A strategy for screening an inhibitor of viral silencing suppressors, which attenuates symptom development of plant viruses

To find out whether we can control plant virus diseases by blocking viral RNA silencing suppressors (RSSs), we developed a strategy to screen inhibitors that block the association of RSSs with siRNAs using a surface plasmon resonance assay. The screened chemicals were tested in competition with RSSs for binding to siRNAs using a mobility shift assay. We then confirmed that tested chemicals actually inhibited the RSS activity in vivo using a protoplast assay which was developed for this purpose. This entire system can be adapted to screening inhibitors of not only plant viruses but also some animal viruses possessing RSSs.     

Food chemical discriminations by an herbivorous lizard, Corucia zebrata

Adjustment of chemosensory response to diet should be apparent in evolutionary changes corresponding to dietary shifts. Because most lizards are generalist predators of small animals, relationships between chemosensory behavior and diet are difficult to detect. Nevertheless, the evolution of herbivory by a small number of lizards provides an opportunity to detect any corresponding change in response to plant chemicals. I studied tongue-flicking and biting by the large, herbivorous scincid lizard Corucia zebrata in response to chemical cues from crickets, romaine lettuce, and control stimuli presented on cotton swabs. The skinks exhibited significantly stronger response to plant and animal chemicals than to controls for several variables: greater number of individuals that bit swabs, shorter latency to bite, greater rate of tongue-flicks, and greater tongue-flick attack score. The clearest differences were observed for tongue-flick attack score, a composite variable that combines the effects of tongue-flicking and biting. An insectivorous member of the same subfamily, Scincella lateralis, shows strong tongue-flicking and biting response to chemical prey cues, but not to plant chemicals. This suggests that response to plant chemicals by C. zebrata may have evolved in tandem with the incorporation of plants into the diet and that response to cricket chemicals has been retained, perhaps due to similarities between plant and animal food. The findings support the hypothesis that dietary shifts induce corresponding changes in chemosensory response, but provide only a single independent contrast for a study of correlated evolution between plant diet and chemosensory response to plants. J. Exp. Zool. 286:372-378, 2000.     

Plant bioactives for ruminant health and productivity

Plants have been used throughout history for their medicinal properties. This use has often focused on human health but plants have also been, and still are, applied in ethnoveterinary practice and animal health management. In recent times, the use of synthetic chemicals has become prevalent. Public awareness of the potential environmental and health risks associated with heavy chemical use has also increased. This has put pressure on regulatory bodies to reduce the use of chemicals in agriculture. The most striking example is the 2006 banning of antibiotics in animal feed by the European Union. Moves such as this have increased the drive to find alternatives to synthetic chemicals and research has again turned to the use of plant bioactives as a means of improving animal health. Current scientific evidence suggests there is significant potential to use plants to enhance animal health in general and that of ruminants (cattle, deer, sheep, etc.) in particular. Active areas of research for plant bioactives (particularly saponin and tannin containing plants) include reproductive efficiency, milk and meat quality improvement, foam production/bloat control and methane production. Nematode control is also a significant area of research and the evidence suggests a much broader range of phytochemicals may be effective. This review presents a summary of the literature and examines international research efforts towards the development of plant bioactives for animal health.     

Responses to chemical cues from animal and plant foods by actively foraging insectivorous and omnivorous scincine lizards

If tongue-flicking is important to lizards to sample chemical cues permitting identification of foods, tongue-flicking and subsequent feeding responses should be adjusted to match diet. This hypothesis can be examined for plant foods because most lizards are insectivores, but herbivory/omnivory has evolved independently in many lizard taxa. Here we present experimental data on chemosensory responses to chemical cues from animal prey and palatable plants by three species of the scincine lizards. When tested with chemical stimuli presented on cotton swabs, the insectivorous Eumeces fasciatus responded strongly to prey chemicals but not to chemicals from plants palatable to omnivorous lizards or to pungent or odorless control stimuli. Two omnivorous species, E. schneideri and Scincus mitranus, responded more strongly to chemical cues from both prey and food plants than to the control chemicals. All available data for actively foraging lizards, including these skinks, show that they are capable of prey chemical discrimination, and insectivores do not exhibit elevated tongue-flicking or biting responses to chemical cues from palatable plants. In all of the several species of herbivores/omnivores tested, the lizards show elevated responses to both animal and plant chemicals. We suggest two independent origins of both omnivory and plant chemical discrimination that may account for the evolution of diet and food chemical discriminations in the eight species of skinks studied, five of which are omnivores. All data are consistent with the hypothesis that acquisition of omnivory is accompanied by acquisition of plant chemical discrimination, but data on a broad diversity of taxa are needed for a definitive comparative test of the evolutionary hypothesis. J. Exp. Zool. 287:327-339, 2000.     

Mechanisms of action and biocontrol potential of Trichoderma against fungal plant diseases - A review

Plant diseases are among the major causes of the low productivity of crops, causing yield losses of up to 30%, heralding an enormous threat to global food security. Indiscriminate use of chemical-based fungicides for controlling fungal diseases has raised severe concerns about ecosystem health. Moreover, pathogens have become insensitive against these chemicals necessitating excessive use of chemicals for adequate control. The resulting accumulation of these chemicals in the food chain has provoked numerous health complications. For combating the adversaries of chemical-based fungicides, biological control of fungal pathogens is proposed as an eco-friendly alternative. Among various biological controls, Trichoderma-based biological control agents (BCAs) are widely used in agriculture for controlling soil-borne pathogens. These BCAs are commercialized and known as; stimulators of resistance in plants, growth enhancers, bio-fertilizers, and bio-pesticides. Biological management of plant pathogens has yielded valuable results in the sustainability of ecosystems and compelling improvements in the quality and quantity of agricultural produce. These BCAs exhibit potential against pathogens, remarkably improve photosynthesis, plant growth, and nutrient use efficiency for impressive crop yields. Despite these peculiarities, Trichoderma's mechanisms against pathogens and their growth promotional effects are not thoroughly investigated, hence formulating the prime objective of the current review. Along with these, Trichoderma-based fungicides marketed in different geographical locations are encompassed in this review. Finally, the knowledge gaps and future research directions for improving the efficacy of Trichoderma-based BCAs are discussed.     

Disease diversity and human fertility

The existence of parasitic constraints on the evolution of life-history traits in free-living organisms has been demonstrated in several plant and animal species. However, the association between different diseases and human traits is virtually unknown. We conducted a comparative analysis on a global scale to test whether the diversity of human diseases, some of them responsible for high incidences of morbidity and mortality, were associated with host life-history characteristics. After controlling for direct confounding effects exerted by historical, spatial, economic, and population patterns and their interactions, our findings show that human fertility increases with the diversity and structure of disease types. Thus, disease control may not only lower the costs associated with morbidity, but could also contribute directly or indirectly to reductions in human population growth.     

Studies on the use of plant extracts in assessing the effects of plant metabolism on the mutagenicity and toxicity of pesticides

We have carried out studies on the effects of plant metabolism on the mutagenicity of agricultural chemicals. Our approach is to use a cell-free plant extract, as a source of metabolic enzymes, in a standard Ames test. Using a number of test compounds, we observe that plant metabolism can alter the mutagenicity of several pesticides, and can in some instances give rise to metabolites apparently unique from those which are formed in animal cells. A number of parameters of the assay have been examined, and we find that the assay temperature and preincubation of the pesticide with the extract can significantly alter the outcome of the test. We also have devised a method of controlling for the effects that natural extracts can have on the spontaneous reversion rate of the Salmonella tester strains, in an effort to distinguish slight mutagenic responses from the effects of nutrients (e.g. histidine for his- bacteria) in the assay.     

Disease Surveillance Strategies in Swine

Preparations derived from plants were the original therapeutic interventions used by man to control diseases (including parasites), both within humans and livestock. Development of herbal products depended upon local botanical flora with the result that different remedies tended to develop in different parts of the world. Nevertheless, in some instances, the same or related plants were used over wide geographic regions, which also was the result of communication and/or the importation of plant material of high repute. Thus, the Nordic countries have an ancient, rich and diverse history of plant derived anthelmintic medications for human and animal use. Although some of the more commonly used herbal de-wormers were derived from imported plants, or their products, many are from endemic plants or those that thrive in the Scandinavian environment. With the advent of the modern chemotherapeutic era, and the discovery, development and marketing of a seemingly unlimited variety of highly efficacious, safe synthetic chemicals with very wide spectra of activities, herbal remedies virtually disappeared from the consciousness – at least in the Western world. This attitude is now rapidly changing. There is a widespread resurgence in natural product medication, driven by major threats posed by multi-resistant pest, or disease, organisms and the diminishing public perceptions that synthetic chemicals are the panacea to health and disease control. This review attempts to provide a comprehensive account of the depth of historical Nordic information available on herbal de-wormers, with emphasis on livestock and to provide some insights on potentially rewarding areas of "re-discovery" and scientific evaluation in this field.     

The role of natural plant products in modulating the immune system: An adaptable approach for combating disease in grazing animals

Plants provide herbivores with an array of chemicals with the potential to improve health and well-being. For instance, phytochemicals, known as secondary compounds, which protect plants from consumers and pests can adversely affect cellular and metabolic processes in herbivores, but at low doses and in appropriate mixtures, they can have beneficial effects on animal nutrition and health, though the latter has not been explored in great breadth or depth. In this review, we summarize the potential impact of natural plant products on immunomodulation and other therapeutic effects in herbivores. Development of preventative strategies to help animals resist disease would be a more economical, ecological and socially effective long-term healthcare strategy than treating diseases. In this realm, immunomodulation promoted by forages emerges as an interesting alternative and complement to chemotherapy. The challenge for feeding systems will be to incorporate mixes of plants with bioactive properties in ways that enhance health without compromising animal production and well-being. A solution to this challenge may involve developing management programs that acknowledge the ability of animals to learn about the beneficial effects of diverse foods.     

Food chemical cues elicit general and population-specific effects on lingual and biting behaviors in the lacertid lizard Podarcis lilfordi

Actively foraging lizards are capable of identifying prey using only chemical cues sampled by tongue-flicking, and the relatively few omnivorous and herbivorous lizards tested similarly can identify both animal and plant foods from chemical cues. Whether lizards that eat plants respond to cues specific to preferred plant types and whether there is geographic variability in responses to cues from various plants correlated with the importance of those plants in local diets is unknown. In three populations of an omnivorous lacertid, the Balearic lizard Podarcis lilfordi, we studied chemosensory sampling and feeding responses to chemical cues from plant and animal foods presented on cotton swabs. Each lizard population is endemic to one islet off the coast of Menorca, Balearic Islands, Spain. Lizards in all three populations discriminated chemical cues from plant and animal foods from control substances. Our results extend findings of prey chemical discrimination and plant chemical discrimination in omnivores, increasing confidence that correlated evolution has occurred between plant diet and chemosensory response to palatable plants. There were no consistent differences among populations in tongue-flicking and biting responses to stimuli from flowers of syntopic and allopatric plant species. The lizards may respond to cues indicative of palatability in a wide range of plant species rather than exhibiting strong responses only to locally available plant species. Nevertheless, tongue-flicking and biting frequencies varied among plant species, perhaps indicating food preferences. In addition, there were differences among populations in tongue-flick rates, latency to bite, and licking behavior. Licking was observed in only one lizard population as a response to floral chemicals from only one of the plants species tested, raising the possibility of a population-specific linkage between identification of a particular plant species and performance of an appropriate feeding response.     

Sustainable agriculture: how to sustain a production system in a changing environment

During the past 10-15 years, sustainable agriculture has progressed from a focus primarily on a low-input, organic farming approach with a major emphasis on small fruit or vegetable production farms, often described as Low Input Sustainable Agriculture, to the current situation where sustainability is an important part of mainstream animal and plant production units. The US Department of Agriculture programmes cover a broad range of activities, including conserving the natural resource base, enhancing environmental quality, and sustaining productivity of the nation's farms. The use of Geographic Information Systems technology to direct application of fertilisers, pesticides, and herbicides is one example of a rapidly emerging technology that can reduce use of external inputs, protect the agricultural environment, and improve economic returns. This Geographic Information Systems technology also is being used to localise animal pest and disease problems, assist in regulatory or control measures, and identify high risk areas that might need different management systems or should be avoided as sites for animal production. Use of intensive grazing systems also has increased markedly over the past 5-6 years. These systems will allow longer grazing seasons in southern parts of the USA, provide more efficient use of the forages being produced and reduce labour costs in the typical dairy operation. Major animal and plant production agriculture-oriented programmes at the US Department of Agriculture focus on integrated production systems, use of Integrated Pest Management techniques, and development of alternative methods to manage pests and diseases that reduce or avoid the use of drugs and chemicals. The US Department of Agriculture has a programme for sustainable agriculture, the Sustainable Agriculture Research and Education programme, which emphasises alternative approaches for animal and plant production systems.     

Sterol biosynthesis in oomycete pathogens

Oomycetes are a diverse group of filamentous eukaryotic microbes comprising devastating animal and plant pathogens. They share many characteristics with fungi, including polarized hyphal extension and production of spores, but phylogenetics studies have clearly placed oomycetes outside the fungal kingdom, in the kingdom Stramenopila which also includes marine organisms such as diatoms and brown algae. Oomycetes display various specific biochemical features, including sterol metabolism. Sterols are essential isoprenoid compounds involved in membrane function and hormone signaling. Oomycetes belonging to Peronosporales, such as Phytophthora sp., are unable to synthesize their own sterols and must acquire them from their plant or animal hosts. In contrast, a combination of biochemical and molecular approaches allowed us to decipher a nearly complete sterol biosynthetic pathway leading to fucosterol in the legume pathogen Aphanomyces euteiches, an oomycete belonging to Saprolegniales. Importantly, sterol demethylase, a key enzyme from this pathway, is susceptible to chemicals widely used in agriculture and medicine as antifungal drugs, suggesting that similar products could be used against plant and animal diseases caused by Saprolegniales.Key words: azoles, fungicides, root rot, elicitin, Saprolegnia, chromoalveolates     

外来入侵植物刺萼龙葵的入侵特征与防治策略

刺萼龙葵是原产于北美洲的恶性外来入侵植物,在世界各地广为分布,已入侵我国北方地区,严重威胁我国农牧业安全,亟待明确刺萼龙葵入侵过程与危害,为我国制定刺萼龙葵防治策略提供参考。本文对刺萼龙葵生物学和生态学特性、传播途径、入侵历史和分布特征、危害、现有防除措施及存在等问题进行综述。刺萼龙葵具有花期长、花粉萌发率和结实率高、果实和种子产量大等高繁殖能力特征;能适应多变气候和异质性生境;具有自体传播、风力传播、水力传播、动物传播和人为传播等多种传播途径;已先后入侵了我国9个省级行政区的55个区县,向华北、华中和华东快速入侵的可能性高;刺萼龙葵的植株及分泌物对人畜安全、动物皮毛质量、草地植被结构、农作物产量等方面造成严重危害,并帮助传播植物病虫害。然而,现有的化学和物理防除措施仍不能彻底遏制传播、消除危害和保障生产。为有效防除刺萼龙葵,应开展刺萼龙葵入侵风险评估,针对不同土地利用类型制定防治措施,加强入侵机制和防控技术研究。     

植物-昆虫间的化学通讯及其行为控制

在植物与昆虫间的化学通讯中植物气味物质起着决定性的作用,它调控着昆虫的多种行为,诸如引诱昆虫趋向寄主植物,刺激昆虫取食,引导昆虫选择产卵场所,进行传粉和防御昆虫等。有些植物则当受到食植性昆虫危害时会释出一些引诱害虫天敌的化学信号。这些化学信号是一些挥发性萜类混合物,天敌昆虫就以此来区分受害和未受害植株。尽管目前在害虫综合治理中,昆虫信息素的应用越来越显得比天然植物气味源更受重视,但是必须指出的是,昆虫信息化合物首次成功地使用于植物保护的却是天然植物气味源。在利用植物气味源作害虫测报和防治中,近年来一种简单价廉的粘胶诱捕器己成为多种害虫的标准测报工具。在害虫综合治理中利用植物气味源的技术显然是具有不可估量的潜力。文中提出了利用基因工程技术来改造植物,使植物能释放特定的驱避剂或其它控制昆虫行为的特殊气味物质的新概念。     

OBPC Symposium: Maize 2004 &; beyond—Plant virus-based vectors in agriculture and biotechnology

Summary The study of plant viruses and their interaction with the plant host has contributed greatly to our understanding of plant biology. The recent development of plant viruses as transient expression vectors has not only enhanced our understanding of virus biology and antiviral defense mechanisms in plants, but has also led to the use of plant viral-based vectors as tools for gene discovery and production of recombinant proteins in plants for control of human and animal diseases. An overview of the state-of-the-art of viral expression systems, is presented, as well as examples from our laboratory on their use in identifying nuclear targeting motifs on viroid molecules and development of therapeutic proteins for control of animal diseases.     

Application of the comet assay to measure DNA damage induced by UV radiation in the hydrophyte, Spirodela polyrhiza

The single-cell gel electrophoresis or comet assay is now widely used to detect DNA damage in animal cells induced by radiation or chemicals. Here, we apply the comet assay to measure ultraviolet (UV)-B-induced DNA damage in plant cells. The accepted animal cell protocol for the comet assay was modified to adapt it to plant cells. The major modifications were conversion of the plant cells to protoplasts and the use of T4 endonuclease V. As a positive control hydrogen peroxide was applied. Significant DNA damage was detected at 100 μ M H₂O₂. This type of DNA damage was not affected by T4 endonuclease V treatment, which implies that the mechanism of H₂O₂-induced DNA damage was different from UV-B-induced DNA damage. Our results also indicate that both UV-A and UV-B radiation can induce DNA single-strand breaks in plant cells, while UV-B was more effective than UV-A for inducing pyrimidine dimer formation.     

植物内生菌及其防治植物病害的研究进展

综述了植物内生菌及其防治植物病害的研究进展.植物内生菌分布广,种类多,几乎存在于所有目前已研究过的陆生及水生植物中,目前全世界至少已在80个属290多种禾本科植物中发现有内生真菌,在各种农作物及经济作物中发现的内生细菌已超过120种.感染内生菌的植物宿主往往具有生长快速、抗逆境、抗病害、抗动物危害等优势,比未感染内生菌的植株更具生存竞争力.植物内生菌的防病机理主要表现在通过产生抗生素类,水解酶类,植物生长调节剂和生物碱类物质,与病原菌竞争营养物质,增强宿主植物的抵抗力以及诱导植物产生系统抗性等途径抑制病原菌生长.另外,对植物内生真菌和内生细菌的分离、筛选和检测方法;利用植物内生菌控制植物病害的途径如人工接种内生菌,利用内生菌代谢产生的抗生素以及将内生菌作为基因工程的载体菌等进行了综述.同时,对植物内生菌作为生物防治因子未来发展前景及存在的问题进行了讨论.利用植物内生菌作为生物防治因子进行大田防病,需要考虑它的病理学、生态学和形态学等方面的影响.     

植物的诱导抗虫性

植物对植食性昆虫的抗性可包括两个方面,即植物的组成抗性（constitutiveresistance）和诱导抗性（inducedresistance）。组成抗性是指植物在遭受植食性昆虫进攻前就已存在的抗虫特性;而诱导抗性是指植物在遭受植食性昆虫进攻后所表现出来的一种抗虫特性[1,2]。根据作用世代的不同,诱导抗性又分为迅速的诱导抗性（rapidlyinducedre－sistance,RIR）和滞后的诱导抗性（delayedinducedresistance,DIR）。前者是指对当前世代的植食性昆虫的影响,而后者是指对后续的1～几个世代的植食性昆虫的影响[2]。研究植物的诱导抗虫性,不仅能在…