Effect of Darkness on Growth and Flowering of Chenopodium rubrum and C. murale Plants in vitro

Chenopodium rubrum, a short-day plant, and C. murale, a long-day plant, were grown in vitro in continuous darkness. Control C. rubrum plants exposed to continuous darkness for 15 d at cotyledonary phase, did not flower, while 80 % of plants flowered on the medium with 5 % glucose and 10 mg dm⁻³ GA₃. Control C. murale plants exposed to continuous darkness for 10 d at the age of 4^th pair of leaves, did not flower, while GA₃ (1 – 5 mg dm⁻³) stimulated flowering up to 65 %. This revised version was published online in July 2006 with corrections to the Cover Date.     

In vitro propagation ofBoswellia serrata Roxb

Anin vitro procedure for large scale multiplication ofBoswellia serrata Roxb. has been developed using cotyledonary node segments. In average 4 shoots per node were obtained on Murashige and Skoog's (MS) medium containing 0.5 mg dm⁻³ 6-benzylaminopurine (BAP) and 0.05 mg dm⁻³ napthaleneacetic acid (NAA) within 22 d. By repeated subculture technique 90–100 shoots per node could be obtained after 88 d of initial culture. Shoots could be rooted on MS medium containing 1/4 salts, 1% saccharose, and a combination of 0.5 mg dm⁻³ indole-3-butyric acid (IBA) and 0.25 mg dm⁻³ indole-3-acetic acid (IAA). Addition of antioxidants like polyvinylpyrrolidone (PVP-50 mg dm⁻³) and ascorbic acid (100 mg dm⁻³) in both multiplication and rooting media prevented browning of cultures. Approximately 80% of shoots rooted within 8–10 d. Rooted plantlets were kept for 15 d in culture bottles containing Soilrite^TM irrigated with a nutrient solution containing 1/4 MS salts and finally transferred to pots containing soil: Soilrite^TM (1∶1), mixture with 70% transplantation success.     

Effects of Some Growth Regulators on Young Iron Deficient Maize Plants

Young maize plants, grown hydroponically, were supplied with 1/10 the optimal amount of iron (0.75 mg dm^–3). Foliar treatments with solutions, containing N⁶-benzyladenine (BA), indole-3-acetic acid (IAA) or (2-chloroethyl)-trimethylammoniumchloride (CCC) were conducted after chlorosis had been well manifested. Changes in growth, chlorophyll content, rate of photosynthesis, catalase and peroxidase activities in leaves, and the contents of Fe, Cu, Zn, Mn, and P in leaves were recorded. Growth regulators improved (CCC, IAA) or aggravated (BA) the physiological state of chlorotic plants. Their effect might be explained by changes in Fe transport towards the leaves, by increased efficiency of Fe utilization, and by effects on plant metabolism not involving Fe.     

Development of Seeded and Seedless Hypanthium of Rosa Canina After Application of Growth Substances

Dog rose (Rosa canina L.) plants in the bloom stages of flowering were sprayed by indole-3-acetic acid (IAA) in concentrations of 0.06 and 0.60 mM and gibberellic acid (GA₃) in concentrations of 0.60 and 1.50 mM. Ascorbic acid, total sugar, reducing sugar and carotenoid contents gradually increased, while the protein content remained unchanged and the content of phenolic substances decreased during hypanthium development. Ascorbic acid, total sugar, reducing sugar and carotenoid contents increased in hypanthium sprayed by GA₃ and IAA. However, IAA and GA₃ applications (except low concentrations) decreased contents of phenolic substances. IAA and GA applications might be a good way to produce the high quality hypanthium in R. canina.     

The effect of selected plant hormones on in vitro proliferation of hyphae of Glomus fistulosum

Effects of N⁶-benzyladenine, kinetin, zeatin, N⁶-benzyladenosine, kinetin riboside, zeatin riboside, jasmonic acid, indole-3-acetic acid, indole-3-butyric acid, indole-3-propionic acid, abscisic acid and gibberellic acid on proliferation of hyphae of arbuscular mycorrhizal (AM) fungus Glomus fistulosum were studied under axenic conditions in vitro. The growth of intraradical hyphae of G. fistulosum was fully suppressed by 30 μM indole-3-acetic acid, but a perceptible decrease in the proliferation of the hyphae was observed already at 3 μM. Because such concentration is near the concentrations common in root tissues in vivo, the effect may be biologically significant. Similar effect was also observed for Glomus mosseae. Inhibitory effects of abscisic acid and cytokinins occurred only at very high, non-physiological concentrations. Ribosylated cytokinins showed stronger inhibition effects than their non-ribosylated counterparts. No stimulation of proliferation of hyphae by any plant hormone tested was observed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Somatic embryogenesis in Chenopodium rubrum and Chenopodium murale in vitro

In order to establish an efficient system for in vitro plant regeneration of a short day plant Chenopodium rubrum L. and a long day plant Chenopodium murale L., optimum culture conditions for somatic embryogenesis were investigated. The effects of different growth regulators, their combination and their concentrations on somatic embryos induction in different explant types (root, hypocotyl, cotyledon and leaf) were tested. Somatic embryogenesis was induced in both plants on Murashige and Skoog (MS) medium supplemented with sucrose (3 %), agar (0.7 %) and 1 - 10 M 2,4-dichlorophenoxyacetic acid (2,4-D) as the sole growth regulator. The largest embryogenic capacity was found in root explants of Chenopodium rubrum on 1 M 2,4-D and in basal parts of cotyledons in C. murale plants on 10 M 2,4-D.     

Induction of Parthenocarpy in Rosa Canina and Diospyros Lotus by the application of growth regulators

The effect of indole-3-acetic acid (IAA) and gibberellic acid (GA₃) applications on parthenocarpic fruit set in Rosa canina and Diospyros lotus was investigated. GA₃ induced parthenocarpic fruit set in both plants, but IAA only in D. lotus. Maturation of seedless fruits was earlier than the seeded fruits. GA₃ caused a decrease in the fresh mass and size of both fruits. IAA induced an increase in the fresh mass and size in parthenocarpic fruit of D. lotus. This revised version was published online in July 2006 with corrections to the Cover Date.     

The effect of indole-3-acetylaspartic acid on adventitious root formation on bean cuttings

The influence of indole-3-acetylaspartic acid (IAAsp) on rooting of stem cuttings from bean plants (Phaseolus vulgaris L.) of different ages, cultivated at different temperatures (17°, 21° and 25°C) was studied and compared to that of indole-3-acetic acid (IAA). At a concentration of 10^–4 M, IAAsp only nonsignificantly stimulated adventitious root formation, approximately to the same level as IAA in all treatments. IAAsp at 5×10^–4 M further enhanced rooting, by up 200% of control values, with little influence of temperature conditions and stock plant age. This concentration of IAA usually stimulated rooting more than the conjugate. The largest differences between the effects of IAAsp and IAA occured at the highest cultivation temperature of 25°C where stock plant age also influenced the response. The number of roots produced in comparison with the control, was enhanced from 350% on cuttings from the youngest plants to more than 600% on cuttings from the oldest. In contrast to the conjugate, 5×10^–4 M IAA induced hypocotyl swelling and injury of the epidermis at the base of cuttings, in all treatments.     

Rapid Axillary Bud Proliferation and ex vitro Rooting of Eupatorium triplinerve

Effective protocol was established for micropropagation of the medicinal plant Eupatorium triplinerve Vahl through rapid axillary bud proliferation and ex vitro rooting. Murashige and Skoog (MS) medium fortified with 8.87 M benzylaminopurine (BAP) and 2.46 M indole-3-butyric acid (IBA) was the best for axillary bud proliferation and developed a mean of 8.1 shoots per node. Excision and culture of the node segments of the in vitro shoots on medium supplemented with the same concentration of growth regulators developed more than 30 shoots within 40 d. Shoot multiplication did not exhibit decrease in the number of shoots even at 7^th subculture. Dipping of the basal end of shoots in 2.46 M IBA solution for 10 d induced roots and its transfer to small pots facilitated the survival of all rooted shoots (100 %). Ex vitro rooting by direct transfer of the shoots from multiplication medium showed 92 % survival.     

Chlorophyll-protein levels and degree of thylakoid stacking in radish chloroplasts from high-light, low-light and bentazon-treated plants

Lemna gibba plants were incubated aseptically on medium containing labelled 10^-7 M indole-3-acetic acid (IAA-1-¹⁴C). Most of the radioactivity disappeared from the culture medium during a 24 h light period. A high percentage of the loss was due to photolysis and only a low percentage of the radioactivity was recovered in the plants. Uptake of ¹⁴C by the plants was strongly stimulated by light. The radioactivity taken up by the plants was the sum of photosynthetically taken up ¹⁴CO₂ and ¹⁴C taken up in IAA. Analyses with the indolo-α-pyrone fluorescence method revealed that the free IAA content was almost the same in plants grown in control and in IAA media for 5 h, whereas the amount of IAA which could be liberated by alkaline hydrolysis was doubled by the presence of IAA in the medium.     

Effect of 2,5-norbornadiene on abscission and ethylene production in citrus leaf explants

Citrus ( Citrus sinensis L. Osbeck) leaf explants completely abscise within 48 h when exposed to saturating amounts of ethylene at 25°C. When 2,5-norbornadiene was added, 2000 μl 1⁻¹ reduced abscission of explants also exposed to 2 μl 1⁻¹ of ethylene to the level of the control, and 8000 μl 1⁻¹ reduced abscission in explants exposed to 10 μl 1⁻¹ of ethylene to the level of the control, but abscission was complete when 1 000 μl 1⁻¹ of ethylene was used in the presence of 8 000 μl 1⁻¹ of 2,5-norbornadiene. When explants were exposed to 2 μl 1⁻¹ of ethylene, 2000 μl 1⁻¹ of 2,5-norbornadiene prevented abscission if applied up to 10 h after exposure to ethylene. After 18 h, applied 2,5-norbornadiene had little effect on abscission at 48 h. A Lineweaver-Burk plot gave a 1/2 maximum value of 0.12 μl 1⁻¹ for ethylene on abscission, 2,5-Norbornadiene gave competitive kinetics with respect to ethylene with a K₁ value of approximately 120 μl 1⁻¹ of 2,5-norbornadiene. The presence of 2,5norbornadiene stimulated ethylene production, which progressively increased as the 2,5-norbornadiene concentration was increased from 250 to 8 000 μl 1⁻¹ 2,5-Norbornadiene also suppressed the induction of cellulase and polygalacturonase by ethylene. Together, 2,5-norbornadiene and 2,4-dichlorophenoxyacetic acid were more effective than either alone in reducing abscission. 2,5-Norbornadiene also was effective in preventing the reduction of indole-3-acetic acid transport induced by ethylene.     

Effects of sodium diethyldithiocarbamate, solvent, temperature and plant extracts on the stability of indoles

A study has been made of the effects of solvent, temperature, and the antioxidant, sodium diethyldithiocarbamate, on the breakdown of indole-3-pyruvic acid to indole-3-acetic acid (IAA). In addition, the degradation of tryptophan, tryptamine, indole-3-pyruvic acid, indole-3-acetaldehyde and indole-3-ethanol to IAA during the purification and analysis of extracts from Pinus sylvestris L. needles, in the presence and absence of sodium diethyldithiocarbamate, has been investigated. The data obtained indicate that if the antioxidant is supplied throughout the analytical sequence there is a marked reduction in the spontaneous formation of IAA from other indolic compounds and, by inference, the stability of indoles in general is enhanced.     

Effects of low nitrogen medium on endogenous changes in ethylene, auxins, and cytokinins in in vitro shoot formation from rhizomes of Cymbidium kanran

Summary Shoot formation from rhizome explants of Cymbidium kanran was promoted on Murashige and Skoog (MS) medium: (1) with 1 mgl⁻¹ (4.4μM) 6-benzyladenine (BA) and 0.1 mgl⁻¹ (0.54μM) α-naphthaleneacetic acid (NAA); (2) with ethylene inhibitor (silver nitrate, AgNO₃); or (3) by reducing ammonium nitrate (NH₄NO₃) and potassium nitrate (KNO₃) to 25 and 50%, respectively, of their original concentrations. Shoot formation by BA and NAA was strongly inhibited with the application of ethephon, an ethylene releaser. The ethylene production from the rhizome explants was reduced 30–55% on low nitrogen medium after 1–3 mo. of culture and 52% on BA and NAA medium after 1 mo. of culture compared with explants on standard MS medium. No difference in endogenous auxin (indole-3-acetic acid, IAA) and cytokinin (isopentenyl adenosine, iPA) contents in the rhizomes was found between the treatments. Low ethylene levels were correlated with higher frequency of shoot formation from the rhizomes.     

Alleviation of salt stress by plant growth regulators in Triticum aestivum L.

Seedlings of the salt sensitive wheat cultivar C-306 evolved more ethylene than the salt tolerant cultivar Kharchia-65 under different levels of both chloride- and sulphate-dominated types of salinity. Pre-sowing seed soaking treatments with kinetin, gibberellic acid and to a lesser extent indole-3-acetic acid alleviated salt stress effects as apparent from seedling dry mass. Treated seedlings also evolved more ethylene both under saline and non-saline conditions. Ethrel did not affect seedling growth as well as ethylene production. Abscisic acid inhibited seedling growth and ethylene production under both types of salinity. This revised version was published online in July 2006 with corrections to the Cover Date.     

Rapid in vitro Regeneration of Sesbania drummondii

This paper describes rapid propagation of Sesbania drummondii using nodal explants isolated from seedlings and young plants. The nodal segments proliferated into multiple shoots on Murashige and Skoog's (MS) medium supplemented with 22.2 M benzyladenine. MS medium containing 2.2 and 4.5 M thidiazuron induced 5 – 6 shoots per stem node from 3-month-old plants. Nodal explants when cultured on MS medium containing combinations of benzyladenine (8.8 and 11.1 M) and indole-3-butyric acid (0.24 – 2.46 M) or indole-3-acetic acid (0.28 – 2.85 M) gave lesser number of shoots. Callus induced on cotyledonary explants when subcultured on 2.2 M thidiazuron containing medium resulted in its mass proliferation having numerous embryoid-like structures. Indole-3-butyric acid (0.24 – 2.46 M) was found suitable for root induction. In vitro regenerated plants were acclimatized in greenhouse conditions.     

Endosperm dormancy breakage in olive seeds

Seeds of Olea europaea L. ssp. oleaster Hoffm. and Link freed from the sclerous endoearp and incubated in water at 15 or 25°C in darkness or in 12:12 h white light:dark conditions, did not germinate, due to dormancy imposed by the endosperm. Seeds also did not germinate when incubated in abscisic acid, gibberellic acid, kinetin or zeatin in darkness and at cither 15 or 25°C. SAN 9789 |4-chloro-5-(methylamine)-2-(a,a,a-trifluoro-m-tolyl)-3-(2H)-pyridazmone] did not promote germination at 15°C but it did to a 75% level at 25°C. This promoting effect of SAN was counteracted by abscisic acid. Cultures of naked embryos grew equally well in the presence or absence of SAN 9789. 6-Benzylaminopurine promoted whole seed germination to a 15% level.     

Effects of Growth Regulators on Direct Flowering of Isolated Ginseng Buds <Emphasis Type="Italic">in vitro</Emphasis>

An in vitro method for obtaining gingseng inflorescences directly from explants of gingseng (Panax ginseng) is reported. Isolated shoot-buds of somatic embryo-derived plantlets ginseng were used as explants and incubated in B5 medium supplemented with 1 mg l⁻¹ benzyladenine (BA) and 1 mg l⁻¹ gibberellic acid (GA₃). About 15% of the buds flowered directly without developing vegetative organs. Cytokinin was found to be the key factor for inducing these isolated buds to proliferate and flower, but both these processes also occurred when benzyladenine (BA) was replaced by thidiazuron (TDZ). The optimal concentration of TDZ for obtaining the best ratios of bud proliferation and total flowering was 0.1 mg l⁻¹, while the highest number of vegetative shoots was obtained in medium supplemented with 1 mg l⁻¹ GA₃ and 0.1 mg l⁻¹ TDZ. The explant elongated abnormally in the presence of 10 mg l⁻¹ GA₃. Although a low concentration (1 mg l⁻¹) of NAA increased the bud proliferation ratio in the medium supplemented with 0.1 mg l⁻¹ TDZ and 1 mg l⁻¹ GA₃, a high concentration (5 mg l⁻¹) of NAA reduced the bud proliferation ratio and inhibited the flowering.     

Enhancement of Lipid Peroxidation by Indole-3-Acetic Acid and Derivatives: Substituent Effects

The peroxidation of liposomes by a haem peroxidase and hydrogen peroxide in the presence of indole-3-acetic acid and derivatives was investigated. It was found that these compounds can accelerate the lipid peroxidation up to 65 fold and this is attributed to the formation of peroxyl radicals that may react with the lipids, possibly by hydrogen abstraction. The peroxyl radicals are formed by peroxidase-catalyzed oxidation of the enhancers to radical cations which undergo cleavage of the carbon-carbon bond on the side-chain to yield CO2 and carbon-centred radicals that rapidly add oxygen. In competition with decarboxylation, the radical cations deprotonate reversibly from the Nl position. Rates of decarboxylation,pK_a values and rate of reaction with the peroxidase compound I indicate consistent substituent effects which, however, can not be quantitatively related to the usual Hammett or Brown parameters. Assuming that the rate of decarboxylation of the radical cations taken is a measure of the electron density of the molecule (or radical), it is found that the efficiency of these compounds as enhancers of lipid peroxidation increases with increasing electron density, suggesting that, at least in the model system, the oxidation of the substrates is the limiting step in causing lipid peroxidation.     

Role of lndole-3-acetic Acid in the Interaction of the Phanerogamic Parasite Cuscuta and Host Plants

Abstract: During infection with the phanerogamic parasite Cuscuta reflexa the incompatible host plant Lycopersicon esculentum shows characteristic anatomical tissue modifications at the infection sites that are exclusively provoked following contact with Cuscuta - striking cell elongation is observed in the hypo-dermis and collenchyma of the host plant. Due to the influence of auxins on the process of cell elongation in plants, the role of indole-3-acetic acid (IAA) in the interaction of Cuscuta and Lycopersicon was studied using a highly specific enzyme-linked im-munosorbent assay for hormone measurement. It was shown that the tissue modifications in the host plant as well as cell elongation in the parasite tissue leading to the formation of an adhesive-secretory epithelium are correlated with increasing IAA levels in the respective tissues. Both anatomical modifications can also be induced artificially by injection of IAA into control tissue. Based on the obtained data, it can be hypothesized that during the parasitic attack IAA is accumulated in the haus-toria-bearing regions of Cuscuta and exuded from the epithelial cells. Due to the close contact between host and parasite, the hormone probably enters the host plant tissue causing the observed anatomical reactions.