Distribution of actin gene isoforms in the Arabidopsis leaf measured in microsamples from intact individual cells

Laval, V, Koroleva, O, Murphy, E, Lu, C ORCID: https://orcid.org/0000-0002-0064-4725, Milner, J, Hooks, M and Tomos, D, 2002. Distribution of actin gene isoforms in the Arabidopsis leaf measured in microsamples from intact individual cells. Planta, 215 (2), pp. 287-292. ISSN 0032-0935

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Abstract

The contents of single plant cells can be sampled using glass microcapillaries. By combining such single-cell sampling with reverse transcription-polymerase chain reaction (RT-PCR), transcripts of individual genes can be identified and, in principle, quantified. This provides a valuable technique for the analysis and quantification of the intercellular distribution of gene expression in complex tissues. In a proof-of-principle study, the cellular locations of the transcripts of the eight isoforms of actin (ACT) expressed in Arabidopsis thaliana (L.) Heynh. were analyzed. Cell sap was extracted from epidermal and mesophyll cells of leaves of 3- to 4-week-old plants. Single-cell (SC)-RT-PCR was used to amplify the actin transcripts using specific primer pairs for ACT1, 2, 3, 4, 7, 8, 11 and 12. Only ACT2 and ACT8 were found in epidermal and in mesophyll cells. In individual trichomes, in addition to ACT2 and ACT8, ACT7 and ACT11 transcripts were detectable. By employing the already well-characterized actin system we demonstrate the practicality and power of SC-RT-PCR as a technique for analyzing gene expression at the ultimate level of resolution, the single cell.

Item Type:	Journal article
Publication Title:	Planta
Creators:	Laval, V., Koroleva, O., Murphy, E., Lu, C., Milner, J., Hooks, M. and Tomos, D.
Publisher:	Springer
Date:	June 2002
Volume:	215
Number:	2
ISSN:	0032-0935
Identifiers:	Number Type 10.1007/s00425-001-0732-y DOI
Divisions:	Schools > School of Architecture, Design and the Built Environment
Record created by:	Linda Sullivan
Date Added:	08 Sep 2016 10:16
Last Modified:	09 Jun 2017 14:05
URI:	https://irep.ntu.ac.uk/id/eprint/28414

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