Difference in hydroxamic acid content in roots and root exudates of wheat (Triticum aestivum L.) and rye (Secale cereale L.): Possible role in allelopathy

Hydroxamic acids (Hx) produced by some cereal crops have been associated with allelopathy. However, the release of Hx to the soil by the producing plant-an essential condition for a compound to be involved in allelopathy-has not been shown. GC and HPLC analysis of roots and root exudates of wheat (Triticum aestivum L.) and rye (Secale cereale L.) cultivars, with high Hx levels in their leaves, demonstrated the presence of these compounds in the roots of all cultivars analyzed and in root exudates of rye. Moreover, bioassays employing root exudates collected from wheat and rye seedlings demonstrated that only rye exudates inhibited root growth of wild oats,Avena fatua L., a weed whose root growth is inhibited by Hx. These results suggest that rye could potentially interfere with the growth ofAvena fatua in nature and that this interference could be due to the release of Hx to the soil by way of roots.     

Allelopathic polyacetylenes fromCentaurea repens (Russian knapweed)

The allelopathic weed Russian knapweed (Centaurea repens) was found to contain polyacetylenes VIII–XIV in the roots. Dose vs. response of the root length elongation against lettuce, alfalfa, barnyard grass, and red millet showed IX to be active. Closely related isomers were not active. Examination of the soil surrounding the Russian knapweed roots revealed the presence of IX in sufficient concentration to have an appreciable effect on the surrounding plant community.     

Evidence for allelopathy by tree-of-heaven (Ailanthus altissima)

Ailanthus altissima (Mill.) Swingle contains one or more phytotoxic compounds in roots and leaves. Activity is higher in roots, where it occurs primarily in the bark. Powdered root bark and leaflets strongly inhibited growth of garden cress (Lepidium sativum L.) when mixed with soil in Petri dishes (ID₅₀ values=0.03 g root bark, 0.6 g leaflet/dish). The toxic material was readily extracted by methanol but not dichloromethane. Pieces of root bark mixed with soil at 2, 1, and 0.5 g/pot reduced cress biomass in the greenhouse, whereas methanol-extracted root bark did not. The inhibitory effect ofAilanthus tissues in soil was short-lived (4 weeks in pots in greenhouse, 3 days in Petri dishes in laboratory). Inhibition by root bark was sometimes superseded by stimulation. FreshAilanthus root segments placed in or on soil reduced growth of nearby cress seedlings. Fine roots were more inhibitory than coarse, and inhibition became more pronounced with increased time of soil exposure to roots. Soil collected nearAilanthus roots in the field supported reduced radicle growth of cress compared to control soil. In contrast, stemflow fromAilanthus trees stimulated cress growth. The results suggest allelopathy caused by toxin exudation from roots may contribute to the aggressiveness and persistence ofAilanthus in certain habitats.     

Distribution and Exudation of Allelochemicals in Wheat Triticum aestivum

Wheat allelopathy has potential for weed suppression. Allelochemicals were identified in wheat seedlings, and they were exuded from seedlings into agar growth medium. p-Hydroxybenzoic, trans-p-coumaric, cis-p-coumaric, syringic, vanillic, trans-ferulic, and cis-ferulic acids and 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) were identified in both the shoots and roots of 17-day-old wheat seedlings and their associated agar growth medium. Wheat accessions with previously identified allelopathic activity tended to contain higher levels of allelochemicals than poorly allelopathic ones. The allelopathic compounds present in the shoots generally also were identified in the roots and in the agar medium. Allelochemicals were distributed differentially in wheat, with roots normally containing higher levels of allelochemicals than the shoots. When the eight allelochemicals were grouped into benzoic acid and cinnamic acid derivatives, DIMBOA, total coumaric, and total ferulic acids, the amount of each group of allelochemicals was correlated between the roots and the shoots. Most of the allelochemicals identified in the shoots and roots could be exuded by the living roots of wheat seedling into the agar growth medium. However, the amounts of allelochemicals in the agar growth medium were not proportional to those in the roots. Results suggest that wheat plants may retain allelochemicals once synthesized. The presence of allelochemicals in the agar growth medium demonstrated that wheat seedlings were able to synthesize and to exude phytotoxic compounds through their root system that could inhibit the root growth of annual ryegrass.     

Allelopathic potential of sorghum-sudangrass hybrid (sudex)

Experiments were conducted under controlled conditions to investigate the apparent allelopathic effects of sudex [Sorghum bicolor (L.) Moench ×Sorghum Sudanese (P.) Stapf, cv. FFR 201] on weed and vegetable species. Allelopathic potential, as measured by radicle elongation of herbaceous indicator species, decreased with increasing sudex age. Greatest potential allelopathic activity of sudex shoot tissue was observed when sudex was collected at 7 days of age. Small-seeded broadleaf species were more inhibited in the presence of sudex shoot tissue than were grass species. Two major phytoinhibitors were isolated from aqueous extracts of sudex shoot material by partitioning with diethyl ether, followed by thin-layer and liquid column chromatography. Phytoinhibitors were identified asp-hydroxybenzoic acid andp-hydroxybenzaldehyde, potentially the enzymatic breakdown products of the cyanogenic glycoside dhurrin. The I₅₀ values of these compounds using a cress (Lepidium sativum L.) seed bioassay were 140 and 113 g/ml for the acid and aldehyde, respectively. Sudex tissue collected at 7 days of age possessed a greater percentage of these phytoinhibitors on a per gram basis than did older sudex tissue. As sudex tissue age increased, the percentage ofp-hydroxybenzaldehyde in ether extracts of tissue also increased, while the percentage ofp-hydroxybenzoic acid decreased.Journal article No. 88-10-163 of the Kentucky Agricultural Experiment Station.     

Allelochemicals fromPolygonum sachalinense Fr. Schm. (Polygonaceae)

The root exudates fromPolygonum sachalinense in a recirculating system significantly inhibited lettuce seedling growth. The rhizomes and roots ofP. sachalinense were extracted with 80% acetone. Bioassay of the neutral-acidic fraction on the TLC agar plate showed the inhibitory activity corresponded to the two yellow pigment bands. Two orange needles were isolated and identified as anthraquinone compounds: emodin and physcion. Both compounds exhibited inhibitory activities against the seedling growth of several testing plant species. Glucosides were isolated fromP. sachalinense and were identified as emodin-1-O-β-D-glucoside and physcion-1-O-β-D-glucoside, respectively. On plant growth bioassay, these glucosides showed no phytotoxic activity against lettuce seedlings. The concentrations of emodin, physcion, and their glucosides from rhizome with roots, aerial parts, fallen leaves, and soil were determined. The rhizome with roots and fallen leaves contained emodin and physcion at relatively high concentrations. Emodin also occurs in the soil of this plant community with effective concentrations in the fall. The results indicate that these anthraquinones are responsible for the observed interference and are potent allelopathic substances.     

Allelopathic Potential of Aquatic Plants Associated with Wild Rice (Zizania palustris): I. Bioassay with Plant and Lake Sediment Samples

The allelopathic potential of eight aquatic plants associated with wild rice was investigated using lettuce and wild rice seedling bioassays. Rhizome aqueous extracts of Scirpus acutus, Potamogeton natans, Nymphaea odorata, Nuphar variegatum; shoot extract of Eleocharis smallii; whole plant extract of Myriophyllum verticillatum; and leaf extract of P. natans significantly reduced the root length of lettuce and wild rice seedlings. The lettuce seedling bioassay was more sensitive than the wild rice bioassay. Shoot growth was less affected than the root growth. Water extract of sediments associated with the aquatic plants had little growth inhibitory effect on wild rice. Our study did not yield any conclusive evidence that the wild rice-associated aquatic plants have allelopathic effects on wild rice. We emphasize the use of target species as a bioassay material in allelopathic studies. Further investigation on allelopathic effects of lake sediments associated with the neighboring plants of wild rice is necessary to evaluate their ecological significance.     

Phytotoxic substances in root exudates of cucumber (Cucumis sativus L.)

The addition of activated charcoal to a nutrient solution for the hydroponic culture of cucumber resulted in significant increases in the dry weight of the plant and fruit yield. Hydrophobic root exudates were collected at different growth stages with Amberlite XAD-4 resin and bioassayed with lettuce seedlings. The exudates at the reproductive stage were more phytotoxic than those at the vegetative stage. The exudates contained organic acids such as benzoic,p-hydroxybenzoic, 2,5-dihydroxybenzoic, 3-phenylpropionic, cinnamic,p-hydroxycinnamic, myristic, palmitic, and stearic acids, as well asp-thiocyanatophenol and 2-hydroxybenzothiazole, all of which, except 2-hydroxybenzothiazole, were toxic to the growth of lettuce.     

Isothiocyanates as alleopathic compounds fromRorippa indica Hiern. (Cruciferae) roots

The ethyl acetate extracts ofRorippa indica Hiern. contained hirsutin, arabin, camelinin, and three novel -methylsulfonylalkyl isothiocyanates (n=8, 9, and 10). These compounds severely inhibited lettuce (Lactucasaliva) hypocotyl and root growth at 0.1 mM or above. The precursor glucosinolates of hirsutin, arabin, and camelinin were isolated. Presence of the three -methylsulfonylalkylglucosinolates, along with other glucosinolates in the roots were verified by the isolation and identification of their desulfoderivatives. Using the continuous root exudate trapping apparatus and GC-MS, hirsutin and the three-methlylsulfonylalkyl isothiocyanates were detected in the root exudates ofR. indica, suggesting that these isothiocyanates are the primary candidate of allelopathic compounds contributing to the aggressiveness of this cruciferous weed.     

Induction and/or Selection of Phenolic Acid-Utilizing Bulk-Soil and Rhizosphere Bacteria and Their Influence on Phenolic Acid Phytotoxicity

Bulk-soil and rhizosphere bacteria are thought to exert considerable influence over the types and concentrations of phytotoxins, including phenolic acids, that reach a root surface. Induction and/or selection of phenolic acid-utilizing (PAU) bacteria within the bulk-soil and rhizosphere have been observed when soils are enriched with individual phenolic acids at concentrations 0.25 mol/g soil. However, since field soils frequently contain individual phenolic acids at concentrations well below 0.1 mol/g soil, the actual importance of such induction and/or selection remains uncertain. Common bacteriological techniques (e.g., isolation on selective media, and plate dilution frequency technique) were used to demonstrate in Cecil A_p soil systems: (1) that PAU bacterial communities in the bulk soil and the rhizosphere of cucumber seedlings were induced and/or selected by mixtures composed of individual phenolic acids at concentrations well below 0.25 mol/g soil; (2) that readily available carbon sources other than phenolic acids, such as glucose, did not modify induction and/or selection of PAU bacteria; (3) that the resulting bacterial communities readily utilize mixtures of phenolic acids as a carbon source; and (4) that depending on conditions (e.g., initial PAU bacterial populations, and phenolic acid concentration) there were significant inverse relationships between PAU bacteria in the rhizosphere of cucumber seedlings and absolute rates of leaf expansion and/or shoot biomass. The decline in seedling growth could not be attributed to resource competition (e.g., nitrogen) between the seedlings and the PAU bacteria in these studies. The induced and/or selected rhizosphere PAU bacteria, however, reduced the magnitude of growth inhibition by phenolic acid mixtures. For a 0.6 mol/g soil equimolar phenolic acid mixture composed of p-coumaric acid, ferulic acid, p-hydroxybenzoic acid, and vanillic acid, modeling indicated that an increase of 500% in rhizosphere PAU bacteria would lead to an approximate 5% decrease (e.g., 20–25%) in inhibition of absolute rates of leaf expansion. As far as we know, this is the first time that such a relationship has been quantified.     

Evaluation of some weed extracts against field dodder on alfalfa (Medicago sativa)

Experiments were conducted to evaluate the effectiveness of some weed extracts against field dodderCuscuta campestris Yunck. on alfalfa (Medicago stiva L.) and to identify and quantify the phytotoxic agents of these extracts. All concentrations of aqueous extract of every weed showed significant effectiveness on dodder when compared to the untreated plant under lath house and field conditions. Control percentages of the (0.5 g) of Bermuda grass (Cynodon dactylon) and wall goosefoot (Chenopodium murale) ranged between 83 and 96, and the same concentration caused injury to alfalfa foliage up to 43% when applied in the field. Phytotoxic agents were identified as phenolic compounds such as chlorogenic, isochlorogenic,p-coumaric acids, and scopoletin. Their quantities varied with species; Bermuda grass had the highest content (32.2 g/g dry weight) followed by Johnson grass (Sorghum halepense), well goosefoot, and tumble pigweed (Amaranthus albus). These results might aid in screening for effective alternate approaches for controlling dodder on alfalfa planted for seeds.     

Allelopathic effects ofChrysanthemum morifolium on germination and growth of several herbaceous plants

Aqueous extracts obtained from young green tops ofChrysanthemum morifolium inhibited the germination of six flowering plants, including chrysanthemum itself, provided for experiments. The same phenomenon was also clearly observed when powder made from young green tops and old leaves of chrysanthemum was used. Moreover, the growth of seedlings planted again in garden soil which was once used for the culture of chrysanthemum was greatly interrupted. Chrysanthemum cultured in used garden soil showed far less dry weight than that cultured in fresh garden soil. The weight of chrysanthemum cultured using its root exudates was also less than that cultured with water leachate of fresh garden soil, and therefore these results may be considered to indicate allelopathic effects. In order to find the allelochemicals related to this phenomenon, benzoic acid and phenolic acids such as salicylic,p-hydroxybenzoic, vanillic, gentisic, protocatechuic, syringic, gallic, ferulic, and caffeic acids were identified by gas chromatography.     

Role of allelopathy as a possible factor associated with the rising dominance ofBunias orientalis L. (Brassicaceae) in some native plant assemblages

Leaf extracts ofBunias orientalis were shown to inhibit seed germination of a variety of cultivar plant species and of species cooccurring withB. orientalis in the field. Root exudate solutions and leaf litter leachates ofB. orientalis were tested for their allelopathic activity using seedling growth assays. Additionally, in comparative seedling growth assays soil cores removed from denseB. orientalis stands were tested bimonthly for elevated allelopathic effects. The impact of root exudates on seedling growth was generally weak and varied between species. Similar results were obtained for the effect ofB. orientalis leaf litter leachates on seedlings grown in sand culture relative to the effect of leaf litter leachates of a plant species mixture. When soil as a growth substrate was used, no consistent differences in seedling growth were obtained between the two litter leachate treatments. In the soil core experiment seedlings grown in soil cores collected from a denseB. orientalis stand unexpectedly showed better performance than seedlings grown in soil cores collected from a nearby mixed plant stand withoutB. orientalis, at least in early spring and late autumn. Predominating nutrient effects are, therefore, assumed to conceal a potentially increased allelopathic effect of soil beneath denseB. orientalis stands. It is concluded that other factors than allelopathy must be investigated to explain the rapid establishment of dense stands of this alien plant species.     

Effects of mixtures of phenolic acids on leaf area expansion of cucumber seedlings grown in different pH portsmouth A1 soil materials

Cucumber seedlings growing in A₁horizon Portsmouth soil material adjusted to pH 5.2, 6.0, or 6.9 were treated with 0, 0.25 or 0.5, mol/ g soil ferulic acid,p-coumaric acid, vanillic acid, or an equal mixture (0.5 mol/g total) of two acids every other day. A total of five treatments was given starting with day 7 from seeding. Absolute rates of leaf expansion were determined for seedlings. The experiment was terminated when seedlings were 17 days old. All three phenolic acids inhibited leaf expansion. The dose required for 50% inhibition of absolute rates of leaf expansion increased as pH of the soil systems increased. The order of toxicity based on 50% dose and relative potency were as follows: ferulic acid > vanillic acid =p-coumaric acid. Effects of mixtures of phenolic acids on absolute rates of leaf expansion, when compared to the effects of individual phenolic acids, were found to be antagonistic for the ferulic-vanillic acid mixture and the ferulic-p-coumaric acid mixture in the pH 5.2 soil systems. Several phenolic acid treatments were required before antagonistic effects of mixtures were evident. In all other instances, when treatment effects were significant, the effects of individual phenolic acids were additive.Paper No. 11875 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, North Carolina 27695-7601. The use of trade names in this publication does not imply endorsement by North Carolina Agricultural Research Service of the product named, nor criticism of similar products not mentioned. This research was partially supported by US-Spain Joint Committee for Scientific and Technological Corporation project CCA-8309/166.     

Autointoxication mechanism ofOryza sativa I. Phytotoxic effects of decomposing rice residues in soil

The aqueous extracts of decomposing rice residues in soil exhibited inhibition on the radicle growth of lettuce and rice seeds and the growth of rice seedlings. The phytotoxicity was found in extracts obtained from the early stage of decomposition (first month), and gradually declined thereafter. The inhibition was also found in extracts obtained from rice fields, and was persistent for 4 months. The root initiation of hypocotyl cuttings of mungbeans was suppressed by extracts of decaying rice residues and extracts obtained from paddy soil. Five phytotoxins,p-hydroxybenzoic,p-coumaric, vanillic, ferulic, ando-hydroxyphenylacetic acids, and several unknowns were found in the decomposing rice residues under waterlogged conditions. At 25 ppm,o-hydroxyphenylacetic acid revealed significant inhibition on the radicle growth of rice and lettuce seeds and suppressed root initiation of mungbean seedlings. It was concluded that the growth of rice seedlings was retarded by decaying rice residues in soil; thus, this appeared to be an autointoxication phenomenon.Paper No. 176 of the Scientific Journal Series, Institute of Botany, Academia Sinica. This study was financially supported by the National Science Council, the Republic of China.     

Effect of gamma irradiation on allelopathic potential ofSorghum bicolor against weeds and nitrification

The effect of low doses of gamma irradiation on the allelopathic potential ofSorghum bicolor against weeds and nitrification was investigated. The results revealed that all test doses (500, 1000 and 1500 rad) significantly increased the allelopathic activity of root exudates, aqueous extracts, and decaying residues aganst seed germination and seedling growth ofAmaranthus retroflexus. The results also indicated that all test doses stimulated the allelopathic potential of aqueous extracts and decaying residues against nitrification activity. The possible application of this approach in biological control is briefly discussed.     

Effects of soil nitrogen level on ferulic acid inhibition of cucumber leaf expansion

It has been suggested that the allelopathic activity of phenolic acids should be primarily important in soils of low fertility. If this is true, then plant growth inhibition by phenolic acids may be unimportant in managed agricultural soils. The objective of this study was to determine how soil nitrogen (N) level might modify phenolic acid inhibition of growth. Cucumber seedlings (Cucumis sativus cv Early Green Cluster) grown in containers in growth chambers under varying N levels (5, 10, 15, 20, and 25 g N/g soil) in Portsmouth B,-horizon soil material were treated with ferulic acid (0 or 10 g/g soil). Nitrogen and ferulic acid (FA) were applied every other day to the soil surface. The amount of FA in the soil solution declined with depth in the containers. A more rapid disappearance of FA from the soil solution was observed for the last FA treatment (0% recovered after 10 hr on day 23) than the first treatment (44% recovered after 10 hr on day 13). Both low N (5 g N/g soil) and FA treatments reduced shoot dry weight, the mean absolute (AGR) and the mean relative (RGR) rates of leaf expansion, and increased the root-shoot ratio. High N treatments reduced shoot dry weight and the AGR. Ferulic acid inhibited cucumber seedling growth over a range of N concentrations, suggesting that the allelopathic activity of phenolic acids may be important in both nutrient limiting and nonlimiting soils for some species.Paper No. 12219 of the journal series of the North Carolina Agricultural Research Service, Raleigh, North Carolina 27695-7601. The use of tradenames in this publication does not imply endorsement by the North Carolina Agricultural Research Service of products named, nor criticism of similar ones not mentioned.     

Defoliation Affects Chemical Defenses in All Plant Parts of Rye Seedlings

The effect of defoliation and consecutive defoliation (condefoliation) of rye seedlings on the allocation patterns of biomass and hydroxamic acids (Hx) was evaluated five days after treatment. Growth of condefoliated seedlings was lower than that of defoliated and nondefoliated ones. Concentration of Hx decreased in shoots of condefoliated seedlings compared to nondefoliated ones, while concentration of Hx in root exudates increased. Allocation of Hx to roots and root exudates increased at the expense of allocation to the shoots in condefoliated seedlings. The ratio of Hx-aglucone to Hx-glucoside was higher in shoots of defoliated and condefoliated seedlings. The decrease in quantity of defense in shoots was accompanied by an increase in its quality, given that aglucones are more toxic than glucosides. The increase in concentration of Hx—an allelopathic compound also involved in mineral uptake by roots—in root exudates of condefoliated seedlings was suggested to lead to an advantage in the acquisition of resources for the regeneration of lost biomass.     

Growth Inhibitory Effects of Nutgrass (Cyperus rotundus) on Rice (Oryza sativa) Seedlings

Growth inhibitory effects of aqueous extracts and leachates of leaves and tubers of Cyperus rotundus L. were investigated by using rice (Oryza sativa L.) seedling as a bioassay material. Both the extracts and leachates of Cyperus were inhibitory to the growth of rice seedlings. Growth inhibition was more pronounced in the presence of aqueous extracts than the leachates. The extract and leachate of leaves had higher total phenolic contents than those of the tubers. Soil amendment with fresh leaves of Cyperus reduced plant height, leaf area, and root and shoot weight of rice seedlings. Total phenolic content was higher in soil amended with fresh Cyperus leaves than the unamended control soil. Nineteen compounds were tentatively identified from the aqueous extracts of leaves and tubers by ethyl acetate extraction followed by gas chromatography–mass spectroscopy (GC-MS). Dicarboxylic, phenolic, and fatty acids were the major compounds. Our results suggest that Cyperus may affect the growth and establishment of rice seedlings after sowing or transplanting, especially when Cyperus plants are mixed in soil during land preparation by ploughing in rain-fed rice culture.     

Inhibitory effect of parthenium (Parthenium hysterophorus L.) residue on growth of water hyacinth (Eichhornia crassipes mart solms.) I. Effect of leaf residue

The allelopathic effect of parthenium (Parthenium hysterophorus L.) leaf residue (dry leaf powder, DLP) on water hyacinth (Eichhornia crassipes Mart Solms.) was studied. The treatment caused wilting starting from the margins of the older leaves and desiccation of above-water plant parts (shoot). Appearance, persistence, and disappearance of symptoms depended on the level and duration of the treatment and recovery of the treated plants, if it occurred. The treatment drastically reduced the number of healthy leaves (HLN) and the plant biomass at 0.25% (w/v) DLP; the treated plants recovered in about one month. At and above 0.50% (w/v) DLP, the plants were killed in about one month, resulting in sinking of the dead mass in water. Physiological effects of the treatment included deterioration of membrane integrity, loss of dehydrogenase activity with concurrent drastic reduction or total failure of water absorption by the roots, and reduction of chlorophyll contents in the leaves. The results indicate that the inhibitors leached out of the DLP affected the water hyacinth plants through changes in macromolecules: protein, lipid, and nucleic acid, resulting in root dysfunction and other inhibitory activities both in the root and shoot. Phenolic and other inhibitors including those found in the parthenium plant (except sesquiterpene lactones which have not been tested) at 50 ppm, exceptp-hydroxybenzoic acid, did not affect the treated plants. Such a high concentration of the allelochemicals is unlikely to be present in the medium at the lethal dose (0.50% w/v) of the DLP. Even withp-hydroxybenzoic acid, the plants recovered subsequently and grew normally. Thus, it appears that other allelochemicals including sesquiterpene lactones were mainly responsible for the inhibitory activity of the DLP on water hyacinth plants.