Auxin-like Growth Activity of 3-Phenylpropionitrile from Water-Cress (Nasturtium officinale R. Br.)

Solutions of 3-phenylpropionitrile (PPN) and 3-phenylpropionicacid (PPA) exhibited auxin-like plant growth activity by stimulatingelongation of sections cut from wheat coleoptiles and garden-cresshypocotyls. Elongation of sugar-beet hypocotyl sections wasstimulated by PPA but not by PPN. Although PPA stimulated elongationof pea-epicotyl sections after 19 h incubation, most concentrationsof PPN did not show activity until 70 h. PPN generally stimulatedadventitious root formation on hypocotyls of garden-cress andsugar-beet, and on epicotyls of pea seedlings whereas dilutesolutions of PPA were inactive and concentrated solutions weretoxic. Steam distillates of water-cress plants contained a substancewith auxin-like growth activity and chromatographic propertiessimilar to PPN. Nasturtium officinale R. Br. water-cress, 3-phenylpropionitrile, 3-phenylpropionic acid, auxins     

Potent endogenous allelopathic compounds in Lepidium sativum seed exudate: effects on epidermal cell growth in Amaranthus caudatus seedlings

Many plants exude allelochemicals--compounds that affect the growth of neighbouring plants. This study reports further studies of the reported effect of cress (Lepidium sativum) seed(ling) exudates on seedling growth in Amaranthus caudatus and Lactuca sativa. In the presence of live cress seedlings, both species grew longer hypocotyls and shorter roots than cress-free controls. The effects of cress seedlings were allelopathic and not due to competition for resources. Amaranthus seedlings grown in the presence of cress allelochemical(s) had longer, thinner hypocotyls and shorter, thicker roots--effects previously attributed to lepidimoide. The active principle was more abundant in cress seed exudate than in seedling (root) exudates. It was present in non-imbibed seeds and releasable from heat-killed seeds. Release from live seeds was biphasic, starting rapidly but then continuing gradually for 24 h. The active principle was generated by aseptic cress tissue and was not a microbial digestion product or seed-treatment chemical. Crude seed exudate affected hypocotyl and root growth at ~25 and ~450 μg ml(-1) respectively. The exudate slightly (28%) increased epidermal cell number along the length of the Amaranthus hypocotyl but increased total hypocotyl elongation by 129%; it resulted in a 26% smaller hypocotyl circumference but a 55% greater epidermal cell number counted round the circumference. Therefore, the effect of the allelochemical(s) on organ morphology was imposed primarily by regulation of cell expansion, not cell division. It is concluded that cress seeds exude endogenous substances, probably including lepidimoide, that principally regulate cell expansion in receiver plants.     

Isolation and identification of an allelopathic substance in Pisum sativum

The residue of peas (Pisum sativum L.) has allelopathic activity and the putative compound causing this inhibitory effect was isolated from a methanol extract of pea shoots. Chemical structure of this compound was determined by high-resolution MS, IR and 1H NMR spectral data as pisatin. Pisatin inhibited growth of cress (Lepidium sativum L.) and lettuce (Lactuca sativa L.) seedlings at concentrations greater than 10 and 30 microM, respectively. The doses required for 50% growth inhibition of roots and hypocotyls of cress were 61 and 91 microM, respectively, and those of lettuce were 78 and 115 microM, respectively. The concentration of pisatin in the pea shoots was 32.7 nmol x g(-1) fresh weight. The effectiveness of pisatin on growth inhibition in cress and lettuce, and its occurrence in pea shoots suggest that it may contribute to the growth inhibitory effect of pea residue, and may play an important role in pea allelopathy.     

Physiological activities of helminthosporol in comparison with those of gibberellin and auxin

Summary Helminthosporol, a metabolic product of Helminthosporium sativum, remarkably stimulated elongation of the second leaf sheath of rice seedlings of various varieties. It also stimulated the elongation of hypocotyls of cucumber seedlings. This substance did not have any effect on the growth of peas, tomato, sunflower and the dwarf mutants of maize. This substance did not have auxin-like activities.     

Aspects of the biological activity of surfactants that are potential eradicants of apple mildew

Among several surface active agents tested, both anionic and cationic materials usually caused greater changes in the microflora of treated soil than non-ionic materials, although the effects of a non-ionic formulated mixture of aliphatic alcohols (Off-Shoot T) were considerable. In vitro tests with micro-organisms and earthworms indicated similar differences in response to types of compound. There was a positive correlation between hydrophobicity and biological activity of a series of alkyl phenol ethoxylates in some tests. Although many surfactants caused gross phytotoxicity to cress (Lepidium) seedlings in the laboratory, the growth of barley and pea seedlings in treated soil was significantly decreased only by one cationic material.     

Specificity patterns indicate that auxin exporters and receptors are the same proteins

A study of transport and action of synthetic auxin analogues can help to identify transporters and receptors of this plant hormone. Both aspects--transportability and action on growth--were tested with 2-naphthoxyacetic acid (2-NOA) and compared across several plant species. 2-NOA stimulates elongation effectively at low concentrations in petioles of the gymnosperm Ginkgo biloba L., in hypocotyls or internodes of the dicot legumes, mung bean (Vigna mungo L.) and pea (Pisum sativum L.), in cotyledons of onion (Allium cepa L.) and in leaf bases of chive (Allium schoenoprasum L.), the latter two of the monocot order Asparagales. In contrast, elongation of coleoptile segments of maize (Zea mays L.) is poorly responsive to 2-NOA. Significant auxin-like transport of 2-NOA was observed in segments of mung bean hypocotyls, pea internodes, and chive leaf bases, but not in segments of the grass coleoptiles. Thus, for the two assays, elongation and polar transportability, the same difference in ligand specificity was observed between the grass and all other species assayed. This finding supports the hypothesis that a common protein mediates auxin efflux as well as auxin action on elongation.     

Effects of Capsaicin on Plant Growth

Rice (Oryza sativa L.) seedlings inhibited the growth of hypocotyls and roots of cress (Lepidium sativum L.) seedlings when both seedlings were grown together. Two growth inhibiting substances were found in the culture solution in which rice seedlings were hydroponically grown for 14 d. One growth inhibitor was further purified. This suggests that the rice seedlings may produce growth inhibiting substances, acting as allelochemicals to other plants, and release them from their roots into the environment.     

Occurrence of osmiophilic particles is correlated to elongation growth of higher plants

Summary The occurrence of elongation growth-related osmiophilic particles (OPs) was investigated in hypocotyls of sunflower, bean, and spruce as well as in pea epicotyls and in cress roots of intact seedlings. In all analyzed species, OPs were found to occur specifically within the periplasmic space between plasma membrane and the outer epidermal cell walls of elongating parts of hypocotyls, epicotyls, and roots, whereas cells of nonelongating parts were devoid of OPs. Auxin (IAA) markedly increased the number of OPs in epicotyl and hypocotyl segments. Treatment of pea epicotyl segments with the lectin concanavalin A inhibited their elongation growth in the presence of IAA. At a subcellular level this effect was characterized by the occurrence of a pronounced osmiophilic layer in the periplasmic space of the outer periclinal and the outer part of the anticlinal epidermal cell walls. Treatment of IAA-incubated segments with the secretion inhibitor brefeldin A inhibited both elongation growth and periplasmic occurrence of OPs. This effect was accompanied by complementary accumulation of OPs in the peripheral cytoplasm of epidermal cells. Together the results indicate that IAA-induced epidermis-specific secretion of OPs is closely related to cell elongation growth not only in organs of monocotyledonous species, but also in dicotyledonous angiosperms as well as in gymnosperms.Abbreviations OPs osmiophilic particles - ConA concanavalin A - BFA brefeldin A - IAA -indolyl acetic acid     

The depression of the synthesis of pea diamine oxidase due to light and the verification of its participation in growth processes using competitive inhibitors

The time courses of the synthesis of diamine oxidase in pea plants grown for 14 days either in the light or in the dark are similar with the highest increase in activity occurring in the cotyledons and in the shoots during the first 6 to 8 days. Plants grown in the dark showed a 2- to 3-fold higher enzyme activity than plants grown in the light. Pea diamine oxidase could bein vivo efficiently inhibited by substrate analogues 1,4-diamino-2-butanone and 1,5-diamino-3-pentanone. The first compound inhibited proportionally to its concentration the growth of etiolated pea plants, but its instability makes an unequivocal interpretation of the results difficult. On the other hand, 1,5-diamino-3-pentanone a stable and more efficient diamine oxidase inhibitor depressed the growth of pea seedlings only at concentrations as high as 5 mM and 10 mM, at which the growth of cress seedlings not containing diamine oxidase was also strongly depressed. The results obtained indicate that tryptamine oxidation catalyzed by diamine oxidase is not involved in the main metabolic pathway leading from tryptophan to indoleacetate in pea plants.     

Biological evaluation of 5-substituted pyrimidine derivatives as inhibitors of brassinosteroid biosynthesis

A series of 5-substituted pyrimidine derivatives was synthesized, and their ability to inhibit brassinosteroid biosynthesis was tested. The biological activity of these compounds was evaluated by the cress stem elongation method. Among the synthesized compounds, alpha-(4-chlorophenyl)-alpha-phenyl-5-pyrimidinemethanol (DPPM 4) exhibited potent inhibitory activity for retarding cress stem elongation in the light. This inhibition was reversed by the application of 10 nM brassinolide, but not by 1 microM GA3. DPPM 4 also affected Arabidopsis growth in the dark. DPPM 4-treated Arabidopsis had phenotypes like those of brassinosteroid-deficient mutants, with short hypocotyls and open cotyledons, in the dark. These biological changes were restored by the co-application of 10 nM brassinolide, but not by 1 microM GA3, suggesting that the primary site of action of DPPM 4 was the brassinosteroid biosynthetic pathway.     

The Geotropic Curvature in Roots of Norway Spruce (Picea abies) Containing Anthocyanins

Seedlings of Norway spruce (Picea abies L.) have been found to synthesize anthocyanins in the root tips as well as in the hypocotyls upon irradiation with white light when kept at 4°C for 6–8 days. In addition, it has also been found that the elongation and the geotropic curvature of spruce roots are dependent on the light conditions. The course of the geotropic curvature in spruce roots containing anthocyanins has been followed during a period of 5 h, in which the seedlings were geotropically stimulated continuously in the horizontal position. When the stimulation was performed in white light and in darkness at 21°C, significantly larger curvatures were observed in the roots pretreated at 4°C in darkness than in the roots containing anthocyanins. The specific curvature (curvature in degrees per mm elongation), however, was approximately the same in both types of roots stimulated in white light. This was due to a retarded elongation of the roots pretreated with light at 4°C and containing anthocyanins. A smaller difference in elongation rate between roots with and without anthocyanins was observed in the dark than in the light, but even in the dark the anthocyanin-containing roots grew more slowly than roots without anthocyanins. In order to find out if it is the anthocyanin content or the illumination which affects the elongation and geotropic curvature in the roots, a series of similar experiments was performed using cress seedlings grown at 4°C in light or darkness. Roots of cress seedlings cultivated under conditions which would induce anthocyanin formation in spruce roots exhibited the highest geotropic responses both in light and darkness as compared to cress seedlings grown at 4°C in darkness. As in the case of spruce roots an increase in elongation was observed in cress roots illuminated during the geotropic stimulation. These similarities in the behaviour made it relevant to compare the development of the geotropic curvature in cress and spruce roots.     

Developmental and environmental regulation of tissue- and cell-specific expression for a pea protein farnesyltransferase gene in transgenic plants

Farnesylation mediates membrane targeting and in vivo activities of several key regulatory proteins such as Ras and Ras-related GTPases and protein kinases in yeast and mammals, and is implicated in cell cycle control and abscisic acid (ABA) signaling in plants. In this study, the developmental expression of a pea protein farnesyl-transferase (FTase) gene was examined using transgenic expression of the β-glucuronidase (GUS) gene fused to a 3.2 kb 5′ upstream sequence of the gene encoding the pea FTase β subunit. Coordinate expression of the GUS transgene and endogenous tobacco FTase β subunit gene in tobacco cell lines suggests that the 3.2 kb region contains the key FTase promoter elements. In transgenic tobacco plants, GUS expression is most prominent in meristematic tissues such as root tips, lateral root primordia and the shoot apex, supporting a role for FTase in the control of the cell cycle in plants. GUS activity was also detected in mature embryos and imbibed embryos, in accordance with a role for FTase in ABA signaling that modulates seed dormancy and germination. In addition, GUS activity was detected in regions that border two organs, e.g. junctions between stems and leaf petioles, cotyledons and hypocotyls, roots and hypocotyls, and primary and secondary roots. GUS is expressed in phloem complexes that are adjacent to actively growing tissues such as young leaves, roots of light-grown seedlings, and hypocotyls of dark-grown seedlings. Both light and sugar (e.g. sucrose) treatments repressed GUS expression in dark-grown seedlings. These expression patterns suggest a potential involvement of FTase in the regulation of nutrient allocation into actively growing tissues.     

Neutral growth inhibitors in light-grown dwarf pea shoots –separation, identification and growth regulation

To correlate endogenous growth inhibitors of dwarf pea to its dwarfism a thorough search of growth inhibitors was made in the neutral fraction of an acetone extract from the shoots of light-grown seedlings of Pisum sativum L. cvs Progress No 9 and Alaska. From both cultivars six inhibitors were separated and named A-1, A-2, A-3, B-1, B-2 and B-3 based on their order of elution from the silica gel column. Their contents as determined by cress root bioassay were: in cv. Progress 0.40, 16.5, 6.36, 1.02, 0.11 and 0.10, and in cv. Alaska 0.33, 2.35, 3.51, 0.95, 0.10 and 0.09 cress units (g fresh weight)⁻¹. Their contributions to growth regulation of the relevant pea plants were estimated as the products of the above-stated contents times the ratios of the specific activities of each standard sample in the cress roots and the relevant pea cultivars, and they were; in cv. Progress A-2, 5.44 and A-3, 2.10, and in cv. Alaska A-2, 0.68 and A-3, 0.88, those of A-2 and A-3 constituting more than 90% of the total contribution of the six inhibitors in either cultivar. The great differences in the content and contribution to growth regulation of A-2 and A-3 between the two cultivars suggest that the higher contents of and greater responsiveness to the two inhibitors in cv. Progress may be causes of dwarfism of this cultivar. A-l and A-3 were spectroscopically identified with pisatin and a mixture of cis, trans– , and trans, trans xanthoxins.     

Slow wave potentials in cucumber differ in form and growth effect from those in pea seedlings

A positive hydraulic signal in the form of a xylem pressure step was applied to the roots of intact seedlings of Cucumis sativus L. and Pisum sativum L. Surface electrodes at three positions along the epicotyl/hypocotyl recorded a propagating depolarization which appeared first in the basal, then the central and sometimes the apical electrode positions and fitted the characteristics of a slow wave potential (SWP). This depolarization differed between pea and cucumber. It was transient in cells of pea epicotyls but sustained in cucumber hypocotyls. It was not associated with a change in cell input resistance in pea epicotyls but preceded an increase in the input resistance of cucumber hypocotyl cells. With the increased xylem pressure the growth rate (GR) of cucumber hypocotyls and pea epicotyls underwent a transient increase, peaking after 5 min. If the depolarization reached the growing upper region, it preceded a sustained decrease in the GR of cucumber hypocotyls but only a transient decrease in the GR of pea epicotyls. A temperature jump in the root medium (heat treatment) induced a steep pressure spike in the xylem of the cucumber hypocotyl which showed similar electric and growth effects as the previously applied, non-injurious pressure steps. We suggest that the observed differences in the electric and growth responses between the species were caused by the closure of ion channels in depolarized cells of cucumber but not pea seedlings.     

Effect of IAA on Growth and Soluble Cell Wall Polysaccharides Centrifuged from Pine Hypocotyl Sections

Auxin-induced elongation and cell wall polysaccharide metabolism were studied in excised hypocotyl sections of ponderosa pine (Pinus ponderosa) seedlings. Sections excised from hypocotyls of ponderosa pine elongate in response to the addition of auxin. The neutral sugar composition of the extracellular solution removed from hypocotyl sections by centrifugation was examined. In cell wall solution from freshly excised sections, glucose, galactose, xylose, and arabinose make up more than 90% of the neutral sugars, while rhamnose, fucose, and mannose are relatively minor components. The neutral sugar composition of the polysaccharides of the pine cell wall solution is both qualitatively and quantitatively similar to that of pea. Following auxin treatment of pine hypocotyls, the neutral sugar composition of the cell wall changes; glucose, xylose, rhamnose, and fucose increase by nearly 2-fold relative to controls in buffer without auxin. These changes in neutral sugars in response to auxin treatment are similar to those found in pea, with the exception that in pea, rhamnose levels decline in response to auxin treatment.     

Antioxidant enzymes and isoflavonoids in chilled soybean (Glycine max (L.) Merr.) seedlings

Changes of activity antioxidant enzymes and of levels of isoflavonoids were studied in the roots and hypocotyls of the etiolated soybean (Glycine max (L.) Merr. var. Essor) seedlings, submitted to cold. Prolonged exposure to 1 degrees C inhibited hypocotyl and root elongation and limited their growth after seedlings were transferred to 25 degrees C. Roots were more sensitive to chilling than hypocotyls. At 1 degrees C a gradual increase in MDA concentration in roots but not in hypocotyls was observed. An increase in catalase (CAT, EC 1.11.1.6) and superoxide dismutase (SOD, EC 1.15.1.1) activity in hypocotyls was observed both at 1 degrees C and after transfer of plants to 25 degrees C. In roots, CAT activity increased after 4 days of chilling, while SOD activity only after rewarming. L-Phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) activity decreased in roots of chilled seedlings, but did not change in hypocotyls until activity increased after transfer to 25 degrees C. The content of genistein and daidzein increased after 24 h of treatment by low temperature and then decreased with prolonged chilling in hypocotyls and remained high in roots. However, it should be noted that genistin level (genistein glucoside) in chilled hypocotyls is 10 times higher than in roots, despite falling tendency. The role of antioxidant enzymes and isoflavonoids in preventing chilling injury in hypocotyls and roots of soybean seedlings is discussed.     

Photoregulation of hypocotyl growth: geotropic evidence for the operation of two photosystems

Abstract. The rate of curvature of etiolated cress ( Lepi-dium sativum L. ) hypocotyls in response to gravity (negative geotropism) was retarded by red or blue light; far-red irradiation was without effect. The timing of the irradiation period in relation to the presentation for geostimulus markedly affected the response. When seedlings were irradiated during the 1–2 h period of geostimulus, blue light was more effective than red at retarding curvature; when seedlings were irradiated prior to geostimulus, only red light affected geocurvature. These results are interpreted as a further example of the kinetically distinct effects of red and blue light on hypocotyl development. Blue light elicited a rapid, immediate response effective only during the period of irradiation; red light induced a response characterized by a lag period and persistence in subsequent darkness. Etiolated mustard seedlings showed similar responses to light and gravity. The results are discussed in relation to the possibility that two photosystems operate in hypocotyl growth.     

Brassinolide, a growth-promoting steroidal lactone

Brassinolide (BR), a naturally-occurring-steroidal lactone from rape ( Brassica napus L.) pollen, was compared with auxin for activity in a number of bioassay systems. Responses similar to IAA were elicited by BR in bioassays based upon bean hypocotyl hook opening, elongation of maize mesocotyl, pea epicotyl and azuki bean epicotyl sections, and fresh weight increase in Jerusalem artichoke (2,4-D used) and pea epicotyl sections. The azuki bean and dwarf pea epicotyl bioassays were much more responsive to BR than IAA (at 10 μ M ). Responses approximately two-fold greater in magnitude were elicited by IAA in the maize mesocotyl, bean hypocotyl hook and Jerusalem artichoke bioassays. Little or no response was elicited by BR (0.01 to μ M ) in the cress root or decapitated pea-lateral bud bioassays. A powerful synergism between BR and IAA was observed in the azuki bean, pea epicotyl and bean hypocotyl hook bioassays. Although, as previously reported, other steroidal substances are active in some of the bioassay systems tested, none compared with BR in magnitude and diversity of elicited responses.     

On the interaction of growth retardants with IAA and kinetin

In the pea test a highly positive response to the treatment with IAA reversed to a negative one or became 5 to 6 times weaker when CCC was applied together with IAA. In cultivating pea seedlings, following their decapitation, for two days in a 0.25 per cent CCC solution and then in water, growth of their cotyledonous axillaries (cotylaries) were inhibited. This inhibitive action of CCC could be made ineffective when the seedlings, following two-days’ cultivation in the CCC solution, were grown further in kinetin solutions (0.37–3 mg per 1). Cotylaries of decapitated pea seedlings, when grown in kinetin solutions were inhibited. With kinetin solutions of 6–12 mg/l a strong inhibition also occured in the growth of roots at the apical parts of which spherical swellings were developing. The CCC supplied to the roots of intact etiolated pea seedlings is translocated acropetally into the stem at a rate of about 5 cm per hour. Decapitation of the plant causes retardation of this transport, yet a coat of 0.00001–1% IAA or kinetin paste produces acceleration of the stream. Existence of an antagonism between CCC and IAA, demonstrated earlier, was found holding true also for B-9 (N, N-dimethyl-aminesuccinamic acid) and IAA, as the inhibitive action of B-9, 0.06% solution on the growth of lettuce hypocotyls was reduced to a highly significant degree when the plants were supplied with B-9 together with IAA at a concentration of 10 mg/l.     

Phytochrome control of β-fructosidase activity in radish

Soluble and cell-wall fractions from radish seedlings show β-fructosidase activity. Light (via phytochrome) induces a transfer of the enzyme from the cytosol to the cell-wall in roots and hypocotyls. Light also increases β-fructosidase activity in hypocotyls. Cycloheximide prevents the increase of β-fructosidase activity but has no effect on the transfer.