Synchronous Arabidopsis suspension cultures for analysis of cell‐cycle gene activity

Synchronous Arabidopsis suspension cultures for analysis of cell‐cycle gene activity 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png The Plant Journal Wiley

Synchronous Arabidopsis suspension cultures for analysis of cell‐cycle gene activity

The Plant Journal, Volume 30 (2) – Apr 1, 2002

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Publisher
Wiley
Copyright
Copyright © 2002 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0960-7412
eISSN
1365-313X
D.O.I.
10.1046/j.1365-313X.2002.01274.x
Publisher site
See Article on Publisher Site

Abstract

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.

Journal

The Plant JournalWiley

Published: Apr 1, 2002

References

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