scholarly journals An Immunocytochemical Procedure for Protein Localization in Various Nematode Life Stages Combined with Plant Tissues Using Methylacrylate-Embedded Specimens

2012 ◽  
Vol 102 (10) ◽  
pp. 990-996 ◽  
Author(s):  
Paulo Vieira ◽  
Mohamed Youssef Banora ◽  
Philippe Castagnone-Sereno ◽  
Marie-Noëlle Rosso ◽  
Gilbert Engler ◽  
...  
Keyword(s):  
Protein Localization ◽  
Plant Morphology ◽  
Cytoskeletal Proteins ◽  
Parasitic Nematodes ◽  
Plant Tissues ◽  
Life Stages ◽  
Plant Parasitic Nematodes ◽  
In Planta ◽  
Tissue Sections ◽  
Double Immunolabeling

Plant-parasitic nematodes possess a large number of proteins that are secreted in planta, allowing them to be successful parasites of plants. The majority of these proteins are synthesized mainly in the nematode subventral and dorsal glands as well as in other organs. To improve the immunovisualization of these proteins, we adapted a methacrylate embedding method for the localization of proteins inside nematode tissues, and extracellularly when secreted in planta or within plant cells. An important advantage is that the method is applicable for all nematode stages: preparasitic as well as parasitic stages, including large mature females. Herein, the method has been successfully applied for the localization of four nematode secreted proteins, such as Mi-MAP-1, Mi-CBM2-bearing proteins, Mi-PEL3, and Mi-6D4. In addition, we could also localize 14-3-3 proteins, as well as two cytoskeletal proteins, by double-immunolabeling on preparasitic juveniles. Superior preservation of nematode and plant morphology, allowed more accurate protein localization as compared with other methods. Besides excellent epitope preservation, dissolution of methacrylate from tissue sections unmasks target proteins and thereby drastically increases antibody access.

scholarly journals Exploitation of FTA Cartridges for the Sampling, Long-Term Storage, and DNA-Based Analyses of Plant-Parasitic Nematodes

2014 ◽  
Vol 104 (3) ◽  
pp. 306-312 ◽  
Author(s):  
Martin Marek ◽  
Miloslav Zouhar ◽  
Ondřej Douda ◽  
Marie Maňasová ◽  
Pavel Ryšánek
Keyword(s):  
Solid Phase ◽  
Complex Structure ◽  
Heterodera Schachtii ◽  
Parasitic Nematodes ◽  
Plant Tissues ◽  
Single Individual ◽  
Plant Parasitic Nematodes ◽  
Important Species ◽  
Plant Parasitic

The use of DNA-based analyses in molecular plant nematology research has dramatically increased over recent decades. Therefore, the development and adaptation of simple, robust, and cost-effective DNA purification procedures are required to address these contemporary challenges. The solid-phase-based approach developed by Flinders Technology Associates (FTA) has been shown to be a powerful technology for the preparation of DNA from different biological materials, including blood, saliva, plant tissues, and various human and plant microbial pathogens. In this work, we demonstrate, for the first time, that this FTA-based technology is a valuable, low-cost, and time-saving approach for the sampling, long-term archiving, and molecular analysis of plant-parasitic nematodes. Despite the complex structure and anatomical organization of the multicellular bodies of nematodes, we report the successful and reliable DNA-based analysis of nematode high-copy and low-copy genes using the FTA technology. This was achieved by applying nematodes to the FTA cards either in the form of a suspension of individuals, as intact or pestle-crushed nematodes, or by the direct mechanical printing of nematode-infested plant tissues. We further demonstrate that the FTA method is also suitable for the so-called “one-nematode-assay”, in which the target DNA is typically analyzed from a single individual nematode. More surprisingly, a time-course experiment showed that nematode DNA can be detected specifically in the FTA-captured samples many years after initial sampling occurs. Collectively, our data clearly demonstrate the applicability and the robustness of this FTA-based approach for molecular research and diagnostics concerning phytonematodes; this research includes economically important species such as the stem nematode (Ditylenchus dipsaci), the sugar beet nematode (Heterodera schachtii), and the Northern root-knot nematode (Meloidogyne hapla).

Mutualistic endophytic fungi and in-planta suppressiveness to plant parasitic nematodes

2008 ◽  
Vol 46 (1) ◽  
pp. 15-23 ◽  
Author(s):  
Richard A. Sikora ◽  
Luis Pocasangre ◽  
Alexandra zum Felde ◽  
Bjoern Niere ◽  
Tam T. Vu ◽  
...  
Keyword(s):  
Endophytic Fungi ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
In Planta ◽  
Plant Parasitic

scholarly journals Marasmius oreades agglutinin enhances resistance of Arabidopsis against plant-parasitic nematodes and a herbivorous insect

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Aboubakr Moradi ◽  
Tina Austerlitz ◽  
Paul Dahlin ◽  
Christelle AM Robert ◽  
Corina Maurer ◽  
...  
Keyword(s):  
Insecticidal Activity ◽  
Negative Impact ◽  
Crop Protection ◽  
Heterodera Schachtii ◽  
Relative Mass ◽  
Parasitic Nematodes ◽  
Herbivorous Insect ◽  
Plant Parasitic Nematodes ◽  
In Planta ◽  
Plant Parasitic

Abstract Background Plant-parasitic nematodes and herbivorous insects have a significant negative impact on global crop production. A successful approach to protect crops from these pests is the in planta expression of nematotoxic or entomotoxic proteins such as crystal proteins from Bacillus thuringiensis (Bt) or plant lectins. However, the efficacy of this approach is threatened by emergence of resistance in nematode and insect populations to these proteins. To solve this problem, novel nematotoxic and entomotoxic proteins are needed. During the last two decades, several cytoplasmic lectins from mushrooms with nematicidal and insecticidal activity have been characterized. In this study, we tested the potential of Marasmius oreades agglutinin (MOA) to furnish Arabidopsis plants with resistance towards three economically important crop pests: the two plant-parasitic nematodes Heterodera schachtii and Meloidogyne incognita and the herbivorous diamondback moth Plutella xylostella. Results The expression of MOA does not affect plant growth under axenic conditions which is an essential parameter in the engineering of genetically modified crops. The transgenic Arabidopsis lines showed nearly complete resistance to H. schachtii, in that the number of female and male nematodes per cm root was reduced by 86–91 % and 43–93 % compared to WT, respectively. M. incognita proved to be less susceptible to the MOA protein in that 18–25 % and 26–35 % less galls and nematode egg masses, respectively, were observed in the transgenic lines. Larvae of the herbivorous P. xylostella foraging on MOA-expression lines showed a lower relative mass gain (22–38 %) and survival rate (15–24 %) than those feeding on WT plants. Conclusions The results of our in planta experiments reveal a robust nematicidal and insecticidal activity of the fungal lectin MOA against important agricultural pests which may be exploited for crop protection.

scholarly journals Plant-Parasitic Nematodes Attacking Chickpea and Their In Planta Interactions with Rhizobia and Phytopathogenic Fungi

Plant Disease ◽  
2008 ◽  
Vol 92 (6) ◽  
pp. 840-853 ◽  
Author(s):  
Pablo Castillo ◽  
Juan A. Navas-Cortés ◽  
Blanca B. Landa ◽  
Rafael M. Jiménez-Díaz ◽  
Nicola Vovlas
Keyword(s):  
Phytopathogenic Fungi ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
In Planta ◽  
Plant Parasitic

scholarly journals Plant metabolism of nematode pheromones mediates plant-nematode interactions

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Murli Manohar ◽  
Francisco Tenjo-Castano ◽  
Shiyan Chen ◽  
Ying K. Zhang ◽  
Anshu Kumari ◽  
...  
Keyword(s):  
Defense Mechanism ◽  
Parasitic Nematodes ◽  
Plant Tissues ◽  
Chemical Signaling ◽  
Plant Parasitic Nematodes ◽  
Soil Organisms ◽  
Rhizosphere Interactions ◽  
Plant Nematode ◽  
Comparative Metabolomics ◽  
Underlying Mechanisms

AbstractMicroorganisms and nematodes in the rhizosphere profoundly impact plant health, and small-molecule signaling is presumed to play a central role in plant rhizosphere interactions. However, the nature of the signals and underlying mechanisms are poorly understood. Here we show that the ascaroside ascr#18, a pheromone secreted by plant-parasitic nematodes, is metabolized by plants to generate chemical signals that repel nematodes and reduce infection. Comparative metabolomics of plant tissues and excretions revealed that ascr#18 is converted into shorter side-chained ascarosides that confer repellency. An Arabidopsis mutant defective in two peroxisomal acyl-CoA oxidases does not metabolize ascr#18 and does not repel nematodes, indicating that plants, like nematodes, employ conserved peroxisomal β-oxidation to edit ascarosides and change their message. Our results suggest that plant-editing of nematode pheromones serves as a defense mechanism that acts in parallel to conventional pattern-triggered immunity, demonstrating that plants may actively manipulate chemical signaling of soil organisms.

In planta PCR-based detection of early infection of plant-parasitic nematodes in the roots: a step towards the understanding of infection and plant defence

2010 ◽  
Vol 128 (3) ◽  
pp. 343-351 ◽  
Author(s):  
Syarifah Aisyafaznim Sayed Abdul Rahman ◽  
Zulqarnain Mohamed ◽  
Rofina Yasmin Othman ◽  
Rony Swennen ◽  
Bart Panis ◽  
...  
Keyword(s):  
Plant Defence ◽  
Early Infection ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
In Planta ◽  
Plant Parasitic

Organic amendments and their influences on plant-parasitic and free-living nematodes: a promising method for nematode management?

Nematology ◽  
2011 ◽  
Vol 13 (2) ◽  
pp. 133-153 ◽  
Author(s):  
Tim C. Thoden ◽  
Gerard W. Korthals ◽  
Aad J. Termorshuizen
Keyword(s):  
Crop Yields ◽  
Organic Amendments ◽  
Plant Morphology ◽  
Organic Soil ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Free Living ◽  
Nematode Management ◽  
Positive Effects ◽  
Plant Parasitic

Abstract The use of organic soil amendments, such as green manures, animal manures, composts or slurries, certainly has many advantageous aspects for soil quality and is suggested as a promising tool for the management of plant-parasitic nematodes. However, during a recent literature survey we also found numerous studies reporting an increase of plant-parasitic nematodes after the use of organic amendments. Therefore, we critically re-evaluated the usefulness of organic amendments for nematode management and suggest possible mechanisms for a stimulation of plant-parasitic nematodes, as well as mechanisms that might be causing a reduction of plant-parasitic nematodes. In addition, we also elucidate a possible mechanism that might be responsible for the observed overall positive effects of organic amendments on crop yields. It is likely that a significant part of this is, inter alia, due to the proliferation of non-pathogenic, free-living nematodes and their overall positive effects on soil microbial populations, organic matter decomposition, nutrient availability, plant morphology and ecosystem stability.

scholarly journals Tapping Into the Cotton Fungal Phytobiome for Novel Nematode Biological Control Tools

2020 ◽  
Vol 4 (1) ◽  
pp. 19-26
Author(s):  
Wenqing Zhou ◽  
Vijay C. Verma ◽  
Terry A. Wheeler ◽  
Jason E. Woodward ◽  
James L. Starr ◽  
...  
Keyword(s):  
Biological Control ◽  
Seed Treatment ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
In Planta ◽  
Root Knot Nematode ◽  
Negative Effects ◽  
Root Knot Nematodes ◽  
Antagonistic Effects

A number of fungi have been shown to have negative effects on plant-parasitic nematodes. Most of these fungi have been isolated from soil, plant roots, or nematodes themselves. Fungi associated with crops can provide a diverse pool of candidates to test for antagonistic effects against plant parasites and other stressors. We used a hierarchical two-tiered approach to evaluate the efficacy and repeatability of 55 strains of fungi originally isolated as foliar facultative endophytes from upland cotton (Gossypium hirsutum) along with one commercial isolate of Beauveria bassiana for in planta antagonistic effects on root-knot nematodes (Meloidogyne incognita). All fungi were inoculated to cotton using a seed treatment. The number of root galls was quantified 3 weeks after egg inoculation of cotton seedlings. The majority of the fungi tested reduced the number of root galls relative to those on untreated control plants. To assess repeatability, 22 strains that exhibited the strongest reductions in gall numbers were further tested in replicate follow-up assays. Ninety-five percent (21/22) of these retested strains significantly reduced galling in the follow-up assay. Strains that reduced galling the most belonged to the genera Alternaria, Chaetomium, Cladosporium, Diaporthe, Epicoccum, Gibellulopsis, and Purpureocillium. On the contrary, three strains in the genera Alternaria and Curvularia significantly increased gall numbers. Our results indicate that a large proportion of the fungal strains originally isolated from cotton as naturally occurring foliar facultative endophytes are capable of reducing root-knot nematode infection when applied back to the plant as a seed treatment. These findings help establish a rich pool of candidate fungi for further evaluation as novel biological control tools against root-knot nematodes in cotton and other plants.

The Use of RNA Interference in Enhancing Plant Resistance against Nematodes

2020 ◽  
Vol 2 (1) ◽  
Author(s):  
Siye Chen
Keyword(s):  
Rna Interference ◽  
Plant Resistance ◽  
Parasitic Nematodes ◽  
Great Success ◽  
Plant Parasitic Nematodes ◽  
In Planta ◽  
Rnai Technology ◽  
Wide Range ◽  
Plant Parasitic

Plant-parasitic nematodes caused severe yield loss in major crops all over the world. The most wild-used strategies to combat the nematodes is the chamical nematicides, but the overuse of synthetic nematicides threaten sustainable agriculture development. Other strategies, like resistance cultivars and crop rotation, have limited efficiency. Thus, the utilization of molecular biotechnology like RNA interference (RNAi) would be one of the alternative ways to enhance plant resistance against nematodes. RNAi has already used as a tool for gene functional analysis in a wide range of species, especially in the non-parasitic nematode, Caenorhabtidis elegans. In plant-parasitic nematodes, RNAi is induced by soaking nematodes with double-strand RNA(dsRNA) solution mixed with neurostimulants, which is called in vitro RNAi delivery method. In another way around, in planta RNAi method, which is Host-mediated RNAi approach also showed a great success in conferring the resistance against root-knock nematodes. Two main advantages of RNAi-based transgenics are RNAi technology do not produce any functional foreign proteins and it target organisms in a sequence-specific way. Even though the development of RNAi-based transgenics against plant-parasitic nematodes is still in the initial phase, it offers the prospect into a novel nematode control strategy in the future.

A Cotton Blue-Lactophenol Technique for Mounting Plant-Parasitic Nematodes

1949 ◽  
Vol 23 (3-4) ◽  
pp. 175-178 ◽  
Author(s):  
Mary T. Franklin ◽  
J. Basil Goodey
Keyword(s):  
General Problem ◽  
Reliable Method ◽  
Parasitic Nematodes ◽  
Plant Tissues ◽  
Acid Fuchsin ◽  
Plant Parasitic Nematodes ◽  
Cotton Blue ◽  
Living Material ◽  
Parasitic Worms ◽  
Plant Parasitic

The lack of a rapid and reliable method of making permanent preparations of nematodes is most strongly felt in the case of the plant-parasitic forms. The method in general use for most plant- and soil-inhabiting nematodes consists in relaxing the worms by gentle heat, fixing in 5% formalin or Ditlivsen's fixative (Thorne, 1925) and then impregnating slowly with glycerine. This procedure is quite reliable for the non-plant-parasitic forms but its results with plant-parasitic worms are very uncertain. With the latter it is necessary to transfer from fixative to very dilute glycerine, usually 1.5% glycerine in 7.5% alcohol with a trace of thymol or copper sulphate to discourage moulds. The glycerine is allowed to concentrate very slowly during at least four weeks until the worms can be transferred to pure glycerine in which they are mounted. Even then the results are often disappointing for the worms very frequently collapse.In considering this general problem recently the authors called to mind the appearance of worms stained within plant tissues by the acid fuchsin-lactophenol method (Goodey, 1987: Franklin, 1949). This method causes no collapse or distortion of the worms and takes but a few minutes from living material to permanently stained preparations. The process has now been developed for use with free specimens of Anguillulina dipsaci and plant-parasitic species of Aphelenchoides, and is as follows:

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