutant was studied. It was shown that not only IAA levels of intact plants and callus culture but also intensity of auxin polar transport were distinguished in abr and wt plants. It was found that abr mutation affected the mechanism of auxin polar transport in inflorescence of A. thaliana.

T.A. Korkina, N.I. Yakushkina. Hormonal balance of wheat ears depending on the stage of organ genesis and kinetin treatment.
In the experiments it is shown that the contents of both zeatin and abscisic acid and their ratio in the ear of wheat depend on phase of development of wheat and kinetin treatment.

E.M. Kof, A.S. Oorzhak. Structural and functional reorganization of semi-leafless Pea plants and phytohormones.
Pea (Pisum sativum L.) varieties with conventional (genotype Af/Af) and semi-leafless (genotype AF/AF) leaf morphotypes were used in this study. The proportions of tendrils and stipules in total dry matter and their total surface per single af/af plant were 1.5 and 2-2.5 times higher than in Af/Af plants respectivly. The chlorophyll contents in tendrils per weight or surface units and per single plant were essentially higher in af/af genotype. A very high IAA and ZR contents in the stipules and tendrils of semi-leaf less form can play an essential role in their active growth and formation of photosynthetical apparatus. A low level of ABA in tendril tissues can be an attribute of a small total transpiration surfaces.

G.R. Kudoyarova, I.R. Teplova, R.A. Dokicheva, I.Yu. Usmanov, S.Yu. Veselov. The effect of benzylaminopurine on the growth and auxin content in wheat and maize seedlings.
The treatment of wheat (Triticum durum Desf., cv. Bezenchukskaya 139) and maize (Zea mays L., hybrid Krasnodarsky 395 TV VL) seedlings with synthetic cytokinin BA taken in different concentrations (0,022, 0,22 and 2,2 m M) decreased content of IAA quantified with immunoassay in roots of treated plants in relation to the control. The effect was accompanied by the inhibition of roots growth. The comparison of the time course of shoot and root growth responses to high BA concentrations suggests that the reduced shoots growth observed after its initial increase may be related to the hormone-induced changes in roots growth and function.

T.I. Puzina, I.G. Kirillova. Key relations of phytohormones and their role in regulation of physiological processes in potato plants.

M.V. Simonyan, S.Yu. Veselov, T.N. Arhipova. The use of immunoassay for cytokinins in estimation of cytokininoxidase activity.
Immunoenzyme test-system for isopentenyladinosine is proposed and new highly sensitive method for estimation of cytokininoxidase activity is developed, which does not demand the use of radioactive isotopes. The use of known activators and inhibitors of the enzyme is recommended for providing the specificity of analysis.

I. Teplova, A. Christmann, B. Nikander. Quantifying and qualifying of cytokinins in silver fir needles Abies alba Mill. in the process of senescence.
Zeatine-9-riboside (9R-Z) has been shown as a major cytokinin in silver fir needles.. Zeatin (Z), isopentenyladenine (IP) and isopentenyladenosine (IPA) have been also identified as native cytokinins of silver fir needles however their contents were extremely lower as compared to that of 9R-Z. Level of 9R-Z has been dramatically declined with increasing needle age. The most pronounced decrease in the hormone content has been observed in needles of 1-5 years of age that might be relevant to initiation of the first slow phase of senescence. The data obtained by means of b -glucosidase treatment of the needle extracts evidences for accelerated conversion of 9R-Z into its O-glucoside due to the senescence. This type of cytokinins has been reported to be tolerance to oxydase degradation of cytokinins. On the other hand O-glucosides are suggested to be a transport form of cytokinins. Thus all this combined supposes that 9R-Z remaining in senescencing needles might be converted into 9R-Z-O-glucoside and then transferred from older needles to younger ones as more chemically stable form.

R.A. Borzenkova, M.P. Borovkova. Phytohormones, starch biosynthesis and storage during potato tuber growth.
The content of endogenous ABA, IAA, cytokinins (zeatin+zeatin-ribozide) and their distribution in different tissues – cortex and core was investigated. Relations between ABA accumulation and the rate of starch synthesis and accumulation, the rate of cell division and concentration of IAA and cytokinine were estimated.

I.I. Ivanov, S.Yu. Veselov, N.L. Anohina, G.R. Kudoyarova, R. Valcke. Ipt-transgenic tobacco plants as model for studying of source-sink relations between shoots and roots.
ELISA has been used for the cytokinin analysis in wild-type and ipt-transgenic (with hs-promotor) tobacco plants. It is shown that daily heat shock (1.5 hours) within 2 weeks is sufficient for induction of allocation to shoot growth. Accumulation of hormones occurs mainly in shoots. It is most probable, that cytokinin render regulatory action on the source-sink relations between shoots and root system through processes located mainly in shoots.

I.S. Kiseleva. Interaction of assimilate usage and endogenous phytohormones in developing leaves of barley.
Utilization of photosynthates in leaves during their development was studied in relation to export activity and concentration of endogenous phytohormones in preflag and flag barley leaves. It is revealed that ontogenetic changes in synthesis of cellulose and hemicellulose and protein relate to IAA and ABA content and ratio.

S.P. Maslova, E.M. Kof. Intensity of respiration and content of abscisic acid in underground shoot of stolon forming plants.

G.A. Ramazanova, Z.F. Rahmankulova, I.Yu. Usmanov. The role of phytohormones in redistribution of material and energy resources under normal and stress conditions.


L.N. Timergalina, L.B. Vysotskaya, G.R. Kudoyarova, M.V. Simonyan, S.Yu. Veselov. Regulation of concentration of hormones in wheat seedlings under partial root pruning.
Excision of 4 roots out of 5 of wheat plants grown in hydroponical culture did not cause a disturbance in their water relations and led to the increase in the root growth rate. In 0,5 h root pruning of seedlings resulted in increased concentration of IAA in remaining root as compared to the control level, which might be responsible for the observed initiation of lateral root growth. Cytokinin concentration in xylem sap of plants with reduced root system was decreased initially as compared to intact plants that is in accordance with the reduction in the hormone-producing organ. Nevertheless cytokinin content in the shoots remained high, which might be due to a decrease in cytokinin decay. Our results demonstrate a high plasticity of the system controlling hormone concentration in plants, which is achieved through an elevated ability of the pruned organ to synthesise the hormone and ability of shoots to metabolise it. Both IAA and cytokinins seem to be important in the restoration of the shoot/root balance disturbed by root pruning.

M.Yu. Yashkov, R.A. Borzenkova. Hormonal status, production and usage of assimilates in wild species with different sink types.
The hormonal status of wild plant species of the Middle Urals, differed in sink-source relations, was studied in the period of flowering. The relation between cytokinine and ABA distribution among leaves, underground part and generative organs and photosynthate translocation, accumulation of storage substances was estimated. Carbon transportation to upper sink organs and the level of nonstructural carbohydrates positively correlated with the ABA/cytokinine ratio in generative organs and higher cytokinine concentration in generative organs compared to leaves. At the same time monosaccharide and nonstructural polysaccharide accumulation was positively correlated with ABA/cytokinine ratio in rhisome. The results are discussed in relation to different types of ecological strategies in plants.

L.B. Vysotskaya, L.N. Timergalina, T.N. Arhipova, A.V. Dedov, G.R. Kudoyarova. Hormones and characteristics of water relations in plants with partially excised root system.
In hydroponical culture excision of 4 roots from 5 don’t led to decrease in transpiration rate and water content in organs as compared to intact plants, but probably cause changes in hydraulic conductivity of remained root. Possibly ABA and/or IAA are the factors which affected root conductivity. Treatment of root system with HgCl2 solution decreased water content in shoots and roots of plants with reduced root system as compared to control plants, thus we assume the possible involvement of aquaporins in regulation of water balance in wheat seedlings. Our results demonstrate the ability of plants to regulate water content in organs by transpiration stream and by changes in hydraulic conductivity of root system.

D.A. Klimachev, V.T. Starikova, N.I. Yakushkina. The changes in hormonal balance of wheat plants depending on the level of mineral nutrition and BAP-treatment.
The influence of cytokinin on hormonal balance in miscellaneous conditions of mineral power supply was studied. It is exhibited that the processing 6-BAP enlarges cytokinin and IAA contents noticeably and reduces ABA content both in pages and ears, increasing efficiency wheat. The efficiency of 6-BAP influence was increased by sufficient supply with nitrogen and phosphorous.

D.A. Klimachev, A.A. Tarasenko. The effect of gibberellin-treatment on phytohormone content under different conditions of mineral nutrition.
The processing by an exogenous gibberelic acid changes a ratio of hormons. The feedforward between hormonal balance and growth rate of wheat plants was scheduled. Thus the influence of gibberelic acid appeares to depend on supply of plants with mineral elements.

T.A. Korkina, G.N. Firsanova. The effect of potassium ions and cytokinins on hormonal status, anatomical and physiological characterisitcs of wheat.
In experiments it is shown that potassium and kinetin change content of hormones and their ratio in wheat. Weight of plant bodies corresponded to change in IAA+ZT/ABA ratio. Physiological action of potassium and kinetin has additive character.

I.B. Sabirzhanova, A.R. Mustafina, D.S. Veselov, A.V. Dedov, G.R. Kudoyarova, S.Yu. Veselov. Fast changes in concentration of cytokinins in shoot, root and xylem sap of wheat seedlings after addition of PEG to the nutrient medium.

Addition of PEG to the nutrient medium decreased total cytokinin content, while concentration of cytokinins in xylem sap was reduced twice. Since transpiration content in shoots of wheat seedlings decreased by factor of three compared to controls 10 min after the start of treatment, the rate of hormone delivery was three times slower in PEG-treated compared to control seedlings. The decrease in cytokinin content in shoots was accompanied by complete cessation of leaf growth 15 minutes after addition of PEG. Thus hormonal response might contribute to reduced transpiration and shoot growth. Acceleration of cytokinin metabolism in shoots induced by osmotic shock is suggested.

E.Yu. Bahtenko, A.V. Platonov. Importance of phytohormones in adaptation of plants to water excess in soil.

A.P. Veselov, L.N. Olyunina, A.G. Orlova, V.P. Lobov. Fast phytohormonal response to heat shock.
The changes in IAA and ABA levels in wheat and pea seedlings at short exposures of a heat shock were studied. It was found that the earliest phytohormone responses to the heat shock was IAA oscillations between the free and the bound forms and the linear increase in free ABA content. The possible mechanisms and physiological role of the quick phytohormonal response to the heat shock are discussed.

L.V. Victorova, N.N. Maksutova. Effect of abscisic acid on protein synthesis in wheat caryopsis under drought.
Exogenous abscisic acid (ABA) has been studied for its effect on the synthesis intensity of various proteins of spring wheat caryopsis Moscovskaya 35 under optimal (70 % FWC) and limited (30 % FWC) water supply. Under normal water supply endogenous content of ABA increased when passing from the caryopsis formation phase to the milk ripeness phase. Plant dehydratation caused the increase in endogenous ABA, it was espesially noticeable at the stage of milk ripeness. Under optimal water supply exogenous ABA inhibited protein synthesis, and under drought it not only restored the suppressed protein synthesis, but also stimulated protein formation in comparison with the control variant. It is assumed that ABA

G.R. Kudoyarova, M.V. Simonyan, A.R. Mustafina, S.Yu. Veselov. The effect of dehydration on polyribosome and cytokinin contents and cytokininoxidase activity in maize seedlings.

A.N. Mitrichenko, S.V. Veselova, Ye.A. Ivanov, D.S. Veselov, A.V. Dedov, R.G. Farhutdinov. The effect of the increase in air temperature on growth, hormone content and water relations of wheat seedlings.
The increase in temperature from 24 up to 29 oN has resulted in reduced growth rate and very fast hormonal response in leaves of wheat seedlings. The growth rate was quickly reduced after heating starts. Transpiration rate elevated and leaf water content decreased under high temperature that suggests discrepancy between water delivery from roots and transpiration. The heating led to accumulation of both ABA and IAA in the leaves which might be related to growth response. Application of HgCl2 resulted in dramatic delay of growth rate recovering. Possible role of the hormones in plant adaptation to the temperature changes is discussed.

T.N. Pustovojtova, N.E. .Zhdanova, V.N. Zholkevich. The importance of phytohomones in adaptation of plants to drought.

Growth inhibition of cucumber leaves (Cucumis sativus L.) is preceded by a significant decrease in IAA, ABA and cytokinins under drought. Drought adaptation generally occurs while growth is inhibited. IAA and cytokinin can significantly rise during the adaptation period. ABA level rose in the beginning and in final step of adaptation process. Obviously, all three studied phytohormones – ABA, IAA and cytokinins are involved in drought adaptation, although, their contributions to the process vary at different stages.

M.V. Simonyan, S.Yu. Veselov. Cytokinin oxidase: biochemical characteristics, mechanism of regulation of activity and physiological importance.
This review deals with characteristics of cytokininoxidase, which belongs to Cu containing aminooxidase This enzyme regulates concentration of cytokinins in plants through oxigene-dependent decay of unsaturated side chain of cytokinins. Substrate specificity of cytokininoxidase, the effect of different factors on its activity, its physiological role and methods of determination are discussed.

V.V. Talanova, A.F. Titov. The effect of stress factors of different nature on content of ABA and IAA in plant tissues.

The studies of the effects of several stresses (low and high temperatures, chloride salinity and heavy metals) on the endogenous ABA and IAA contents in cucumber, wheat and barley seedlings were carried out. It was found that the stresses of various nature similarly change the ABA and IAA levels in the leaves at the initial stage of their action. So, the change in these hormone levels in leaves can be expected to be one of the possible inductors of the metabolism reorganization directed to the unspecific (common) resistance.

R.G. Farhutdinov, A.N. Mitrichenko, I.R. Teplova, A.V. Dedov, G.R. Kudoyarova. The changes in cytokinin content in wheat plants under heating of nutrient solution up to 35 °C.

Heating of nutrient solution up to 35° C resulted in accelerated shoot growth rate of wheat seedlings. Cytokinin content was rapidly increased by the heating. The return to initial temperature (24 °C) caused short-time decline in cytokinin level followed by increase in it. The first peak of cytokinins are supposed to contain ZR and Z while the second one – ZG, ZN and Z. Heating of wheat roots led to sharp increase in shoot ZR content that might be relevant to both promoted synthesis and transpiration rates of shoot cytokinins or conversion Z into ZR in shoots. The increase in cytokinin concentration caused by the return to initial temperature probably related to accumulation of ZG, ZN and Z accompanied by sharp decrease in ZR level.

D.S. Veselov, K. Ullrich. Dynamics of the content of different forms of cytokinins in shoot tumors of plants of Ricinus communis, induced by Agrobacterium tumefaciens.

Sh.Ya. Gilyazetdinov, N.A. Rozhnova, G.A. Geraschenkov. Hormonal system of plants under action of inductors of system resistance and phytoviruses.

I.V. Maksimov, R.M. Ganiyev, R.M. Hajrullin. The change of the level of IAA, ABA and cytolinins on the initial stages of infection with phytopatogenic fungi.
The content of phytohormones IAA, ABA, cytokinin and their balance was studied in seedlings of two wheat species, differing in resistance to Tilletia caries Tull. The pathogen changed IAA/ABA balance in susceptible wheat Triticum aestivum L. (variety Zhnitsa). These changes resulted in both primary accumulation of IAA at the opening period of pathogenesis, and increased in content of ABA during next period of plant-fungi interaction. In resistant plants (T. timopheevii Zhuk. (e-58666)) content of IAA and decreased ABA but content of cytokinin and ratio cytokinin/ABA increased contrary.

L.N. Olyunina, S.V. Kulikova, G.N. Ladygina, V.N. Shabayev. Production of indolyl-3-acetic acid by Pseudomonas putida, Azotobacter chroococcum.
It has been found that rhizobactera Pseudomonas putida and Azotobacter chroococcum are active producent of IAA. Hypothesis has been suggested that production of IAA is a mechanism of detoxication of triptophan in procaryotic cell. Toxicity of triptophan was proved by adding of high concentration of this aminoacid into culture medium.

N.A. Rozhnova, G.A. Geraschenkov. Hormonal system of tobacco plants under the action of inductors of resistance and VTM-infection.

L.G. Yarullina. Phytohormones in the interaction of plant host and pathogene.
The content of phytohormones in plants of resistant and susceptible wheat varieties under infestation by root rots was investigated by means of immuno-enzyme assay. In response to pathogene invasion the cytokinin levels increased in resistant plants at the stable IAA/ABA balance background. The ABA concentration was increased in infected plants of susceptible varieties. The participation of phytohormones in establishing of resistance or susceptibility mechanisms under fungous pathogenesis is discussed.

E.A. Goncharova. Plant growth regulators in the function of the system of attraction in plants and problems of diagnostics of stress resistance.

The role of hormone regulation of the sourse-sink relation during the period of feeding and its investment to the formation of productivity development and stress resistance was discussed.

T.S. Makarova, D.A. Klimachev. The effect of epibrassinolide on the content of hormones in leaves of different position of tomato plants.

The research has shown that under influence of processing brassins the contents of IAA and zeatin in pages of tomatoes are increased. The content of ABA is reduced. The pages of the upper circle appear to be more sensitive to processing with the regulator.

A.R. Sahabutdinova, M.V. Bezrukova, R.A. Fathutdinova, F.M. Shakirova. Analysis of the balance of IAA and ABA in plants treated with salicilic acid under salinity in regard to their growth.
Pretreatment with salicylic acid of wheat grains prevents changes in IAA/ABA ratio and reduces a degree of damage of grown seedlings.

L.V. Sidorova, L.B. Vysotskaya, A.R. Mustafina, N.I. Fedorov. Phytohormone contents and growth of Aconitum septentrionale shoots treated with diterpenoid alkaloids.
It is shown that growing of Aconitum septentrionale shoots on nutrient medium containing diterpenoid alkaloid lappaconitin caused changes in morphological and biochemical characteristics. Possible relations between direrpenoid alkaloid contents, growth rate and cytokinin contents are discussed.

I.A. Yahin, O.I. Yahin, F.M. Shakirova. The effect of preparation of stifun on the balance of phytohormones and productivity of spring wheat.
The effect of the preparation stifun on the growth, balance of hormones and productivity of wheat was studied. Both presowing of seeds and treatment of plants with stifun stimulate the development of plants, and finally, produce a positive effect on productivity.

V.Yu. Gorbunova. The role of phytohormones in regeneration of plants of genotypes of summer weak wheat.

A.A. Zaripova, R.K. Baiburina. The content of endogenous hormones of Paeonia anomala (L.), Valeriana officinalis (L.) and Polemonium caeruleum (L.) before introduction in vitro culture.


E.P. Ivanova, T.Yu. Mityushkina, S.V. Dolgov. The effect of transformation of Chrysanthemum morifolium plants with the help of rol C gene of Agrobacterium rhizogenes on the content of endogenous phytohormones in their leaves. Chrosanthemum morifolium plants transformed by the rol C gene from Agrobacterium rhizogenes showed compact habitus of the aerial part. Immunoassay of the endogenous phytohormone contents of transgenic plants showed 12-fold increase in cytokinin and 4-fold increase in abscisic acid with 6-fold decrease in auxin levels at the budding phase. The phytohormone ratios varied at any certain stage of development.

N.P. Karsunkina, I.V. Skorobogatova, Ye.V. Zaharova. Peculiarities of hormonal balance of tubers of potato cultivar and somaclones under different physiological status.
Hormonal balance in potato tubers of variety and two its somaclones was studied. Phytohormone contents (ABA, IAA, GA3, cytokinins) in tubers were very different and depended on physiological conditions of tubers (dormancy period or begining of germination) too.

E.A. Osipova, Z.B. Shamina. The use of immunoenzyme assay for the study of the regulation of the synthesis of protoberberine alkaloides in cell culture of Thalictrum minus.
The contents of endogenous IAA and ABA were determined in cell lines of Thalictrum minus – differed in production of protoberberine alkaloids. The interrelation was shown between high level of ABA and activity of alkaloid biosynthesis.

I.F. Shayahmetov, M.V. Bezrukova, R.R. Ahmetov. Interaction of accumulation of lectin and abscisic acid in tissue culture of wheat.
Levels of wheat germ agglutinin (WGA) and abscisic acid (ABA) have been determined by immunoassay in morphogenic and nonmorphogenic callus tissues cultured in the presence of IAA and 2,4-D in growth media. Our results show that both embryogenic and nonembryogenic callus contained WGA and ABA; the levels were higher in the embryogenic callus especially in active growing ones. ABA stimulated, 2,4-D inhibited lectin synthesis irrespective of the nature of callus. Changes in lectin content that occur during normal growth and in response to applied agents (IAA, 2,4-D) are correlated with alterations in endogenous hormone levels. On the whole, lectin content depended on the growth intensity of callus tissue.

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