Flow Cytometric Analysis and Sorting of Heterodera glycines Eggs

dc.contributor.author Tylka, G. L.
dc.contributor.author Niblack, T. L.
dc.contributor.author Tylka, Gregory
dc.contributor.author Walk, T. C.
dc.contributor.author Harkins, K. R.
dc.contributor.author Barnett, L.
dc.contributor.author Baker, N. K.
dc.contributor.department Plant Pathology and Microbiology
dc.date 2018-02-18T19:52:37.000
dc.date.accessioned 2020-06-30T06:22:45Z
dc.date.available 2020-06-30T06:22:45Z
dc.date.copyright Fri Jan 01 00:00:00 UTC 1993
dc.date.issued 1993-12-01
dc.description.abstract <p>A nondestructive technique was developed to characterize and separate eggs of soybean cyst nematode, Heterodera glycines, by developmental stage using flow cytometry. Eggs from cysts cultured on susceptible soybean roots were suspended in 0.1% xanthan gum or 59% sucrose and loaded into either a Coulter EPICS 752 or EPICS 753 flow cytometer. Eggs were analyzed and sorted according to forward angle and 90º light scatter, flow cytometric parameters that are relative measures of object size and granularity, respectively. Mature eggs containing vermiform juveniles were less granular and slightly larger than eggs in earlier stages of embryogeny, allowing for separation of mature eggs from immature eggs. The effectiveness of flow cytometric sorting was evaluated by comparing the developmental stages of subpopulations of unsorted and sorted eggs. Of a subpopulation of unsorted eggs, 62% contained vermiform juveniles, whereas 85 to 95% of sorted subpopulations of larger, less granular eggs contained vermiform juveniles. Suspending H. glycines eggs in 0.1% xanthan gum or 59% sucrose for flow cytometric analysis had no effect on subsequent egg hatch in vitro. This technique is an efficient and effective means to collect large, relatively homogeneous quantities of H. glycines eggs in early or late embryogeny, and would likely be useful for analyzing and sorting eggs of other nematode species for use in developmental, genetic, or physiological research, or for identification and collection of parasitized eggs. Key words: egg development, embryogeny, flow cytometry, Heterodera glycines, method, nematode, nematode egg sorting, soybean cyst nematode.</p>
dc.description.comments <p>This article is published as Tylka, G. L., T. L. Niblack, T. C. Walk, K. R. Harkins, L. Barnett, and N. K. Baker. "Flow Cytometric Analysis and Sorting of Heterodera glycines Eggs." <em>Journal of Nematology</em> 25, no. 4: 596-602. Posted with permission.</p>
dc.format.mimetype application/pdf
dc.identifier archive/lib.dr.iastate.edu/plantpath_pubs/192/
dc.identifier.articleid 1189
dc.identifier.contextkey 10584530
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath plantpath_pubs/192
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/57637
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/plantpath_pubs/192/0-Tylka_Permission_from_Jnl_of_Nematology.pdf|||Fri Jan 14 21:53:26 UTC 2022
dc.source.bitstream archive/lib.dr.iastate.edu/plantpath_pubs/192/1993_Tylka_FlowCytometric.pdf|||Fri Jan 14 21:53:27 UTC 2022
dc.subject.disciplines Agricultural Science
dc.subject.disciplines Agriculture
dc.subject.disciplines Plant Pathology
dc.subject.keywords egg development
dc.subject.keywords embryogeny
dc.subject.keywords flow cytometry
dc.subject.keywords Heterodera glycines
dc.subject.keywords method
dc.subject.keywords nematode
dc.subject.keywords nematode egg sorting
dc.subject.keywords soybean cyst nematode
dc.title Flow Cytometric Analysis and Sorting of Heterodera glycines Eggs
dc.type article
dc.type.genre article
dspace.entity.type Publication
relation.isAuthorOfPublication 1113743f-89dc-4805-8212-529b30642102
relation.isOrgUnitOfPublication a26b5928-54bb-4a0b-a973-95d649d1ad83
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