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Synchronous Arabidopsis suspension cultures for analysis of cell‐cycle gene activity
Author(s) -
Menges Margit,
Murray James A.H.
Publication year - 2002
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.2002.01274.x
Subject(s) - arabidopsis , aphidicolin , cell cycle , biology , microbiology and biotechnology , cyclin dependent kinase , cell synchronization , gene , cell division , gene expression , cell , genetics , mutant
Summary Synchronized suspension cultures are powerful tools in plant cell‐cycle studies. However, few Arabidopsis cell cultures are available, and synchrony extending over several sequential phases of the cell cycle has not been reported. Here we describe the first useful synchrony in Arabidopsis , achieved by selecting the rapidly dividing Arabidopsis cell suspensions MM1 and MM2d. Synchrony may be achieved either by removing and re‐supplying sucrose to the growth media or by applying an aphidicolin block/release. Synchronization with aphidicolin produced up to 80% S‐phase cells and up to 92% G2 cells, together with clear separation of different cell‐cycle phases. These synchronization procedures can be used for analysis of gene expression and protein activity. We show that representatives of three CDK gene classes of Arabidopsis ( CDKA , CDKB1 and CDKB2 ) show differential expression timing, and that three CDK inhibitor genes show strikingly different expression patterns during cell‐cycle re‐entry. We propose that ICK2 (KRP2) may have a specific role in this process.