A cytometry microparticle platform approach for screening tobacco microRNA changes after agrobacterium delivery

Joshua D. Powell, Qiang Chen, Hugh S. Mason

Research output: Contribution to journalArticle

Abstract

MicroRNAs are a class of non-coding regulatory RNAs that can modulate development as well as alter innate antiviral defenses in plants. In this study we explored changes in Nicotiana benthamiana tobacco microRNA expression as it relates to expression of a recombinant anti-Ebola GP1 antibody. The antibody was delivered to tobacco leaves through a bacterial Agrobacterium tumefaciens “agroinfiltration” expression strategy. A multiplex microparticle-based cytometry assay tracked the expression changes of 53 host tobacco microRNAs. Our results revealed that the most abundant microRNAs in actively growing leaves corresponded to nanoparticle probes specific to nta-mir-6149 and nta-miR-168b. After agroinfiltration, probes specific for nta-mir-398, and nta-mir-482d were significantly altered in their respective expression levels, however changes were partially attributed to the infiltration broth medium used in the antibody delivery process. Confirmation of nta-mir-398 and nta-mir-482d expression changes was also verified through RT-qPCR. To our knowledge this study is the first to profile medium and Agrobacterium injection at the microRNA level through a multiplex microparticle approach.

Original languageEnglish (US)
Pages (from-to)230-235
Number of pages6
JournalJournal of Microbiological Methods
Volume127
DOIs
StatePublished - Aug 1 2016

Fingerprint

Agrobacterium
MicroRNAs
Tobacco
Antibodies
Agrobacterium tumefaciens
Untranslated RNA
Nanoparticles
Antiviral Agents

Keywords

  • Agrobacterium tumefaciens
  • Ebola
  • Nicotiana benthamiana
  • Tobacco

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Microbiology (medical)

Cite this

@article{efe777c108ef44a884289061e21df958,
title = "A cytometry microparticle platform approach for screening tobacco microRNA changes after agrobacterium delivery",
abstract = "MicroRNAs are a class of non-coding regulatory RNAs that can modulate development as well as alter innate antiviral defenses in plants. In this study we explored changes in Nicotiana benthamiana tobacco microRNA expression as it relates to expression of a recombinant anti-Ebola GP1 antibody. The antibody was delivered to tobacco leaves through a bacterial Agrobacterium tumefaciens “agroinfiltration” expression strategy. A multiplex microparticle-based cytometry assay tracked the expression changes of 53 host tobacco microRNAs. Our results revealed that the most abundant microRNAs in actively growing leaves corresponded to nanoparticle probes specific to nta-mir-6149 and nta-miR-168b. After agroinfiltration, probes specific for nta-mir-398, and nta-mir-482d were significantly altered in their respective expression levels, however changes were partially attributed to the infiltration broth medium used in the antibody delivery process. Confirmation of nta-mir-398 and nta-mir-482d expression changes was also verified through RT-qPCR. To our knowledge this study is the first to profile medium and Agrobacterium injection at the microRNA level through a multiplex microparticle approach.",
keywords = "Agrobacterium tumefaciens, Ebola, Nicotiana benthamiana, Tobacco",
author = "Powell, {Joshua D.} and Qiang Chen and Mason, {Hugh S.}",
year = "2016",
month = "8",
doi = "10.1016/j.mimet.2016.06.023",
volume = "127",
pages = "230--235",
journal = "Journal of Microbiological Methods",
issn = "0167-7012",
publisher = "Elsevier",

}

TY - JOUR

T1 - A cytometry microparticle platform approach for screening tobacco microRNA changes after agrobacterium delivery

AU - Powell,Joshua D.

AU - Chen,Qiang

AU - Mason,Hugh S.

PY - 2016/8/1

Y1 - 2016/8/1

N2 - MicroRNAs are a class of non-coding regulatory RNAs that can modulate development as well as alter innate antiviral defenses in plants. In this study we explored changes in Nicotiana benthamiana tobacco microRNA expression as it relates to expression of a recombinant anti-Ebola GP1 antibody. The antibody was delivered to tobacco leaves through a bacterial Agrobacterium tumefaciens “agroinfiltration” expression strategy. A multiplex microparticle-based cytometry assay tracked the expression changes of 53 host tobacco microRNAs. Our results revealed that the most abundant microRNAs in actively growing leaves corresponded to nanoparticle probes specific to nta-mir-6149 and nta-miR-168b. After agroinfiltration, probes specific for nta-mir-398, and nta-mir-482d were significantly altered in their respective expression levels, however changes were partially attributed to the infiltration broth medium used in the antibody delivery process. Confirmation of nta-mir-398 and nta-mir-482d expression changes was also verified through RT-qPCR. To our knowledge this study is the first to profile medium and Agrobacterium injection at the microRNA level through a multiplex microparticle approach.

AB - MicroRNAs are a class of non-coding regulatory RNAs that can modulate development as well as alter innate antiviral defenses in plants. In this study we explored changes in Nicotiana benthamiana tobacco microRNA expression as it relates to expression of a recombinant anti-Ebola GP1 antibody. The antibody was delivered to tobacco leaves through a bacterial Agrobacterium tumefaciens “agroinfiltration” expression strategy. A multiplex microparticle-based cytometry assay tracked the expression changes of 53 host tobacco microRNAs. Our results revealed that the most abundant microRNAs in actively growing leaves corresponded to nanoparticle probes specific to nta-mir-6149 and nta-miR-168b. After agroinfiltration, probes specific for nta-mir-398, and nta-mir-482d were significantly altered in their respective expression levels, however changes were partially attributed to the infiltration broth medium used in the antibody delivery process. Confirmation of nta-mir-398 and nta-mir-482d expression changes was also verified through RT-qPCR. To our knowledge this study is the first to profile medium and Agrobacterium injection at the microRNA level through a multiplex microparticle approach.

KW - Agrobacterium tumefaciens

KW - Ebola

KW - Nicotiana benthamiana

KW - Tobacco

UR - http://www.scopus.com/inward/record.url?scp=84976328955&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84976328955&partnerID=8YFLogxK

U2 - 10.1016/j.mimet.2016.06.023

DO - 10.1016/j.mimet.2016.06.023

M3 - Article

VL - 127

SP - 230

EP - 235

JO - Journal of Microbiological Methods

T2 - Journal of Microbiological Methods

JF - Journal of Microbiological Methods

SN - 0167-7012

ER -