Plant growth regulatory activities of artemisinin and its related compounds

Artemisinin, a sesquiterpene lactone produced in the leaves ofArtemisia annua, was evaluated for its phytotoxicity in mono- and dicotyledonous plants. Artemisinin inhibited seed germination, seedling growth, and root induction in all species tested. The concentration of artemisinin required for 50% inhibition ofLemna minor growth was 5 M. Inhibitory plant responses appeared to require the endoperoxide moiety of this compound since similar chemicals without endoperoxide, deoxyartemisinin, arteannuic acid, and arteannuin B, were less phytotoxic. InL. minor, artemisinin and arteannuic acid caused the leakage of proteins into the growth medium, suggesting the site of activity was at the plant cell membrane.     

Synthesis and antifungal activity of analogues of naturally occurring botrydial precursors

Analog compounds of the proposed intermediates of the biogenetic pathway to botrydial have been synthesized. These compounds were tested for their potential antifungal activity against the phytopathogenBotrytis cinerea. Our results showed a fungistatic effect of some compounds on mycelium growth. The most significant effect was exerted by 2--hydroxy-2,3-dihydro-1-epiprobotrydial, which inhibited growth ofB. cinerea. Some aspects of structure-activity relationships are discussed.     

Allelopathic and autotoxic effects ofAnastatica hierochuntica L.

Laboratory experiments were undertaken to investigate the autotoxic effects ofAnastatica hierochuntica and possible effects on five other desert plants:Rumex cyprius, Trigonella stellata, Diplotaxis harra, Cleome droserifolia, andFarsetia aegyptia. Seed germination, seedling growth, and cell division of all species tested were inhibited by the shoot aqueous extract ofA. hierochuntica. A gradual increase in the percentage of prophase and decrease in the other mitotic stages as well as the mitotic index were observed with increasing extract concentration. At an extract concentration of 8% the mitotic index was reduced from the control by 55% inC. droserifolia, 54% inT. stellata, 45% inF. aegyptia, 43% inA. hierochuntica, and 35% inR. cyprius. The inhibitory substances are apparently released onto soil by repeated washing of the standing plants by rain and dew interception.     

Experiments on bioassay sensitivity in the study of allelopathy

The purpose of these experiments was to study the effects of various experimental procedures and conditions on bioassay sensitivity in allelopathic studies. The following factors were considered: bioassay methods, choice of test species, the effect of osmotic potential on germination and growth of the test species, growth in dark or in light and the effect of concentrating the extracts in vacuo. Extracts from rape and rye were used in the studies to act as allelopathic agents. Ryegrass appeared as the most sensitive test species when grown horizontally on quartz sand, while radish was the most sensitive test species when grown on filter paper in transparent boxes at a 45° angle. Root length was the most sensitive growth variable measured. Percent germination appeared a more sensitive measure than the speed of germination index, S, when germinated seeds were counted after one and two days for radish and ryegrass, respectively. A rise in the osmotic potential affected germination more than root length, and radish appeared more sensitive to a rise in osmotic potential than ryegrass. A confounding of germination and root length inhibition may therefore, give misleading results.     

Natural products phytotoxicity A bioassay suitable for small quantities of slightly water-soluble compounds

A large variety of secondary metabolites that can inhibit germination and/or seedling growth are produced by plants in low quantities. The objective of this study was to develop a bioassay capable of reliably assessing reductions in germination percentage and seedling length of small-seeded plant species caused by exposure to minute quantities of these compounds. The germination and growth of alfalfa (Medicago saliva), annual ryegrass (Lolium multiflorum), and velvetleaf (Abutilon theophrasti) were evaluated against six known phytotoxins from five chemical classes; cinmethylin (a herbicidal cineole derivative) was selected as a comparison standard. Each phytotoxin, dissolved in a suitable organic solvent, was placed on water-agar in small tissue culture wells. After the solvent evaporated, imbibed seeds were placed on the agar; after three days, germination percentages and seedling lengths were measured. Compared to a commonly used filter paper procedure, this modified agar bioassay required smaller quantities of compound per seed for comparable bioassay results. This bioassay also readily permitted the measurement of seedling length, a more sensitive indicator of phytotoxicity than germination. Seedling length decreased sigmoidally as the toxin concentration increased logarithmically. Phytotoxicity was a function of both compound and plant species. Cinmethylin, a grass herbicide, reduced the length of annual ryegrass seedlings by 90–100%, whereas that of alfalfa and velvetleaf was inhibited slightly. The agar bioassay facilitated the rapid and reliable testing of slightly water-soluble compounds, requiring only minute quantities of each compound to give reproducible results.The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

Allelopathic properties of α-terthienyl and phenylheptatriyne,naturally occurring compounds from species of asteraceae

Alpha-terthienyl (-T), a naturally occurring polyacetylene derivative from roots ofTagetes erecta L., and phenylheptatriyne (PHT), from leaves ofBidens pilosa L., were tested as possible allelopathic agents against four seedling species (Asclepias syriaca L.,Chenopodium album L.,Phleum pratense L.,Trifolium pratense L.).Asclepias was the most sensitive of the species. Allelopathic activity was enhanced in the presence of sunlight or sources of near-UV, with LC_50s forA. syriaca of 0.15 ppm and 0.66 ppm with -T and PHT, respectively; 0.27 and 0.85 forC. album; 0.79 and 1.43 forP. pratense, and 1.93 and 1.82 forT. pratense. Near-UV exposure was saturating but never more than found in summer sunlight at Ottawa, Canada. Growth inhibition was observed with seedlings treated with -T and PHT but without near-UV irradiation. Germination of seedlings was also sensitive to -T and PHT with or without near-UV treatment. -T was extracted from soil surrounding the roots ofTagetes. Concentrations calculated for the soil (0.4 ppm) indicate that seedling growth could be significantly hindered. The activity and specificity of -T was sufficiently high to warrant future field trials to assess its potential as a natural weed-control agent.     

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.     

Phytotoxicity of <Emphasis Type="Italic">Phytolacca americana</Emphasis> Leaf Extracts on the Growth,and Physiological Response of <Emphasis Type="Italic">Cassia mimosoides</Emphasis>

We examined the allelochemical effects of control soil, native soil (treated soil), and leaf extracts of Phytolacca americana (pokeweed) on the germination rate and seedling growth of Cassia mimosoides var. nomame. We also studied the resulting changes in root-tip ultrastructure and peroxidase isozyme biochemistry. P. americana leaf extract inhibited seed germination, seedling growth, and biomass when compared to control and treated soil. Root and shoot growth in treated soil was stimulated relative to control soil, but root growth was inhibited by 50% in the leaf extract treatment. Biomass of C. mimosoides seedlings grown on leaf extract was reduced sevenfold when compared to the control seedlings. The amounts of total phenolic compounds in the leaf extract, treated soil, and control soil were 0.77, 0.14, and 0.03 mg l⁻¹, respectively. The root tips of C. mimsoides treated with leaf extracts of P. americana showed amyloplasts and large central vacuoles with electron-dense deposits inside them when compared to control root tips. The activity of guaiacol peroxidase (GuPOX) in whole plant, roots, and shoots of C. mimosoides increased as leaf extract increased; maximum activity was observed in extract concentrations of 75% and higher. Root GuPOX activity was three times higher than in shoots. Therefore, we conclude that inhibition of C. mimosoides growth is related to the phenolic compounds in the P. americana leaf extract and the ultrastructure changes in root-tip cells and increased GuPOX activity is a response to these allelochemicals.     

Identification of volatile allelochemicals fromAmaranthus palmeri S. Wats

Allelopathic volatiles associated with the weed Palmer amaranth (Amaranthus palmeri S. Wats.; AMAPA) were trapped on Tenax GC, thermally desorbed, and identified by gas chromatography-mass spectroscopy. Methyl ketones and alcohols (C₄–C₁₁) were the principal components of the volatiles mixture. Seedheads, stems, or roots were placed in a glass container and incubated at 31 °C (10 hr)/21 °C (14 hr) for three days prior to trapping the volatiles. Seedheads were rich in 2-heptanone which was consistently found, together with 2-heptanol, in all AMAPA tissues. Vapors of authentic 2-heptanone and (±)-2-heptanol at concentrations of 1 ppm or higher strongly inhibited the germination of onion and carrot and almost completely suppressed the germination of tomato and AMAPA seeds.Names 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.     

Allelopathic effects of giant foxtail on germination and radicle elongation of loblolly pine seed

Water extracts of giant foxtail grass inhibited germination and radicle elongation of loblolly pine seed in a laboratory test. Most of the toxic effects came from extracts of dried foxtail tops, with lesser amounts from fresh tops and roots. Soil texture affected the phytotoxicity of the extracts. Eight chemical compounds known to be phytotoxins inSetaria were identified in extracts of foxtail leaves but could not be found in soil samples under the leaves.     

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.     

Toxicity studies of analogs of 2-tridecanone,a naturally occurring toxicant from a wild tomato

Toxicity bioassays of the naturally occurring insecticide 2-tridecanone and a group of structural analogs against the tomato fruitworm (Heliothis zea) reveal a significant influence of chain length on toxicity of methyl ketones. This effect may be due to the differential ability of these compounds to penetrate lipid barriers and reach the active site, since a close relationship is seen between toxicity and lipophilicity. Congeners of chain lenths 15 and over are less active than predicted by this relationship, possibly due to steric effects.Received for publication June 29, 1981, paper no. 6973 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, North Carolina 27650.     

Quantification of insect growth and its use in screening of naturally occurring insect control agents

In studies of the effects of allelochemicals or other factors on the development of different insect species, comparison of growth-inhibiting activities is difficult using the parameters currently employed. We introduce two new parameters, growth index (GI) and relative growth index (RGI), which can unify the quantification of insect development. This quantification can also eliminate the effects of different growth characteristics due to the genetic differences between insect species. By measuring growth-inhibiting effects of two phytochemicals, chaparrin and chaparrinone, on the tobacco budworm,Heliothis virescens, and the beet armyworm,Spodoptera exigua, bioassay procedures and GI and RGI calculations are demonstrated.     

Bioassay approaches to assessing behavioral responses of plum curculio adults (Coleoptera: Curculionidae) to host fruit odor

We evaluated several approaches to developing a simple, sensitive, and reliable laboratory bioassay of responses of overwintered adult plum curculios (PCs),Conotrachelus nenuphar (Herbst), to host fruit odor or its attractive components. A high proportion of assayed PCs responded positively to odor of wild plums under no-choice, moving-air conditions in a wind tunnel and under dual-choice, still-air conditions in enclosed Petri dishes. Positive response to controls lacking host odor, however, was much greater in the wind tunnel, arguing in favor of bioassays under dual-choice conditions in still air to provide greater PC discrimination. Response to host odor (from wild plums or hexane extract of wild plums or Liberty apples) in Petri dish bioassay chambers proved greatest: (1) during the scotophase of PCs under total dark or dim red light conditions, (2) when Petri dishes were completely enclosed, (3) when PCs were starved for 24 or 48 hr, and (4) when PCs were tested within seven weeks after apple tree petal fall. Neither the sex of a PC nor the direction in which a PC was obliged to move to find the source of host odor (upward through a port in the Petri dish lid or downward through a port in the base) had a substantial effect on level of response to host odor or discrimination of host odor from a nonodorous control. We conclude that an enclosed Petri dish bioassay chamber of the type described here should be a valuable asset in the process of chemically identifying components of host fruit odor attractive to PCs.     

Berberine: A naturally occurring phototoxic alkaloid

The isoquinoline alkaloid berberine, present in nine different plant families was found to be phototoxic to mosquito larvae. In the presence of near UV the LC₅₀ for acute 24-hr toxicity was 8.8 ppm compared to 250 ppm for dark controls. Mosquito larvae that were treated with 10 ppm berberine plus near UV for 24 hr and then transferred to berberine-free water showed decreased larval survival and resulted in a smaller cumulative number of pupae and adults as compared to controls, during a subsequent 4-week development period. Berberine was found to be a singlet O₂ generator in experiments with the chemical trap 2,5-dimethyl furan. A slight increase in chromosome aberrations in Chinese hamster cells was also observed with berberine plus near UV treatment. The significance of the phototoxicity of berberine is discussed in relation to plant-insect relations.     

Biologically active secondary metabolites of barley. II. Phytotoxicity of barley allelochemicals

The release of alkaloids by barley was quantified by HPLC. Hordenine was released from the roots of barley in a hydroponic system for up to 60 days. The amount reached a maximum, 2g/plant/day, at 36 days, then declined. Effects on white mustard by hordenine and gramine included reduction of radicle length and apparent reduction in health and vigor of radicle tips. Transmission electron microscopic examination of white mustard radicle tips exposed to hordenine and gramine showed damage to cell walls, increase in both size and number of vacuoles, autophagy, and disorganization of organelles. The evidence of the morphological and primary effects of barley allelochemicals at the levels released by living plants indicates that the biologically active secondary metabolites of barley may lead to a significant role in selfdefense by the crop.     

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.     

穿心莲内酯对小麦种子萌发与幼苗生长的影响

探究穿心莲内酯对小麦种子萌发和幼苗生长的影响。采用不同质量浓度穿心莲内酯浸种处理,以清水和生物源植物生长调节剂油菜素内酯为对照,测定发芽率、发芽势、根长、苗高、鲜质量、干质量等指标。结果表明,随着穿心莲内酯质量浓度的升高,其促进作用先升高后降低,质量浓度150 mg/L的穿心莲内酯溶液对小麦种苗生长促进作用明显,为研究绿色农业植物生长调节物质提供基础。     

Light-mediated allelochemical effects of naturally occurring polyacetylenes and thiophenes from asteraceae on herbivorous insects

Polyacetylenes and their thiophene derivatives, characteristic secondary metabolites of the Asteraceae, were examined for their effects on herbivorous insects. Three thiophenes (a monothiophene, a bithiophene, and -terthienyl) and four polyacetylenes (phenylheptatriyne, phenylheptadiynene, phenylheptadiyene acetate, and matricaria lactone) were studied for their phototoxicity and light-independent toxicity to (1) a polyphagous lepidopteran,Ostrinia nubilalis, whose host range includes a number of phototoxic Asteraceae, (2) a polyphagous lepidoteran,Euxoa messoria, whose host range includes very few species of Asteraceae, and (3) an oligophagous lepidopteran,Manduca sexta, which is a specialist on Solanaceae. Several compounds were phototoxic toM. sexta andE. messoria even at very low irradiance levels, but behavioral adaptations, including spinning silk and boring into diet, allowedO. nubilalis to avoid photosensitization. Light-independent activity of the compounds to all three species involved feeding deterrence increasing in the orderO. nubilalis, E. messoria, andM. sexta, and longterm metabolic toxicity in the form of impaired nutrient utilization. The biosynthetically derived thiophenes were more toxic than their acetylenic precursors, and toxicity increased with increasing number of thiophene rings. The results are discussed in terms of plant-insect coevolution.     

苦参碱对大麦种子萌发及幼苗生长的影响

采用室内培养皿法,以清水和油菜素内酯作对比,分析不同浓度的苦参碱制剂浸种大麦种子对大麦的种子发芽率、苗高、根长等形态指标,以及叶绿素和可溶性糖含量等的影响。结果表明,苦参碱对大麦种子萌发及幼苗生长有促进作用,且随着苦参碱质量浓度的增大,大麦的种子发芽率、苗高、根长、叶绿素和可溶性糖含量呈先升高后降低的趋势,以100 mg/L处理各指标最好。此外,100 mg/L苦参碱浸种处理对大麦幼苗各项指标的影响与25 mg/L油菜素内酯相似或比25 mg/L油菜素内酯好。