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Trichome isolation with and without fixation using laser microdissection and pressure catapulting followed by RNA amplification: Expression of genes of terpene metabolism in apical and sub-apical trichome cells of Artemisia annua L.
Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.
Linnaeus University, Faculty of Science and Engineering, School of Natural Sciences.ORCID iD: 0000-0001-8899-5046
2012 (English)In: Plant Science, ISSN 0168-9452, E-ISSN 1873-2259, Vol. 183, p. 9-13Article in journal (Refereed) Published
Abstract [en]

The aim of this project was to evaluate the effect of fixation on plant material prior to Laser Microdissection and Pressure Catapulting (LMPC) and to identify an appropriate method for preserving good RNA quality after cell isolation. Therefore, flower buds from Artemisia annua L. were exposed to either the fixative formaldehyde or a non-fixative buffer prior to cell isolation by LMPC. Proteinase K was used after cell isolation from fixed plant tissue, in an attempt to improve the RNA yield. The ability to detect gene expression using real-time quantitative PCR with or without previous amplification of RNA from cells isolated by LMPC was also evaluated. Conclusively, we describe a new technique, without fixation, enabling complete isolation of intact glandular secretory trichomes and specific single trichome cells of A. annua. This method is based on LMPC and preserves good RNA quality for subsequent RNA expression studies of both whole trichomes, apical and sub-apical cells from trichomes of A. annua. Using this method, expression of genes of terpene metabolism was studied by real-time quantitative PCR. Expression of genes involved in artemisinin biosynthesis was observed in both apical and sub-apical cells.

Place, publisher, year, edition, pages
2012. Vol. 183, p. 9-13
Keywords [en]
LMPC; Artemisia annua; Trichomes; qPCR; Gene expression; Artemisinin biosynthesis
National Category
Biochemistry and Molecular Biology
Research subject
Natural Science, Biochemistry
Identifiers
URN: urn:nbn:se:lnu:diva-18066DOI: 10.1016/j.plantsci.2011.10.019Scopus ID: 2-s2.0-82155203073OAI: oai:DiVA.org:lnu-18066DiVA, id: diva2:511114
Available from: 2012-03-20 Created: 2012-03-20 Last updated: 2017-12-07Bibliographically approved

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Publisher's full textScopushttp://www.sciencedirect.com/science/article/pii/S0168945211003062

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Olofsson, LindaLundgren, AnneliBrodelius, Peter E.

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