Application of antimicrobial peptides in agriculture and food industry

Antimicrobial peptides have captured the attention of researchers in recent years because of their efficiency in fighting against pathogens. These peptides are found in nature and have been isolated from a wide range of organisms. Furthermore, analogs or synthetic derivatives have successfully been developed on the basis of natural peptide patterns. Long use of pesticides and antibiotics has led to development of resistance among pathogens and other pests as well as increase of environmental and health risks. Antimicrobial peptides are under consideration as new substitutes for conventional pesticides and antibiotics. Many plants and animals have been manipulated with antimicrobial peptide-encoding genes and several pesticides and drugs have been produced based on these peptides. Such strategies and products may still have a long way to go before being confirmed by regulatory bodies and others need to surmount technical problems before being accepted as applicable ones. In spite of these facts, several cases of successful use of antimicrobial peptides in agriculture and food industry indicate a promising future for extensive application of these peptides. In this review, we consider the developing field of antimicrobial peptide applications in various agricultural activities.     

Microbial and viral chitinases: Attractive biopesticides for integrated pest management

The negative impact of the massive use of synthetic pesticides on the environment and on human health has stimulated the search for environment-friendly practices for controlling plant diseases and pests. Among them, biocontrol, which relies on using beneficial organisms or their products (bioactive molecules and/or hydrolytic enzymes), holds the greatest promise and is considered a pillar of integrated pest management. Chitinases are particularly attractive to this purpose since they have fungicidal, insecticidal, and nematicidal activities. Here, current knowledge on the biopesticidal action of microbial and viral chitinases is reviewed, together with a critical analysis of their future development as biopesticides.     

Osmoadaptation mechanisms in prokaryotes: distribution of compatible solutes

Microorganisms respond to osmotic stress mostly by accumulating compatible solutes, either by uptake from the medium or by de novo synthesis. These osmotically active molecules preserve the positive turgor pressure required for cell division. The diversity of compatible solutes is large but falls into a few major chemical categories; they are usually small organic molecules such as amino acids or their derivatives, and carbohydrates or their derivatives. Some are widely distributed in nature while others seem to be exclusively present in specific groups of organisms. This review discusses the diversity and distribution of known classes of compatible solutes found in prokaryotes as well as the increasing knowledge of the genes and pathways involved in their synthesis. The alternative roles of some archetypal compatible solutes not subject to osmoregulatory constraints are also discussed.     

Highly specific gene silencing by artificial microRNAs in the unicellular alga Chlamydomonas reinhardtii

MicroRNAs (miRNAs) are small RNAs, 21 to 22 nucleotides long, with important regulatory roles. They are processed from longer RNA molecules with imperfectly matched foldback regions and they function in modulating the stability and translation of mRNA. Recently, we and others have demonstrated that the unicellular alga Chlamydomonas reinhardtii , like diverse multicellular organisms, contains miRNAs. These RNAs resemble the miRNAs of land plants in that they direct site-specific cleavage of target mRNA with miRNA-complementary motifs and, presumably, act as regulatory molecules in growth and development. Utilizing these findings we have developed a novel artificial miRNA system based on ligation of DNA oligonucleotides that can be used for specific high-throughput gene silencing in green algae.     

In vitro uses of biological cryoprotectants

Ice can be anything from a highly destructive agent in agriculture to a useful building material. Established industries are based on the known rules of physics and chemistry which allow some control of amounts of ice or ice crystal geometry. However, organisms have much more subtle requirements to maintain their delicate internal structure if they are to survive freezing. As a result they have selected specific molecules for freezing-point depression, osmotic regulation, ice nucleation and crystal growth inhibition. All these active species may have potential commercial use once they are identified, understood and produced at economic scales. We examine the progress made so far in extending biological subtlety into commercial processes, and look for prospects for further innovation.     

Pesticides in the Cagayan valley (Philippines): usage, drift patterns and the exposure of farmers differing in income and market access

Over the past ten years, the amount and number of different types of pesticides have increased significantly, which led to a growing concern about the possible adverse effects on human health and the environment. This is particularly true for countries where regulations are not strictly implemented and farmers' knowledge of safe handling is often inadequate. This paper discusses the results of a series of spray experiments to determine drift patterns along field boundaries and the exposure of farmers during their usual spraying exercises. Moreover, farmers' pesticide usage and methods of application will be described, and the effects of income and market accessibility on pesticide use patterns will be investigated. It is based on a study conducted in four villages located at increasing distance from the national highway leading to regional markets and connecting the Cagayan Valley in Northeast Luzon with Manila. The 20 pesticides encountered in this study cover 18 different active ingredients, 9 of which are classified by the WHO as 'highly hazardous' or 'moderately hazardous'. The EPA has classified at least 6 of the encountered pesticide formulations as Restricted Use Pesticides. Nevertheless, all pesticides are freely sold in stores or on markets and applied by farmers without personal protection in an unsafe manner. The farmers living nearest to the highway have the highest income and largest farms. Yet they are most at risk, having easiest access to pesticides and spraying the largest quantities of pesticides per hectare, compared to the farmers living at greater distance from the highway. It is recommended to review the list of pesticides approved for use in the Philippines and discern between Restricted and General Use Pesticides. Several recommendations for improving the implementation of pesticide policies and the IPM program are given.     

Midbodies and phragmoplasts: analogous structures involved in cytokinesis

Cytokinesis is an event common to all organisms that involves the precise coordination of independent pathways involved in cell-cycle regulation and microtubule, membrane, actin and organelle dynamics. In animal cells, the spindle midzone/midbody with associated endo-membrane system are required for late cytokinesis events, including furrow ingression and scission. In plants, cytokinesis is mediated by the phragmoplast, an array of microtubules, actin filaments and associated molecules that act as a framework for the future cell wall. In this article (which is part of the Cytokinesis series), we discuss recent studies that highlight the increasing number of similarities in the components and function of the spindle midzone/midbody in animals and the phragmoplast in plants, suggesting that they might be analogous structures.     

Models in palaeontological functional analysis

Models are a principal tool of modern science. By definition, and in practice, models are not literal representations of reality but provide simplifications or substitutes of the events, scenarios or behaviours that are being studied or predicted. All models make assumptions, and palaeontological models in particular require additional assumptions to study unobservable events in deep time. In the case of functional analysis, the degree of missing data associated with reconstructing musculoskeletal anatomy and neuronal control in extinct organisms has, in the eyes of some scientists, rendered detailed functional analysis of fossils intractable. Such a prognosis may indeed be realized if palaeontologists attempt to recreate elaborate biomechanical models based on missing data and loosely justified assumptions. Yet multiple enabling methodologies and techniques now exist: tools for bracketing boundaries of reality; more rigorous consideration of soft tissues and missing data and methods drawing on physical principles that all organisms must adhere to. As with many aspects of science, the utility of such biomechanical models depends on the questions they seek to address, and the accuracy and validity of the models themselves.     

Genetic analysis of nicotinic signaling in worms and flies

The nicotinic acetylcholine receptor is among the most thoroughly characterized molecules in the nervous system, and its role in mediating fast cholinergic neurotransmission has been broadly conserved in both vertebrates and invertebrates. However, the accessory molecules that facilitate or regulate nicotinic signaling remain mostly unknown. One approach to identify such molecules is to use molecular genetics in a simple, experimentally accessible organism to identify genes required for nicotinic signaling and to determine the molecular identity of the mutant genes through molecular cloning. Because cellular signaling pathways are often highly conserved between different animal phyla, the information gained from studies of simple organisms has historically provided many critical insights into more complex organisms, including humans. Genetic screens essentially make no prior assumptions about the types of molecules involved in the process being studied; thus, they are well suited for identifying previously unknown components of cell signaling pathways. The sophisticated genetic tools available in organisms such as the nematode Caenorhabditis elegans and the fruit fly Drosophila melanogaster have also proven extremely powerful in elucidating complex biologic pathways in the absence of prior biochemical information and for assessing a molecule's in vivo function of in the context of an intact nervous system. This review describes how genetic analysis has been used to investigate nicotinic signaling mechanisms in worms and flies, and the prospects for using these studies to gain insight into nicotinic receptor function and regulation in humans.     

A class of plant glycosyltransferases involved in cellular homeostasis

Many small lipophilic compounds in living cells can be modified by glycosylation. These processes can regulate the bioactivity of the compounds, their intracellular location and their metabolism. The glycosyltransferases involved in biotransformations of small molecules have been grouped into Family 1 of the 69 families that are classified on the basis of substrate recognition and sequence relatedness. In plants, these transfer reactions generally use UDP-glucose with acceptors that include hormones such as auxins and cytokinins, secondary metabolites such as flavonoids, and foreign compounds including herbicides and pesticides. In mammalian organisms, UDP-glucuronic acid is typically used in the transfer reactions to endogenous acceptors, such as steroid and thyroid hormones, bile acids and retinoids, and to xenobiotics, including nonsteroidal anti-inflammatory drugs and dietary metabolites. There is widespread interest in this class of enzyme since they are known to function both in the regulation of cellular homeostasis and in detoxification pathways. This review outlines current knowledge of these glycosyltransferases drawing on information gained from studies of plant and mammalian enzymes.     

Gene expression within a dynamic nuclear landscape

Molecular imaging in living cells or organisms now allows us to observe macromolecular assemblies with a time resolution sufficient to address cause-and-effect relationships on specific molecules. These emerging technologies have gained much interest from the scientific community since they have been able to reveal novel concepts in cell biology, thereby changing our vision of the cell. One main paradigm is that cells stochastically vary, thus implying that population analysis may be misleading. In fact, cells should be analyzed within time-resolved single-cell experiments rather than being compared to other cells within a population. Technological imaging developments as well as the stochastic events present in gene expression have been reviewed. Here, we discuss how the structural organization of the nucleus is revealed using noninvasive single-cell approaches, which ultimately lead to the resolution required for the analysis of highly controlled molecular processes taking place within live cells. We also describe the efforts being made towards physiological approaches within the context of living organisms.     

生物源农药在苹果蠹蛾防治中的应用

苹果蠹蛾是世界性检疫害虫,对我国苹果优势产区构成了巨大威胁。长期依赖化学防治使该虫抗性问题变得十分严峻。为了保障食品安全,以环境友好的生物源农药替代化学农药已成为当前苹果蠹蛾防治的热点。本文对国内外现有的生物源农药,如寄生蜂、不育昆虫、颗粒体病毒、病原线虫、Bt、病原真菌、微孢子虫、性信息素、斑蝥素、多杀菌素等,在苹果蠹蛾防治中的最新应用及其存在的问题进行论述,讨论了生物源农药凭借其种类多、来源广且在用药时期上选择性强等特点,在该虫综合治理中的重要地位及面临的挑战。     

Prospective role of insecticides of fungal origin: Review

Advances in the application of microbial‐based technology in insect pest management assist us to counter problems created by the application of chemical pesticides. These are mainly strong environmental effects, resistance development and high costs. Among the microbial pesticides, fungal pesticides are now preferred as they are target specific, ecofriendly, lacking in toxic residue and are economical. Being numerous with great diversification, entomopathogenic fungi therefore have great potential to control a large variety of insect pests. Fungi are applied directly in form of spores, mycelia or blastospores or by their metabolites (mycotoxins). Both approaches have very promising roles in insect pest management. However, there are three main obstacles in the development of fungal pesticides: (i) scant production of mycotoxins; (ii) carcinogenic mycotoxicosis in non‐target organisms; and (iii) slow effectiveness. Therefore, to eliminate these problems, attention has recently been paid to a synergistic approach to combating insecticide resistance. Next to synergism, genetic manipulation is also used to enhance the pathogenicity and virulence of fungal insecticides. However, the key risk associated with the release of recombinant microorganisms into the environment is that the novel organism may have unforeseen undesirable characteristics. Therefore, the introduction of synergists in pest control could have great benefit both economically and ecologically. An ideal synergistic approach is the mixing of more than two accelerating components together, i.e. tripartite or multiple synergism to enhance effectiveness. Thus, synergistic approaches have a bright future and require further research and financial support.     

Chemical conditionality: a genetic strategy to probe organelle assembly

The assembly of the Escherichia coli outer membrane (OM) is poorly understood. Although insight into fundamental cellular processes is often obtained from studying mutants, OM-defective mutants have not been very informative because they generally have nonspecific permeability defects. Here we show that toxic small molecules can be used in selections employing strains with permeability defects to create particular chemical conditions that demand specific suppressor mutations. Suppressor phenotypes are correlated with the physical properties of the small molecules, but the mutations are not in their target genes. Instead, mutations allow survival by partially restoring membrane impermeability. Using "chemical conditionality," we identified mutations in yfgL, and, here and in the accompanying paper by Wu et al. published in this issue of Cell (Wu et al., 2005), we show that YfgL is part of a multiprotein complex involved in the assembly of OM beta barrel proteins. We posit that panels of toxic small molecules will be useful for generating chemical conditionalities that enable identification of genes required for organelle assembly in other organisms.     

Steroid conjugates: Synthesis and preliminary biological testing of pro-juvenoids

A series of 10 new pro-juvenoids (juvenogens, insect hormonogenic compounds, pro-drug-like agents) was synthesized using isomeric synthetic juvenoids (insect juvenile hormone analogs) and steroid molecules as patterns modifying parts of the complex hormonogenic molecules. In addition, several new synthons were prepared, which were required by the designed synthetic protocol to achieve the target molecules. These pro-juvenoids were subjected to the topical screening tests and to the drinking assays on the red firebug (Pyrrhocoris apterus), a convenient model laboratory phytophagous insect. Simple and efficient synthetic procedures for the preparation of the target pro-juvenoids and their synthons are presented. Furthermore, the biological activity of the pro-juvenoids in comparison with the activity of their parent juvenoids and that of several commercially available agents is demonstrated. Juvenoids and pro-juvenoids may replace toxic insecticides persistent in the insect pest control because they have no adverse effects on non-target organisms and/or human.     

Health benefits of microalgae and their microbiomes

Microalgae comprise a phylogenetically very diverse group of photosynthetic unicellular pro- and eukaryotic organisms growing in marine and other aquatic environments. While they are well explored for the generation of biofuels, their potential as a source of antimicrobial and prebiotic substances have recently received increasing interest. Within this framework, microalgae may offer solutions to the societal challenge we face, concerning the lack of antibiotics treating the growing level of antimicrobial resistant bacteria and fungi in clinical settings. While the vast majority of microalgae and their associated microbiota remain unstudied, they may be a fascinating and rewarding source for novel and more sustainable antimicrobials and alternative molecules and compounds. In this review, we present an overview of the current knowledge on health benefits of microalgae and their associated microbiota. Finally, we describe remaining issues and limitation, and suggest several promising research potentials that should be given attention.     

Recombinant viral insecticides: Delivery of environmentally safe and cost-effective products

Biological control agents such as baculovirus insecticides have many attributes which make them attractive alternatives to synthetic chemical pesticides. However, there have been several economic and agronomic barriers to their widespread use. Among the obstacles to commercialization of viral insecticides have been high production costs, the lack of efficacious formulation and application technologies, and a slow speed of action. Biotechnology has contributed several advances toward overcoming these obstacles. The high cost ofin vivo production can be reduced significantly using a newly developed high-density rearing system termed HeRD. The HeRD technology can be used to rear many different species of lepidopterous larvae for production of viral insecticides, as hosts for parasitoid production, or for sterile-male release programs. Using this technology, the baculovirus production costs are equivalent to sprayable Bt toxins. The field efficacy of viral pesticides and other biological control agents requires cost effective, biologically based formulation and application technologies. Based on current field efficacy evaluations of baculovirus pesticides, formulation/application technologies must be improved for viral pesticides to compete effectively and consistently in most pesticide markets. Through recombinant DNA technology, it is now possible to insert foreign pesticidal genes into viral pesticides, resulting in faster time to death or, more importantly, time to cessation of feeding of the target pests. However, the commercial use of recombinant pesticides has raised several potential environmental issues, including possible effects on non-target organisms, ecological interactions, mitigation and genetic stability. Genetic strategies have been developed to mitigate most of the potential problems associated with recombinant baculovirus pesticides. Five field tests have been conducted in the U.S. to evaluate these strategies. The laboratory and field results illustrate that the genetic strategies employed ensure environmental safety while also reducing production costs.     

Can microbial-based insecticides replace chemical pesticides in agricultural production?

Extensive use of chemical insecticides to control insect pests in agriculture has improved yields and production of high-quality food products. However, chemical insecticides have been shown to be harmful also to beneficial insects and many other organisms like vertebrates. Thus, there is a need to replace those chemical insecticides by other control methods in order to protect the environment. Insect pest pathogens, like bacteria, viruses or fungi, are interesting alternatives for production of microbial-based insecticides to replace the use of chemical products in agriculture. Organic farming, which does not use chemical pesticides for pest control, relies on integrated pest management techniques and in the use of microbial-based insecticides for pest control. Microbial-based insecticides require precise formulation and extensive monitoring of insect pests, since they are highly specific for certain insect pests and in general are more effective for larval young instars. Here, we analyse the possibility of using microbial-based insecticides to replace chemical pesticides in agricultural production.     

Progress in enzyme inhibition based detection of pesticides

The previous few decades have seen the development of biosensors and their use in monitoring of pesticides in food and environmental samples. Although inhibition‐based biosensors have been subject of several recent research works, their performance characteristics greatly depend on the type of immobilization and the presence of interfering compounds in the samples. Moreover, sensitivity, detection limits, and rapidity of the response are few of the other major features that need to be investigated further if they are to become operationally user‐friendly. This review will highlight research carried out in the past on biosensors that are based on enzyme inhibition for determination of organophosphorus compounds and carbamate pesticides.