Mustard biofumigation disrupts biological control by Steinernema spp. nematodes in the soil

Mustard green manures or seed meal high in glucosinolates, which produce a natural biofumigant upon incorporation into the soil, form an alternative to synthetic fumigants. However, the non-target impacts of these biofumigants in the field are unclear. We examined the effectiveness of soil incorporation of Brassica carinata seed meal both in controlling the plant-parasitic Columbia root-knot nematode (Meloidogyne chitwoodi), and on the biological control exerted by the entomopathogenic nematodes Steinernema feltiae and Steinernema riobrave on root-knot nematodes and the Colorado potato beetle (Leptinotarsa decemlineata). Singly, both the seed meal and Steinernema spp. reduced root-knot nematode damage to potato tubers and increased marketable tuber yields. However, there was a negative interaction between the two bioagents such that their combination did not further improve suppression of plant-parasitic nematodes. Thus, mustard seed meal applications harmful to the target root-knot nematode also disrupted the ability of Steinernema spp. to act as biocontrol agents. Further, we observed modest disruption of the biological control of potato beetles following biofumigation. But, the potato beetles were less likely to lay eggs on potato plants grown in mustard-amended soil, suggesting a counteracting benefit of mustard application. Multiple, complementary controls must be integrated to replace the very effective pest suppression typical of synthetic soil fumigants. Our study suggests significant interference between biofumigation and biocontrol agents in the soil, presenting challenges in combining these two environmentally friendly approaches to managing plant-parasitic nematodes and other pests.     

Entomopathogenic nematodes,a potential microbial biopesticide: mass production and commercialisation status – a mini review

Parasitic nematodes have several important attributes that make them excellent candidates for biological control of soil insects. These nematodes can be produced by in vivo by baiting technique on insects and commercially by in vitro solid/liquid culturing. Numerous insect pests on many different crops are being controlled by these insect parasitic nematodes, including root weevils, flea beetles, mint root borer, colorado potato beetle, white grubs, caterpillars and plant parasitic root nematode, e.g. root-knot nematodes. Utilisation of entomopathogenic nematodes (EPN) has raised intense interest and has been a growing concern globally mainly because of its potential efficiency, exemption from registration and other impressive attributes for utilising against the control of soil dwelling pests. This review highlights the mass production, commercialisation and utilisation of EPN as microbial biopesticide in bio-intensive pest management programmes.     

Resistance and tolerance of potato varieties to potato rot nematode (Ditylenchus destructor) and stem nematode (Ditylenchus dipsaci)

Ditylenchus destructor and Ditylenchus dipsaci are economically important plant‐parasitic nematodes, affecting potato production mostly in temperate climates. Management through crop rotation is not feasible because of their wide host range. These nematodes are listed as quarantine pests in many countries. Limited information exists on the resistance and tolerance of currently cultivated potatoes to D. destructor and D. dipsaci. Two greenhouse experiments were conducted to screen 25 potato varieties for resistance to and tolerance for D. destructor and D. dipsaci infections. Reproduction factor (RF) and relative susceptibility (RS) were used to evaluate resistance, while potato tuber damage and tuber weight reduction was used to evaluate tolerance. Based on the RF, 16 varieties were evaluated as susceptible (S) while 5 varieties were evaluated as resistant (R) to D. destructor; varieties ‘Innovator’, ‘Aveka’ and ‘Spunta’ were resistant to D. dipsaci based on RF. ‘Désirée’ was observed to be highly susceptible to D. destructor and D. dipsaci in both experiments and was used as the standard susceptible control variety for the calculation of RS. A scale of 1–9 was used to classify RS of the potato varieties to D. destructor and D. dipsaci, where 9 indicated the highest level of resistance. All classes of resistance to D. destructor and D. dipsaci were observed in the potato varieties tested in the experiments. Six varieties had significantly lower RS to D. dipsaci than the standard susceptible control variety. Tolerant to highly sensitive potato varieties to both nematodes were also observed. RS and external potato tuber damage were identified as suitable methods for resistance and tolerance determination, respectively. This study provides essential information on the status of resistance and tolerance in potato varieties against D. destructor and D. dipsaci but needs to be confirmed under field conditions.     

Histopathology of Meloidogyne chitwoodi (Golden et al.) on Russett Burbank Potato

Pathogenesis of M. chitwoodi associated with potato (Solanum tuberosura cv. Russet Burbank) followed a pattern characteristic of root-knot nematodes. Giant ceils developed in the phloem tissues of roots, stolons, and tubers and appeared to arise by hypertrophy and karyokinesis rather than cellular fusion. Gall formation was a function of parasite density and developed by hypertrophy of cortical cells. Brownish lesions which are symptomatic of tuber infection resulted from lignification of cortical cell walls in contact with egg matrix.     

Biocontrol of Meloidogyne incognita inciting disease in tomato by using a mixed compost inoculated with Paenibacillus ehimensis RS820

Meloidogyne spp. causes root-knot disease in tomato plants. Biological control of the disease may present economically feasible, agronomically durable and environmentally safe alternative of nematicides. A chitinolytic bacterial strain, Paenibacillus ehimensis RS820, previously isolated from the soil in Korea, produced lytic enzymes in higher amounts and inhibited the growth of phytopathogenic root-knot nematodes. Moreover, the juveniles and eggs of root-knot nematodes induced secretion of lytic enzymes by RS820 including chitinases, gelatinases and collagenases. Furthermore, mixed compost containing increased amounts of chitin and inoculated with RS820 was prepared in the present study. Use of the mixed compost not only reduced the disease caused by root-knot nematodes but also improved the plant growth. The extent of inoculation of the mixed compost with RS820 significantly influenced its ability to control the root-knot disease in tomato. The mixed compost also significantly altered the activity and density of the rhizosphere bacteria. Chitinase and gelatinase producing soil bacteria, as well as their enzyme activities, were significantly influenced by the mixed compost. The mixed compost proposed in the present study may represent a viable alternative to nematicides against the root-knot nematodes in tomato.     

<Emphasis Type="Italic">CaMi</Emphasis>, a root-knot nematode resistance gene from hot pepper (<Emphasis Type="Italic">Capsium annuum</Emphasis> L.) confers nematode resistance in tomato

Several root-knot nematode (Meloidogyne spp.) resistance genes have been discovered in different pepper (Capsium annuum L.) lines; however, none of them has yet been cloned. In this study, a candidate root-knot nematode resistance gene (designated as CaMi) was isolated from the resistant pepper line PR 205 by degenerate PCR amplification combined with the RACE technique. Expression profiling analysis revealed that this gene was highly expressed in roots, leaves, and flowers and expressed at a lower level in stems and was not detectable in fruits. To verify the function of CaMi, a sense vector containing the genomic DNA spanning the full coding region of CaMi was constructed and transferred into root-knot nematode susceptible tomato plants. Sixteen transgenic plants carrying one to five copies of T-DNA inserts were generated from two nematode susceptible tomato cultivars. RT-PCR analysis revealed that the expression levels of CaMi gene varied in different transgenic plants. Nematode assays showed that the resistance to root-knot nematodes was significantly improved in some transgenic lines compared to untransformed susceptible plants, and that the resistance was inheritable. Ultrastructure analysis showed that nematodes led to the formation of galls or root knots in the susceptible lines while in the resistant transgenic plants, the CaMi gene triggered a hypersensitive response (HR) as well as many necrotic cells around nematodes. Rugang Chen and Hanxia Li are contributed equally to this work.     

Techniques for image analysis of movement of juveniles of root-knot nematodes encumbered with Pasteuria penetrans spores

Root-knot nematodes (Meloidogyne spp.) are the most significant plant-parasitic nematodes that damage many crops all over the world. The free-living second stage juvenile (J2) is the infective stage that enters plants. The J2s move in the soil water films to reach the root zone. The bacterium Pasteuria penetrans is an obligate parasite of root-knot nematodes, is cosmopolitan, frequently encountered in many climates and environmental conditions and is considered promising for the control of Meloidogyne spp. The infection potential of P. penetrans to nematodes is well studied but not the attachment effects on the movement of root-knot nematode juveniles, image analysis techniques were used to characterize movement of individual juveniles with or without P. penetrans spores attached to their cuticles. Methods include the study of nematode locomotion based on (a) the centroid body point, (b) shape analysis and (c) image stack analysis. All methods proved that individual J2s without P. penetrans spores attached have a sinusoidal forward movement compared with those encumbered with spores. From these separate analytical studies of encumbered and unencumbered nematodes, it was possible to demonstrate how the presence of P. penetrans spores on a nematode body disrupted the normal movement of the nematode.     

Effects of Zygorrhynchus moelleri on the growth of potato and reproduction of potato cyst nematodes

Laboratory, pot and field experiments investigated the effects of the fungus Zygorrhynchus moelleri on the growth of potato and on the reproduction of the potato cyst nematodes (PCN), Globodera pallida and G rostochiensis. Preliminary laboratory tests showed that Z. moelleri growth was favoured by temperatures and pH ranges commonly present in field soils. The fungus colonised potato roots in vitro and in compost or field soil. It also stimulated in vitro root growth of three potato cultivars. In pot experiments Z. moelleri stimulated potato growth, particularly in the presence of PCN attack. In field plots infested with a mixture of G pallida and G. rostochiensis, tuber yields were not increased after application of the fungus but, in G pallida‐infested plots, yields were significantly increased after drills were inoculated with Z. moelleri. The application of Z. moelleri had no apparent effects on nematode reproduction. Factors influencing the interactions between Z. moelleri, potato and potato cyst nematodes are discussed and the potential role of the fungus as a plant growth promoter in organic potato production considered.     

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.     

Histopathology of Root-knot Nematode (Meloidogyne incognita) Infection on White Yam (Dioscorea rotundata) Tubers

White yam tissues naturally and artificially infected with root-knot nematodes were fixed, sectioned, and examined with a microscope. Infective second-stage juveniles of Meloidogyne incognita penetrated and moved intercellularly within the tuber. Feeding sites were always in the ground tissue layer where the vascular tissues are distributed in the tubers. Giant cells were always associated with xylem tissue. They were thin walled with dense cytoplasm and multinucleated. The nuclei of the giant cells were only half the size of those found in roots of infected tomato plants. Normal nematode growth and development followed giant cell formation. Females deposited eggs into a gelatinous egg mass within the tuber, and a necrotic ring formed around the female after eggs had been produced. Second-stage juveniles hatched, migrated, and re-infected other areas of the tuber. No males were observed from the tuber.     

Survey report on occurrence of root knot disease in winged bean

Winged bean (Psophocarpus tetragonolobus) is of rapidly increasing interest as a high-protein multipurpose crop; in the future the winged bean could become as important as the soybean in world agriculture, with the added bonus of yielding substantial quantities of edible, high-protein root tubers. During the survey of root-knot nematodes (Meloidogyne species) on vegetable crops in Uttar Pradesh (India), it was observed that winged bean was restricted to home gardens and backyard cultivation where it was found severe galling of roots and reduces tuber production and may affect pod and seed yield. Sometimes leaf anthracnose disease was also observed but it does not cause severe loss to the crop.     

植物罹病组织中南方、爪哇、花生根结线虫的LAMP快速检测

【目的】建立一种基于环介导等温扩增(loop-mediated isothermal amplification,LAMP)技术,从植物罹病组织中直接检测3种常见的根结线虫,为根结线虫的监测和防治提供技术支持。【方法】分别采用3种根结线虫的种类特异性引物对所选择的根结线虫的DNA片段进行PCR扩增,扩增产物纯化、回收并测序。根据3种根结线虫的测序结果,针对种类特异区段,采用PrimerExplorerV4软件,分别设计3种根结线虫的LAMP引物。设计的引物组人工合成后,以提取的纯化种群线虫DNA为模板,分别进行引物组的特异性测试,筛选出分别针对3种根结线虫的最佳引物组。【结果】研究设计的3种根结线虫的LAMP特异性引物能够直接从植物根结中检测出南方、花生、爪哇3种常见根结线虫,LAMP快速检测体系为:dNTPS浓度为1 mmol·L~(-1),Mg~(2+)的浓度为5 mmol·L~(-1),不添加甜菜碱,反应时间为45 min。【结论】本实验建立的南方、花生、爪哇根结线虫LAMP快速分子检测方法,具有特异性强、灵敏度高、简单、快速、经济等特征,能够从罹病植物组织中快速准确地检测出南方、花生和爪哇根结线虫,具有极高的实践应用价值。     

Interaction of vesicular arbuscular mycorrhiza with root knot nematodes in tomato

Summary The interaction between the VA mycorrhizal fungus,Glomus fasciculatus and the root-knot nematodes,Meloidogyne incognita andM. javanica, and their effects on the growth and phosphorus nutrition of tomato was studied in a red sandy loam soil of pH 6.0. Inoculation of tomato roots with root-knot nematodes enhanced infection and spore production byG. fasciculatus. Inoculation of tomato plants withG. fasciculatus significantly reduced the number and size of the root-knot galls produced byM. incognita andM. javanica. Inoculation withG. fasciculatus although improved plant growth and its total phosphorus content compared to the uninoculated plants, the difference were not statistically significant.     

Bacillus-based bionematicides: development,modes of action and commercialisation

Agricultural crops are severely damaged by root-knot nematodes causing extensive financial losses globally. Historically, agrochemicals have been the preferred method to combat these pests; however, threats to humans and the environment posed by these agrochemicals led to the need for developing new biocontrol agents. Importantly, the latter should adhere to biosafety regulations while being highly effective. Root-knot nematodes live in soil and thus the use of rhizobacteria such as Bacillus for biocontrol development have shown potential. Although various Bacillus species have been tested in this capacity, little is known about their secondary metabolites and the mechanisms of action responsible for their nematicidal activity. If these secondary metabolites can be qualitatively and quantitatively characterised, metabolic features could be synthetically engineered and used to combat root-knot nematodes. Although there is great potential for bionematicides, the commercialisation and development of such products can be difficult. This review summarises the importance of Bacillus species as natural antagonists of root-knot nematodes through the production of secondary metabolites. It provides an overview of the significance of root-knot nematodes in agriculture and the advances of chemical nematicides in recent years. The potential of Bacillus species as biocontrol agents, the known mechanisms of action responsible for the nematicidal activity demonstrated by Bacillus species, non-target effects of biocontrol agents and the commercialisation of Bacillus-based bionematicides are discussed.     

The effect of soil- and tuber-borne inoculum on the incidence of potato gangrene

Under optimum growing conditions neither tuber- nor soil-borne Phoma exigua var. foveata inoculum appreciably affected stand or yield of the subsequent potato crop. Seed tubers with gangrene rots caused high levels of stem and tuber symptoms when planted in var. foveata contaminated or uncontaminated land; contaminated seed tubers with no rots also produced progeny with a high gangrene potential. Sufficient soil-borne inoculum was carried over in land that produced a gangrene affected crop in the previous year to override the effect of tuber disinfection. Effective gangrene control was achieved by a combination of tuber disinfection shortly after harvest over successive years with a 1 in 5 yr potato crop rotation. Gangrene rots usually developed through injuries to the tuber periderm, rots in other tubers being associated with pustules of powdery scab (Spon-gospora subterranea).     

Screening of non-tuber bearing Solanaceae for resistance to and induction of juvenile hatch of potato cyst nematodes and their potential for trap cropping

Ninety accessions of non‐tuber bearing Solanaceae were screened for (i) resistance to and (ii) stimulatory effect on juvenile hatch of potato cyst nematodes, and (iii) their growth under temperate climatic conditions. All plant species belonging to the genus Solanum tested induced hatching but this effect was most pronounced for plant species of the Solanum nigrum complex. Hatching of juveniles was hardly or not stimulated by other plant genera of the Solanaceae. Solanum sisymbriifolium combined a high hatching effect with complete resistance to both Globodera rosiochiensis and G pallida. Two S. nigrum varieties showed full resistance to G rostochiensis and a high level of resistance to G pallida. Moreover, S. sisymbriifolium and the two varieties of S. nigrum performed very well under Dutch field conditions and, therefore, they are suggested as candidate trap crops for the control of potato cyst nematodes.     

New approaches to control plant parasitic nematodes

Plant parasitic nematodes are a serious threat for crop production worldwide. This review summarizes our understanding of plant nematode interactions and presents new alternatives for nematode control in the field. Breeding for resistance has been a major goal for many important crop species like soybean, potato, tomato and sugar-beet. As a result numerous nematode-resistance genes have been identified, two of which have been cloned recently, Hs1 ^pro-1 from sugar-beet, giving resistance to the beet cyst nematode Heterodera schachtii, and Mi from tomato, giving resistance to the root-knot nematode Meloidogyne incognita. Also artificial resistance genes, coding for nematotoxic proteins or causing rapid death of feeding cells, have been elucidated. In the future, genetic engineering of nematode resistance will become more and more important for plant breeding. Transformation techniques will allow genes to be quickly introduced into susceptible breeding lines and then combined with each other to produce plant varieties with durable resistance. Received: 26 August 1998 / Received revision: 16 December 1998 / Accepted: 21 December 1998     

Evaluation of the parasitoid Copidosoma koehleri for biological control of the potato tuber moth,Phthorimaea operculella,in Israeli potato fields

The potato tuber moth Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae) is a major agricultural pest of solaneceous crops in warm countries worldwide. The encyrtid polyembryonic parasitoid Copidosoma koehleri (Blanchard) has been successfully introduced for biological control of the moth in potato fields in South Africa and Australia; however, augmentative releases of the parasitoid in trial plots and in commercial potato fields in Israel did not reduce pest populations or infestation levels more than chemical treatment. Copidosoma koehleri accounted for 4–5% of parasitism on tuber moth caterpillars, while most parasitism was due to local species of larval parasitoids. The abundance and composition of local parasitoids did not differ between C. koehleri release plots and conventionally treated control plots. These findings can be interpreted as failure of the introduced parasitoids to survive and locate their hosts, or as mortality of C. koehleri within hosts in the field. Sentinel hosts, placed in trial plots and collected after 24 h, were rarely parasitised by C. koehleri, supporting the first interpretation. To test the second hypothesis, hosts parasitised by C. koehleri were placed in field plots for a week, collected, and reared out in the laboratory. The emergence rates of C. koehleri from these hosts resembled those of lab-reared controls, suggesting that mortality within hosts in the field is not a major cause of C. koehleri's poor biocontrol performance.     

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.