Phytotoxic properties of nordihydroguaiaretic acid,a lignan fromLarrea tridentata (Creosote bush)

The phytotoxic properties of nordihydroguaiaretic acid (NDGA) isolated from creosote bush,Larrea tridentata (Zygophyllaceae), were examined. NDGA dramatically reduces the seedling root growth of barnyard grass, green foxtail, perennial ryegrass, annual ryegrass, red millet, lambsquarter, lettuce, and alfalfa, and reduces the hypocotyl growth of lettuce and green foxtail. It has no effect on the germination of lettuce seeds. NDGA almost certainly contributes to the observed allelopathic nature of creosote bush.Work performed during sabbatical year leave at Western Regional Research Center.     

1-Cyano-2-Hydroxy-3-Butene, A Phytotoxin From Crambe (Crambe abyssinica) Seedmeal

Crambe (Crambe abyssinica) seedmeal was found to suppress seedling emergence and biomass accumulation when added to a sandy loam soil containing wheat (Triticum aestivum Cardinal) and hemp sesbania (Sesbania exahata) seeds. Hexane, CH₂Cl₂, MeOH, and water extracts of the seedmeal were prepared and bioassayed against wheat and velvetleaf (Abutilon theophrasti) radicle elongation. The CH₂Cl₂ extract was the most inhibitory, while the other extracts inhibited the bioassay species only slightly (MeOH) or not at all (hexane, water). Fractionation of the CH₂Cl₂ extract identified the major phytotoxin as l-cyano-2-hydroxy-3-butene (CHB), comprising 96.1% of the active CH₂Cl₂ fraction. Radicle elongation of wheat and velvetleaf were inhibited by CHB with I₅₀ values of 2.1 × 10^–4 M for wheat and 2.7 × l0^–3 M for velvetleaf.     

Allelopathic effect of parthenium (Parthenium hysterophorus L.) extract and residue on some agronomic crops and weeds

Allelopathic effects of entire shoot extract, plant part extracts, and shoot residue of parthenium (Parthenium hysterophorus L.) on corn (Zea mays L.), ryegrass (Lolium multiflorum Lam.), wheat (Triticum aestivum L.), velvetleaf (Abutilon theophrasti Medik.), and soybean [Glycine max (L) Merr.] growth were examined. Parthenium shoot contained water-soluble materials that were toxic to root growth of velvetleaf and wheat. At 4% (w/ v) concentration, root growth of velvetleaf and wheat were reduced by 60 and 75%, respectively. The order of increasing sensitivity to parthenium was ryegrass, corn, wheat, and velvetleaf. There was a strong correlation between extract concentration and increased toxicity to test species. The toxicity of plant part extracts was also concentration dependent. At 1 and 2% (w/v), the inflorescence and leaves caused more root inhibition than stem extract. Parthenium shoot incorporated in soil at 1% (w/w) caused significantly more root inhibition of wheat than soybean, corn, and ryegrass. At 4% (w/w), root growth of all the test species was inhibited compared to the control. Toxicity of parthenium residue to wheat diminished with increasing periods of decomposition. Residue decomposed for four weeks was less toxic than the undecomposed residue.Florida Agricultural Experiment Stations Journal Series No. 7506.     

Physiological Assessment and Path Coefficient Analysis to Improve Evaluation of Alfalfa Autotoxicity

Reseeding of alfalfa is affected until autotoxic chemicals break down or are dispersed, often requiring a year or more. Bioassays of seed germination and early seedling growth, on agar medium in petri dishes, were conducted to evaluate autotoxic responses of 20 alfalfa germplasms to water-soluble extracts of alfalfa leaf tissue. Root length, 120 hr after placing imbibed seed on agar, was more sensitive to the autotoxin(s) than was hypocotyl length, germination speed, and final germination percentage. Path coefficient analyses showed variation in root length had 7–17 times more effect than variation in hypocotyl length in determining autotoxic effects on total seedling length. Although variations in seed size and germination rate were negatively associated (P < 0.05) with final root length, the autotoxin had little effect on these factors relative to that on root length. Germplasms in the control differed (P < 0.05) in root length, requiring tolerance to be evaluated as percent of control. Germplasms, as percent of control, differed significantly (P<0.05) at extract concentrations of 1.0 and 4.0 g l^–1, but the range and LSD were more favorable for selection at 1.0 g l^–1. Root length is appropriate for genetic assessments of tolerance to the autotoxin when expressed as percent of control.     

Biological activity and tentative identification of flavonoid components in velvetleaf (Abutilon theophrasti Medik.) seed coats

The biological activity of aqueous extracts of velvetleaf (Abutilon theophrasti Medik.) seed coats and their flavonoid components against three plant species and five soil fungi was investigated. Aqueous extracts slightly inhibited germination and significantly inhibited radicle growth of all plant species tested. Fungal growth was inhibited or not affected, depending on species, by aqueous extracts. The aqueous extracts were extracted into methanol and separated using paper chromatography for identification of compounds responsible for inhibition of seedling and fungal growth. Six flavonoid compounds were isolated and tentatively identified as delphinidin, cyanidin, quercetin, myricetin, (+)-catechin, and (–)-epicatechin. Bioassays revealed that the flavonoid compounds significantly inhibited germination and radicle growth of all test species at a concentration of 1.0 mM. Individual flavonoids had variable effects on fungi but appeared to inhibit growth and sporulation of potential seed-decomposing fungi rather than beneficial fungi. These results indicate that the flavonoid complement of velvetleaf seed coats may function in a dual defensive role against competing seedlings and seed-attacking fungi.Journal article No. 9878 of the Missouri Agricultural Experiment Station.     

Isolation of substance from sweet potato (Ipomoea batatas) periderm tissue that inhibits seed germination

Chromatographic procedures were used to isolate inhibitors of seed germination from sweet potato root periderm tissue. The inhibitory activity of all fractions was monitored using a proso millet seed germination bioassay. A single HPLC peak, representing approximately 1.2% of the periderm dry weight, accounted for most of the inhibitory activity. The active fraction was labile in methanolic solution. Further fractionation of this peak by HPLC methods was not successful. In vitro seed germination dose-response relationships were established for the peak. The various seed species exhibited an extremely wide range of sensitivity. The I₅₀ values were 0.16, 0.013 and 0.011 mg/ml for redroot pigweed, velvetleaf, and proso millet, respectively. Tall morning glory was not inhibited by any concentration tested.     

Phytotoxicity of water-soluble substances from alfalfa and barley soil extracts on four crop species

Problems associated with continuously planting alfalfa (Medicago saliva L.) or seeding to thicken depleted alfalfa stands may be due to autotoxicity, an intraspecific form of allelopathy. A bioassay approach was utilized to characterize the specificity and chemical nature of phytotoxins in extracts of alfalfa soils as compared to fallow soil or soil where a cereal was the previous crop. In germination chamber experiments, water-soluble substances present in methanol extracts of soil cropped to alfalfa or barley (Hordeum vulgare L.) decreased seedling root length of alfalfa L-720, winter wheat (Triticum aestivum L. Nugaines) and radish (Raphanus sativa L. Crimson Giant). Five days after germination, seedling dry weights of alfalfa and radish in alfalfa soil extracts were lower compared to wheat or red clover (Trifolium pralense L. Kenland). Growth of red clover was not significantly reduced by soil extracts from cropped soil. Extracts of crop residue screened from soil cropped to alfalfa or barley significantly reduced seedling root length; extracts of alfalfa residue caused a greater inhibition of seedling dry weight than extracts of barely residue. A phytotoxic, unidentified substance present in extracts of crop residue screened from alfalfa soil, which inhibited seedling root length of alfalfa, was isolated by thin-layer chromatography (TLC). Residues from a soil cropped continuously to alfalfa for 10 years had the greatest phytotoxic activity.     

A quantitative feeding bioassay forPissodes strobi Peck (Coleoptera: Curculionidae)

A feeding bioassay forPissodes strobi Peck was developed that consisted of plastic petri dishes containing paired agar disks immersed in paraffin wax. Lens paper covering the top surface of the disks provided 1, 2, or 3 weevils in each replicate with a surface through which they made regular feeding punctures which could be easily counted. The number of feeding punctures was correlated with the amount of agar ingested. Candidate feeding stimulants or deterrents were applied to the paper covering one of the disks in the dishes, while the other served as a solvent control. Feeding stimulants were tested using disks of pure agar, and deterrents were assayed on agar disks that contained 2% ground, dried Sitka spruce bark,Picea sitchensis Carr. The weevils exhibited a concentration-dependent response to the amount of spruce bark in the agar disk. Feeding stimulation by Sitka spruce bark extracts, and deterrency by leaf oil of western red cedar,Thuja plicata Donn, was demonstrated. The bioassay would be useful in chemical isolation programs aimed at identification of feeding stimulants and deterrents.Research supported by National Research Council of Canada Cooperative Grant A0243 and Operating Grants A3881 and A3706.     

Allelopathic potentials of alfalfa (Medicago sativa) saponins: Their relation to antifungal and hemolytic activities

A wheat seedling bioassay was used to indicate the relationship between the chemical structure of alfalfa saponins and their allelopathic activity. None of the tested saponins significantly influenced wheat germination. Seedling growth bioassays indicated significant differences among the activities of individual glycosides. The most active were medicagenic acid, its glycosides substituted at the C-3 position with glucose, and hederagenin monoglycoside. Medicagenic acid glycosides, substituted at the C-3 position with glucuronic acid, and zahnic acid tridesmoside were less active. No significant correlation was found among the allelopathic, hemolytic, and antifungal activities of the individual glycosides.     

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.     

Correlation Between Phytotoxicity on Annual Ryegrass (Lolium rigidum) and Production Dynamics of Allelochemicals Within Root Exudates of an Allelopathic Wheat

An improved allelopathic correlation between phytotoxicity measured in root growth bioassay upon annual ryegrass (Lolium rigidum Gaud.) and the concentrations of a selection of dynamically produced allelochemicals quantified in the root exudates of cv. Khapli wheat (Triticum turgidum ssp. durum (Desf.) Husn.) monitored during the first 15 days of wheat seedling growth in a sterile, agar–water medium, has been established. Changes over the 15-day growth period in the quantities of five exuded benzoxazinones and seven phenolic acids were measured simultaneously using GC/MS/MS. Substantiating pure compound dose–response measurements were conducted over a range of concentrations for the putative allelochemicals within the wheat exudates. One synergism-based proposal using the monitored compounds to explain the observed low-exudate-concentration phytotoxicity was explored, but was found to be experimentally inadequate.     

HPLC identification of allelopathic compounds fromLantana camara

Aqueous extracts ofLantana camara L. leaves inhibited ryegrass (Lolium multiflorum Lam.) germination and seedling growth. Phytotoxic compounds were fractionated from crude aqueous extracts and fractions were evaluated for their phytotoxicity. Inhibition was most pronounced with the alkaline and acid hydrolysates. Plant inhibition by the crude extract reflected a complex interaction of numerous individual components of diverse chemical compositions and potencies. Presumptive identification of the individual components was accomplished with high-performance liquid chromatography (HPLC). Thirteen phenolic compounds were identified, and most of these compounds were phytotoxic to ryegrass seedlings. Radicle elongation was more sensitive to the toxins than shoot elongation.Florida Agricultural Experiment Station Journal Series No. 7906.     

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.     

Lack of chemical factors in host-plant resistance of alfalfa towards alfalfa weevil: Ovicidal activity of some coumarin derivatives

A resistant line of alfalfa was studied in an attempt to identify possible chemical factors responsible for alfalfa weevil (Hypera brunneipennis) resistance. Bioassays were developed to measure ovicidal, larval development, pupation, and adult development rates. Serial alfalfa extracts of a weevil-resistant line showed no effect in the bioassays when compared to those of a weevil-susceptible line. It was concluded that resistance substances are not present but that resistant lines are simply more tolerant to weevil attack. Alfalfa constitutents and synthetic analogs were also screened in the egg hatch bioassay. Among these were a series of 3-acyl-4-hydroxycoumarins. 3-Acyl-4-hydroxycoumarins with short-chain acyl groups showed good ovicidal rates, but activity decreased with increasing length of the acyl group.     

Allelopathy in agroecosystems: Wheat phytotoxicity and its possible roles in crop rotation

The germination rates of cotton and wheat seeds were significantly affected by various extracts of wheat mulch and soils collected from the wheat field. This toxicity was even more pronounced against seedling growth. Five allelochemics: ferulic,p-coumaric,p-OH benzoic, syringic, and vanillic acids, were identified from the wheat mulch and its associated soil. Quantitatively, ferulic acid was found at higher concentrations thanp-coumaric acid in the soil. Various concentrations of ferulic andp-coumaric acids were toxic to the growth of radish in a bioassay. The functional aspects of allelochemic transfer from decaying residue to soil and the subsequent microbial degradation within agroecosystems are discussed, particularly as they relate to wheat crop rotation, with wheat and cotton, in Pakistan.     

Allelopathic effect of alfalfa (Medicago sativa) on bladygrass (Imperata cylindrica)

Greenhouse and laboratory experiments were conducted at the Agricultural and Water Resources Research Center Station, Baghdad, in 1985 and 1986 to investigate the possible allelopathic potential of alfalfa (Medicago saliva L.) and its decomposed residues on bladygrass (Imperata cylin-drica L. Beauv.), a noxious weed in Iraq, and to isolate, characterize, and quantify possible allelopathic agents in alfalfa residues and root exudates. Results indicated that decomposed alfalfa roots and their associated soil produced a 51–56% reduction in bladygrass seed germination. Root and shoot length of bladygrass seedlings were reduced by an average of 88%. Decayed and undecayed mixtures of alfalfa roots and soil at 0.0151 (w/w) inhibited bladygrass seedlings reproduced from rhizomes by 30 and 42%. It was found that root exudates of alfalfa seedlings caused significant reduction in shoot and root dry weights of bladygrass seedlings when alfalfa and bladygrass were grown together in nutrient culture. Caffeic, chlorogenic, isochloro-genic,p-coumaric,p-OH-benzoic, and ferulic acids were detected in alfalfa root exudates and residues. The highest amount (126 fig phenolic acids/g soil) of these compounds was found in alfalfa root residues after six months of decomposition in soil.     

Bioassay of naturally occurring allelochemicals for phytotoxicity

The bioassay has been one of the most widely used tests to demonstrate allelopathic activity. Often, claims that a particular plant species inhibits the growth of another are based entirely on the seed germination response to solvent extracts of the suspected allelopathic plant; few of these tests are of value in demonstrating allelopathy under natural conditions. The veracity of the bioassay for evaluating naturally occurring compounds for phytotoxicity depends upon the physiological and biochemical response capacity of the bioassay organism and the mechanism(s) of action of the allelochemicals. The possibility that more than one allelochemical, acting in concert at very low concentrations, may be responsible for an observed allelopathic effect makes it imperative that bioassays be extremely sensitive to chemical growth perturbation agents. Among the many measures of phytotoxicity of allelochemicals, the inhibition (or stimulation) of seed germination, radicle elongation, and/or seedling growth have been the parameters of choice for most investigations. Few of these assays have been selected with the view towards the possible mechanism of the allelopathic effect.     

Isolation of feeding deterrents against argentine stem weevil from ryegrass infected with the endophyteAcremonium loliae

Infection of ryegrass (Lolium perenne) by an endophytic fungus (Acremonium loliae) confers resistance against the Argentine stem weevil (Listronotus bonariensis). Extracts from ryegrass clones, infected and uninfected withA. loliae, were compared in a feeding choice bioassay, and several fractions were identified which affected stem weevil feeding behavior. One stem weevil feeding deterrent, peramine C₁₂H₁₇N₅O, has been isolated from infected ryegrass and partially characterized as a basic indole derivative. Extracts from culturedA. loliae had no effect on stem weevil feeding behavior nor was peramine detected in the fungal cultures examined. Peramine and the other active substances are hydrophilic in contrast to the lipophilic properties reported for the neurotoxic lolitrems also isolated from ryegrass infected withA. loliae and associated with causing ryegrass staggers disorder in livestock. It is suggested that ryegrass staggers and stem weevil feeding deterrency may arise by different biochemical mechanisms.     

Allelopathic effects of water extracts ofArtemisia princeps var.orientalis on selected plant species

The allelopathic effects of wormwood plants (Artemisia princeps var.orientalis) and their possible phytotoxicity on receptor species were investigated. The aqueous extracts of mature leaf, stem, and root of wormwood plants caused significant inhibition in germination and decreased seedling elongation of receptor plants, whereas germination of some species was not inhibited by extracts of stems and roots. Dry weight growth was slightly increased at lower concentrations of the extract, whereas it was proportionally inhibited at higher concentrations. The calorie value of the organic matter in receptor plants measured by bomb calorimeter was reduced proportionally to the extract concentration. However, results with extracts of juvenile leaf did not correlate with inhibition or promotion of elongation and dry weight.     

Seed-germination inhibition by volatile alcohols and other compounds associated withAmaranthus palmeri residues

Effects of 3-pentanone and eight low molecular weight aliphatic alcohols and aldehydes identifed in the mixtures of volatiles released byAmaranthus palmeri S. Wats. (AMAPA) residues were determined on germination of onion, carrot, AMAPA, and tomato seeds. Three-day exposures to these volatiles significantly inhibited germination of these assay seeds, and the inhibition was dependent upon exposure time and concentration. Based on the degree of inhibition observed in both time- and concentration-dependent assays, the following activity series was obtained: 2-heptanol > 3-methyl-1-butanol, 1-hexanol > hexanal, 1-pentanol, 3-pentanone, acetaldehyde > ethanol, 2-methyl-1-propanol. The activities of this group of compounds with that of 2-heptanone appear to be additive and related to test compound volatility and hydrophilicity. 2-Heptanol and 2-heptanone also significantly inhibited the germination of other species, including shepherdspurse, soybean, lettuce, alfalfa, common purslane, oats, and lovegrass.Name of companies or commercial products are given solely for the purpose of providing specific information; their mention does not imply recommendation or endorsement by the U.S. Department of Agriculture over others not mentioned.