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JXB Advance Access published online on June 20, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri204
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved.
Received November 24, 2004
Accepted April 19, 2005

RESEARCH PAPER

Identification and characterization of mutants capable of rapid seed germination at 10 °C from activation-tagged lines of Arabidopsis thaliana

Louai Salaita 1, Rup K. Kar 2, Manoj Majee 3, and A. Bruce Downie 3*

1 Department of Horticulture, 434 Plant Science Building, University of Kentucky, Lexington, KY 40546-0312, USA; Present address: College of Medicine, Department of Physiology, University of Arizona, Ina Gittings Room 101, PO Box 210093, Tucson, AZ 85721, USA
2 Department of Horticulture, 434 Plant Science Building, University of Kentucky, Lexington, KY 40546-0312, USA; Present address: Department of Botany, Visva-Bharati University, Santiniketan 731 235 West Bengal, India
3 Department of Horticulture, 434 Plant Science Building, University of Kentucky, Lexington, KY 40546-0312, USA

* To whom correspondence should be addressed.
A. Bruce Downie, E-mail: adownie{at}uky.edu


  Abstract

Five cold temperature germinating (ctg) mutants, completing germination at 10 °C faster than wild type, have been recovered from activation-tagged populations of Arabidopsis thaliana. Three (ctg10-D, 41-D, and 144-D) were tagged and segregated 3:1 for BASTA resistance in the F2 when crossed with wild type. None of the tagged ctg mutants was disturbed in sensitivity to abscisic acid or glucose but all were less sensitive to GA4+7 and osmoticum. The other two mutants (ctg156 and ctg225) were recessive, BASTA sensitive, and exhibited a transparent testa (tt) phenotype. They were more sensitive to abscisic acid, paclobutrazol, and GA4+7 than wild type but had similar sensitivity to osmoticum. Dimethylaminocinnamaldehyde staining of seeds from the two tt mutants, compared with stained seeds from the publicly available tt lines 1-10, suggested that ctg156 was a new allele of tt1, while ctg225 was similar to tt7-1. However, reciprocal crosses determined that ctg156 was not allelic to tt1 while ctg225 was a new allele of tt7. When the gene was sequenced from ctg225 it was missing 10 bp in the second exon, resulting in the incorporation of two spurious amino acids (G282E and D283A) followed by a stop. The screen successfully recovered mutants completing germination faster than wild type at 10 °C.

Keywords: Activation tag; mean germination time; seed germination; transparent testa.
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