Abstract
A cDNA clone, pBN115, encoding a low-temperature-regulated transcript in winter Brassica napus has been isolated. Northern blot analyses show that levels of transcripts hybridizing to pBN115 increase within 24 h of exposure of B. napus to low temperature, peak at 3 d, and then remain at an elevated level for the duration of the cold treatment (up to 10 weeks). Transferring plants from 2 degrees C to room temperature results in the loss of detectable transcripts hybridizing to pBN115 within 1 d. The transcript was not detected in RNA isolated from roots of cold-acclimated B. napus. Results of in vivo labeling of nascent RNA in leaf discs of B. napus with thiouridine suggest that regulation of expression may be transcriptional, at least at the onset of cold temperature. Although drought stress leads to a slight increase in transcript level at room temperature, neither a brief exposure to elevated temperatures nor exogenous application of abscisic acid resulted in the appearance of the transcript represented by pBN115. Furthermore, transcripts hybridizing to pBN115 were present at the same levels whether the plants were acclimated in the light or dark. Hybridization experiments show that pBN115 hybridizes strongly to cold-regulated transcripts in Arabidopsis thaliana, Descurania sophia, and spring B. napus, all of which are cruciferous plants capable of cold acclimation. No hybridizing transcript could be detected in cold-acclimated Spinacea oleracea, winter Secale cereale, or cold-grown Nicotiana tabacum. DNA sequence analysis of pBN115 reveals a single open reading frame that potentially encodes a protein of 14.8 kD. This size closely approximates that of a polypeptide produced by in vitro transcription/translation experiments. Two additional cDNA clones, pBN19 and pBN26, with divergent 5'-and 3'-untranslated regions, were also isolated and found to encode similar, but not identical, polypeptides.
Full Text
The Full Text of this article is available as a PDF (2.3 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Cattivelli L., Bartels D. Molecular cloning and characterization of cold-regulated genes in barley. Plant Physiol. 1990 Aug;93(4):1504–1510. doi: 10.1104/pp.93.4.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Daie J., Wyse R. ABA Uptake in Source and Sink Tissues of Sugar Beet. Plant Physiol. 1983 Jun;72(2):430–433. doi: 10.1104/pp.72.2.430. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gilmour S. J., Artus N. N., Thomashow M. F. cDNA sequence analysis and expression of two cold-regulated genes of Arabidopsis thaliana. Plant Mol Biol. 1992 Jan;18(1):13–21. doi: 10.1007/BF00018452. [DOI] [PubMed] [Google Scholar]
- Gilmour S. J., Hajela R. K., Thomashow M. F. Cold Acclimation in Arabidopsis thaliana. Plant Physiol. 1988 Jul;87(3):745–750. doi: 10.1104/pp.87.3.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
- Hajela R. K., Horvath D. P., Gilmour S. J., Thomashow M. F. Molecular Cloning and Expression of cor (Cold-Regulated) Genes in Arabidopsis thaliana. Plant Physiol. 1990 Jul;93(3):1246–1252. doi: 10.1104/pp.93.3.1246. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Houde M., Danyluk J., Laliberté J. F., Rassart E., Dhindsa R. S., Sarhan F. Cloning, characterization, and expression of a cDNA encoding a 50-kilodalton protein specifically induced by cold acclimation in wheat. Plant Physiol. 1992 Aug;99(4):1381–1387. doi: 10.1104/pp.99.4.1381. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kurkela S., Franck M. Cloning and characterization of a cold- and ABA-inducible Arabidopsis gene. Plant Mol Biol. 1990 Jul;15(1):137–144. doi: 10.1007/BF00017731. [DOI] [PubMed] [Google Scholar]
- Lin C., Guo W. W., Everson E., Thomashow M. F. Cold acclimation in Arabidopsis and wheat : a response associated with expression of related genes encoding ;boiling-stable' polypeptides. Plant Physiol. 1990 Nov;94(3):1078–1083. doi: 10.1104/pp.94.3.1078. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lin C., Thomashow M. F. DNA Sequence Analysis of a Complementary DNA for Cold-Regulated Arabidopsis Gene cor15 and Characterization of the COR 15 Polypeptide. Plant Physiol. 1992 Jun;99(2):519–525. doi: 10.1104/pp.99.2.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mohapatra S. S., Wolfraim L., Poole R. J., Dhindsa R. S. Molecular cloning and relationship to freezing tolerance of cold-acclimation-specific genes of alfalfa. Plant Physiol. 1989 Jan;89(1):375–380. doi: 10.1104/pp.89.1.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nordin K., Heino P., Palva E. T. Separate signal pathways regulate the expression of a low-temperature-induced gene in Arabidopsis thaliana (L.) Heynh. Plant Mol Biol. 1991 Jun;16(6):1061–1071. doi: 10.1007/BF00016077. [DOI] [PubMed] [Google Scholar]
- Skriver K., Mundy J. Gene expression in response to abscisic acid and osmotic stress. Plant Cell. 1990 Jun;2(6):503–512. doi: 10.1105/tpc.2.6.503. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stetler G. L., Thorner J. Molecular cloning of hormone-responsive genes from the yeast Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1144–1148. doi: 10.1073/pnas.81.4.1144. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Woodford T. A., Schlegel R., Pardee A. B. Selective isolation of newly synthesized mammalian mRNA after in vivo labeling with 4-thiouridine or 6-thioguanosine. Anal Biochem. 1988 May 15;171(1):166–172. doi: 10.1016/0003-2697(88)90138-8. [DOI] [PubMed] [Google Scholar]