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).

scholarly journals Toward genetic modification of plant-parasitic nematodes: delivery of macromolecules to adults and expression of exogenous mRNA in second stage juveniles

2021 ◽  
Vol 11 (2) ◽  
Author(s):  
Olaf Kranse ◽  
Helen Beasley ◽  
Sally Adams ◽  
Andre Pires-daSilva ◽  
Christopher Bell ◽  
...  
Keyword(s):  
Food Security ◽  
Genetic Modification ◽  
The Body ◽  
Forward Genetics ◽  
Heterodera Schachtii ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Root Knot Nematode ◽  
Cyst Nematodes ◽  
Plant Parasitic

Abstract Plant-parasitic nematodes are a continuing threat to food security, causing an estimated 100 billion USD in crop losses each year. The most problematic are the obligate sedentary endoparasites (primarily root knot nematodes and cyst nematodes). Progress in understanding their biology is held back by a lack of tools for functional genetics: forward genetics is largely restricted to studies of natural variation in populations and reverse genetics is entirely reliant on RNA interference. There is an expectation that the development of functional genetic tools would accelerate the progress of research on plant-parasitic nematodes, and hence the development of novel control solutions. Here, we develop some of the foundational biology required to deliver a functional genetic tool kit in plant-parasitic nematodes. We characterize the gonads of male Heterodera schachtii and Meloidogyne hapla in the context of spermatogenesis. We test and optimize various methods for the delivery, expression, and/or detection of exogenous nucleic acids in plant-parasitic nematodes. We demonstrate that delivery of macromolecules to cyst and root knot nematode male germlines is difficult, but possible. Similarly, we demonstrate the delivery of oligonucleotides to root knot nematode gametes. Finally, we develop a transient expression system in plant-parasitic nematodes by demonstrating the delivery and expression of exogenous mRNA encoding various reporter genes throughout the body of H. schachtii juveniles using lipofectamine-based transfection. We anticipate these developments to be independently useful, will expedite the development of genetic modification tools for plant-parasitic nematodes, and ultimately catalyze research on a group of nematodes that threaten global food security.

Identification of cytokinins produced by the plant parasitic nematodes Heterodera schachtii and Meloidogyne incognita

2003 ◽  
Vol 4 (4) ◽  
pp. 271-277 ◽  
Author(s):  
Jan De Meutter ◽  
Tom Tytgat ◽  
Erwin Witters ◽  
Greetje Gheysen ◽  
Henri Van Onckelen ◽  
...  
Keyword(s):  
Meloidogyne Incognita ◽  
Heterodera Schachtii ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
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.

Nematode management through genome editing.

2021 ◽  
pp. 408-413
Author(s):  
Shahid Siddique ◽  
Sebastian Eves-van den Akker
Keyword(s):  
Rna Interference ◽  
Recent Progress ◽  
Plant Pathogens ◽  
Low Cost ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Cyst Nematodes ◽  
Root Knot Nematodes ◽  
Plant Parasitic

Abstract Plant parasitic nematodes are among the most destructive plant pathogens, causing an estimated US$78 billion yield losses globally. Although approximately 3000 species of plant parasitic nematodes have been described, most of the damage is caused by a small group of root-infecting sedentary endoparasitic nematodes that include root-knot nematodes (Meloidogyne spp.) and cyst nematodes (Heterodera spp.). Given that previous literature amply reviews the breadth of biotechnological methods for the control of plant parasitic nematodes, this chapter will briefly touch on long-standing biotechnological methods but focus on recent progress in, and long-term promise of, the use of CRISPR technology for introducing targeted modifications into host genomes with the goal of enhancing resistance against plant parasitic nematodes. It is predicted that expanding reverse genetic approaches beyond RNA interference, using low-cost, technically simple and efficient transformation (transient or stable) will be the single most important advance in the field in some years.

Bioprospecting Compost for Long-Term Control of Plant Parasitic Nematodes

2020 ◽  
pp. 35-50
Author(s):  
Judy Rouse-Miller ◽  
Ezra S. Bartholomew ◽  
Chaney C. G. St. Martin ◽  
Piterson Vilpigue
Keyword(s):  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Plant Parasitic

THE INFLUENCE OF MINERAL FERTILIZER APPLICATION AND PLANT NUTRITION ON PLANT-PARASITIC NEMATODES IN UPLAND AND LOWLAND RICE IN CÔTE D'IVOIRE AND ITS IMPLICATIONS IN LONG TERM AGRICULTURAL RESEARCH TRIALS

2004 ◽  
Vol 40 (2) ◽  
pp. 245-256 ◽  
Author(s):  
D. L. COYNE ◽  
K. L. SAHRAWAT ◽  
R. A. PLOWRIGHT
Keyword(s):  
Upland Rice ◽  
Mineral Fertilizer ◽  
Nematode Species ◽  
Fertilizer Application ◽  
Lowland Rice ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
P Application ◽  
Plant Parasitic

Mineral fertilizer application and consequent plant nutrition has long been observed to influence associated plant-parasitic nematode population densities, offering the potential as a nematode management option. Observations were made on the influence of mineral fertilizer application on nematode populations on three separate long-term rice experiments, (differential mineral application on upland and on lowland rice, and P application on upland rice) undertaken between 1994 and 1997 in Côte d'Ivoire. In 1995, on upland rice, treatments with K or N withheld from the comprehensive mineral application treatment (range of elements including N, P, K, Ca, Mg and Zn) led to lower densities of Pratylenchus zeae at harvest than the comprehensive mineral application. By withholding K or Mg, Helicotylenchus pseudorobustus densities were greater than with either the control (no mineral application) or comprehensive mineral application in the same year. No differences were observed between treatments in 1994, or between treatments for densities of other nematode species present (Meloidogyne incognita, Criconemella tescorum) or for total nematode density. In the lowland rice trial, no treatment effects on nematode species (Hirschmanniella oryzae and Uliginotylenchus palustris) were observed. In the P application trial on a P-deficient Ultisol, Heterodera sacchari densities were lower in treatments receiving 180 kg P ha−1, than untreated in 1995; in 1996 no differences were observed between untreated and 135 kg P ha−1, while in 1997 higher densities of H. sacchari were present in 135 kg P ha−1 than untreated. Regression analysis of nematode densities against the mineral straw content in the P application trial revealed a negative correlation between M. incognita and Mn and Ca, and between P. zeae and Zn or Fe. A positive correlation was observed between Helicotylenchus spp. and Mg. This study provides strong arguments for taking plant parasitic nematodes into account when planning and executing long-term research trials.

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:

scholarly journals Anatomical Alterations in Plant Tissues Induced by Plant-Parasitic Nematodes

2017 ◽  
Vol 8 ◽  
Author(s):  
Juan E. Palomares-Rius ◽  
Carolina Escobar ◽  
Javier Cabrera ◽  
Alessio Vovlas ◽  
Pablo Castillo
Keyword(s):  
Parasitic Nematodes ◽  
Plant Tissues ◽  
Plant Parasitic Nematodes ◽  
Plant Parasitic

scholarly journals Towards genetic modification of plant-parasitic nematodes: delivery of macromolecules to adults and expression of exogenous mRNA in second stage juveniles

2020 ◽  
Author(s):  
Olaf Kranse ◽  
Helen Beasley ◽  
Sally Adams ◽  
Andre Pires da Silva ◽  
Chris Bell ◽  
...  
Keyword(s):  
Food Security ◽  
Genetic Modification ◽  
The Body ◽  
Forward Genetics ◽  
Heterodera Schachtii ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Root Knot Nematode ◽  
Cyst Nematodes ◽  
Plant Parasitic

AbstractPlant-parasitic nematodes are a continuing threat to food security, causing an estimated 100 billion USD in crop losses each year. The most problematic are the obligate sedentary endoparasites (primarily root knot nematodes and cyst nematodes). Progress in understanding their biology is held back by a lack of tools for functional genetics: forward genetics is largely restricted to studies of natural variation in populations, and reverse genetics is entirely reliant on RNA interference. There is an expectation that the development of functional genetic tools would accelerate the progress of research on plant-parasitic nematodes, and hence the development of novel control solutions. Here, we develop some of the foundational biology required to deliver a functional genetic tool kit in plant-parasitic nematodes. We characterise the gonads of male Heterodera schachtii and Meloidogyne hapla in the context of spermatogenesis. We test and optimise various methods for the delivery, expression, and/or detection of exogenous nucleic acids in plant-parasitic nematodes. We demonstrate that delivery of macromolecules to cyst and root knot nematode male germlines is difficult, but possible. Similarly, we demonstrate the delivery of oligonucleotides to root knot nematode gametes. Finally, we develop a transient expression system in plant-parasitic nematodes by demonstrating the delivery and expression of exogenous mRNA encoding various reporter genes throughout the body of H. schachtii juveniles using lipofectamine-based transfection. We anticipate these developments to be independently useful, will expedite the development of genetic modification tools for plant-parasitic nematodes, and ultimately catalyze research on a group of nematodes that threaten global food security.

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