Microaspiration of Esophageal Gland Cells and cDNA Library Construction for Identifying Parasitism Genes of Plant-Parasitic Nematodes

Author(s):  
Richard S. Hussey ◽  
Guozhong Huang ◽  
Rex Allen
2012 ◽  
Vol 1 (1) ◽  
pp. 81-87
Author(s):  
Ajit K. Ngangbam ◽  
Nongmaithem B. Devi

Plant parasitic nematodes which are highly successful parasites evolved a very specialized feeding relationship with the host plant to cause the destructive root-knot disease. They initiate their parasitic relationship with the host by releasing their secretions into root cells which in turn stimulate the root cells of the host to become specialized feeding cells which are considered as the single source of nutrients essential for the nematode's survival. The parasitism genes expressed in nematode's esophageal gland cells encode secretory proteins that are released through its stylet to direct the interactions of the nematode with its host plants.


2013 ◽  
Vol 26 (1) ◽  
pp. 9-16 ◽  
Author(s):  
Tarek Hewezi ◽  
Thomas J. Baum

A key feature of sedentary plant-parasitic nematodes is the release of effector proteins from their esophageal gland cells through their stylets into host roots. These proteinaceous stylet secretions have been shown to be crucial for successful parasitism by mediating the transition of normal root cells into specialized feeding sites and by negating plant defenses. Recent technical advances of purifying mRNA from esophageal gland cells of plant-parasitic nematodes coupled with emerging sequencing technologies is steadily expanding our knowledge of nematode effector repertoires. Host targets and biological activities of a number of nematode effectors are continuously being reported and, by now, a first picture of the complexity of sedentary nematode parasitism at the molecular level is starting to take shape. In this review, we highlight effector mechanisms that recently have been uncovered by studying the host–pathogen interaction. These mechanisms range from mediating susceptibility of host plants to the actual triggering of defense responses. In particular, we portray and discuss the mechanisms by which nematode effectors modify plant cell walls, negate host defense responses, alter auxin and polyamine signaling, mimic plant molecules, regulate stress signaling, and activate hypersensitive responses. Continuous molecular characterization of newly discovered nematode effectors will be needed to determine how these effectors orchestrate host signaling pathways and biological processes leading to successful parasitism.


2013 ◽  
Vol 26 (1) ◽  
pp. 31-35 ◽  
Author(s):  
Tom R. Maier ◽  
Tarek Hewezi ◽  
Jiqing Peng ◽  
Thomas J. Baum

Esophageal glands of plant-parasitic nematodes are highly specialized cells whose gene expression products include secreted effector proteins, which govern nematode parasitism of host plants. Therefore, elucidating the transcriptomes of esophageal glands with the goal of identifying nematode effectors is a promising avenue to understanding nematode parasitism and its evolutionary origins as well as to devising nematode control strategies. We have developed a method to separate and isolate individual esophageal gland cells from multiple species of plant-parasitic nematodes while preserving RNA quality. We have used such isolated gland cells for transcriptome analysis via high-throughput DNA sequencing. This method relies on the differential histochemical staining of the gland cells after homogenization of phytonematode tissues. Total RNA was extracted from whole gland cells isolated from eight different plant-parasitic nematode species. To validate this approach, the isolated RNA from three plant-parasitic nematode species—Globodera rostochiensis, Pratylenchus penetrans, and Radopholus similis—was amplified, gel purified, and used for 454 sequencing. We obtained 456,801 total reads with an average read length of 409 bp. Sequence analyses revealed the presence of homologs of previously known nematode effectors in these libraries, thus validating our approach. These data provide compelling evidence that this technical advance can be used to relatively easily and expediently discover effector repertoires of plant-parasitic nematodes.


Parasitology ◽  
2014 ◽  
Vol 142 (S1) ◽  
pp. S71-S84 ◽  
Author(s):  
DAVID McK. BIRD ◽  
JOHN T. JONES ◽  
CHARLES H. OPPERMAN ◽  
TAISEI KIKUCHI ◽  
ETIENNE G. J. DANCHIN

SUMMARYPlant-parasitic nematodes cause considerable damage to global agriculture. The ability to parasitize plants is a derived character that appears to have independently emerged several times in the phylum Nematoda. Morphological convergence to feeding style has been observed, but whether this is emergent from molecular convergence is less obvious. To address this, we assess whether genomic signatures can be associated with plant parasitism by nematodes. In this review, we report genomic features and characteristics that appear to be common in plant-parasitic nematodes while absent or rare in animal parasites, predators or free-living species. Candidate horizontal acquisitions of parasitism genes have systematically been found in all plant-parasitic species investigated at the sequence level. Presence of peptides that mimic plant hormones also appears to be a trait of plant-parasitic species. Annotations of the few genomes of plant-parasitic nematodes available to date have revealed a set of apparently species-specific genes on every occasion. Effector genes, important for parasitism are frequently found among those species-specific genes, indicating poor overlap. Overall, nematodes appear to have developed convergent genomic solutions to adapt to plant parasitism.


2005 ◽  
Vol 18 (7) ◽  
pp. 615-620 ◽  
Author(s):  
M.-N. Rosso ◽  
M. P. Dubrana ◽  
N. Cimbolini ◽  
S. Jaubert ◽  
P. Abad

Plant parasitic nematodes have been, so far, refractory to transformation or mutagenesis. The functional analysis of nematode genes relies on the development of reverse genetic tools adapted to these obligate parasites. Here, we describe the application of RNA interference (RNAi) to the root-knot nematode Meloidogyne incognita for the knock-down of two genes expressed in the subventral esophageal glands of the nematode and potentially involved in parasitism, the calreticulin (Mi-crt) and the polygalacturonase (Mi-pg-1) genes. Incubation in 1% resorcinol for 4 h induced double-stranded RNA uptake through the alimentary track of the nematodes and led to up to 92% depletion of Mi-crt transcripts. Timecourse analysis of the silencing showed different temporal patterns for Mi-crt and Mi-pg-1. The silencing of Mi-crt was optimal 20 h after soaking, whereas the silencing of Mi-pg-1 was optimal 44 h after soaking. For the two genes, the silencing effect was highly time-limited, since no transcript depletion was detectable 68 h after soaking.


2001 ◽  
Vol 14 (10) ◽  
pp. 1247-1254 ◽  
Author(s):  
Bingli Gao ◽  
R. Allen ◽  
Tom Maier ◽  
Eric L. Davis ◽  
Thomas J. Baum ◽  
...  

Cloning parasitism genes encoding secretory proteins expressed in the esophageal gland cells is the key to understanding the molecular basis of nematode parasitism of plants. Suppression subtractive hybridization (SSH) with the microaspirated contents from Heterodera glycines esophageal gland cells and intestinal region was used to isolate genes expressed preferentially in the gland cells of parasitic stages. Twenty-three unique cDNA sequences from a SSH cDNA library were identified and hybridized to the genomic DNA of H. glycines in Southern blots. Full-length cDNAs of 21 clones were obtained by screening a gland-cell long-distance polymerase chain reaction cDNA library. Deduced proteins of 10 clones were preceded by a signal peptide for secretion, and PSORT II computer analysis predicted eight proteins as extracellular, one as nuclear, and one as plasmalemma localized. In situ hybridization showed that four of the predicted extracellular clones were expressed specifically in the dorsal gland cell, one in the subventral gland cells, and three in the intestine in H. glycines. The predicted nuclear clone and the plasmalemma-localized clone were expressed in the subventral gland cells and the dorsal gland cell, respectively. SSH is an efficient method for cloning putative parasitism genes encoding esophageal gland cell secretory proteins that may have a role in H. glycines parasitism of soybean.


EDIS ◽  
2017 ◽  
Vol 2017 (2) ◽  
pp. 8
Author(s):  
Zane Grabau

This 8-page fact sheet written by Zane J. Grabau and published in January 2017 by the UF Department of Entomology and Nematology explains how to diagnose and manage nematode problems in cotton production.­http://edis.ifas.ufl.edu/ng015


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