Plant Growth Inhibitory Compounds from Aqueous Leachate of Wheat Straw

When seedlings of lettuce, cress, rice and wheat were incubated with the leachate of wheat straw, the roots growth of lettuce and garden cress were particularly inhibited. The leachate of wheat straw (100 g eq./l) showed 80.5 and 79.4% inhibition for lettuce and cress roots, respectively. The inhibitory activity was stronger as the concentration of wheat straw leachate was greater. This result indicates that allelochemical(s) inhibiting the roots growth of lettuce and cress are leached from the wheat straw into the water. Two potent compounds were isolated from the leachate of the wheat straw and identified as syringoylglycerol 9-O-β-d-glucopyranoside and l-tryptophan by spectral analyses. Syringoylglycerol 9-O-β-d-glucopyranoside inhibited the roots growth of lettuce and cress at concentrations greater than 0.1 and 10.0 μM, respectively. On the other hand, l-tryptophan inhibited the roots growth of lettuce and cress at concentrations greater than 0.1 and 1.0 μM, respectively. The content of syringoylglycerol 9-O-β-d-glucopyranoside and l-tryptophan in the leachate of wheat straw (100 g eq./l) was 18.4 ± 0.7 and 6.2 ± 0.6 μM, respectively. Syringoylglycerol 9-O-β-d-glucopyranoside (18.4 μM) showed 21.5 and 13.5% inhibition in the lettuce and cress roots assay, respectively. On the other hand, 6.2 μM of l-tryptophan showed 47.5 and 35.0% inhibition in the lettuce and cress roots assay, respectively. These results suggested that l-tryptophan may be a major contributor to the allelopathy in aqueous leachate of wheat straw and syringoylglycerol 9-O-β-d-glucopyranoside may be a minor contributor.     

Effects of some natural and synthetic growth regulators on sex-expression of Myrica esculenta Buch-Ham. ex D. Don

Gibberellic acid (GA,), chlorflurenol and ethrel were applied at different concentrations to either male or female trees of Myrica esculenta. GA₃ induced intersexual flowers both on the male and female trees whereas chlorflurenol and ethrel induced similar (intersexual) flowers but only on the male plants. When GA₃ was applied in combination with chlorflurenol, fewer intersexual inflorescences appeared on the male plants along with a significant decline in the number of flowers per inflorescence. In combination, chlorflurenol and ethrel induced copious female and intersexual inflorescences per male plant, especially when two successive treatments of 100 mg/I of chlorflurenol and 1920 mg/l of ethrel were applied. The induced female inflorescences later bore fruits.     

Salt stress in Thellungiella halophila activates Na+ transport mechanisms required for salinity tolerance

Salinity is considered one of the major limiting factors for plant growth and agricultural productivity. We are using salt cress (Thellungiella halophila) to identify biochemical mechanisms that enable plants to grow in saline conditions. Under salt stress, the major site of Na+ accumulation occurred in old leaves, followed by young leaves and taproots, with the least accumulation occurring in lateral roots. Salt treatment increased both the H+ transport and hydrolytic activity of salt cress tonoplast (TP) and plasma membrane (PM) H(+)-ATPases from leaves and roots. TP Na(+)/H+ exchange was greatly stimulated by growth of the plants in NaCl, both in leaves and roots. Expression of the PM H(+)-ATPase isoform AHA3, the Na+ transporter HKT1, and the Na(+)/H+ exchanger SOS1 were examined in PMs isolated from control and salt-treated salt cress roots and leaves. An increased expression of SOS1, but no changes in levels of AHA3 and HKT1, was observed. NHX1 was only detected in PM fractions of roots, and a salt-induced increase in protein expression was observed. Analysis of the levels of expression of vacuolar H(+)-translocating ATPase subunits showed no major changes in protein expression of subunits VHA-A or VHA-B with salt treatment; however, VHA-E showed an increased expression in leaf tissue, but not in roots, when the plants were treated with NaCl. Salt cress plants were able to distribute and store Na+ by a very strict control of ion movement across both the TP and PM.     

Suppression of gravitropic response of primary roots by submergence

Primary roots of six plant species were placed horizontally either in humid air or under water, and their growth and gravitropic responses were examined. In air, all the roots showed a normal gravitropic curvature. Under water without aeration, roots of rice (Oryza sativa L.), oat (Avena sativa L.), azuki bean (Vigna angularis Ohwi et Ohashi), and cress (Lepidium sativum L.) curved downward at almost same rate as in air, whereas the curvature of roots of maize (Zea mays L.) and pea (Pisum sativum L.) was strongly suppressed. Submergence did not cause a decrease in growth rate of these roots. When roots of maize and pea were placed horizontally under water without aeration and then rotated in three dimensions on a clinostat in air, they showed a significant curvature, suggesting that the step suppressed by submergence is not graviperception but the subsequent signal transmission or differential growth process. Constant bubbling of air through the water partly restored the gravitropic curvature of maize roots and completely restored that of pea roots. The curvature of pea roots was also partly restored by the addition of an inhibitor of ethylene biosynthesis, aminooxyacetic acid. In air, ethylene suppressed the gravitropic curvature of roots of maize and pea. Furthermore, the level of ethylene in the intercellular space of the roots was increased by submergence. These results suggest that the accumulation of ethylene in the tissue is at least partly involved in suppression of transmission of the gravity signal or of differential growth in maize and pea roots under conditions of submergence.Abbreviations AOA aminooxyacetic acid - 3-D three-dimensional Dedicated to Professor Andreas Sievers on the occasion of his retirementWe thank Professor H. Suge and Drs. H. Takahashi and H. Kataoka, Tohoku University and Dr. T. Suzuki, Yamagata University, for helpful suggestions. The present study was supported in part by a Grant for Basic Research in Space Station Utilization from the Institute of Space and Astronautical Science, Japan.     

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.     

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.     

Identification of two phytotoxins, blumenol A and grasshopper ketone, in the allelopathic Japanese rice variety Awaakamai

Aqueous methanol extracts of the traditional rice (Oryza sativa) variety Awaakamai, which is known to have the greatest allelopathic activity among Japanese traditional rice varieties, inhibited the growth of roots and shoots of cress (Lepidium sativum), lettuce (Lactuca sativa), timothy (Phleum pratense), Digitaria sanguinalis, Lolium multiflorum and Echinochloa crus-galli. Increasing the extract concentration increased the inhibition, suggesting that the extract of Awaakamai contains growth inhibitory substances. The extract of Awaakamai was purified and two main growth inhibitory substances were isolated and determined by spectral data as blumenol A and grasshopper ketone. Blumenol A and grasshopper ketone, respectively, inhibited the growth of cress shoots and roots at concentrations greater than 10 and 30 μmol/L. The concentrations required for 50% growth inhibition on cress roots and shoots were 84 and 27 μmol/L, respectively, for blumenol A, and 185 and 76 μmol/L, respectively, for grasshopper ketone. These results suggest that blumenol A and grasshopper ketone may contribute to the growth inhibitory effect of Awaakamai and may play an important role in the allelopathy of Awaakamai.     

The effects of a morphactin,chlorflurenol, on organ redifferentiation from tobacco calluses culturedin vitro

A certain morphactin, chlorflurenol (methyl-2-chloro-9-hydroxyfluorene-9-carboxylic acid) was found to be effective in inducing organ redifferentiation (shoot formation) from calluses derived from the pith parenchyma ofNicotiana tabacum ‘Wisconsin No. 38.’ The regenerative ability of chlorflurenol was strongly intensified in combination with kinetin. A combination of 1 mg/l of chlorflurenol and 2 mg/l of kinetin resulted in shoot formation in 70% of inoculated calluses. One reason for this high frequency of regeneration seems to be the promotion of tracheary elements, probably due to some synergism of chlorflurenol and kinetin. Chromosome numbers of the root-tip meristematic cells in regenerated plants varied widely over a range of 6 to 98. One regenerated plant showed normal chromosome constitution in the first metaphase of meiosis, and showed normal pollen and seed fertility; but the others remained unstable and had low pollen and seed fertility.     

An allelopathic substance in red pine needles (Pinus densiflora)

Aqueous methanol extracts of red pine (Pinus densiflora) needles inhibited the growth of roots and shoots of cress (Lepidium sativum), lettuce (Lactuca sativa), alfalfa (Medicago sativa), ryegrass (Lolium multiflorum), timothy (Pheleum pratense), Digitaria sanguinalis and Echinochloa crus-galli. Increasing the extract concentration increased inhibition, suggesting that the pine needles may have growth inhibitory substances and possess allelopathic potential. The aqueous methanol extract of the pine needles was purified, and a main inhibitory substance was isolated and determined by spectral data as 9alpha,13beta-epidioxyabeit-8(14)en-18-oic acid. This substance inhibited root and shoot growth of cress and Echinochloa crus-galli seedlings at concentrations greater than 0.1 mM. The endogenous concentration of the substance was 0.13 mmol/kg pine needle. These results suggest that 9alpha,13beta-epidioxyabeit-8(14)en-18-oic acid may contribute to the growth inhibitory effect of the pine needles and may play an important role in the allelopathy of red pine.     

Organ-specific-active allelopathic substance in red pine needles

An organ-specific-growth inhibitory substance was isolated from an aqueous methanol extract of red pine needles and determined by spectral data as 1-mono(16-hydroxyhexadecanoyl)glycerol. This substance inhibited root growth of cress (Lepidium sativum L.) and barnyard grass (Echinochloa crus-galli (L.) Beauv) seedlings at concentrations greater than 0.01 and 0.03???M, respectively. The concentrations required for 50?% growth inhibition on roots of cress and barnyard grass were 0.16 and 3.4???M, respectively. However, the inhibitory activity of the substance on shoots of cress and barnyard grass was very weak. The endogenous concentration of 1-mono(16-hydroxyhexadecanoyl)glycerol in the pine needles was 4.6???mol?kg^?1. Two related compounds, 1-monohexadecanoylglycerol and 16-hydroxyhexadecanlic acid had no activity up to 1,000???M on cress roots and shoots. The effectiveness of 1-mono(16-hydroxyhexadecanoyl)glycerol on root growth inhibition and the occurrence of 1-mono(16-hydroxyhexadecanoyl)glycerol in pine needles suggest the substance may play an important role in the allelopathy of red pine. Root-specific-growth-inhibition by the substance may be one of the strategies for red pine to compete with neighboring plants for nutrients and space because root growth of competitive plants may be very important for their whole plant development.     

The effects of 3,5-diiodo-4-hydroxybenzoic acid on the oxidation of IAA and auxin-induced ethylene production by cress root segments

Summary In previous research here, 3,5-diiodo-4-hydroxybenzoic acid (DIHB) was shown to promote the elongation of roots of cress (Lepidium sativum) seedlings growing in light, and to inhibit the auxin-induced production of ethylene in this tissue. Although DIHB is a cofactor for the oxidation of indole-3-acetic acid (IAA) by horse-radish peroxidase, it inhibits the decarboxylation of [1-¹⁴C]IAA by segments excised from cress roots. The inhibition by DIHB of ethylene production by this tissue does not, therefore, arise from a reduction of IAA levels. These findings are discussed in relation to the effects of DIHB on cress root growth.Abbreviations IAA indole-3-acetic acid - DIHB 3,5-diiodo-4-hydroxybenzoic acid - DCP 2,4-dichlorophenol - 2,4-D 2,4-dichlorophenoxyacetic acid This study forms part of a research project to be submitted by M.L.R. for PhD degree and supported by a grant from Consejo Nacional de Ciencia y Tecnología (México).     

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 a potent allelopathic substance in Bangladesh rice

Aqueous methanol extracts of Bangladesh rice (Oryza sativa L. cv. BR17) inhibited the growth of roots and shoots of cress (Lepidium sativum), lettuce (Lactuca sativa), alfalfa (Medicago sativa), timothy (Phleum pratense), Digitaria sanguinalis, Echinochloa crus-galli and Echinochloa colonum. Increasing the extract concentration increased the inhibition, suggesting that the BR17 may have growth inhibitory substances and possess allelopathic potential. The aqueous methanol extract of the BR17 was purified and a main inhibitory substance was isolated and determined by spectral data as 2,9-dihydroxy-4-megastigmen-3-one. This substance inhibited root and shoot growth of cress and E. crus-galli seedlings at concentrations greater than 0.03 and 3 μM, respectively. The concentrations required for 50% growth inhibition on cress roots and shoots were 0.22 and 0.47 μM, respectively, and on E. crus-galli roots and shoots were 36 and 133 μM, respectively. These results suggest that 2,9-dihydroxy-4-megastigmen-3-one may contribute to the growth inhibitory effect of BR17 and may play an important role in the allelopathy of BR17. Thus, Bangladesh rice BR17 may be potentially useful for weed management in a field setting. An erratum to this article can be found at     

Changes in plant growth processes under microgravity conditions simulated by a three-dimensional clinostat

We developed a three-dimensional (3-D) clinostat to simulate a microgravity environment and studied the changes in plant growth processes under this condition. The rate of germination of cress (Lepidium sativum), maize (Zea mays), rice (Oryza sativa), pea (Pisum sativum), or azuki bean (Vigna angularis) was not affected on the clinostat. The clinostat rotation did not influence the growth rate of their roots or shoots, except for a slight promotion of growth in azuki roots and epicotyls. On the contrary, the direction of growth of plant organs clearly changed on the 3-D clinostat. On the surface of the earth, roots grow downward while shoots upward in parallel to the gravity vector. On the 3-D clinostat, roots of cress elongated along the direction of the tip of root primordia after having changed the direction continuously. Rice roots also grew parallel to the direction of the tip of root primordia. On the other hand, roots of maize, pea, and azuki bean grew in a random fashion. The direction of growth of shoots was more controlled even on the 3-D clinostat. In a front view of embryos, shoots grew mostly along the direction of the tip of primordia. In a side view, rice coleoptiles showed an adaxial (toward the caryopsis) while coleoptiles of maize and epicotyls of pea and azuki bean an abaxial curvature. The curvature of shoots became larger with their growth. Such an autotropism may have an important role in regulation of life cycle of higher plants under a microgravity environment.     

Plant-growth inhibitory activity of heliannuol derivatives

In addition to (+)-, (-)- and (+/-)-heliannuol E, growth-inhibitory activities of five synthetic chromans and four tetrahydrobenzo[b]oxepins were examined against oat and cress. All heliannuol E isomers exhibited similar biological activities against cress, whereas when tested against oat roots, the unnatural optical isomer (+) showed no inhibitory activity. Four brominated chromans and two tetrahydrobenzo[b]oxepin derivatives also showed apparent inhibition against both cress and oat.     

Ethylene production by cress roots and excised cress root segments and its inhibition by 3,5-diiodo-4-hydroxybenzoic acid

Summary 3,5-Diiodo-4-hydroxybenzoic acid (DIHB) has been shown to exert an inhibitory effect on the formation of ethylene by the roots of intact cress Lepidium sativum seedlings in light, and by excised cress root segments. Adding IAA to the culture solution greatly promoted ethylene production, which was suppressed by DIHB. The findings together with results obtained with dinitrophenol (DNP), L-methionine and L-ethionine and also the horseradish peroxidase/methional system of Yang are discussed.The results indicate that the effect of DIHB in promoting the root growth of cress seedlings in nutrient solution in the light operates, at least in part, by suppressing the formation of the root growth inhibitor ethylene.Abbreviation GLC gas-liquid chromatography - dIHB 3,5-diiodo-4-hydroxybenzoic acid This study forms part of research to be submitted for PhD degree and supported by a grant from Consejo Nacional de Ciencia y Tecnología (México).     

THE PHYSIOLOGICAL BASIS OF MORPHOLOGICAL POLARITY IN REGENERATION. II

1. The experiments show that the mass of air roots formed in a stem increases with the mass of the leaf attached to the stem, though it has not been possible to establish an exact mathematical relation between the two masses, owing to unavoidable sources of error. 2. Darkened leaves do not increase the mass of roots formed. 3. In stems suspended horizontally air roots appear on the lower side of the stem, with the exception of the cut end where they usually appear around the whole circumference of the stem. When the lower half of a stem suspended horizontally is cut off, roots are formed on the upper side. It is shown by experiments on leaves suspended horizontally that the more rapidly growing roots and shoots on the lower side inhibit the root and shoot formation in the upper half of such a leaf; and likewise the more rapid formation of roots on the lower side of a horizontally suspended stem seems to account for the inhibition of root formation on the upper side of such a stem. Likewise the more rapid growth of shoots on the upper side of a stem suspended horizontally is likely to inhibit the growth of shoots on the lower side. 4. Each leaf contains in its axil a preformed bud capable of giving rise to a root, which never grows out in the normal stem on account of the inhibitory influence of the normal roots at the base of the plant. These dormant root buds are situated above (apically from) the dormant shoot bud. The apical root buds can be caused to develop into air roots when a piece of stem is cut out from a plant from which the leaves except those in the basal node of the piece are removed. The larger these basal leaves the better the experiments succeed. 5. These apical air roots grow out in a few days, while the roots at the basal end of the stem (which in our experiments dip into water) grow out about a week later. As soon as the basal roots grow out in water they cause the air roots in the more apical region of the stem to dry out and to disappear. 6. In addition to the basal roots, basal nodes have also an inhibitory effect on the growth of the dormant root buds in the apical region of a stem. This is indicated by the fact that a stem with one pair of leaves near the base will form apical air roots more readily when no node is situated on the stem basally from the leaf than if there is a node basally from the leaf.     

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.     

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.     

Rice seedlings release momilactone B into the environment

Since the growth inhibitor momilactone B was found recently in root exudates of rice (Oryza sativa L.), 3-day-old rice seedlings were transferred to hydroponic culture and the level of momilactone B released into the environment from the seedlings was measured. At day 15 after transfer, the level of momilactone B in the culture solution was 1.8 nmol per seedling compared with endogenous levels of 0.32 and 0.63 nmol per root and shoot, respectively, suggesting that rice seedlings actively releases momilactone B into the culture solution. This release must occur from the roots because only rice roots were immersed in the culture solution. Momilactone B inhibited the growth of ten cress (Lepidium sativum L.) seedlings at concentrations greater than 3 microM. Ten rice seedlings were incubated with ten cress seeds in a Petri dish containing 1 ml of medium, the medium contained 18 nmol of momilactone B, which came to 18 microM. This level of momilactone B was enough to reveal growth inhibition of the cress seedlings. Release level of momilactone B and its effectiveness as a growth inhibitor suggest that it may play an important role in rice allelopathy.