The Surface Coat of Plant-Parasitic Nematodes: Chemical Composition,Origin, and Biological Role—A Review

Chemical composition, origin, and biological role of the surface coat (SC) of plant-parasitic nematodes are described and compared with those of animal-parasitic and free-living nematodes. The SC of the plant-parasitic nematodes is 5-30 nm thick and is characterized by a net negative charge. It consists, at least in part, of glycoproteins and proteins with various molecular weights, depending upon the nematode species. The lability of its components and the binding of human red blood cells to the surface of many tylenchid plant-parasitic nematodes, as well as the binding of several neoglycoproteins to the root-knot nematode Meloidogyne, suggest the presence of carbohydrate-recognition-domains for host plants and parasitic or predatory soil microorganisms (Pasteuria penetrans and Dactylaria spp., for example). These features may also assist in nematode adaptations to soil environments and to plant hosts with defense mechanisms that depend on reactions to nematode surfaces. Surface coat proteins can be species and race specific, a characteristic with promising diagnostic potential.     

Shift from morphological to recent advanced molecular approaches for the identification of nematodes

Nematodes are the most diverse but most minor studied microorganisms found in soil, water, animals, or plants. Either beneficial or pathogenic, they significantly affect human and animal health, plant production and ultimately affect the environmental equilibrium. Knowledge of their taxonomy and biology are the main issues to answer the different challenges associated with these microorganisms. The classical morphology-based nematode taxonomy and biodiversity studies have proved insufficient to identify closely related taxa and have challenged most biologists. Several molecular approaches have been used to supplement morphological methods and solve these problems with markable success. The molecular techniques range from enzyme analysis, protein-based information to DNA sequence analysis. For several decades, efforts have been made to integrate molecular approaches with digital 3D image-capturing technology to improve the identification accuracy of such a taxonomically challenging group and communicate morphological data. This review presents various molecular techniques and provides examples of recent advances in these methods to identify free-living and plant-parasitic nematodes.     

Plant-parasitic Nematode Acetylcholinesterase Inhibition by Carbamate and Organophosphate Nematicides

The sensitivity of acetylcholinesterases (ACHE) isolated from the plant-parasitic nematodes Meloidogyne arenaria, M. incognita, and Heterodera glycines and the free-living nematode Caenorhabditis elegans to carbamate and organophosphate nematicides was examined. The AChE from plant-parasitic nematode species were more sensitive to carbamate inhibitors than was AChE from C. elegans, but response to the organophosphates was approximately equivalent. The sulfur-containing phosphate nematicides were poor inhibitors of nematode acetylcholinesterase, but treatment with an oxidizing agent greatly improved inhibition. Behavioral bioassays with living nematodes revealed a poor relationship between enzyme inhibition and expression of symptoms in live nematodes.     

Computational and phylogenetic validation of nematode horizontal gene transfer

Sequencing of expressed genes has shown that nematodes, particularly the plant-parasitic nematodes, have genes purportedly acquired from other kingdoms by horizontal gene transfer. The prevailing orthodoxy is that such transfer has been a driving force in the evolution of niche specificity, and a recent paper in BMC Evolutionary Biology that presents a detailed phylogenetic analysis of cellulase genes in the free-living nematode Pristionchus pacificus at the species, genus and family levels substantiates this hypothesis.     

Dynamics of free-living and plant-parasitic nematodes on vegetables in Dhapa municipal waste of Kolkata,India

We investigated areas at the eastern fringe of Kolkata, Dhapa–a land filled with Municipal Solid Waste (MSW) land of West Bengal (India) for exploring nematode species diversity, abundance and dynamic at three sites practising different vegetable-based cropping sequences. The area sampled at monthly interval and the soil and root populations estimated at laboratory. The plant-parasitic nematode (PPN) species and free-living group (saprozoics) were estimated from soil. No differences in the diversity of nematode species and feeding groups among the sites in the MSW land were found but considerable variations in abundance and dynamics of PPNs were noted. All three sites at MSW, total PPNs outnumbered free-living nematodes in most of the samples but mononchids were less abundant. Saprozoic nematode index (SNI) was determined; SNI value was low (0.33–0.40) in this organic rich production system.     

The rDNA Internal Transcribed Spacer Region as a Taxonomic Marker for Nematodes

The ITS region from a wide taxonomic range of nematodes, including secernentean and adenophorean taxa, and free-living, entomopathogenic, and plant-parasitic species, was evaluated as a taxonomic marker. Size of the amplified product aided in the initial determination of group membership, and also suggested groups that may require taxonomic reevaluation. Congeneric species often displayed identically sized ITS regions, but genera such as Pratylenchus and Tylenchorhynchus had species with large differences in size. ITS heterogeneity in individuals and populations was identified in several nematode taxa. PCR-RFLP of ITS1 is advocated as a method of taxonomic analysis in genera such as Helicotylenchus that contain numerous species with few diagnostic morphological characteristics.     

Reproductive mode evolution in nematodes: insights from molecular phylogenies and recently discovered species

The Phylum Nematoda has long been known to contain a great diversity of species that vary in reproductive mode, though our understanding of the evolutionary origins, causes and consequences of nematode reproductive mode change have only recently started to mature. Here we bring together and analyze recent progress on reproductive mode evolution throughout the phylum, resulting from the application of molecular phylogenetic approaches and newly discovered nematode species. Reproductive mode variation is reviewed in multiple free-living, animal-parasitic and plant-parasitic nematode groups. Discussion ranges from the model nematode Caenorhabditis elegans and its close relatives, to the plant-parasitic nematodes of the Meloidogyne genus where there is extreme variation in reproductive mode between and even within species, to the vertebrate-parasitic genus Strongyloides and related genera where reproductive mode varies across generations (heterogony). Multiple evolutionary transitions from dioecous (obligately outcrossing) to hermaphroditism and parthenogenesis in the phylum are discussed, along with one case of an evolutionary transition from hermaphroditism to doioecy in the Oscheius genus. We consider the roles of underlying genetic mechanisms in promoting reproductive plasticity in this phylum, as well as the potential evolutionary forces promoting transitions in reproductive mode.     

A Free-living Panagrolaimus sp. from Armenia Can Survive in Anhydrobiosis for 8.7 Years

Although the ability of plant-parasitic nematodes to survive in a dehydrated or anhydrobiotic state for long periods of time has been well documented, the ability of free-living nematodes has not. Here we report on the survival of a free-living nematode, Panagrolaimus sp., from Armenia in the anhydrobiotic state for 8.7 years. This Panagrolaimus sp. can be cultured and maintained readily and may provide a good system for studying anhydrobiosis in nematodes.     

Soil Organic Matter and Management of Plant-Parasitic Nematodes

Organic matter and its replenishment has become a major component of soil health management programs. Many of the soil''s physical, chemical, and biological properties are a function of organic matter content and quality. Adding organic matter to soil influences diverse and important biological activities. The diversity and number of free-living and plant-parasitic nematodes are altered by rotational crops, cover crops, green manures, and other sources of organic matter. Soil management programs should include the use of the proper organic materials to improve soil chemical, physical, and biological parameters and to suppress plant-parasitic nematodes and soilborne pathogens. It is critical to monitor the effects of organic matter additions on activities of major and minor plant-parasitic nematodes in the production system. This paper presents a general review of information in the literature on the effects of crop rotation, cover crops, and green manures on nematodes and their damage to economic crops.     

First record of a free-living dalyellioid turbellarian from the pacific:Balgetia pacifica nov. spec.

Balgetia pacifica nov. spec. from coarse sand beaches along the west coast of San Juan Island (Washington) is described and its taxonomy discussed. The species is characterized by its large bursal apparatus with a strengthened copulatory bursa.B. pacifica is the first free-living species of the Turbellaria Dalyellioida recorded from the Pacific Ocean.     

棕囊藻属(Phaeocystis)的分类与生活史(综述)

棕囊藻属Phaeocystis（定鞭藻纲Prymnesiophyceae)的分类问题目前还有争论。其种的分类标准是以初始的群体形态、地理分布、细胞特征等以及分子生物学特征,如染色体倍性,基因组大小等为依据。基于以上各种分类特征,目前比较确定的棕囊藻属藻类有四种：一种是只观察到单细胞形态的凹孔棕囊藻（P.scrobiculata),另外三种是能够形成群体的波切棕囊藻（P.pouchetii)、球形棕囊藻（P.globosa)和南极棕囊藻（P.antarctica)。棕囊藻具有一个复杂的异形生活史,介于几种游离的单细胞（不动的细胞,具有鞭毛的动细胞,小游动孢子以及可能存在的大游动孢子）和群体之间的形态交替。但其生活史中仍有许多不确定的问题。     

Sensitive PCR Detection of Meloidogyne arenaria, M. incognita, and M. javanica Extracted from Soil

We have developed a simple PCR assay protocol for detection of the root-knot nematode (RKN) species Meloidogyne arenaria, M. incognita, and M. javanica extracted from soil. Nematodes are extracted from soil using Baermann funnels and centrifugal flotation. The nematode-containing fraction is then digested with proteinase K, and a PCR assay is carried out with primers specific for this group of RKN and with universal primers spanning the ITS of rRNA genes. The presence of RKN J2 can be detected among large numbers of other plant-parasitic and free-living nematodes. The procedure was tested with several soil types and crops from different locations and was found to be sensitive and accurate. Analysis of unknowns and spiked soil samples indicated that detection sensitivity was the same as or higher than by microscopic examination.     

Functional characterisation of a cyst nematode acetylcholinesterase gene using Caenorhabditis elegans as a heterologous system

Migration of plant-parasitic nematode infective larval stages through soil and invasion of roots requires perception and integration of sensory cues culminating in particular responses that lead to root penetration and parasite establishment. Components of the chemoreceptive neuronal circuitry involved in these responses are targets for control measures aimed at preventing infection. Here we report, to our knowledge, the first isolation of cyst nematode ace-2 genes encoding acetylcholinesterase (AChE). The ace-2 genes from Globodera pallida (Gp-ace-2) and Heterodera glycines (Hg-ace-2) show homology to ace-2 of Caenorhabditis elegans (Ce-ace-2). Gp-ace-2 is expressed most highly in the infective J2 stage with lowest expression in the early parasitic stages. Expression and functional analysis of the Globodera gene were carried out using the free-living nematode C. elegans in order to overcome the refractory nature of the obligate parasite G. pallida to many biological studies. Caenorhabditis elegans transformed with a GFP reporter construct under the control of the Gp-ace-2 promoter exhibited specific and restricted GFP expression in neuronal cells in the head ganglia. Gp-ACE-2 protein can functionally complement its C. elegans homologue. A chimeric construct containing the Ce-ace-2 promoter region and the Gp-ace-2 coding region and 3′ untranslated region was able to restore a normal phenotype to the uncoordinated C. elegans double mutant ace-1;ace-2. This study demonstrates conservation of AChE function and expression between free-living and plant-parasitic nematode species, and highlights the utility of C. elegans as a heterologous system to study neuronal aspects of plant-parasitic nematode biology.     

The effects of Brassica green manures on plant parasitic and free living nematodes used in combination with reduced rates of synthetic nematicides

Brassica plants once incorporated into soil as green manures have recently been shown to have biofumigant properties and have the potential of controlling plant-parasitic nematodes. In Washington State, plant-parasitic nematodes are successfully managed with synthetic nematicides. However, some of the synthetic nematicides became unavailable recently or their supply is limited leaving growers with few choices to control plant-parasitic nematodes. The objective of this project was to evaluate the effects of Brassica green manures on their own and in combination with reduced rates of synthetic nematicides on plant-parasitic nematodes and free living nematodes. In a greenhouse experiment and field trials in three seasons, Brassica green manures in combination with half the recommended rate of 1,3-dichloropropene (1,3-D, Telone) reduced root knot nematode, Meloidogyne chitwoodi to below detection levels, and reduced lesion nematodes, Pratylenchus penetrans and stubby root nematodes, Paratrichodorus allius, to below economic thresholds. The combination treatments did not affect the beneficial free-living nematode populations and the non-pathogenic Pseudomonas. The total cost of growing and soil-incorporating Brassica crops as green manures in combination with reduced rates of 1,3-D was approximately 35% lower than the present commercial costs for application for the full rate of this fumigant. Integrating conventional management practices with novel techniques fosters sustainability of production systems and can increase economic benefit to producers while reducing chemical input.     

Occurrence of Plant-Parasitic Nematodes of Turfgrass in Korea

Plant-parasitic nematodes are not only an important constraint on agricultural crop production, but also cause both direct and indirect damage to turfgrass, which is a ground cover plant. However, studies on plant-parasitic nematodes of turfgrass in Korea are scarce. A survey for plant-parasitic nematodes was carried out on 13 golf courses in Korea. The results yielded 28 species/taxa belonging to 16 genera and 12 families of plant-parasitic nematodes. Among the isolated species, Helicotylenchus microlobus, Mesocriconema nebraskense, Tylenchorhynchus claytoni, Mesocriconema sp., and Meloidogyne graminicola were the most prevalent species in all management zones. Twelve species were new records of plant-parasitic nematodes in Korea. Highest maximum densities were showed by T. claytoni, Paratylenchus nanus, M. nebraskense, M. graminicola, and H. microlobus. Diversity (H′), was significantly higher in fairways compared to tees and greens, though species evenness (J′) and dominance (D) showed no statistically significant differences. This information is crucial in nematode problem diagnosis, and the subsequent formulation of management strategies.     

Overview of organic amendments for management of plant-parasitic nematodes, with case studies from Florida

Organic amendments have been widely used for management of plant-parasitic nematodes. Relatively rapid declines in nematode population levels may occur when decomposing materials release toxic compounds, while longer-term effects might include increases in nematode antagonists. Improved crop nutrition and plant growth following amendment use may lead to tolerance of plant-parasitic nematodes. Results depend on a great variety of factors such as material used, processing/composting of material, application rate, test arena, crop rotation and agronomic practices, soil type, climate, and other environmental factors. Reasons for variable performance and interpretation of results from amendment studies are discussed. Case studies of amendments for nematode management are reviewed from Florida, where composts and crop residues are the most frequently used amendments. Plant growth was often improved by amendment application, free-living nematodes (especially bacterivores) were often stimulated, but suppression of plant-parasitic nematodes was inconsistent. Amendments were generally not as effective as soil fumigation with methyl bromide for managing root-knot nematodes (Meloidogyne spp.), and often population levels or galling of root-knot nematodes in amended plots did not differ from those in non-amended control plots. While amendments may improve plant growth and stimulate soil food webs, additional study and testing are needed before they could be used reliably for management of plant-parasitic nematodes under Florida conditions.     

Imbroglios of viral taxonomy: genetic exchange and failings of phenetic approaches

The practice of classifying organisms into hierarchical groups originated with Aristotle and was codified into nearly immutable biological law by Linnaeus. The heart of taxonomy is the biological species, which forms the foundation for higher levels of classification. Whereas species have long been established among sexual eukaryotes, achieving a meaningful species concept for prokaryotes has been an onerous task and has proven exceedingly difficult for describing viruses and bacteriophages. Moreover, the assembly of viral "species" into higher-order taxonomic groupings has been even more tenuous, since these groupings were based initially on limited numbers of morphological features and more recently on overall genomic similarities. The wealth of nucleotide sequence information that catalyzed a revolution in the taxonomy of free-living organisms necessitates a reevaluation of the concept of viral species, genera, families, and higher levels of classification. Just as microbiologists discarded dubious morphological traits in favor of more accurate molecular yardsticks of evolutionary change, virologists can gain new insight into viral evolution through the rigorous analyses afforded by the molecular phylogenetics of viral genes. For bacteriophages, such dissections of genomic sequences reveal fundamental flaws in the Linnaean paradigm that necessitate a new view of viral evolution, classification, and taxonomy.     

Abiotic Soil Factors and Plant-parasitic Nematode Communities

A natural community of plant-parasitic nematodes is usually polyspecific. The host plant is the most important driving force in nematode populations, but abiotic factors are important in maintaining the steady state. Nematode communities often separate by abiotic soil factors. In any continuous habitat, including crop plants, generally there is a consistency of the most abundant species, which are largely predictable. Data on single species provide little information about community patterns. Although certain nematode species might be indicators of certain environments, only when we discuss such aspects as diversity and ordination do we relate to communities irrespective of any interactions among component species. Only if plant-parasitic nematodes act independently of each other do autecological studies have validity in polyspecific communities.     

Morphological Comparison and Taxonomic Utility of Copulatory Structures of Selected Nematode Species

Spicules of 9 Meloidogyne, 2 Heterodera, 3 Globodera, and 12 other plant-parasitic, insect-parasitic, and free-living nematodes were excised and examined using scanning electron microscopy (SEM). Gubernacula of some of the species were also excised, and their structure was determined. The two spicules of all species examined were symmetrically identical in morphology. The spicule typically consisted of three parts: head, shaft, and blade with dorsal and ventral vela. The spicular nerve entered through the cytoplasmic core opening on the lateral outer surface of the spicule head and generally communicated with the exterior through one or two pores at the spicule tip. Spicules of Xiphinema sp. and Aporcelaimellus sp. were not composed of three typical parts, were less sclerotized, and lacked a cytoplasmic core opening and distal pores. Spicules of Aphelenchoides spp. had heads expanded into apex and rostrum and had very arcuate blades with thick dorsal and ventral edges (limbs). Gubernaculum shapes were stable within a species, but differed among species examined. The accessory structures of Hoplolaimus galeatus consisted of a tongue-shaped gubernaculum with two titillae at its distal end and a plate-like capitulum terminating distally in two flat, wing-like structures. A comparison of spicules of several species of Meloidogyne by SEM and light microscopy revealed no striking morphological differences.