Chang-Hung Chou

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.

Identification and phytotoxic activity of compounds produced during decomposition of corn and rye residues in soil

Residues from corn and rye plants were allowed to decompose in soil for periods up to 30 days at 22–23 °C, and the identity of some of the compounds produced as well as their relative phytotoxicity to lettuce seed and seedlings were determined. Paper, thin-layer, and gas chromatography were the principal methods used to identify the various compounds formed. The identities were confirmed by comparison with known synthetic compounds. Eighteen compounds were identified in the decomposing corn residues. Of these, salicylaldehyde, and butyric, phenylacetic, and 4-phenylbutyric acids were “volatile”, and benzoic,p-hydroxybenzoic, vanillic, ferulic,o-coumaric,o-hydroxyphenylacetic, salicylic, syringic,p-coumaric,trans-cinnamic, and caffeic acids were “not volatile”. Resorcinol,p-hydroxybenzaldehyde, and phloroglucinol were also found. In the decomposing rye residues, nine compounds were identified, including vanillic, ferulic, phenylacetic, 4-phenylbutyric,p-coumaric,p-hydroxybenzoic, salicylic, ando-coumaric acids, and salicylaldehyde. In the lettuce seed bioassay, most of the above compounds from corn and rye decomposition products exhibited some phytotoxicity. Phenylacetic, 4-phenylbutyric, salicylic, benzoic, ando-hydroxyphenylacetic acids were highly inhibitory to the growth of lettuce at concentrations between 25 and 50 ppm. The others reduced growth significantly at 100 ppm. Most of the phototoxic spots were located in theRf0.37–0.97 zone when developed in 2% acetic acid solvent.

Allelopathic research of subtropical vegetation in Taiwan : III. Allelopathic exclusion of understory byLeucaena leucocephala (Lam.) de Wit.

Leucaena leucocephala plantations in Kaoshu, southern Taiwan, exhibit, after several years of growth, a unique pattern of weed exclusion beneathLeucaena canopy. The pattern has been observed in manyLeucaena plantations in Taiwan and is particularly pronounced in the area where a substantial amount ofLeucaena litter has accumulated on the ground. Field data showed that the phenomenon was primarily not due to physical competition involving light, soil moisture, pH, and nutrients. Instead, aqueous extracts ofLeucaena fresh leaves, litter, soil, and seed exudate showed significantly phytotoxic effects on many test species, including rice, lettuce,Acacia confusa, Alnus formosana, Casuarina glauca, Liquidambar formosana, andMimosa pudica. However, the extracts were not toxic to the growth ofLeucaena seedlings. The decomposing leaves ofLeucaena also suppressed the growth of the aforementioned plants grown in pots but did not inhibit that ofLeucaena plants. By means of paper and thin-layer chromatography, UV-visible spectrophotometry, and high-performance liquid chromatography, 10 phytotoxins were identified. They included mimosine, quercetin, and gallic, protocatechuic,p-hydroxybenzoic,p-hydroxyphenylacetic, vanillic, ferulic, caffeic, andp-coumaric acids. The mature leaves ofLeucaena possess about 5% dry weight of mimosine, the amount varying with varieties. The seed germination and radicle growth of lettuce, rice, and rye grass were significantly inhibited by aqueous mimosine solution at a concentration of 20 ppm, while that of the forest species mentioned was suppressed by the mimosine solution at 50 ppm or above. However, the growth ofMiscanthus floridulus andPinus taiwanensis was not suppressed by the mimosine solution at 200 ppm. The seedlings ofAgeratum conzoides died in mimosine solution at 50 ppm within seven days and wilted at 300 ppm within three days. It was concluded that the exclusion of understory plants was evidently due to the allelopathic effect of compounds produced byLeucaena. The allelopathic pattern was clearly shown in the area with a heavy accumulation ofLeucaena leaf litter, which was a result of drought and heavy wind influence.

Allelopathic research of subtropical vegetation in Taiwan. IV. Comparative phytotoxic nature of leachate from four subtropical grasses.

The phytotoxicity of plant leachates was evaluated from four subtropical grasses:Brachiaria mutica, Digitaria decumbens, Imperata cylindrica var. Major, andPanicum repens. The aqueous leachate of each grass was used to water the growth of the four grasses in pots. The leachates exhibited variable inhibition of grass growth as compared to the tap water control. By the 41st day after treatment, the leachate ofD. decumbens significantly suppressed the growth of itself and retarded that ofB. mutica andP. repens. The growth ofB. mutica was inhibited by its own leachate, but that ofI. cylindrica was not affected by any of the grass leachates. In crop growth rate (CGR) analysis, the four grass leachates exhibited a similar inhibition pattern. In laboratory bioassays, the leachates showed a significant phytotoxic effect on the radicle growth of ryegrass and lettuce. Six phytotoxic phenolics were quantitatively compared by high-performance liquid chromatography, and the amount of compounds varied with species. The highest total amount of phytotoxic phenolics occurred inD. decumbens, followed, in decreasing order, byP. repens, B. mutica, andI. cylindrica. These findings show that the leachates of four grasses possess phytotoxic compounds that may play a significant role in grass dominance in the field.

Phytotoxic substances in twelve subtropical grasses

Aqueous extracts of 12 subtropical grasses inhibited seed germination and radicle growth ofLactuca sativa var. Great Lakes at osmotic concentrations as low as 10 milliosmol.Acroceras macrum,Chloris gayana, Digitaria decumbens, andPanicum maximum exhibited the highest inhibition, whileCortaderia selloana revealed the least. Toxic spots were found on chromatograms of the ether fraction of aqueous extracts.Cynodon dactylon, Setaria sphacelata, andTripsacum laxum showed more than six toxic spots, whileAndropogon nodosum,Bracharia mutica, andChloris gayana gave less than three toxic spots. The phytotoxins ferulic, syringic,p-coumaric, vanillic,p-hydroxybenzoic, ando-hydroxyphenylacetic acids were identified. These compounds are differentially distributed in the 12 grasses studied. Additionally, most of these compounds were also found in the associated soils; the control (nonherb-growth) soil provided the toxic compounds in significantly less amount than did the grass soils.

Autointoxication mechanism ofOryza sativa

The phytotoxicity produced during decomposition of rice straw in soil was evaluated under both constant and changing temperature conditions. Bioassay tests showed that the aqueous extract from a soilstraw mixture after incubation at constant temperature was more than twice as phytotoxic as the extract from soil incubated alone. The phytotoxicity was highest at 20–25 ° C. Temperatures above 25 ° C enhanced rice straw decomposition and also degraded the phytotoxic substances more rapidly. After incubation of soil mixtures under changing temperature regimes in a phytotron, the phytotoxicy of the soil aqueous extracts increased in the following order: soil alone < soil + fertilizer < soil + straw < soil + straw + fertilizer. Growth inhibition of lettuce or rice seedlings was also at the highest at the temperature range of 25–30 ° C irrespective of the direction of temperature changes from either low to high or vice versa. Five phytotoxic phenolics,p-hydroxybenzoic, vanillic,p-coumaric, syringic, and ferulic acids, were obtained from both the aqueous extract and residue of the incubated soil samples and were quantitatively estimated by chromatography. The amount of phytotoxins found in various soil mixtures followed the same increasing order as that found by the seed bioassay test. Although no definite distribution pattern of the phenolics in the incubated soil samples can be attributed to temperature variations, the amount of the phenolics was likely higher in the samples incubated at 25 ° C than at either 15 ° C or 35 ° C. The quantity of toxins released during decomposition of rice straw in soil reached highest levels six weeks after incubation and gradually disappeared after twelve weeks.

Possible allelopathic constituents ofCoffea arabica

The aqueous extracts of leaves, stems, and roots ofCoffea arabica significantly inhibited the seed germination and radicle growth of rye grass, lettuce, and fescue. When the extracts were diluted to 1% solution, significant suppression of lettuce growth was still found and was particularly pronounced in the extract of young seedlings. The paper chromato-gram of the ether fraction of an aqueous extract of coffee leaves was bioassayed with lettuce seeds and revealed a remarkable inhibition throughout the chromatogram except for the segment ofRf 0.00–0.12. Paper without spotting extract was used as a standard. The phytotoxins present in coffee tissue were identified by paper and thin-layer chro-matography and mass spectrometry. The compounds include caffeine, theobromine, theophylline, paraxanthine, scopoletin, and chlorogenic, ferulic,p-coumaric,p-hydroxybenzoic, caffeic, and vanillic acids. All compounds except caffeic acid exhibited significant phytotoxicity to lettuce growth at a concentration of 100 ppm.

Allelopathic dominance ofMiscanthus transmorrisonensis in an alpine grassland community in Taiwan

A study site located at 2600 m elevation in Tartarchia Anpu, Nantou county, Taiwan, exhibits a unique grassland community composed of two principal species,Miscanthus transmorrisonensis andYushinia niitakayamensis, and 35 other species. The relative frequencies of the two species are 12% and 11%, while their relative coverages are 25% and 19.5%, respectively. The values for the remaining 35 species are lower than4% each, while species diversity of the community is −3.04839, indicating great diversity. To elucidate the mechanism of dominance ofM. transmorrisonensis, allelopathic evaluation of the plant was conducted. Aqueous extracts of M.Transmorrisonensis plant parts with two ecotypes were bioassayed. The extracts showed significant phytotoxic effects on seed germination and radicle growth of four tested plants: rye grass, lettuce, and two varieties of Chinese cabbage. In addition, rhizosphere soils underMiscanthus also exhibited significant phytotoxicity, indicating that allelopathic interaction was involved. Some responsible phytotoxic phenolics, namely, p-coumaric, ferulic, vanillic, protocatechuic, o-hydroxyphenylacetic, andm-hydroxyphenylacetic acids, and 4-hydroxycoumarin and phloridzin were identified. Allelopathy thus can play an important role in regulating plant diversity in the field.

Allelopathic Potential of Acacia confusa and Related Species in Taiwan

Acacia confusa (an endemic species) and other introduced species, namely A. aulacocarpa, A. auricumiformis, A. cincinnata, A. crassicarpa, A. leptocarpa, A. margium, A. polystachya, and A. torfilis were evaluated for allelopathic potential. Among these, A. confusa is widely distributed on the hills and lowlands of Taiwn and often exhibits a unique pattern of weed exclusion under stands. Four study sites were selected. Field observations and measurements were carried out at sites that exhibited relatively pure stands of A. confusa. Although the diversity of understory species was comparatively higher in Acacia stands than in adjacent weedy sites, the total coverage and biomass of understory plants was significantly lower than in control sites. Aqueous extracts (0.5%, 1%, 2%, 3%, 4%, and 5%) of dry leaves and litter of Acacia confusa and other species collected from various sites and dates were bioassayed using lettuce, alfalfa, and Chinese cabbage to examine their phytotoxicity. Most extracts exhibited phytotoxicity even at a concentration as low as 0.5%. Inhibition of radicle growth of test plants varied with sampling sites and dates. Bioassay with 5% extracts or above produced more than 85% inhibition of test plants regardless of habitat. In addition, surface soils collected from the upper 20 cm layer of Acacia stands revealed significant inhibition as compared with adjacent grassland control soils. The most inhibitory compounds isolated from the ether fraction of aqueous extracts were identified as: ferulic, vanillic, caffeic, gallic, m-hydroxybenzoic, and m-hydroxyphenylacetic acids. Unidentified flavonoids were also found. Chromatographic bioassays of compounds isolated from both ethyl acetate and water fractions of methanolic extracts of Acacia leaves also showed significant phytotoxicity but none was found in the fractions of chloroform and hexane, suggesting the phytotoxic compounds present in Acacia plants are water soluble.

Allelopathic substances and interactions of Delonix regia (Boj) Raf

A unique pattern of weed exclusion was found under the canopy ofDelonix regia, which was planted in many places as an ornamental tree in the south of Taiwan. A quadrat method was employed to examine the botanical composition between the area underneathD. regia and its adjacent control grassland. The number of species and coverage of understory species were significantly lower in the area of theD. regia than that of the grassland, indicating the growth of understory species was suppressed byD. regia. A series of aqueous extracts of leaves, flowers, and twigs ofD. regia were bioassayed against three species to determine their phytotoxicity, and the results showed highest inhibition in the flowers. A water-culture experiment indicated the aqueous extract of flowers ofD. regia on two local understory species (Isachne nipponensis andCentella asiatica) inhibited growth of both species by more than 70%. The phytotoxicities of fallen leaves and flowers ofD. regia were not significantly affected by temperature. When the plant material was subjected to temperatures above 70°C, however, phytotoxicity was decreased, indicating that the allelopathic nature ofD. regia could easily be decomposed by fire. By means of paper, thin-layer, and high-performance liquid chromatography, and UV-visible spectrophotometry, responsible phytotoxins present in leaves, flowers, and twigs ofD. regia were identified as 4-hydroxybenzoic, chlorogenic, 3,4-dihydroxybenzoic, gallic, 3,4-dihydroxycinnamic, 3,5-dinitrobenzoic, and L-azetidine-2-carboxylic acids, and 3,4-dihydroxybenzaldehyde. The findings of bioassays and the number and amount of responsible allelopathic compounds found inD. regia are well correlated, thus permitting the conclusion that the exclusion of understory plants under the canopy ofD. regia trees was due primarily to the allelopathic effect of the fallen flower, leaves, and twigs of theD. regia. A possible mechanism of action is discussed.