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A rapid method for detection of plant gene transcripts from single epidermal, mesophyll and companion cells of intact leaves
Author(s) -
Brandt Stephan,
Kehr Julia,
Walz Christina,
Imlau Astrid,
Willmitzer Lothar,
Fisahn Joachim
Publication year - 1999
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1999.00583.x
Subject(s) - biology , gene , plant cell , complementary dna , gene expression , microbiology and biotechnology , cell , botany , genetics
Summary A protocol for the detection of gene transcripts from single plant cells in living, undamaged plant tissue is described. Samples of leaf epidermal, mesophyll and companion cells were extracted by using glass microcapillaries and directly subjected to RT–PCR without any purification steps nor time consuming construction of cDNA libraries. The procedure is not restricted to surface cells or outer cell layers. Even cells from the central region of leaves could be harvested. For identification, companion cells were labelled by expression of the green fluorescent protein under control of a companion cell specific promoter. The described method is applicable to a wide range of plants and genes with different expression levels.