The genome and lifestage-specific transcriptomes of a plant-parasitic nematode and its host reveal susceptibility genes involved in trans-kingdom synthesis of vitamin B5

2021 ◽  
Author(s):  
Shahid Siddique ◽  
Zoran S. Radakovic ◽  
Clarissa Hiltl ◽  
Clement Pellegrin ◽  
Thomas J. Baum ◽  
...  
Keyword(s):  
Metabolic Pathways ◽  
Crop Production ◽  
Susceptibility Genes ◽  
Heterodera Schachtii ◽  
Parasitic Nematodes ◽  
Cyst Nematodes ◽  
Novel Approach ◽  
Vitamin B5 ◽  
Dual Analysis ◽  
Plant Parasitic

AbstractPlant-parasitic nematodes are a major, and in some cases a dominant, threat to crop production in all agricultural systems. The relative scarcity of classical resistance genes highlights a pressing need to identify new ways to develop nematode-resistant germplasm. Here, we sequence and assemble a high-quality genome of the model cyst nematode Heterodera schachtii to provide a platform for the first system-wide dual analysis of host and parasite gene expression over time, covering all major stages of the interaction. This novel approach enabled the analysis of the hologenome of the infection site, to identify metabolic pathways that were incomplete in the parasite but complemented by the host. Using a combination of bioinformatic, genetic, and biochemical approaches, we show that the highly atypical completion of vitamin B5 biosynthesis by the parasitic animal, putatively enabled by a horizontal gene transfer from a bacterium, is critically important for parasitism. Knockout of either the plant-encoded or the now nematode-encoded steps in the pathway blocks parasitism. Our experiments establish a reference for cyst nematodes, use this platform to further our fundamental understanding of the evolution of plant-parasitism by nematodes, and show that understanding congruent differential expression of metabolic pathways represents a new way to find nematode susceptibility genes, and thereby, targets for future genome editing-mediated generation of nematode-resistant crops.

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.

scholarly journals Plant Parasitic Nematodes and their management in crop production: a review

2021 ◽  
Vol 4 (2) ◽  
pp. 327-338
Author(s):  
Honey Raj Mandal ◽  
Shambhu Katel ◽  
Sudeep Subedi ◽  
Jiban Shrestha
Keyword(s):  
Crop Production ◽  
Bilateral Symmetry ◽  
Cultural Control ◽  
Crop Damage ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Cyst Nematodes ◽  
The World ◽  
The Many ◽  
Plant Parasitic

Plant Parasitic Nematodes are small worm like transparent, bilateral symmetry, pseudocoelomate, multicellular, free living or parasitic microorganism which are predatory, aquatic, terrestrial, entopathogenic, ectoparasite, endoparasite, semi-endoparasite or sedentary. They cause substantial problems to major crops throughout the world, including vegetables, fruits, and grain crops. The root knot and cyst nematodes are economically important pests in numerous crops. Crop damage from nematodes is not readily apparent in most cases, and it often remains hidden by the many other factors limiting plant growth. In the past, the control of the nematodes has been based on the synthetic nematicides, the number of which has been drastically restricted in the EU because of their environmental side effects and subsequent restriction in European Union (EU) rules and regulations. Many other methods like cultural control, biological control, use of biotechnological tools and methods, use of resistant cultivars are tested and proven successful in controlling different species of nematodes all over the world. Alternatively, combinations of the different methods are proven to be highly effective both economically and environmentally.

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.

scholarly journals Marine Organisms for the Sustainable Management of Plant Parasitic Nematodes

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 369
Author(s):  
Pasqua Veronico ◽  
Maria Teresa Melillo
Keyword(s):  
Crop Production ◽  
Control Strategies ◽  
Marine Organisms ◽  
New Drugs ◽  
Effective Control ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Wide Range ◽  
Plant Growth Stimulants ◽  
Plant Parasitic

Plant parasitic nematodes are annually responsible for the loss of 10%–25% of worldwide crop production, most of which is attributable to root-knot nematodes (RKNs) that infest a wide range of agricultural crops throughout the world. Current nematode control tools are not enough to ensure the effective management of these parasites, mainly due to the severe restrictions imposed on the use of chemical pesticides. Therefore, it is important to discover new potential nematicidal sources that are suitable for the development of additional safe and effective control strategies. In the last few decades, there has been an explosion of information about the use of seaweeds as plant growth stimulants and potential nematicides. Novel bioactive compounds have been isolated from marine cyanobacteria and sponges in an effort to find their application outside marine ecosystems and in the discovery of new drugs. Their potential as antihelmintics could also be exploited to find applicability against plant parasitic nematodes. The present review focuses on the activity of marine organisms on RKNs and their potential application as safe nematicidal agents.

How do nematodes get their sweets? Solute supply to sedentary plant-parasitic nematodes

Nematology ◽  
2007 ◽  
Vol 9 (4) ◽  
pp. 451-458 ◽  
Author(s):  
Julia Hofmann ◽  
Florian Grundler
Keyword(s):  
Nutrient Supply ◽  
Heterodera Schachtii ◽  
Transport Systems ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Cyst Nematodes ◽  
Root Knot Nematodes ◽  
Feeding Sites ◽  
Inorganic Solutes ◽  
Nematode Development

AbstractSedentary cyst and root-knot nematodes withdraw large amounts of solutes from feeding structures induced in host roots. The feeding structures are specialised cells with a high metabolic activity and a tremendous capacity in translocation of nutrients. The required nutrients are provided by the plant transport systems – water and inorganic solutes from the xylem, assimilates such as sugars and amino acids from the phloem. Here we discuss the available data on the mechanisms by which nutrients are translocated into the nematode feeding sites. The interaction between Heterodera schachtii and Arabidopsis thaliana serves as a model system for cyst nematodes. In this case sufficient data are available to propose a conclusive concept for the mechanisms of nutrient flow: basically, in the early stages of nematode development syncytia are symplasmically isolated, so that transport proteins are responsible for the nutrient supply. Later, connections to the phloem via plasmodesmata are established, so that developing females are well supplied with assimilates. The interactions of root-knot nematodes with their hosts share a number of similarities but the data currently available are not sufficient to draw similar conclusions. As nutrient supply and functionality of feeding structures are the basis of biotrophic parasitism of sedentary nematodes, it is tempting to unravel the mechanisms by which both plant and nematodes influence each other via nutrient fluxes.

scholarly journals A New Class of Ubiquitin Extension Proteins Secreted by the Dorsal Pharyngeal Gland in Plant Parasitic Cyst Nematodes

2004 ◽  
Vol 17 (8) ◽  
pp. 846-852 ◽  
Author(s):  
Tom Tytgat ◽  
Bartel Vanholme ◽  
Jan De Meutter ◽  
Myriam Claeys ◽  
Marjolein Couvreur ◽  
...  
Keyword(s):  
Syncytium Formation ◽  
Cyst Nematode ◽  
Heterodera Schachtii ◽  
Regulatory Function ◽  
Cyst Nematodes ◽  
New Class ◽  
Genes Encoding ◽  
Pharyngeal Gland ◽  
Positively Charged ◽  
Plant Parasitic

By performing cDNA AFLP on pre- and early parasitic juveniles, we identified genes encoding a novel type of ubiquitin extension proteins secreted by the dorsal pharyngeal gland in the cyst nematode Heterodera schachtii. The proteins consist of three domains, a signal peptide for secretion, a mono-ubiquitin domain, and a short C-terminal positively charged domain. A gfp-fusion of this protein is targeted to the nucleolus in tobacco BY-2 cells. We hypothesize that the C-terminal peptide might have a regulatory function during syncytium formation in plant roots.

scholarly journals Changes in mRNA Abundance within Heterodera schachtii-Infected Roots of Arabidopsis thaliana

2000 ◽  
Vol 13 (3) ◽  
pp. 309-315 ◽  
Author(s):  
Dieter Hermsmeier ◽  
Jennifer K. Hart ◽  
Marina Byzova ◽  
Steven R. Rodermel ◽  
Thomas J. Baum
Keyword(s):  
Arabidopsis Thaliana ◽  
Differential Display ◽  
Tissue Sample ◽  
Mrna Abundance ◽  
Heterodera Schachtii ◽  
Parasitic Nematodes ◽  
Cdna Clones ◽  
Cyst Nematodes ◽  
Tissue Samples ◽  
Feeding Site

Gene expression changes in plant roots infected by plant-parasitic cyst nematodes are involved in the formation of nematode feeding sites. We analyzed mRNA abundance changes within roots of Arabidopsis thaliana during the early compatible interaction with Heterodera schachtii, the sugarbeet cyst nematode. Approximately 1,600 root sections, each containing a single parasitic nematode and its feeding site, and 1,600 adjacent, nematode-free root sections were excised from aseptic A. thaliana cultures 3 to 4 days after inoculation with H. schachtii. These tissue samples were termed infected and uninfected, respectively. Preparasitic nematodes were added to the uninfected tissue sample to maintain the nematode to plant tissue proportion. mRNA extracted from these two tissue samples was subjected to differential display analysis. Thirty-six cDNA clones corresponding to mRNA species with different abundance between both tissue samples were isolated. Of these clones, 24 were of A. thaliana origin and 12 were from H. schachtii. Differential display data predicted that the A. thaliana cDNA clones corresponded to 13 transcripts that were more abundant in the infected root sections and 11 transcripts that were more abundant in the uninfected root sections. H. schachtii cDNA clones were predicted to correspond to four transcripts that were more abundant in parasitic nematodes and to eight transcripts that were more abundant in preparasitic nematodes. In situ hybridization experiments confirmed the mRNA abundance changes in A. thaliana roots predicted by the differential display analyses for two A. thaliana clones.

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).

Plant-parasitic nematode associated with wheat in central, eastern, and south-eastern Kazakhstan

Plant Disease ◽  
2021 ◽  
Author(s):  
İmren Mustafa ◽  
Göksel Özer ◽  
Timothy Paulitz ◽  
Alexei Morgounov ◽  
Abdelfattah A. Dababat
Keyword(s):  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Cyst Nematodes ◽  
Pratylenchus Neglectus ◽  
Wheat Production ◽  
South Eastern ◽  
Heterodera Filipjevi ◽  
Central Eastern ◽  
Plant Parasitic ◽  
Eastern Kazakhstan

Kazakhstan is one of the biggest wheat producers, however, its wheat production is far below the average international wheat production standard due to biotic and abiotic stressors. Plant-parasitic nematodes are devastating for cereal production systems worldwide. A comprehensive survey was conducted in 2019 to identify plant-parasitic nematodes associated with wheat in different locations of central, eastern, and south-eastern Kazakhstan. The results revealed 33 root-lesion and 27 cyst nematode populations from the 77 localities sampled. These two genera occurred in separate or in mixed populations. The root-lesion populations were identified as Pratylenchus neglectus and P. thornei while all cyst nematodes were identified as Heterodera filipjevi. The identification of nematodes was firstly performed based on morphological and morphometric features and confirmed by BLAST and phylogenetic analyses based on the internal transcribed spacer and the D2-D3 expansion located in the 28S gene of ribosomal DNA for CCN and RLN populations, respectively. Pratylenchus neglectus and P. thornei populations from Kazakhstan showed a high similarity with the American, European, and Asian populations. Heterodera filipjevi populations formed a well-supported cluster with the corresponding populations from different countries and showed a slightly intraspecific polymorphism. Kazakhstan populations of H. filipjevi may have multiple introductions in Kazakhstan due to the divergence among them. The results of this study are of great importance for breeding programs and will enable awareness to extension advisors to develop measures to control these nematodes in cereal cropping areas in Kazakhstan.

Humic acid – a potential bioresource for nematode control

Nematology ◽  
2021 ◽  
pp. 1-10
Author(s):  
Seenivasan Nagachandrabose ◽  
Richard Baidoo
Keyword(s):  
Humic Acid ◽  
Crop Production ◽  
Trichoderma Viride ◽  
Foliar Spray ◽  
Crop Productivity ◽  
Systemic Resistance ◽  
Parasitic Nematodes ◽  
Nematode Control ◽  
Plant Parasitic Nematodes ◽  
Plant Parasitic

Summary There is a growing interest in the use of natural products for crop production and protection. Humic acid is a well-known bioresource that intensifies soil functions and improves crop productivity. This review article provides a synopsis of the humic acid-plant-nematode association and the prospects for using humic acid as an alternative to chemical control of nematodes. Humic acid is known to have toxic and antagonistic effects against many plant-parasitic nematodes, including Meloidogyne spp., Rotylenchulus reniformis, Radopholus similis and Helicotylenchus multicinctus. The required dose for getting significant nematode control ranges from 0.04 to 2.0% concentration. Delivery methods like soil application or drenching, seedling root dip treatment and foliar spray on leaves are effective for nematode control. Humic acid controls plant-parasitic nematodes through various mechanisms including killing juveniles, inhibiting hatching, reducing nematode infectivity and reproduction, and inducing systemic resistance. Humic acid is compatible with bio-inoculants such as Azospirillum spp., phosphobacterium, Bacillus megaterium, Pseudomonas fluorescens, Trichoderma viride, Glomus spp., Pochonia chlamydosporia, Purpureocillium lilacinum and T. asperellum. These attributes of humic acid show a promising potential for use in nematode control. However, further work on bio-efficacy against a broad spectrum of plant-parasitic nematodes is needed.

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