Fluorescent imaging of GUS activity and RT-PCR analysis of gene expression in the shoot apical meristem

Fleming, Andrew J; Manzara, Thiande; Gruissem, Wilhelm; Kuhlemeier, Cris (1996). Fluorescent imaging of GUS activity and RT-PCR analysis of gene expression in the shoot apical meristem. The Plant Journal, 10(4), pp. 745-754. Blackwell Science 10.1046/j.1365-313X.1996.10040745.x

[img] Text
1996_PlantJ_10_745.pdf - Published Version
Restricted to registered users only
Available under License Publisher holds Copyright.

Download (1MB) | Request a copy

The use of promoter-reporter gene constructs in transgenic plants is a powerful tool in the analysis of gene expression which can, however, be limited in the resolution of small structures, such as the apical meristem. This paper reports on a fluorescent imaging technique for the analysis of GUS reporter gene expression to cellular resolution in the apical meristem of tomato. Using this technique in combination with an RT-PCR analysis of RES gene-specific transcript levels, it is shown that: 5' upstream sequences of RBCS genes are sufficient to mimic the pattern of transcripts revealed by in situ hybridisation (no expression in the apical meristem, high expression in the initiated leaf primordial; the genes RBCS2, RBCS3A and RBCS3B are transcriptionally activated upon primordium initiation with transcripts for RBCS1 and RBCS3C accumulating later in leaf development; and that RBCS promoter activity cannot be induced in the apical meristem by light, an environmental signal which elevates RBCS transcript level in other aerial parts of the plant. These data provide a detailed picture of the complex transcriptional events occurring on leaf initiation and the establishment of the photosynthetic machinery; they describe two complementary techniques which allow the analysis of such complex events at the tissue and cellular level; and they characterize an in vivo assay system which can be used to analyse the factors involved in the initiation and maintenance of gene expression patterns in the apical meristem.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Department of Biology > Institute of Plant Sciences (IPS) > Plant Development
08 Faculty of Science > Department of Biology > Institute of Plant Sciences (IPS)

UniBE Contributor:

Kuhlemeier, Cris

Subjects:

500 Science > 580 Plants (Botany)

ISSN:

0960-7412

Publisher:

Blackwell Science

Language:

English

Submitter:

Peter Alfred von Ballmoos-Haas

Date Deposited:

06 Jul 2017 09:09

Last Modified:

06 Jul 2017 09:09

Publisher DOI:

10.1046/j.1365-313X.1996.10040745.x

BORIS DOI:

10.7892/boris.97860

URI:

https://boris.unibe.ch/id/eprint/97860

Actions (login required)

Edit item Edit item
Provide Feedback