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 ◽  
...  

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.

Author(s):  
Olaf Kranse ◽  
Helen Beasley ◽  
Sally Adams ◽  
Andre Pires da Silva ◽  
Chris Bell ◽  
...  

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.


Plant Disease ◽  
2019 ◽  
Vol 103 (7) ◽  
pp. 1621-1630 ◽  
Author(s):  
Oliver Chitambo ◽  
Solveig Haukeland ◽  
Komi K. M. Fiaboe ◽  
Florian M. W. Grundler

Plant-parasitic nematodes, particularly root-knot nematodes (RKN: Meloidogyne spp.) and cyst nematodes (CN: Globodera and Heterodera spp.) cause severe yield reduction in most cultivated crops and are of high economic importance. African nightshade (Solanum spp.) and African spinach (Amaranthus spp.) are important African indigenous vegetables (AIV) and are rich sources of nutrition and income. However, their host status to plant-parasitic nematodes remains largely speculative. Therefore, a survey was conducted which revealed that S. villosum exhibited high root galling, whereas on S. scabrum, A. cruentus, and A. dubius root galling was rare or very low. Additionally, soil collected from the rhizosphere of S. villosum and S. scabrum contained few cysts of potato cyst nematodes (PCN), and no developing PCN females were observed on the roots of growing plants. Therefore, we studied the dynamics of RKN and PCN on A. dubius, A. cruentus, S. scabrum, and S. villosum over 2 years in a field experiment. The effects of AIV crop species on RKN and PCN soil infestation were evaluated using susceptible S. lycopersicum or S. tuberosum. After first, second, and third cultivation of A. dubius, A. cruentus, and S. scabrum, RKN infestation of the soil decreased by more than 85%, whereas S. scabrum and S. villosum decreased PCN densities by more than 80%. When cropping susceptible crops, after three seasons of successive cultivation of these AIV, galling index and number of developing PCN females measured on susceptible crops decreased by more than 75%. Wilting and RKN-PCN coinfection incidences also decreased significantly. Here, we present data that support the development of a novel cropping system including African spinach and African nightshade, which reveals a high potential to manage RKN and PCN in an environmentally friendly, effective, and productive way.


Nematology ◽  
2007 ◽  
Vol 9 (4) ◽  
pp. 451-458 ◽  
Author(s):  
Julia Hofmann ◽  
Florian Grundler

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.


2007 ◽  
Vol 47 (5) ◽  
pp. 620 ◽  
Author(s):  
B. L. Blair ◽  
G. R. Stirling

Damage to sugarcane caused by root-knot nematode (Meloidogyne spp.) is well documented in infertile coarse-textured soils, but crop losses have never been assessed in the fine-textured soils on which more than 95% of Australia’s sugarcane is grown. The impact of nematodes in these more fertile soils was assessed by repeatedly applying nematicides (aldicarb and fenamiphos) to plant and ratoon crops in 16 fields, and measuring their effects on nematode populations, sugarcane growth and yield. In untreated plant crops, mid-season population densities of lesion nematode (Pratylenchus zeae), root-knot nematode (M. javanica), stunt nematode (Tylenchorhynchus annulatus), spiral nematode (Helicotylenchus dihystera) and stubby-root nematode (Paratrichodorus minor) averaged 1065, 214, 535, 217 and 103 nematodes/200 mL soil, respectively. Lower mean nematode population densities were recorded in the first ratoon, particularly for root-knot nematode. Nematicides reduced populations of lesion nematode by 66–99% in both plant and ratoon crops, but control of root-knot nematode was inconsistent, particularly in ratoons. Nematicide treatment had a greater impact on shoot and stalk length than on shoot and stalk number. The entire community of pest nematodes appeared to be contributing to lost productivity, but stalk length and final yield responses correlated most consistently with the number of lesion nematodes controlled. Fine roots in nematicide-treated plots were healthier and more numerous than in untreated plots, and this was indicative of the reduced impact of lesion nematode. Yield responses averaged 15.3% in plant crops and 11.6% in ratoons, indicating that nematodes are subtle but significant pests of sugarcane in fine-textured soils. On the basis of these results, plant-parasitic nematodes are conservatively estimated to cost the Australian sugar industry about AU$82 million/annum.


EDIS ◽  
2018 ◽  
Vol 2018 (4) ◽  
Author(s):  
Mary Ann D. Maquilan ◽  
Ali Sarkhosh ◽  
Donald W. Dickson

One of the production issues that peach growers in Florida must contend with is plant-parasitic nematodes. One such species is the more recently discovered peach root-knot nematode, Meloidogyne floridensis, which is the subject of this 5-page publication. Written by Mary Ann D. Maquilan, Ali Sarkhosh, and Donald Dickson and published by the UF/IFAS Horticultural Sciences Department, July 2018. http://edis.ifas.ufl.edu/hs1320


Author(s):  
Pranaya Pradhan ◽  
Dhirendra Kumar Nayak ◽  
Manaswini Mahapatra

The significant constraints in Chickpea (Cicer arietinum L.) production hampers a bit more than 14% global yield loss due to plant-parasitic nematodes. Root-knot nematode (Meloidogyne sp.) is an endoparasite and a significant species affecting the chickpea plant. So, the chemical basis of management is more cost-effective, and pest resurgence building is enhanced in the pathogen. So, ecological-based nematode management is requisite, which also is got hampered due to breeding for resistance against such plant-parasitic nematodes. This was the primary reason to conduct this experiment to enhance resistance in the chickpea plants based on Zinc uptake by using bioagent, Pseudomonas fluorescens alone or in combination. where Different treatments including nematode, bacterium, and chemicals were used sustaining the enhancement of disease resistance in chickpea cultivars, RSG 974, GG 5, GNG 2144. Zinc content of chickpea variety GNG 2144 was found the highest in treatment, when only bacterium (P. fluorescens) was inoculated, i.e., 3.14 mg/100g of root followed by GG 5, i.e., 2.79 mg/100g of root and RSG 974 was, i.e., 2.35 mg/100g of root respectively in a descending order. Application of P. fluorescence combined or alone gradually increased the Zn concentration in roots of chickpea plants compared to healthy check followed by chemical treated plants.


2021 ◽  
Author(s):  
Shahid Siddique ◽  
Zoran S. Radakovic ◽  
Clarissa Hiltl ◽  
Clement Pellegrin ◽  
Thomas J. Baum ◽  
...  

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.


1993 ◽  
Vol 33 (2) ◽  
pp. 197 ◽  
Author(s):  
GR Stirling ◽  
A Nikulin

Twelve pineapple fields with various densities of root-knot nematode were selected during a ratoon crop. Nematode populations were monitored regularly after the crop was ploughed out. Regardless of the original population density, rootknot nematodes were almost nondetectable at the end of the subsequent 3-6-month fallow intercycle period. In the absence of nematicide treatment there were marked differences between sites in the manner in which rootknot nematodes increased in the newly planted crop. At some sites, they were detectable 9-15 months after planting, whereas at other sites, nematodes were not observed at 15 months. Increases in ratoon crop yield following application of ethylene dibromide or fenamiphos were related to root-knot nematode population density. Significant increases in yield were not obtained at sites where the nematode was not detectable at 15 months. The results suggest that some pineapple growers are needlessly applying nematicides and that nematode diagnostic services should be developed to provide growers with advice on their nematode management programs.


1989 ◽  
Vol 29 (1) ◽  
pp. 129 ◽  
Author(s):  
M Edwards

Three grapevine rootstock trials in North-East Victoria, Australia, were sampled over several summers to determine nematode populations and the resistance or the tolerance of some commonly used winegrape varieties and rootstocks to plant parasitic nematodes. Rootstocks on which nematodes failed to reproduce or reproduced poorly were considered resistant, rootstocks which supported high populations of nematodes with no apparent effect on yield were considered tolerant. Susceptible rootstocks supported high numbers of nematodes and yielded poorly. The varieties studied were: Shiraz, Chardonnay, Cabernet Sauvignon; and the rootstocks were: Harmony, Richter 110, Schwarzmann, ARG No. 1, 5A Teleki, Rupestris du Lot, 5BB Kober, SO4, Ramsey, K5 1-32 and 1202. The plant parasitic nematodes found on the trial sites were root-knot (Meloidogyne javanica (Treub) Chitwood) and citrus (Tylenchulus semipenetrans Cobb) nematodes. Cabernet Sauvignon, Chardonnay, Shiraz and ARG No. 1 were susceptible to the root-knot nematode. Harmony exhibited tolerance to the citrus nematode and was a good host, allowing a large population to build up. Ramsey appeared to be resistant to the citrus nematode, at least in the Cabernet Sauvignon rootstock trial at Wahgunyah.


2014 ◽  
Vol 104 (3) ◽  
pp. 306-312 ◽  
Author(s):  
Martin Marek ◽  
Miloslav Zouhar ◽  
Ondřej Douda ◽  
Marie Maňasová ◽  
Pavel Ryšánek

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).


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