Isolation of Whole Esophageal Gland Cells from Plant-Parasitic Nematodes for Transcriptome Analyses and Effector Identification

dc.contributor.author Maier, Tom
dc.contributor.author Baum, Thomas
dc.contributor.author Hewezi, Tarek
dc.contributor.author Peng, Jiqing
dc.contributor.author Baum, Thomas
dc.contributor.department Plant Pathology and Microbiology
dc.date 2018-02-18T18:43:26.000
dc.date.accessioned 2020-06-30T06:22:20Z
dc.date.available 2020-06-30T06:22:20Z
dc.date.copyright Tue Jan 01 00:00:00 UTC 2013
dc.date.issued 2013-01-01
dc.description.abstract <p>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—<em>Globodera rostochiensis, Pratylenchus penetrans</em>, and <em>Radopholus similis</em>—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.</p>
dc.description.comments <p>This article is published as Maier, Tom R., Tarek Hewezi, Jiqing Peng, and Thomas J. Baum. "Isolation of whole esophageal gland cells from plant-parasitic nematodes for transcriptome analyses and effector identification." <em>Molecular Plant-Microbe Interactions</em> 26, no. 1 (2013): 31-35, doi: <a href="https://doi.org/10.1094/MPMI-05-12-0121-FI">10.1094/MPMI-05-12-0121-FI</a>. Posted with permission.</p>
dc.format.mimetype application/pdf
dc.identifier archive/lib.dr.iastate.edu/plantpath_pubs/137/
dc.identifier.articleid 1147
dc.identifier.contextkey 10532782
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath plantpath_pubs/137
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/57580
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/plantpath_pubs/137/2013_Baum_IsolationWhole.pdf|||Fri Jan 14 19:59:02 UTC 2022
dc.source.uri 10.1094/MPMI-05-12-0121-FI
dc.subject.disciplines Agricultural Science
dc.subject.disciplines Agriculture
dc.subject.disciplines Genetics
dc.subject.disciplines Plant Pathology
dc.title Isolation of Whole Esophageal Gland Cells from Plant-Parasitic Nematodes for Transcriptome Analyses and Effector Identification
dc.type article
dc.type.genre article
dspace.entity.type Publication
relation.isAuthorOfPublication ea8d53ba-1a8f-4f5c-9b4a-1ecbd67e2f43
relation.isOrgUnitOfPublication a26b5928-54bb-4a0b-a973-95d649d1ad83
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