Evaluation of Putative Allelochemicals in Rice Root Exudates for Their Role in the Suppression of Arrowhead Root Growth

In previous studies, 15 putative allelopathic compounds detected in rice root exudates were quantified by GC/MS/MS. In this study, multiple regression analysis on these compounds determined that five selected phenolics, namely caffeic, p-hydroxybenzoic, vanillic, syringic, and p-coumaric acids, from rice exudates were best correlated with the observed allelopathic effect on arrowhead (Sagittaria montevidensis) root growth. Despite this positive association, determination of the phenolic acid dose-response curve established that the amount quantified in the exudates was much lower than the required threshold concentration for arrowhead inhibition. A similar dose-response curve resulted from a combination of all 15 quantified compounds. Significant differences between the amounts of trans-ferulic acid, abietic acid, and an indole also existed between allelopathic and non-allelopathic rice cultivars. The potential roles of these three compounds in rice allelopathy were examined by chemoassay. Overall, neither the addition of trans-ferulic acid nor 5-hydroxyindole-3-acetic acid to the phenolic mix significantly contributed to phytotoxicity, although at higher concentrations, trans-ferulic acid appeared to act antagonistically to the phytotoxic effects of the phenolic mix. The addition of abietic acid also decreased the inhibitory effect of the phenolic mix. These studies indicate that the compounds quantified are not directly responsible for the observed allelopathic response. It is possible that the amount of phenolic acids may be indirectly related to the chemicals finally responsible for the observed allelopathic effect.     

Identification and Quantitation of Compounds in a Series of Allelopathic and Non-Allelopathic Rice Root Exudates

An investigation of the chemical basis for rice allelopathy to the rice weed arrowhead (Sagittaria montevidensis) was undertaken using GC/MS and GC/MS/MS techniques. Twenty-five compounds were isolated and identified from the root exudates of both allelopathic and non-allelopathic rice varieties. Phenolics, phenylalkanoic acids, and indoles were among the chemical classes identified. Two indoles previously unreported in rice were detected in the exudates, 5-hydroxy-2-indolecarboxylic acid and 5-hydroxyindole-3-acetic acid. Several other compounds identified in this study have not previously been reported in rice root exudates, namely mercaptoacetic acid, 4-hydroxyphenylacetic acid, and 4-vinylphenol. The levels of 15 compounds present in the exudates were quantified using GC/MS/MS. Six of the seven most abundant compounds were phenolic acids. Significant differences exist between the allelopathic and non-allelopathic cultivars in their production of three of these six compounds. Greater amounts of trans-ferulic acid, p-hydroxybenzoic acid, and caffeic acid were detected in the exudates of allelopathic cultivars. The seventh compound, abietic acid, was significantly higher in the non-allelopathic cultivars.     

Changes in Rice Allelopathy and Rhizosphere Microflora by Inhibiting Rice Phenylalanine Ammonia-lyase Gene Expression

Gene expression of phenylalanine ammonia-lyase (PAL) in allelopathic rice PI312777 was inhibited by RNA interference (RNAi). Transgenic rice showed lower levels of PAL gene expression and PAL activity than wild type rice (WT). The concentrations of phenolic compounds were lower in the root tissues and root exudates of transgenic rice than in those of wild type plants. When barndyardgrass (BYG) was used as the receiver plant, the allelopathic potential of transgenic rice was reduced. The sizes of the bacterial and fungal populations in rice rhizospheric soil at the 3-, 5-, and 7-leaf stages were estimated by using quantitative PCR (qPCR), which showed a decrease in both populations at all stages of leaf development analyzed. However, PI312777 had a larger microbial population than transgenic rice. In addition, in T-RFLP studies, 14 different groups of bacteria were detected in WT and only 6 were detected in transgenic rice. This indicates that there was less rhizospheric bacterial diversity associated with transgenic rice than with WT. These findings collectively suggest that PAL functions as a positive regulator of rice allelopathic potential.     

Root Distribution and Potential Interactions Between Allelopathic Rice,Sprangletop (<Emphasis Type="Italic">Leptochloa</Emphasis> spp.), and Barnyardgrass (<Emphasis Type="Italic">Echinochloa crus-galli</Emphasis>) based on <Superscript>13</Superscript>C Isotope Discrimination Analysis

Weed-suppressive rice cultivars hold promise for improved and more economical weed management in rice. Interactions between roots of rice and weeds are thought to be modulated by the weed-suppressive activity of some rice cultivars, but these phenomena are difficult to measure and not well understood. Thus, above-ground productivity, weed suppression, and root distribution of 11 rice cultivars and two weed species were evaluated in a drill-seeded, flood-irrigated system at Stuttgart, Arkansas, USA in a two-year study. The allelopathic cultivars, PI 312777 and Taichung Native 1 (TN-1), three other weed-suppressive cultivars, three indica-derived breeding selections, and three non-suppressive commercial cultivars were evaluated in field plots infested with barnyardgrass (Echinochloa crus-galli (L.) Beauv.) or bearded sprangletop (sprangletop, Leptochloa fusca (L.) Kunth var. fascicularis (Lam.) N. Snow). The allelopathic cultivars produced more tillers and suppressed both weed species to a greater extent than did the breeding selections or the non-suppressive cultivars. ¹³C isotope discrimination analysis of mixed root samples to a depth of 15 cm revealed that the allelopathic cultivars typically produced a greater fraction of their total root mass in the surface 0–5 cm of soil depth compared to the breeding selections or the non-suppressive cultivars, which tended to distribute their roots more evenly throughout the soil profile. These trends in root mass distribution were apparent at both early (pre-flood) and late-season stages in weed-free and weed-infested plots. Cultivar productivity and root distribution generally responded similarly to competition with the two weed species, but barnyardgrass reduced rice yield and root mass more than did sprangletop. These findings demonstrate for the first time that roots of the allelopathic cultivars PI 312777 and TN-1 explore the upper soil profile more thoroughly than do non-suppressive cultivars under weed-infested and weed-free conditions in flood-irrigated U.S. rice production systems. They raise the interesting prospect that root proliferation near the soil surface might enhance the weed-suppressive activity of allelochemical exudates released from roots. Plant architectural design for weed suppressive activity should take these traits into consideration along with other proven agronomic traits such as high tillering and yield.     

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.     

Inhibition of Water Fern Salvinia minima by Cattail (Typha domingensis) Extracts and by 2-Chlorophenol and Salicylaldehyde

The allelopathic potential of a cattail species (Typha domingensis) towards the common water fern (Salvinia minima) was studied in a short-term bioassay. Aqueous extracts of cattail plants (roots, stems, and leaves) as well as two of its phenolic compounds (2-chlorophenol and salicylaldehyde) were found to inhibit the growth of Salvinia when incorporated in the growth medium. After a period of seven days, chlorophyll concentration, fresh weight, dry weight, and number of fronds of Salvinia were compared and correlations were found among the different parameters. The results suggest an inhibitory effect by components of Typha, especially of the root fractions. Results from pure chemicals are in qualitative and quantitative agreement with those obtained from the cattail extracts.     

The Role of Momilactones in Rice Allelopathy

Large field screening programs and laboratory experiments in many countries have indicated that rice is allelopathic and releases allelochemical(s) into its environment. A number of compounds, such as phenolic acids, fatty acids, phenylalkanoic acids, hydroxamic acids, terpenes, and indoles, have been identified as potential rice allelochemicals. However, the studies reviewed here demonstrate that the labdane-related diterpenoid momilactones are the most important, with momilactone B playing a particularly critical role. Rice plants secrete momilactone B from their roots into the neighboring environments over their entire life cycle at phytotoxic levels, and momilactone B seems to account for the majority of the observed rice allelopathy. In addition, genetic studies have shown that selective removal of the momilactones only from the complex mixture found in rice root exudates significantly reduces allelopathy, demonstrating that these serve as allelochemicals, the importance of which is reflected in the presence of a dedicated momilactone biosynthetic gene cluster in the rice genome.     

Allelopathic potential ofErica vagans,calluna vulgaris,andDaboecia cantabrica

In laboratory tests the allelopathic potential ofErica vagans, Calluna vulgaris, andDaboecia cantabrica was determined. Aqueous extracts of flowers ofD. cantabrica and leaves ofC. vulgaris inhibit root and hypocotyl growth of red clover, the former causing 51% inhibition of germination. Intact aerial parts of the Ericaceae here studied drastically reduced the growth of red clover and 100% inhibition of germination was caused by flowers ofD. cantabrica. Inhibition of aqueous extracts remains after Chromatographic separation, and two well-defined inhibition zones may be observed. Hydrosoluble organic compounds (phenol-like compounds) could probably be responsible for the inhibitions detected.     

Role of Methyl Salicylate on Oviposition Deterrence in Arabidopsis thaliana

Plants attacked by herbivores have evolved different strategies that fend off their enemies. Insect eggs deposited on leaves have been shown to inhibit further oviposition through visual or chemical cues. In some plant species, the volatile methyl salicylate (MeSA) repels gravid insects but whether it plays the same role in the model species Arabidopsis thaliana is currently unknown. Here we showed that Pieris brassicae butterflies laid fewer eggs on Arabidopsis plants that were next to a MeSA dispenser or on plants with constitutively high MeSA emission than on control plants. Surprisingly, the MeSA biosynthesis mutant bsmt1-1 treated with egg extract was still repellent to butterflies when compared to untreated bsmt1-1. Moreover, the expression of BSMT1 was not enhanced by egg extract treatment but was induced by herbivory. Altogether, these results provide evidence that the deterring activity of eggs on gravid butterflies is independent of MeSA emission in Arabidopsis, and that MeSA might rather serve as a deterrent in plants challenged by feeding larvae.     

Carnivore Attractant or Plant Elicitor? Multifunctional Roles of Methyl Salicylate Lures in Tomato Defense

Synthetic plant volatile lures attract natural enemies, but may have non-target effects due to the multifunctional nature of volatile signals. For example, methyl salicylate (MeSA) is used to attract predators, yet also serves as a signaling hormone involved in plant pathogen defense. We investigated the consequences of deploying MeSA lures to attract predators for tomato (Solanum lycopersicum) defense against herbivores. To understand the spatial distribution of the lure’s effect, we exposed tomatoes in the field to MeSA along a linear distance gradient and induced defenses by simulating feeding by hornworm caterpillars in a fully crossed factorial design (+/? MeSA, +/? herbivory). Subsequently, we analyzed activity of several defensive proteins (protease inhibitors, polyphenol oxidase, peroxidase), development of hornworm larvae (Manduca sexta), growth of fungal pathogens (Cladosporium and Alternaria), and attractiveness to herbivores and predators. Overall, MeSA-exposed plants were more resistant to both insects and pathogens. Secondary pathogen infection was reduced by 25% in MeSA exposed plants, possibly due to elevated polyphenol oxidase activity. Interestingly, we found that lures affected plant pathogen defenses equivalently across all distances (up to 4 m away) indicating that horizontal diffusion of a synthetic volatile may be greater than previously assumed. While thrips avoided colonizing hornworm– damaged tomato plants, this induced resistance was not observed upon pre-exposure to MeSA, suggesting that MeSA suppresses the repellant effect induced by herbivory. Thus, using MeSA lures in biological control may inadvertently protect crops from pathogens, but has mixed effects on plant resistance to insect herbivores.     

Use of a comparative approach to identify allelopathic potential and relationship between allelopathy bioassays and “competition” experiments for ten grassland and plant species

Various allelopathy bioassays were used to evaluate the allelopathic potential of 10 grassland forage species against a common test (phytometer) species,Carduus nutans L. Aqueous extracts did not influenceC. nutans germination, although radicle elongation was often severely inhibited.C. nutans was strongly affected by shoot, but not root, leachates. Decomposing ground tissue had mixed effects, and often stimulated shoot production ofC. nutans. Calculation ofR ² (coefficient of determination) values between these results, and the results of previous experiments investigating the effects of the same 10 species onC. nutans emergence and development in field plots and glasshouse competition experiments frequently revealed strong, statistically significant relationships. Our results therefore provide correlative evidence for the importance of allelopathy in field conditions.     

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.     

Mechanism of Selective Phytotoxicity of l-3,4-Dihydroxyphenylalanine (l-Dopa) in Barnyardglass and Lettuce

l-3,4-Dihydroxyphenylalanine (l-dopa) is one of the few allelochemicals in which the phytotoxic action mechanism has been studied. Excess exogenous l-dopa suppresses root elongation in some plant species, and the inhibitory action is species-selective. The main factor of phytotoxicity of l-dopa is considered to be oxidative damage by reactive oxygen species (ROS) and/or free radical species (FRS). This study was performed to elucidate the mechanism of species-selective phytotoxicity. The involvement of ROS/FRS and polyphenol oxidase (PPO) in species-selective phytotoxicity was examined with barnyardgrass (Echinochloa crus-galli L.) and lettuce (Lactuca sativa L.), tolerant and susceptible species, respectively. Lipid peroxidation and melanin accumulation correlated with growth inhibition by L-dopa. Antioxidants, ascorbic acid and α-tocopherol, decreased lipid peroxidation and melanin accumulation and rescued lettuce root from growth inhibition. The oxidation of L-dopa by PPO was much greater in lettuce than in barnyardgrass. From these results, the phytotoxicity of L-dopa is considered due to the oxidative damage caused by ROS/FRS generated from the melanin synthesis pathway. PPO activity might be involved in the mechanism of species-selective phytotoxicity between barnyardgrass and lettuce.     

Macroalgal Defense against Competitors and Herbivores

Macroalgae are the source of many harmful allelopathic compounds, which are synthesized as a defense strategy against competitors and herbivores. Therefore, it can be predicted that certain species reduce aquaculture performance. Herein, the allelopathic ability of 123 different taxa of green, red, and brown algae have been summarized based on literature reports. Research on macroalgae and their allelopathic effects on other animal organisms was conducted primarily in Australia, Mexico, and the United States. Nevertheless, there are also several scientific reports in this field from South America and Asia; the study areas in the latter continents coincide with areas where aquaculture is highly developed and widely practiced. Therefore, the allelopathic activity of macroalgae on coexisting animals is an issue that is worth careful investigation. In this work, we characterize the distribution of allelopathic macroalgae and compare them with aquaculture locations, describe the methods for the study of macroalgal allelopathy, present the taxonomic position of allelopathic macroalgae and their impact on coexisting aquatic competitors (Cnidaria) and herbivores (Annelida, Echinodermata, Arthropoda, Mollusca, and Chordata), and compile information on allelopathic compounds produced by different macroalgae species. This work gathers the current knowledge on the phenomenon of macroalgal allelopathy and their allelochemicals affecting aquatic animal (competitors and predators) worldwide and it provides future research directions for this topic.     

Inhibition of salvinia (Salvinia molesta Mitchell) by parthenium (Parthenium hysterophorus L.). II. Relative effect of flower,leaf, stem,and Root residue on Salvinia and Paddy

The relative effect of parthenium (Parthenium hysterophorus L.) plant residue on growth of salvinia and paddy seedlings was studied. The inhibitory activity of the residue as shown by its effect on the number of healthy fronds (HFN) and biomass was in the order: flower and leaf > stem and root. The flower (FP) and leaf (LP) residue was lethal at and above 0.75% (w/v, the convention used throughout), and inhibitory at lower doses. The stem (SP) and root (RP) residue supported growth of salvinia at lower doses and were slightly inhibitory at higher (1.25%) dose. All the above residue types supported the growth of paddy seedlings except at 1.25%, the highest concentration tested, which was slightly inhibitory. The amounts of chlorophylla, b, total chlorophyll, and carotenoid pigments in the leaves of the paddy seedlings grown in the medium were comparable to the amounts in the leaves of seedlings grown in distilled water. This demonstrates beneficial effects of the treatments. The study shows that salvinia is more sensitive to allelochemicals released by FP and LP into the aqueous medium. Both salvinia and paddy responded similarly to SP and RP by supporting growth at lower doses, probably due to lower levels of inhibitors. The results are discussed with reference to the possible role of allelopathy by parthenium on the population dynamics of aquatic weeds in natural ecosystems.     

Volatile Emissions from an Odorous Plant in Response to Herbivory and Methyl Jasmonate Exposure

Induced volatile terpenes have been commonly reported among diverse agricultural plant species, but less commonly investigated in odorous plant species. Odorous plants synthesize and constitutively store relatively large amounts of volatiles, and these may play a role in defense against herbivores. We examined the effect of herbivory and methyl jasmonate (MeJA) exposure on the release of volatile organic compounds (VOCs) in the marsh elder, Iva frutescens, which contains numerous constitutive VOCs, mainly mono- and sesquiterpenes. Our specific goal was to test for the presence of inducible VOCs in a naturally occurring plant already armed with VOCs. The abundant, native specialist leaf beetle Paria aterrima was used in herbivore induction trials. VOCs were sampled from herbivore wounded and unwounded, and from MeJA treated and untreated I. frutescens. Total VOC emissions were significantly greater in response to herbivory and MeJA treatment compared to unwounded controls. Herbivore wounding caused a substantial shift in the emission profile (42 VOCs from wounded, compared to 8 VOCs from unwounded I. frutescens), and MeJA had a similar yet less substantial influence on the emission pattern (28 VOCs from MeJA treated compared to 8 VOCs from untreated I. frutescens). Constitutive VOC emissions predominated, but some VOCs were detected only in response to herbivory and MeJA treatment, suggesting de novo synthesis. Several VOCs exhibited a delayed emission profile in contrast to the rapid release of constitutive VOCs, and principal components analysis revealed they were not associated with constitutive emissions. While I. frutescens contains many constitutive VOCs that are released immediately in response to herbivory, it also produces novel VOCs in response to feeding by the specialist P. aterrima and MeJA treatment.     

Field-Testing of Methyl Salicylate for Recruitment and Retention of Beneficial Insects in Grapes and Hops

Evidence for recruitment and retention of beneficial insects in grapes and hops using controlled-release dispensers of methyl salicylate (MeSA), a component of herbivore-induced volatile blends, is presented. In a replicated experiment conducted in a juice grape vineyard, sticky cards in blocks baited with MeSA captured significantly greater numbers of five species of predatory insects (Chrysopa nigricornis, Hemerobius sp., Deraeocoris brevis, Stethorus punctum picipes, Orius tristicolor) than unbaited blocks. Four insect families (Syrphidae, Braconidae, Empididae, Sarcophagidae) were also significantly more abundant in the MeSA-baited blocks, as indicated by sticky card captures. Canopy shake samples and sticky card monitoring conducted in a MeSA-baited, unsprayed hop yard indicated development and maintenance of a beneficial arthropod population that was nearly four times greater than that present in an unbaited reference yard. Four times as many S. punctum picipes and six times as many O. tristicolor were sampled in the MeSA yard. Similar contrasts in abundance of these predators and others were apparent when compared with levels recorded in the yard in previous years. The large population of predatory insects in the MeSA-baited hop yard was associated with a dramatic reduction in spider mite numbers, the major arthropod pest of hops, in late June, and subeconomic populations were maintained for the rest of the season. The evidence presented here is highly suggestive that the use of controlled-release MeSA in a crop could increase recruitment and residency of populations of certain beneficial insects. This strategy may have the potential to enhance the efficacy and reliability of conservation biological control in crop pest management.     

Methyl jasmonate as an allelopathic agent: sagebrush inhibits germination of a neighboring tobacco,Nicotiana attenuata

Artemisia tridentata ssp. tridentata is the dominant and defining shrub in the Great Basin Desert, with well-documented allelopathic tendencies that have generally been ascribed to its most abundantly released secondary metabolites. However, as a minor component, sagebrush releases a highly biologically active substance, methyl jasmonate (MeJA), which is known to function as both a germination inhibitor and promoter in laboratory studies. Nicotiana attenuata is a tobacco species native to the Great Basin Desert and grows in newly burned juniper–sagebrush habitats for 2–3 yr following a fire. With a combination of field and laboratory studies, we examined the role of MeJA release from sagebrush by both air and water transport in inhibiting N. attenuata seed germination. We demonstrated that sagebrush interacts allelopathically with the seed bank of N. attenuata through its release of MeJA. In the field, seeds buried 0–40 cm from sagebrush plants for 4 months in net bags had significantly reduced germination compared to seeds buried similarly but protected in plastic bags. Moreover, germination on soils collected from underneath sagebrush plants was reduced by 60% compared to seeds placed on soils collected between sagebrush plants or outside of the sagebrush population. Exposure to A. tridentata seeds and seedlings did not affect N. attenuata germination, suggesting that established sagebrush plants only influence the tobacco's seed bank. In the laboratory, exposure of seeds to sagebrush emissions resulted in germination delays of up to 6 d. Exposure to volatile and aqueous MeJA also inhibited germination of N. attenuata seeds at quantities that are released naturally by sagebrush: 3.5 g/hr and 1.12 g/seed cup (56 ng/seed), respectively. A. tridentata seeds were significantly more resistant to MeJA, being inhibited at 336 g MeJA (16.8 g/seed), 300 times greater than the level of aqueous MeJA required to inhibit N. attenuata seeds. MeJA inhibited N. attenuata germination regardless of the seed's dormancy state and the specific epimer (trans- or cis-) of MeJA. Germination on sagebrush chaff that had been heated to reduce MeJA content was negatively correlated with the amount of MeJA remaining in the chaff. Germination of a nondormant, conspecific tobacco, N. trigonophylla, which grows in the same area but is not associated with fire, is less sensitive than N. attenuata to the extracts of sagebrush litter, but similarly sensitive to MeJA. Additionally, four of five other tobacco species that are not known to be associated with sagebrush are less sensitive to MeJA, suggesting an evolved sensitivity to MeJA. To determine the proportion of germination inhibition of a sagebrush extract that could be attributed to MeJA, we serially diluted sagebrush extracts with water and restored the quantity of MeJA of the original extract by adding appropriate quantities of synthetic MeJA; 16–60% of the inhibitory activity of the original extract could be attributed to the MeJA. We conclude that MeJA release from sagebrush plays an allelopathic role for N. attenuata seed banks, but other unidentified compounds are also involved.