Effects of two sesquiterpene lactones isolated fromArtemisia annua on physiology ofLemna minor.

The effects of artemisinin and arteannuic acid extracted fromArtemisia annua on the physiology ofLemna minor were evaluated. Changes in frond production, growth, dry weight, and chlorophyll content ofL. minor were determined. Photosynthesis and respiration were evaluated with a differential respirometer. Artemisinin (5 µM) inhibitedL. minor frond production and dry weight 82 and 83%, relative to methanol controls. Chlorophyll content was reduced 44% by artemisinin (2.5 µM). Arteannuic acid (10 µM) was less active, inhibiting frond production 61% and reducing chlorophyll content 66% at 5 µM. Artemisinin (1 µM) reducedL. minor photosynthesis 30% and 2.5 µM reduced respiration 39%. Arteannuic acid had no significant effect on photosynthesis or respiration at the levels tested.

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 µM. 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.

Effects of three phenolic compounds onLemna gibba G3.

Lemna gibba L. G3, (duckweed) was used as a bioassay organism to test the allelochemical effects of salicylic acid (SA), ferulic acid (FA), and umbelliferone (UM). Growth rate (K), dry weight (DW) and total chlorophyll (CHL) production were measured after seven days of growth. The bioassay procedure used 50 ml of E medium with and without sucrose in 125-ml Erlenmeyer flasks plus the selected concentration of allelochemical. At concentrations of 50 µM and greater, SA caused inhibition of K and DW production inL. gibba G3, while the threshold for CHL reduction was 20 µM. FA inhibited the DW and CHL production at 100 µM when the compound was auto-claved in E medium containing sucrose. Treatments of UM were least toxic with an inhibition threshold of 500 µM for K and DW production in medium without sucrose. UM did not reduce CHL production until 750 µM. In some cases, different thresholds of inhibition were observed depending on the presence or absence of sucrose and tartaric acid in the medium, and whether or not the chemicals were autoclaved with the medium.

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.

Potentials for exploiting allelopathy to enhance crop production.

Strategies for utilizing allelopathy as an aid in crop production include both avoidance and application protocols. There are immediate opportunities for management of weed and crop residues, tillage practices, and crop sequences to minimize crop losses from allelopathy and also to use allelopathic crops for weed control. Varieties of grain and forage sorghums (Sorghum Spp.), sunflower (Helianthus annuus L.), oats (Avena sativa L.), wheat (Triticum sativum L.),rye (Secale cereale L.), and others may provide weed control and in some instances crop stimulation from their residues. Our four-year field study with cultivated sunflower resulted in no differences in weed biomass between plots with and without herbicide (EPTC) applications. Strip cropping that included sorghum showed that in the subsequent year weed density and biomass were significantly lower in the previous-year sorghum than in soybean strips. Possibilities exist for modification of crop plant metabolism to alter production of allelochemicals. Allelochemical-environmental interactions must be considered in efforts to benefit from allelopathy. Under greenhouse conditions, joint application of low levels of atrazine, trifluralin, alachlor, or cinmethylin with a phenolic allelochemical showed that these two categories of inhibitors acted in concert to reduce plant growth. Allelochemicals may also be adapted as yield stimulants or environmentally sound herbicides, such as cinmethylin and methoxyphenone. Isolation of bialophos, tentoxin, and others shows that bacteria and fungi are good sources of biologically active compounds.

Allelopathic influence ofSorghum bicolor on weeds during germination and early development of seedlings.

The allelopathic interaction between sorghum [Sorghum bicolor (L.) Moench] and 10 species of grass and broadleaf weeds was investigated. Germination of weed seeds was slightly inhibited or stimulated, depending on species, when incubated in closed Petri dishes with germinating sorghum. Subsequent radicle and hypocotyl or coleoptile elongation of weeds was significantly inhibited by the germinating sorghum. For weeds interplanted with sorghum and grown under greenhouse conditions. The inhibitory effect on some weed species was still evident after 2 months of growth. Significant differences were found in the dry matter per weed plant grown in pots in proximity to sorghum vs. weeds grown in monoculture. Aqueous leachates from pots planted with sorghum alone or from a system in which sorghum roots protruded into water had strong allelopathic activity. These results indicate that water-soluble allelochemicals are produced by germinating sorghum seeds and that production of these substances continues during seedling growth.

Use ofLemna minor L. as a bioassay in allelopathy.

Investigations in allelopathy often require the use of a bioassay for evaluating limited quantities of potentially active growth regulators. A bioassay procedure was developed usingL. minor grown in 1.5-ml aliquots of nutrient medium with and without allelochemicals in wells of 24-well tissue culture cluster dishes with loose-fitting lids. Tests using six replications per treatment with several flavonoid compounds and derivatives of coumarin, benzoic acid, and cinnamic acid demonstrated that the bioassay was capable of measuring inhibition at levels of compound ranging from 50 to 1000 µmol. Strongly inhibitory treatments were visible after 1 or 2 days. After 7 days of growth, frond number, growth rate, and dry weight were used to evaluate effects. The bioassay system is relatively simple, very sensitive, reproducible, and can be used for testing small amounts and dilute concentrations of unknowns which have been separated by chromatography.

Weed control using allelopathic crop plants.

The concept that some crop plants may be allelopathic to common weeds of agricultural lands is receiving greater attention as an alternative weed control strategy. Several crops showing promise are: grain and forage species such as barley (Hordeum), oat (A vena), fescue (Festuca), and sorghum (Sorghum), and the agronomic species of corn (Zea) and sunflower (Helianthus). Among the problems that hinder the conclusive demonstration of allelopathic effects of crop plants are the loss of that capacity through selection and the variability among cultivars. Recent studies to evaluate the allelopathic potential of crop plants have shown that several sunflower varieties inhibit the germination and growth of associated weeds and to a greater extent than found in several biotypes of native sunflower. Aqueous extracts of dried sunflower and rape tissue inhibited or stimulated germination and growth of weeds, and the response depended upon the source of extract, the extract concentration, and the weed species tested. The validity of bioassay results was tested in a 5-year field study with sunflower and oat grown in rotation. Weed density increased in all plots but the extent of increase was significantly less in plots of sunflower than in control plots. The use of crop plants with increased allelochemical production could limit the need for conventional herbicides to early season application with late season control provided by the crop.

Increased Ethylene Production during Clinostat Experiments May Cause Leaf Epinasty.

Ethylene production from tomato (Lycopersicum esculentum L. cv. Rutgers) plants based on a clinostat doubled during the first 2 hours of rotation. Carbon dioxide blocked the appearance of leaf epinasty normally associated with plants rotated on a clinostat. These results support the idea that epinasty of clinostated plants was due to increased ethylene production and not to the cancellation of the gravitational pull on auxin transport in the petiole.

Fate of air pollutants: removal of ethylene, sulfur dioxide, and nitrogen dioxide by soil.

The ultimate sink for many air pollutants is unknown. Data are presented here in support of the idea that reaction with soil, through microbial or chemical means, can remove ethylene, other hydrocarbons, sulfur dioxide, and nitrogen dioxide from the air.

Abscission: the phytogerontological effects of ethylene.

The role of ethylene in the aging of bean (Phaseolus vulgaris L. cv. Red Kidney) petiole abscission zone explants was examined. The data indicate that ethylene does accelerate aging in addition to inducing changes in break strength. Application of ethylene during the aging stage (stage 1) promoted abscission when followed by a second ethylene treatment during the cell separating stage (stage 2). The half-maximal effective concentration of ethylene to induce aging was around 0.3 microliter per liter; 10 microliters per liter was a saturating dose. CO(2) reversal of ethylene action during stage 1 was incomplete and gave ambiguous results. CO(2) (10%) reversed the effect of 10 microliters per liter ethylene but not 1 microliter per liter ethylene. The possibility that ethylene not only accelerated aging but was also a requirement for it was tested, and experimental evidence in favor of this idea was obtained. It was concluded that ethylene plays a dual role in the abscission of bean petiole explants: a phytogerontological effect and a cellulase-inducing effect.

Abscission: Control of cellulase secretion by ethylene.

Ethylene was found to be required for the release or secretion of cellulase from the cytoplasm to the cell wall in bean-petiole abscission-zone explants. This is an addition to its previously known action in accelerating senescence and cellulase synthesis in abscission.

Abscission: movement and conjugation of auxin.

A 1-hour application of indole-3-acetic acid to bean (Phaseolus vulgaris L. cv. Red Kidney) explants inhibited abscission for an 8-hour aging period. Use of indole-3-acetic acid-(14)C showed that the applied indole-3-acetic acid was conjugated within explant tissue and that this conjugation mechanism accounts for loss of effectiveness of indole-3-acetic acid in inhibiting abscission after 8 hours. Reapplication of indole-3-acetic acid to an explant at a later time, before the induced aging requirement was completed reinhibited abscission. 2,4-Dichlorophenoxyacetic acid, which is not destroyed or conjugated by this system, did not lose its ability to inhibit abscission. It was concluded that indole-3-acetic acid destruction is one of the processes involved in the aging stage of abscission in explants.