Fungal chitinases and their biological role in the antagonism onto nematode eggs. A review

Chitin, the most abundant aminopolysaccharide in nature, is a rigid and resistant structural component that contributes to the mechanical strength of chitin-containing organisms. Chemically, it is a linear cationic heteropolysaccharide composed of N-acetyl-D-glucosamine and D-glucosamine units. The enzymatic degradation of chitin is performed by a chitinolytic system with synergistic and consecutive action. Diverse organisms (containing chitin or not) produce a great variety of chitinolytic enzymes with different specificities and catalytic properties. Their physiological roles involve nutrition, parasitism, chitin recycling, morphogenesis, and/or defense. Microorganisms, as the main environmental chitin degraders, constitute a very important natural source of chitinolytic enzymes. Nowadays, the most used method for pest and plant diseases control is the utilization of chemical agents, causative of significant environmental pollution. Social concern has generated the search for alternative control systems (i.e., biological control), which contribute to the generation of sustainable agricultural development. Interactions among the different organisms are the natural bases of biological control. Interest in chitinolytic enzymes in the field of biological control has arisen due to their possible involvement in antagonistic activity against pathogenic chitin-containing organisms. The absence of chitin in plants and vertebrate animals allows the consideration of safe and selective “target” molecules for control of chitin-containing pathogenic organisms. Fungi show appropriate characteristics as potential biological control agents of insects, fungi, and nematodes due to the production of fungal enzymes with antagonistic action. The antagonistic interactions between fungi and plant nematode parasites are among the most studied experimental models because of the high economic relevance. Fungi which target nematodes are known as nematophagous fungi. The nematode egg is the only structural element where the presence of chitin has been demonstrated. In spite of being one of the most resistant biological structures, eggs are susceptible to being attacked by egg-parasitic fungi. A combination of physical and chemical phenomena result in their complete destruction. The contribution of fungal chitinases to the in vitro rupture of the eggshell confirms their role as a pathogenic factor. Chitinases have been produced by traditional fermentation methods, which have been improved by optimizing the culture conditions for industrial processes. Although wild-type microorganisms constitute an alternative source of chitinolytic enzymes, the advances in molecular biology are allowing the genetic transformation of fungi to obtain strains with high capability as biocontrol agents. Simultaneously, a better understanding of rhizosphere interactions, additional to the discovery of new molecular biology tools, will allow the choosing of better alternatives for the biological control of nematodes in order to achieve an integrated management of the soil ecosystem.     

Compatibility Between Pasteuria penetrans Isolates and Meloidogyne Populations from Spain

Several populations of Pasteuria isolated from fields in Spain were compared with other Pasteuria populations, held in collections at the Institute de Recerca i Tecnologia, Agroalimentaries (IRTA), Cabrils or IACR-Rothamsted, for their ability to adhere to and infect root-knot nematodes ( Meloidogyne spp.) grown on host plants differing in their susceptibility to root-knot nematodes. The results showed a high level of variation in both the ability of a population of Pasteuria to adhere to a particular population of nematode and vice versa. In particular the isolates of Pasteuria originating from M. hapla retained a high level of specificity for the species from which they originated. The infection of the nematodes by the bacteria was generally low, even when nematodes were encumbered with relatively high levels of spores. It is suggested that prolonged storage (6 years) may reduce the ability of spores to infect nematodes independently of adhesion.     

Nematode feeding sites: unique organs in plant roots

Although generally unnoticed, nearly all crop plants have one or more species of nematodes that feed on their roots, frequently causing tremendous yield losses. The group of sedentary nematodes, which are among the most damaging plant-parasitic nematodes, cause the formation of special organs called nematode feeding sites (NFS) in the root tissue. In this review we discuss key metabolic and cellular changes correlated with NFS development, and similarities and discrepancies between different types of NFS are highlighted.     

Natural genetic and induced plant resistance, as a control strategy to plant-parasitic nematodes alternative to pesticides

Plant-parasitic nematodes are pests of a wide range of economically important crops, causing severe losses to agriculture. Natural genetic resistance of plants is expected to be a valid solution of the many problems nematodes cause all over the world. Progress in resistance applications is particularly important for the less-developed countries of tropical and subtropical regions, since use of resistant cultivars may be the only possible and economically feasible control strategy in those farming systems. Resistance is being considered of particular importance also in modern high-input production systems of developed countries, as the customary reliance on chemical nematicides has been restricted or has come to an end. This review briefly describes the genetic bases of resistance to nematodes in plants and focuses on the chances and problems of its exploitation as a key element in an integrated management program. Much space is dedicated to the major problem of resistance durability, in that the intensive use of resistant cultivars is likely to increasingly induce the selection of virulent populations able to “break” the resistance. Protocols of pest-host suitability are described, as bioassays are being used to evaluate local nematode populations in their potential to be selected on resistant germplasm and endanger resistant crops. The recent progress in using robust and durable resistances against nematodes as an efficient method for growers in vegetable cropping systems is reported, as well as the possible use of chemicals that do not show any unfavorable impact on environment, to induce in plants resistance against plant-parasitic nematodes.     

Overexpression of Wild Arachis Lipocalin Enhances Root-Knot Nematode Resistance in Peanut Hairy Roots

Plant Molecular Biology Reporter - Plant parasitic root-knot nematodes (RKN) have evolved sophisticated strategies for exploiting plants, and they cause devastating yield losses in many susceptible...     

Strategies for transgenic nematode control in developed and developing world crops

Nematodes cause an estimated $118b annual losses to world crops and they are not readily controlled by pesticides or other control options. For many crops natural resistance genes are unavailable to plant breeders or progress by this approach is slow. Transgenic plants can provide nematode resistance for such crops. Two approaches have been field trialled that control a wide range of nematodes by either limiting use of their dietary protein uptake from the crop or by preventing root invasion without a direct lethality. In addition, RNA interference increasingly in tandem with genomic studies is providing a range of potential resistance traits that involve no novel protein production. Transgenic resistance can be delivered by tissue specific promoters to just root tissues where most economic nematodes invade and feed rather than the harvested yield. High efficacy and durability can be provided by stacking nematode resistance traits including any that natural resistance provides. The constraints to uptake centre on market acceptance and not the availability of appropriate biotechnology. The need to deploy nematode resistance is intensifying with loss of pesticides, an increased need to protect crop profit margins and in many developing world countries where nematodes severely damage both commodity and staple crops.     

The relationship of potato yield with and without nematicide to density of potato cyst nematodes, lobodera rostochiensis and G. pallida

Trials relating response to nematicide to potato cyst nematode density were conducted initially in the West Midlands and later in other Regions. Thirteen trials, only three of which were within the intensive potato growing areas, conformed to a general pattern with yield losses being largely recouped by nematicide treatment. Five trials, four within the intensive areas, gave no correlation between potato yield and nematode density and an unpredictable response to nematicide. The control of the nematode appeared to be poorer in the latter trials but other factors affecting yield and nematode multiplication may have been involved. It is speculated the main factor might be interaction with fungal organisms e.g. Rhizoctonia or Verticillium. Evidence is presented to show that on several trials the nematicide has an effect other than by controlling potato cyst nematodes. Yield losses caused by potato cyst nematodes are more variable than previous work indicated, probably due to varietal, seasonal or environmental influences.     

Protein proteinase inhibitor genes in combat against insects, pests, and pathogens: natural and engineered phytoprotection

The continual need to increase food production necessitates the development and application of novel biotechnologies to enable the provision of improved crop varieties in a timely and cost-effective way. A milestone in this field was the introduction of Bacillus thuringiensis (Bt) entomotoxic proteins into plants. Despite the success of this technology, there is need for development of alternative strategies of phytoprotection. Biotechnology offers sustainable solutions to the problem of pests, pathogens, and plant parasitic nematodes in the form of other insecticidal protein genes. A variety of genes, besides (Bt) toxins that are now available for genetic engineering for pest resistance are genes for vegetative insecticidal proteins, proteinase inhibitors, alpha-amylase inhibitors, and plant lectins. This review presents a comprehensive summary of research efforts that focus on the potential use and advantages of using proteinase inhibitor genes to engineer insect- and pest-resistance. Crop protection by means of PI genes is an important component of Integrated Pest Management programmes.     

Use of endophytes as biocontrol agents

Plant diseases, caused by various microorganisms, including viruses, bacteria, fungi, protozoa and nematodes, affect agricultural practices and result in significant crop losses. Fungal pathogens are the major cause of plant diseases and infect most plants. Agrochemicals play a significant role in plant disease management to ensure a sustainable and productive agricultural system. However, the intensive use of chemicals has adverse effects on humans and ecosystem functioning and also reduces agricultural sustainability. A sustainable agriculture is achieved through reduction or elimination of fertilizers and agrochemicals, resulting in minimal impact to the environment. Recently, the use of antagonistic endophytes as biocontrol agents is drawing special attention as an attractive option for management of some plant diseases, resulting in minimal impact to the environment. Endophytes that resides asymptomatically within a plant, have the potential to provide a source of candidate strains for potential biocontrol applications. This review addresses biocontrol methods using endophytic fungi such as Colletotrichum, Cladosporium, Fusarium, Pestalotiopsis and Trichoderma species as an attractive option for management of some plant diseases. Potential endophytes are screened in vitro and in vivo to test their antagonistic actions by different mechanisms, including mycoparasitism, production of lytic enzymes and/or antibiotics and induction of plant defenses. Currently, efforts are being made to commercialize these biocontrol agents. A continued research pipeline consisting of screening, in vitro and in vivo testing, biomass production and commercialization of endophytes as biocontrol agents may contribute to sustainable agriculture.     

Targeting Biocontrol with the Slug-Parasitic Nematode Phasmarhabditis hermaphrodita in Slug Feeding Areas: a Model Study

The nematode Phasmarhabditis hermaphrodita was applied to soil in an outdoor miniplot experiment to protect Chinese cabbage seedlings from damage by the field slug Deroceras reticulatum. The aim was to investigate the possibility of reducing the numbers of nematodes applied by only partially spraying soil in the area where slug control was needed. Nematodes sprayed as overall applications were compared with band applications along plant rows and spot applications around individual plants, in plots with nine or 18 plants. Band and spot applications were applied at two rates, designated the full rate (same number of nematodes per plot as in the overall application) and the area rate (same number of nematodes per unit area comprising 43% (band) and 18% (spot) of the overall application). In plots with 18 plants, where spot-treated plant alternated with untreated plants, no significant difference in damage was found between spot-treated plants and untreated plants. This indicates that slugs were not repelled from nematode-treated areas and that any effects in reducing slug damage were not due to repellency. All nematode treatments resulted in a significant reduction in the mean level of slug damage to seedlings from six or more days after treatment. However, there were significant interactions between nematode treatment, the number of plants per plot, the position of plants within plots (edge or middle) and time after treatment. The effect of time after treatment was modelled. The log time to 50% reduction in slug damage (t 50 ) was related to the area treated and the dose applied. In plots with band or spot treatments at the full dose, there was a relatively small increase in t 50 with declining area treated. In plots treated with band or spot treatments at the area dose, t 50 increased consistently with declining relative area treated. The final level of damage, expressed as a percentage of damage on untreated plots (P), was influenced by both the dose and area treated. Final damage was greatest on spoti treated plots where half the plants were untreated. We conclude that partial treatment of soil around all plants to be protected from slug damage is a potentially valuable method of reducing the overall nematode dose required for control of slug damage, provided that some damage can be tolerated.     

Production losses and control of helminths in ruminants of temperate regions

In this review of research on production losses and control principles in ruminant helminthiasis, the discussion primarily refers to nematode infections under conditions in Europe.Among the loss-producing effects of nematodes much interest has recently been centered on the possible reduction in milk yield of cows brought about by Ostertagia ostertagi. Another important factor of new interest is the inappetence associated with trichostrongyle infections. Furthermore, an example is given of hidden losses in association with the adoption of less profitable production systems to avoid parasitism.It is imperative that scientists and advisors are aware of newer management practices which engender risks of loss-producing infections. High stocking rates and the spreading of slurry onto pastures are examples of procedures which may increase the hazards of parasitism. To implement practices of routine control it is important to examine the regularity with which the herbage infectivity patterns occur. Principles of controlling nematode infections by systems of grazing management and by strategic anthelmintic treatments are discussed and illustrated with examples. Finally, problems of integrating control in farm enterprises are discussed.     

Management of root-knot nematode (Meloidogyne spp.) on sweet pepper (Capsicum annuum L.) with moringa (Moringa oleifera Lam.) leaf powder

A major constraint facing sweet pepper production is infestation by nematodes leading to reduced yields. Field studies were conducted during the 2012 cropping season at the Experimental Farms of the University for Development Studies, Nyankpala, Northern region, Ghana, to determine efficacy of various levels of moringa leaf powder for the control of root-knot nematodes in sweet pepper (Capsicum annuum L.) in the savanna ecology of Ghana. Treatments consisted of three levels of moringa leaf powder (40, 60 and 80?g/L) per plot and 0?g/L (control). The experiment was laid out in a randomised complete block design with each treatment replicated four times. The infestations of root-knot nematodes were significantly lower in the moringa leaf powder-treated plots than the control. Although significant differences were not observed in all the parameters evaluated among the moringa leaf powder treatments, sweet pepper plants treated with 80?g/L of moringa leaf powder per plot recorded the highest mean value of plant height, number of leaves, number of fruits per plant, fruit weight per plant total yield per plot and the thickest plant girth. Similarly, the sweet pepper plants treated with 80?g/L of moringa leaf powder had the lowest infection index (root gall) and nematode population. Application of moringa leaf powder at 40, 60 and 80?g/L increased sweet pepper yield and decreased nematode population confirming their potential in management of root-knot nematodes.     

Rice root-knot nematode (Meloidogyne graminicola) infestation in rice

Rice (Oryza sativa) is an important staple food crop for majority of human population in the world in general and in Asia in particular. However, among various pests and diseases which constitute important constraints in the successful crop production, plant parasitic nematodes play an important role and account for yield losses to the extent of 90%. The major nematode pests associated with rice are Ditylenchus angustus, Aphelenchoides besseyi, Hirschmanniella spp., Heterodera oryzicola and Meloidogyne graminicola. However, rice root-knot nematode (M. graminicola) happens to be the most important pest and is prevalent in major rice producing countries of the world. In India, the distribution of M. graminicola in rice growing areas of different states has been documented in nematode distribution atlas prepared by All India Coordinated Research Project (Nematodes) and published by Directorate of Information and Publications of Agriculture, Indian Council of Agricultural Research, New Delhi, India during 2010. M. graminicola affected rice plants show stunting and chlorosis due to the characteristic terminal swellings/galls on the roots which ultimately result in severe reduction in growth and yield. Number of eco-friendly management technologies against M. graminicola have been developed and demonstrated, including the use of bioagents for minimising the losses due to rice root-knot nematode. This review is focused on collating information to understand the current scenario of rice root-knot nematodes with greater emphasis on its ecological requirements, damage symptoms, biology, morphology, host range and management strategies.     

The sensory amphidial structures of Caenorhabditis elegans are involved in macrocyclic lactone uptake and anthelmintic resistance

Parasitic nematodes represent formidable pathogens of humans, livestock and crop plants. Control of these parasites is almost exclusively dependent on a small group of anthelmintic drugs, the most important of which belong to the macrocyclic lactone class. The extensive use of these drugs to control the ubiquitous trichostrongylid parasites of grazing livestock has resulted in the emergence of both single and multi-drug resistance. The expectation is that this resistance will eventually occur in the human parasites such as the common and debilitating soil transmitted nematodes and vector-borne filarial nematodes. While the modes of action of anthelmintics such as ivermectin, have been elucidated, notably in the model nematode Caenorhabditis elegans, the molecular nature of this resistance remains to be fully determined. Here we show that the anterior amphids play a key role in ivermectin uptake and mutations in these sensory structures result in ivermectin resistance in C. elegans. Random genetic mutant screens, detailed analysis of existing amphid mutants and lipophilic dye uptake indicate that the non-motile ciliated amphid neurons are a major route of ivermectin ingress; the majority of the mutants characterised in this study are predicted to be involved in intraflagellar transport. In addition to a role in ivermectin resistance, a subset of the amphid mutants are resistant to the non-related benzimidazole class of anthelmintics, raising the potential link to a multi-drug resistance mechanism. The amphid structures are present in all nematodes and are clearly defined in a drug-sensitive strain of Haemonchus contortus. It is predicted that amphidial drug uptake and intraflagellar transport may prove to be significant in the development of single and multi-drug resistance in the nematode pathogens of veterinary and human importance.     

Modelling the spatial configuration of refuges for a sustainable control of pests: a case study of Bt cotton

The 'high-dose-refuge' (HDR) strategy is widely recommended by the biotechnology industry and regulatory authorities to delay pest adaptation to transgenic crops that produce Bacillus thuringiensis (Bt) toxins. This involves cultivating nontoxic plants (refuges) in close proximity to crops producing a high dose of Bt toxin. The principal cost associated with this strategy is due to yield losses suffered by farmers growing unprotected, refuge plants. Using a population genetic model of selection in a spatially heterogeneous environment, we show the existence of an optimal spatial configuration of refuges that could prevent the evolution of resistance whilst reducing the use of costly refuges. In particular, the sustainable control of pests is achievable with the use of more aggregated distributions of nontransgenic plants and transgenic plants producing lower doses of toxin. The HDR strategy is thus suboptimal within the context of sustainable agricultural development.     

Naz, a resistant cultivar on bean root rot disease in Zanjan province, northwest Iran

Field bean is a major crops in different parts of northwest Iran especially Zanjan province. Recently the bean plants were severely subjected to damping off or decline disease which caused yield losses in bean growing regions. A regional research was done from 2003 to 2005 to get general information on the causal agent of disease and its control management. Infected plants were collected from different studied areas and transferred to laboratory. Crown and plant roots were cultured in PDA as common media and PPA as selective media for Fusarium species after surface sterilization with sodium hypochlorite. Plates were incubated in standard culture room then isolated fungi were identified. Different Fusarium species were isolated, however the main pathogen isolated from plant samples and soil around the roots was F. sambucium Fuckel. The disease caused up to 50% yield losses in some fields in studied areas. Study showed the "Naz" cultivar was the main resistant race to the disease and had the most yield production in the field.     

Microbial control of plant-parasitic nematodes: a five-party interaction

Plant-parasitic nematodes cause significant economic losses to a wide variety of crops. Chemical control is a widely used option for plant-parasitic nematode management. However, chemical nematicides are now being reappraised in respect of environmental hazard, high costs, limited availability in many developing countries or their diminished effectiveness following repeated applications. This review presents progress made in the field of microbial antagonists of plant-parasitic nematodes, including nematophagous fungi, endophytic fungi, actinomycetes and bacteria. A wide variety of microorganisms are capable of repelling, inhibiting or killing plant-parasitic nematodes, but the commercialisation of these microorganisms lags far behind their resource investigation. One limiting factor is their inconsistent performance in the field. No matter how well suited a nematode antagonist is to a target nematode in a laboratory test, rational management decision can be made only by analysing the interactions naturally occurring among “host plant–nematode target–soil–microbial control agent (MCA)–environment”. As we begin to develop a better understanding of the complex interactions, microbial control of nematodes will be more fine-tuned. Multidisciplinary collaboration and integration of biological control with other control methods will␣also contribute to more successful control practices.