Stephen O. Duke

Invited Paper: Chemicals from nature for weed management

Abstract Natural products represent a vast repository of materials and compounds with evolved biological activity, including phytotoxicity. Some of these compounds can be used directly or as templates for herbicides. The molecular target sites of these compounds are often unique. Strategies for the discovery of these materials and compounds are outlined. Numerous examples of individual phytotoxins and crude preparations with weed management potential are provided. An example of research to find a natural product solution of a unique pest management problem (blue-green algae in aquaculture) is described. Finally, the problems associated with natural products for pest control are discussed.

Somatic mutation‐mediated evolution of herbicide resistance in the nonindigenous invasive plant hydrilla (Hydrilla verticillata)

Hydrilla (Hydrilla verticillata L.f. Royle) was introduced to the surface water of Florida in the 1950s and is today one of the most serious aquatic weed problems in the USA. As a result of concerns associated with the applications of pesticides to aquatic systems, fluridone is the only USEPA‐approved chemical that provides systemic control of hydrilla. After a decrease in fluridones efficacy at controlling hydrilla, 200 Florida water bodies were sampled to determine the extent of the problem and the biological basis for the reduced efficacy. Our studies revealed that hydrilla phenotypes with two‐ to six‐fold higher fluridone resistance were present in 20 water bodies. Since fluridone is an inhibitor of the enzyme phytoene desaturase (PDS), the gene for PDS (pds) was cloned from herbicide‐susceptible and ‐resistant hydrilla plants. We report for the first time in higher plants three independent herbicide‐resistant hydrilla biotypes arising from the selection of somatic mutations at the arginine 304 codon of pds. The three PDS variants had specific activities similar to the wild‐type enzyme but were two to five times less sensitive to fluridone. In vitro activity levels of the enzymes correlated with in vivo resistance of the corresponding biotypes. As hydrilla spread rapidly to lakes across the southern United States in the past, the expansion of resistant biotypes is likely to pose significant environmental challenges in the future.

Strategies for Using Transgenes to Produce Allelopathic Crops1

Crop allelopathy has seldom been used effectively by farmers in weed management. Traditional breeding methods have not been successful in producing highly allelopathic crops with good yields. Genetic engineering may have the potential for overcoming this impasse. Crops have been made resistant to insects, pathogens, and herbicides with transgenes, but biotechnology has not produced crops that control weeds with allelochemicals. The strategies for producing allelopathic crops by biotechnology are relatively complex, usually involving multiple genes. One can choose to enhance production of allelochemicals already present in a crop or to impart the production of new compounds. The first strategy involves identification of the allelochemical(s), determination of the enzymes and genes encoding them, and the use of genetic engineering to enhance their production. The latter strategy employs altering existing biochemical pathways by insertions of transgenes to produce new allelochemicals. With either strategy, there are potential problems with tissue-specific promoters, autotoxicity, metabolic imbalances, and proper movement of the allelopathic compound to the rhizosphere. Nomenclature: Glufosinate; barley, Hordeum vulgare L.; celery, Apium graveoens L. var. Dulce (Miller) Pers.; cucumber, Cucumis sativa L.; diffuse knapweed, Centaurea diffusa Lam #3 CENDI.; maize, Zea mays L.; potato, Solanum tuberosum L.; rice, Oryza sativa L.: sorghum, Sorghum bicolor (L.) Moench # SORVU; sudangrass, Sorghum sudanese (Piper) Stapf; tomato, Lypersicon esculentum L.; wheat, Triticum aestivum L. Additional index words: Allelochemical, genetic engineering, phytotoxin, transgene. Abbreviations: DIMBOA, 2,4-dihydroxy-7-methoxy-1, 4-benzoxazin-3-one; PCR, polymerase chain reaction.

Challenges, achievements and opportunities in allelopathy research

Abstract Allelopathy is defined as the suppression of any aspect of growth and/or development of one plant by another through the release of chemical compounds. Although allelopathic interference has been demonstrated many times using in vitro experiments, few studies have clearly demonstrated allelopathy in natural settings. This difficulty reflects the complexity in examining and demonstrating allelopathic interactions under field conditions. In this paper we address a number of issues related to the complexity of allelopathic interference in higher plants: These are: (i) is a demonstrated pattern or zone of inhibition important in documenting allelopathy? (ii) is it ecologically relevant to explain the allelopathic potential of a species based on a single bioactive chemical? (iii) what is the significance of the various modes of allelochemical release from the plant into the environment? (iv) do soil characteristics clearly influence allelopathic activity? (v) is it necessary to exclude other plant interference mechanisms?, and (vi) how can new achievements in allelopathy research aid in solving problems related to relevant ecological issues encountered in research conducted upon natural systems and agroecosystems? A greater knowledge of plant interactions in ecologically relevant environments, as well as the study of biochemical pathways, will enhance our understanding of the role of allelopathy in agricultural and natural settings. In addition, novel findings related to the relevant enzymes and genes involved in production of putative allelochemicals, allelochemical persistence in the rhizosphere, the molecular target sites of allelochemicals in sensitive plant species and the influence of allelochemicals upon other organisms will likely lead to enhanced utilization of natural products for pest management or as pharmaceuticals and nutraceuticals. This review will address these recent findings, as well as the major challenges which continue to influence the outcomes of allelopathy research.

Transcriptional responses to cantharidin, a protein phosphatase inhibitor, in Arabidopsis thaliana reveal the involvement of multiple signal transduction pathways.

Cantharidin is a natural compound isolated from the blister beetle (Epicauta spp.). It is a potent inhibitor of protein serine/threonine phosphatases (PPPs), especially PP2A and PP4. Protein phosphatases and kinases maintain a sensitive balance between dephosphorylated and phosphorylated forms of appropriate proteins, thereby playing important roles in signal transduction pathways and regulation of gene expression, cellular proliferation, cell differentiation, apoptosis and other processes. The foliage of 12-day-old Arabidopsis thaliana seedlings was treated with 200 µM (IC(30) ) of the PPP inhibitor cantharidin, and the entire transcriptome profile was determined by microarray analysis at 2, 10 and 24 h after treatment. The transcription of approximately 10% (2577) of the 24 000 genes of Arabidopsis changed significantly (P≤ 0.05 and signal log ratios: ≥1 or ≤-1) after treatment. Inhibition of PPPs significantly reduced transcription of genes associated with auxin and light signaling and induced expression of genes involved in the hypersensitive response and in flagellin and abscisic acid signaling. The great variety of up- and downregulated genes in this microarray experiment implied that cantharidin interfered with the activities of PPPs that interact directly or indirectly with receptors or are located near the beginning of signal transduction pathways. In many cases, PPPs interact with protein complexes of various receptors such as ethylene or light sensors localized in different cell compartments. They function as negative regulators modifying receptor functions, thus altering signaling that influences transcriptional responses.

INTERACTIONS OF SYNTHETIC HERBICIDES WITH PLANT DISEASE AND MICROBIAL HERBICIDES

Synthetic herbicides have the potential to influence plant disease by several mechanisms. They can enhance disease or protect plants from pathogens due to direct effects on the microbe, to effects on the plant, or to effects on both organisms. The particular effect is a function of many factors including the herbicide class and its formulation, the disease species, the plant species, timing of herbicide application and infection, and environmental factors. These secondary effects of herbicides have not been sufficiently studied to fully understand their environmental toxicology implications or their potential for enhanced integrated pest management. Furthermore, understanding these interactions can sometimes be critical in the success of biocontrol of weeds with plant pathogens.

Potential Ecological Roles of Artemisinin Produced by Artemisia annua L.

Artemisia annua L. (annual wormwood, Asteraceae) and its secondary metabolite artemisinin, a unique sesquiterpene lactone with an endoperoxide bridge, has gained much attention due to its antimalarial properties. Artemisinin has a complex structure that requires a significant amount of energy for the plant to synthesize. So, what are the benefits to A. annua of producing this unique compound, and what is the ecological role of artemisinin? This review addresses these questions, discussing evidence of the potential utility of artemisinin in protecting the plant from insects and other herbivores, as well as pathogens and competing plant species. Abiotic factors affecting the artemisinin production, as well as mechanisms of artemisinin release to the surroundings also are discussed, and new data are provided on the toxicity of artemisinin towards soil and aquatic organisms. The antifungal and antibacterial effects reported are not very pronounced. Several studies have reported that extracts of A. annua have insecticidal effects, though few studies have proven that artemisinin could be the single compound responsible for the observed effects. However, the pathogen(s) or insect(s) that may have provided the selection pressure for the evolution of artemisinin synthesis may not have been represented in the research thus far conducted. The relatively high level of phytotoxicity of artemisinin in soil indicates that plant/plant allelopathy could be a beneficial function of artemisinin to the producing plant. The release routes of artemisinin (movement from roots and wash off from leaf surfaces) from A. annua to the soil support the rationale for allelopathy.

Hormesis with glyphosate depends on coffee growth stage

Weed management systems in almost all Brazilian coffee plantations allow herbicide spray to drift on crop plants. In order to evaluate if there is any effect of the most commonly used herbicide in coffee production, glyphosate, on coffee plants, a range of glyphosate doses were applied directly on coffee plants at two distinct plant growth stages. Although growth of both young and old plants was reduced at higher glyphosate doses, low doses caused no effects on growth characteristics of young plants and stimulated growth of older plants. Therefore, hormesis with glyphosate is dependent on coffee plant growth stage at the time of herbicide application.

Autofluorescence: Biological functions and technical applications

Chlorophylls are the most remarkable examples of fluorophores, and their fluorescence has been intensively studied as a non-invasive tool for assessment of photosynthesis. Many other fluorophores occur in plants, such as alkaloids, phenolic compounds and porphyrins. Fluorescence could be more than just a physicochemical curiosity in the plant kingdom, as several functional roles in biocommunication occur or have been proposed. Besides, fluorescence emitted by secondary metabolites can convert damaging blue and UV into wavelengths potentially useful for photosynthesis. Detection of the fluorescence of some secondary phytochemicals may be a cue for some pollinators and/or seed dispersal organisms. Independently of their functions, plant fluorophores provide researchers with a tool that allows the visualization of some metabolites in plants and cells, complementing and overcoming some of the limitations of the use of fluorescent proteins and dyes to probe plant physiology and biochemistry. Some fluorophores are influenced by environmental interactions, allowing fluorescence to be also used as a specific stress indicator.

Overview of glyphosate‐resistant weeds worldwide

Glyphosate is the most widely used and successful herbicide discovered to date, but its utility is now threatened by the occurrence of several glyphosate-resistant weed species. Glyphosate resistance first appeared in Lolium rigidum in an apple orchard in Australia in 1996, ironically the year that the first glyphosate-resistant crop (soybean) was introduced in the USA. Thirty-eight weed species have now evolved resistance to glyphosate, distributed across 37 countries and in 34 different crops and six non-crop situations. Although glyphosate-resistant weeds have been identified in orchards, vineyards, plantations, cereals, fallow and non-crop situations, it is the glyphosate-resistant weeds in glyphosate-resistant crop systems that dominate the area infested and growing economic impact. Glyphosate-resistant weeds present the greatest threat to sustained weed control in major agronomic crops because this herbicide is used to control weeds with resistance to herbicides with other sites of action, and no new herbicide sites of action have been introduced for over 30 years. Industry has responded by developing herbicide resistance traits in major crops that allow existing herbicides to be used in a new way. However, over reliance on these traits will result in multiple-resistance in weeds. Weed control in major crops is at a precarious point, where we must maintain the utility of the herbicides we have until we can transition to new weed management technologies.