Cold induced promoter from winter Brassica napus

Details Cold induced promoter from winter Brassica napus Cold induced promoter from winter Brassica napus

1995-04-12

1998-12-08

Fox, David T.

Organic compounds -- part of the class 532-570 series

Organic compounds

Carbohydrates or derivatives

536 236, 4351723, 4353201, 800205, 800DIG17, C12N 1529, C12N 1582, C12N 1511, A01H 100

Patent

active

058471022

ABSTRACT:
A 1.2 kb fragment of the 5' regulatory region (from bp -1107 to +100) was fused to the GUS (.beta.-glucuronidase) reporter gene and BN115-promoted GUS expression was observed in green tissues of transgenic Brassica napus plants only after incubation at 2.degree. C. No expression was observed after incubation at 22.degree. C., either in the presence or absence of abscisic acid. Microprojectile bombardment of winter B. napus leaves with a BN115 promoter/GUS construct yielded similar results and was used to analyze a series of deletions from the 5' end of the promoter. Results obtained from transient expression studies showed that the low temperature regulation of BN115 expression involves a possible enhancer region between bp -1107 and -606 and a second positive regulatory region located between bp -302 and -274. Deletion analyses and results from replacement with a truncated CaMV (cauliflower mosaic virus) 35S promoter suggest that the minimal size required for any maintenance of low temperature GUS expression is a -300 bp fragment. Within this fragment are two 8 bp elements with the sequence TGGCCGAC, which are identical to those present in the positive regulatory region of the promoter of the homologous Arabidopsis cor15a gene and to a 5 bp core sequence in the low temperature- and dehydration-responsive elements (LTREs and DREs) identified in the promoter regions of several cold-responsive Arabidopsis thaliana genes. Mutation of either one or both of the GGCAC core sequence of the putative LTRE's to AATTC resulted in loss of cold-inducible gene expression, providing for the first time, direct evidence that CCGAC is required for low temperature expression. Furthermore, replacement of an enhancer region (-605 to -1107) of the promoter with a more active enhancer from the 35S constitutive cauliflower mosaic virus (CaMV) promoter resulted in a "hybrid" promoter with increased low temperature induced activity several fold over that of the native promoter.

REFERENCES:
White et al, Regulation of BN115, a Low-Temperature-Responsive Gene from Winter Brassica napus. Plant Physiol., vol. 106 (1994) pp. 917-928.
Singh et al, Biochemical and Cellular Mehcanisms of Stress Tolerance in Plants, NATO ASI Series, vol. H 86, Biochemical and Cellular Mechanisms of Stress Tolerance in Plants (1994) pp. 557-567.
Weretilnyk et al Characterization of Three Related Low-Temperature-Regulated cDNAs from Winter Brassica napus, Plant Physiol., vol. 101 (1993) pp. 171-177.
Baker et al, The 5'-region of Arabidopsis thaliana cor15a has cis-acting elements that confer cold-, drought-and ABA-regulated gene expression. Plant Molecular Biology, vol. 24, (1994) pp. 701-713.
Yamaguchi-Shinozaki et al, A novel cis-acting element in an arabidopsis gene is involved in responsiveness to drought, low-temperature, or high-salt stress. The Plant Cell, vol. 6 (1994), pp. 251-264.
Orr et al, Complementary DNA sequence of a low temperature-induced Brassica napus gene with homology to the Arabidopsis thaliana kin1 gene. Plant Physiol., vol. 98 (1992), pp. 1532-1534.
Plant Physiol, vol. 99, No. 1 (Suppl.), p. 78 (Abstract No. 467). (1992) White, TC, Simmonds, D, Singh, J. Annual Meeting of the American Society of Plant Physiologists, Pittsburg, Pennsylvania, USA, Aug. 1-5.

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