Β-indolylacetic Acid Research Articles

МИКРОКЛОНАЛЬНОЕ РАЗМНОЖЕНИЕ GLADIOLUS × HYBRIDUS СОРТА ‘ПЕРМСКИЙ СУВЕ НИР ’ СЕЛЕКЦИИ УЧЕБНОГО БОТАНИЧЕСКОГО САДА ПГНИУ

The article presents data on micropropagation of new Gladiolus × hybridus cultivar ‘Пермский Сувенир’. It is found that for the sterilization of corms and cormels better use the 7% sodium hypochlorite solution (15 min or 20-25 min) and then 96% ethanol (30 sec or 60 sec). Sterile culture was more than 56% for corms and 76% for cormels. At the micropropagation good results were obtained when vitamins (0.1 mg/l thiamine, 0.5 mg/l pyridoxine, 0.5 mg/l nicotinic acid) and β-indolylacetic acid (0.1-1 mg/l) were added to the Murashige and Skoog medium. The breeding rate for cormels was 2-3, for corm fragments was 2–4. The survival rate of plants transferred from in vitro to in vivo conditions was 70%. The diameter of the obtained juvenile corms from in vitro culture and the number of cormels formed on them did not have statistically significant differences from those planted in vivo (t = [1.16; 1.22] <t05 = 1.96).

In vitro micropropagation and ex vitro rooting of some potato varieties

The article presents the results on selection of optimal concentrations of nutrient media components and nutrient solution at the stages of clonal micropropagation of potato varieties Lyubava, Kemerovochanin, and Tuleevskiy (actual reproduction, rooting in vitro, adaptation to ex vitro conditions). The influence of some components of the nutrient medium (sucrose, agar-agar, growth regulators, namely α-naphthyl acetic acid, β-indolyl acetic acid, and β-indolyl propionic acid) was studied at the stages of reproduction and rooting in order to obtain regenerants of the studied potato varieties. The best development of plants on nutrient medium with addition of 4 g L-1 of agar-agar was revealed. The addition of sucrose in the concentration of 3-5% contributed to the formation of more internodes. The influence of naphthyl acetic acid, β-indolyl oil, and β-indolyl propionic acids in different concentrations on the rhizogenesis of regenerating plants of three potato varieties at the stages of reproduction and establishment of different types and concentrations of auxins was studied. The researches have shown that a one-stage method of adaptation of Solanum tuberosum L. varieties of Lyubava, Kemerovochanin, and Tuleevskiy to the conditions of ex vitro cultivation with the use of hydroponic installation is characterized by efficiency and allows to receive plants with well-developed root system. The efficiency of using the hydroponic plant for adaptation of potato regenerants at the final stage of micropropagation is shown.

Optimization of clonal micropropagation of Chrysanthemum

Micropropagation of Chrysanthemum is an alternative to the traditional method of reproduction. Thanks to this method, the Chrysanthemum reproduction time is reduced to 3-4 months. For clonal micropropagation, sterile microshoots of Chrysanthemums of the varieties Snow White, Stranger, and Baltica White were used. At the stage of the micropropagation, the Murashige-Skoog (MS) medium with the full and half composition of mineral salts and growth regulators (kinetin, 6-benzylaminopurine, β-indolylacetic acid) were used. A universal culture medium for clonal micropropagation of all varieties of Chrysanthemum and optimal mediums, taking into account variety-specificity were established. It was noted that under in vitro conditions, high proliferative activity was observed in Neznakomka variety.

Catharanthus roseus (L.) G. Don - plant regeneration and alkaloids content

We describe here a regeneration of plantlets of <i>Catharanthus roseus</i> (L.) G. Don from shoot tips and axillary buds. Shoot tips were excised from 7-day-old seedlings and were incubated in solid Nitsch and Nitsch (NN) medium supplemented with kinetin, benzyladenine (BA), indole-3-butyric acid (IBA) and β-indolylacetic acid (IAA) in various combinations. After two months in culture, regenerated rooted plantlets were cut and transferred to a new medium; the explants contained shoot tips or axillary buds. Four passages were done. We obtained about 200 rooting plantlets from one seedling. Then the plantlets were transferred to the soil and they grew under a foil tent. After five months of vegetation they were collected, dried and weighed. Chemical investigations of leaves of these plants were done. The vindoline and catharanthine were dominant alkaloids in the juvenile stage of plants (before blooming). Total amount of alkaloids, equal 2.95%, was gravimetrically determined in leaves of plants, after 4th passage, regenerated <i>in vitro</i> on NN medium supplemented with kinetin and IBA.

Synthesis of tritium-labeled phytohormones

A method has been developed for obtaining tritium-labeled analogues of β-indolylacetic acid and of 6-benzylaminopurine by the method of isotopic catalytic exchange. The localization of cytokinin- and auxin-binding proteins in cotton seedlings has been investigated.

The Effect of paclobutrazol on the Level of endogenous IAA in Relation to the Rooting of Cuttings and Abscission of Petioles

The relationship between paclobutrazol (i.e. Cultar) and β-indolylacetic acid (IAA) was studied. Immersion of the bases of Ligustrum vulgare cuttings into a 0.05% solution of Cultar stimulated rhizogenesis similarly as when applying a 100mgl -1 concentration of IAA. However, the degree of rhizogenesis was much higher if the cuttings were first immersed in a 0.05% solution of Cultar and then in a 100mg1 -1 solution of IAA. Abscission of leafless petioles of Malus sylvestris was significantly inhibited when applying paclobutrazol (Cultar) to the distal cut surface of the excised leaf. The fact that the immersion of the bases of Ligustrum vulgare cuttings into a Cultar solution resulted in a statistically significant increase in the level of endogenous IAA in the basal parts of the cuttings could explain the above given effects on rhizogenesis and abscission.

Induction of plantlets in axenically cultivated rhizoids of Fucus spiralis

Rhizoids of Fucus spiralis were cultivated axenically in the artificial seawater ASP6 F2. Experiments were made to increase the filamental growth as well as to induce adventive primordia (plantlets). Additions of such carbon compounds as glucose, acetate, and formate had no favourable effects even in concentrations as low as 1∙10−4 M. Mannitol killed the rhizoids in higher concentrations and inhibited growth even in a concentration as low as 1∙10−5 M. Higher concentrations of glycerol also inhibited growth, but 1∙10−4 M was an exception as it initiated plantlets. Many simple phenolic compounds induced plantlets. Among the most active substances were phenylacetic acid, p-hydroxyphenylacetic acid, 3,4-dihydroxybenzoic acid, o-hydroxybenzoic acid, and o-acetoxybenzoic acid, with optimal effects in the concentration range of 1∙10−7 to 1∙10−6 M. β-Indolylacetic acid strongly influenced the dry weight as well as plantlet formation at concentrations of 1∙10−8 to 1∙10−7 M, with 1∙10−8 M favouring plantlet induction. It is obvious that β-indolylacetic acid plays an important role in the earlier stages of the development of Fucus.

Effects of Mycotoxins on the Development of Epiphyllous Buds of Kalanchoe daigremontiana

Summary In Kalanchoe daigremontiana aflatoxin B 1 (100 μg/ml) and rubratoxin (1 μg/ml) inhibit the elongation of roots, patulin (100 μg/ml) reduces particularly strongly the average number of newly formed roots, and rubratoxin B (100 and 10 μg/ml) totally suppresses the development of roots. Citrinin (100 μg/ml) strongly affects the formation of roots. Sterigmatocystin (100 μg/ml) and diacetoxyscirpenol (100 μg/ml) have only moderate effects. It is assumed that the inhibitory action of aflatoxin B 1 , rubratoxin B and diacetoxyscirpenol may be the result of an antagonism to β-indolylacetic acid and that the toxicity of patulin is due to an interaction with sulfhydryl-containing enzymes.

Untersuchungen über den Einfluß von Mycotoxinen auf die Aktivität der β-Indolylessigsäure und die Keimung von Kressesamen (Lepidium sativum)

Aflatoxin B 1 rubratoxin B and diacetoxyscirpenol in low concentrations (≤ 1 μg/ml) act as synergists, in higher doses (> 1 μg/ml) as antagonists of β-indolylacetic acid. Patulin is without influence on this auxin. The germination of seeds of Lepidium sativum is inhibited by patulin and diacetoxyscirpenol, the latter toxin being the most toxic. The development of the hypocotyl is stronger influenced than that of the radicula.

Stoffwechsel von Isoflavonen und Cum�stanen in Zell- und Callussuspensionskulturen von Phaseolus aureus Roxb.

The isoflavone daidzein, the coumestanes coumestrol and soyagol as well as 2′,4,4′-trihydroxychalcone were isolated from callus and cell suspensions of root tip tissue from Phaseolus aureus Roxb. Upon prolonged culturing callus suspensions gradually became cell suspensions, a process which was accompanied by a decrease in the accumulation of phenolics. Upon transfer of the cells into 3 different media containing β-indolyl acetic acid, kinetin or α-naphthalene acetic acid, a drastic increase in the amount of coumestrol was measured.

Effect of beta-indolylacetic acid on the growth and the auxin catabolism of Lactobacillus bulgaricus.

ENDOGENOUS auxin may act in the regulation of bacteria growth1, although its precise role is as yet poorly understood2. For example, Lactobacillus can grow without auxin3; however, in the presence of β-indolylacetic acid (IAA), its multiplication can be strongly modified4. In the following experiments, Lactobacillus bulgaricus (strain 1416, Liebefeld Institute of Dairy Research) was used and the medium was Lactobacillus broth AOAC (Difco). For each series of growth measurements, tubes containing 10 ml. of medium (with IAA) were inoculated with 1 ml. of a young culture of Lactobacillus (standard: 0.416 ml. of protein). The cultures were incubated at 37° C and the growth was measured after 6 and 12 h. Two methods were found most satisfactory for the growth determinations. The first is related to a direct measurement of the increase in turbidity5, using the ‘Lumetron 402 E’ with a double photoelectric cell. The second is based on the analysis of the total protein content6. The protein was determined by precipitating the washed bacteria cells with trichloroacetic acid. Comparing the turbidity with a series of standard solutions of serum-albumin, precipitated with trichloroacetic acid, a good correlation was found. Further examination of this technique will be discussed elsewhere.

Synthesis of a Plant Hormone by the Salivary Apparatus of Plant-sucking Hemiptera

As a result of investigations into the galls and other plant deformations caused by Hemiptera, the salivary glands of aphids have been claimed to convert tryptophan to β-indolyl acetic acid (IAA) in vitro1, and their saliva has been shown to contain varying amounts of this plant hormone2. Experiments have demonstrated the ability of a mirid and a leafhopper to transfer ingested IAA to the salivary glands and saliva3, but the salivary system of all the phytophagous Hemiptera may have the requirements for the synthesis of IAA from endogenous metabolites4. Thus tryptophan is known to be converted to IAA by quinones5 such as would arise by the action of the insects' own salivary polyphenol oxidase6; and both tryptophan and a substrate for the enzyme have been found in the secretions of the salivary glands7. The following is an account of experiments designed to test the possible synthesis of salivary IAA in Hemiptera. Two insects were used: Eumecopus punctiventris Stal (Pentatomidae) and Elasmolomus sordidus (F.).

Action of Gibberellic Acid on Auxin Transport

THE polar transport of β-indolylacetic acid (IAA) in plant organs has been known since the observations of Went1 and van der Weij2. Most of the direct investigations of IAA movement have used bioassays for detecting auxin3. But these analyses have been limited by the sensitivity of the method4. The recent use of IAA labelled with carbon-14 (IAA*) facilitates the study of the auxin translocation5.

Colorimetric Determination of 3,6-Anhydrogalactose with the Indolyl-3-acetic Acid Reagent

THE sugar 3,6-anhydrogalactose is a major constituent of the agarose fraction of agar, of kappa carrageenin, and of some other polysaccharides from marine algae1. This sugar can be determined spectrophotometrically as 5-hy-droxymethyl-2-furaldehyde2 or by an adaptation of the rosorcinol test for ketoses3. The rate of degradation of 3,6-anhydrogalactose to 5-hydroxymethyl-2-furaldehyde suggests that any method used for the colorimetric determination of fructose should be applicable to 3,6-anhydrogalactose. Steinitz4 compared six methods for the determination of fructose in body fluids, and favoured the method of Heyrovsky5, who used β-indolylacetic acid as the colour reagent. The present communication describes the use of this reagent for the determination of 3,6-anhydrogalactose.

Studies on Extension Growth in Coleptile Sections

Wheat coleoptile sections were grown at various temperatures from 1–40°C. in water and solutions of the sodium salt of β-indolylacetic acid (NaIAA) ranging from low to very high concentrations (0.001 p.p.m. to 10,000 p.p.m.). Growth-time curves are sigmoid, obviously so under conditions unfavourable to growth, less clearly as conditions either of temperature or growth-substance supply are improved, the point of inflection moving nearer to zero time. The growth-rate is reduced by temperatures below the optimum, and although growth continues for a longer time, final section length is also reduced. Such reductions are also brought about by sub-optimal levels of NaIAA and by supra-optimal levels of both factors. In the test method employed, sections are grown in closed tubes of small volume, and after a time lose turgidity and become waterlogged; the onset of this condition is accelerated by high temperature or growth-substance concentration, and greatly delayed by low temperature. Under comparable conditions, but with the tubes uncorked, flaccidity is delayed even at high temperature or high concentration of NaIAA. If sections are transferred from one temperature to another the change in growth-rate is abrupt; effects of transfer on final length are complex. A contour diagram represents the idealized relationships between section growth and temperature and NaIAA supply over their whole range of effectiveness. Relations between temperature and growth in other systems are briefly discussed.

Effect of Gibberellic Acid on Corolla Size

IN a previous communication1 I reported that emasculation of Glechoma hederacea flower buds resulted in a reduction in corolla size similar to that found in naturally occurring female flowers, and I suggested that the anthers produced a hormone causing enlargement of the corolla. An attempt to replace this by the application of β-indolylacetic acid to emasculated buds was ineffective—low concentrations having no effect and high concentrations (500 and 1,000 p.p.m.) causing inhibition of corolla growth.

Acidity of sprays used in detecting ketohexoses on paper chromatograms.

SEVERAL spray reagents are available for the specific detection of ketoses and ketose-containing sugars on paper chromatograms. Probably the most widely used are resorcinol and naphthoresorcinol, originally prepared as sprays in 2 N hydrochloric acid by Forsyth1. Afterwards, Partridge2 replaced the hydrochloric acid with trichloracetic acid in order to minimize attack on the filter paper. Heyrovsky3 has recently reported that β-indolyl acetic acid in trichloracetic acid is also a very sensitive ketose-specific reagent. In work in this laboratory on two fructose-containing disaccharides it has been found that all three of the above reagents, when prepared in trichloracetic acid, failed completely to detect these sugars on paper chromatograms. When the trichloracetic acid was replaced with 2 N hydrochloric acid strong positive tests were obtained. The results are listed in Table 1.

Interaction of Hormone and Photoperiod in an Early Variety of Rice

THE interaction of hormone and photoperiod has been studied in cocklebur1–5, Biloxi soybean5 and Orchad grass6. In the present investigation the effect of the hormones in relation to photoperiodism has been studied in an early variety of rice, N136, which has been found to be intermediate in its photoperiodic behaviour7. Graded seeds of this pure strain were soaked on December 10, 1956, with 500 p.p.m. of the three plant-growth substances, β-indolylacetic acid (IAA), α-naphthylacetic acid (NAA) and 2,3,5-tri-iodobenzoic acid (TIBA), for 72 hr., a treatment which was found to be effective in earlier studies8,9. The grains were next thoroughly washed in water and sown in earthenware pots containing a mixture of loamy garden soil and cow-dung manure. The pots were divided into two series. In series 1 the plants simply grew after the initial hormone seed-treatment. In series 2 the plants also received an 8-hr, photoperiod (6 a.m.–2 p.m.) from the twenty-first day until panicle emergence. Two other series of unsoaked seeds were used. In series 3 the plants were given the short photoperiod as in series 2. In series 4, plants remained as untreated controls. The results are shown in Tables 1 and 2.

Factors affecting the Abscission of Reproductive Organs in Yellow Lupms (Lupinus luteusL.)

Abscission of flowers in Lupinus luteus L. (var. Weiko II) without growth of ovaries is followed by abscission of small pods (15–20 mm. long). Normally flower abscission is much more pronounced than pod abscission. Abscission was delayed on plants from which laterals or their terminal and axifliary buds were removed. Flower abscission was not affected, but pod abscission increased as a result of defoliation. When flowers at the base of the main inflorescence were replaced by auxins and anti-auxins flower abscission was induced in either an auxin pattern in which most of the flowers near the site of application dropped, and pods developed on the apical whorls, or an anti-auxin pattern in which pods developed on basal whorls near the site of application but not higher up. The anti-auxin pattern was similar to the pattern of abscission normally induced by developing pods on basal whorls. α-Naphthylacetic acid (NAA) was much more effective in inducing abscission than β-indolylacetic acid (IAA). 2:3:5-triiodobenzoic acid (TIBA), NAA, and IAA applied in mixtures at various concentrations acted mainly antagonistically, i.e. the abscission-inducing effect of NAA and LAA was depressed in basal whorls, and in apical whorls the effect of TIBA was less prevalent. Consequently the effect of the mixtures on the total number of pods was about equal to that of the most active component by itself. All growth substances seemed to move much more efficiently in acropetal direction than in basipetal direction in the flower stalk. Transport in lateral direction was very limited. The effect of growth substances applied on laterals was enhanced by defoliating the main 8tem. The influence of assimilates on flower and pod abscission and transport of growth substances is discussed.

Studies on Extension Growth in Coleoptile Sections

One of the problems involved in carrying out large-scale coleoptile cylinder straight growth assays is the effect of coleoptile age upon the behaviour of the sections. This effect, as measured both by the length of coleoptile and the time from sowing the seeds, has been investigated by an examination of the growth of sections cut from coleoptiles of several length grades on five occasions 6–10 hours apart. The response of these sections to a range of concentrations of β-indolylacetic acid (IAA) was followed during the period of growth to ascertain the most suitable time for measurement of sections in a routine test. A number of general trends associated with coleoptile age (whether measured by length, or time from planting) were observed in amount of growth, growth-rate, period of growth, and sensitivity. It is suggested that these characteristics are associated with the amount of the total growth already made by the parent coleoptile at the time of taking the section, and also that the section exhibits an ‘intrinsic growth-rate’ related to the growth-rate of the coleoptile at the time the section is cut. A period of growth of 17–20 hours seems satisfactory for routine tests except where very young coleoptiles are used; in this case growth less than that in water is sometimes observed in very low concentrations of IAA.