Isolation and Characterisation of Carotenoid Pathway Genes and Promoters from Vitis Vinifera as Resources Towards Stress-Tolerant Grapes with Superior Quality


Project Number
IWBT 05-09A

Project title
Isolation and characterisation of carotenoid pathway genes and promoters from Vitis vinifera as resources towards stress-tolerant grapes with superior quality

Project leader
Vivier, M A

Institution
University of Stellenbosch. Faculty of AgriSciences. Institute for Wine Biotechnology

Project description
To develop grapevines with enhanced capabilities to grow under adverse conditions, we are targeting the carotenoid pathway of grapevine as a central pathway that is linked to stress-tolerance (specifically light-stress), pigment development as well as the formation of the stress-hormone, abscisic acid.

Seven full-length carotenoid (and precursor) biosynthetic genes from Vitis vinifera (DXS, IPI, BCH, LBCy, NCED, PSY, and ZEP) have been isolated and characterised. This includes:
– cDNA- and genomic copies of the listed genes have been isolated, cloned, sequenced and analysed;
– the putative promoters have been isolated, cloned, sequenced and analysed (all except IPI);
– transient activity of the putative promoters using a reporter gene assay has been completed;
– preliminary expression profiles of candidate genes in the pathway have been established in grapevine,
– the functionality of the isolated genes has been confirmed in a heterologous bacterial system (IPI, BCH, LBCy, PSY, and ZEP).

Presentation(s)
Young, P R, Vivier, M A and Pretorius, I S. 1998. Isolation and characterisation of a ß-carotene hydroxylase from Vitis vinifera. Paper presented at the 2nd Federation of African Societies of Biochemistry and Molecular Biology Congress and the 15th Congress of the South African Society of Biochemistry and Molecular Biology. 30 September – 3 October 1998, Potchefstroom, South Africa

Young, P R, Vivier, M A and Pretorius, I S. 1999. Sequence analysis of a ß-carotene hydroxylase from Vitis vinifera. Paper presented at the 23rd National Congress of the South African Society for Enology and Viticulture. 12-13 November, Cape Town, South Africa.

Young, P R, Vivier, M A, Pretorius, I S. 2000. The structural analysis of a ß-carotene hydroxylase-encoding gene from Vitis vinifera. Paper presented at BIOY2K Combined Millennium Meeting. 23-28 January 2000, Grahamstown, South Africa.

Young, P R, Taylor, K L, Chen, S W, Pretorius, I S and Vivier, M A. 2001. Elucidation and manipulation of carotenoid biosynthesis in Vitis vinifera. Paper presented at the 26th World Congress of the Vine and Wine and the 81st General Assembly of the OIV. 11-17 October 2001, Adelaide, Australia.

Young, P R, Chen, S W and Vivier, M A. 2003. The photoprotective capability of tobacco is improved by the overexpression of the grapevine (Vitis Vinifera L.) ß-carotene hydroxylase-encoding gene. Paper presented at the 29th International Congress of Biochemistry and Molecular Biology. 20-24 July, Toronto, Canada.

Young, P R, Taylor, K L and Vivier, M A. 2004. Isolation and characterisation of candidate genes in the isoprenoid- and carotenoid biosynthetic pathways of Vitis vinifera. Paper presented at the 7th International Symposium on Grapevine Physiology and Biotechnology. 21-25 June 2004, Davis, California, United States.

Young, P R, Chen, S W, Pretorius, I S and Vivier, M A. 2004. Overexpression of the Vitis vinifera ß-carotene hydroxylase gene improves the photoprotective ability of tobacco. Paper presented at the 13th Conference of the South African Society for Microbiology. 2-5 April, Stellenbosch, South Africa.

Young, P R, Chen, S W, Pretorius, I S and Vivier, M A. 2004 Overexpression of the Vitis vinifera ß-carotene hydroxylase gene improves the photoprotective ability of tobacco. Paper presented at the 7th International Symposium on Grapevine Physiology and Biotechnology. 21-25 June 2004, Davis, California, United States.

Taylor, K, Young, P, Brackenridge, A and Vivier M A. 2004. Overexpression of grapevine carotenoid biosynthetic genes in Arabidopsis thaliana. Paper presented at the 13th Conference of the South African Society for Microbiology. 2-5 April 2004, Stellenbosch, South Africa.

Vivier, M A, Joubert, D A, Becker, J v W, Wentzel, L, Taylor, K and Young, P R. 2005. Functional analysis of grapevine genes. Paper presented at the 1st International Grape Genomics Symposium. 12-14 July 2005, St. Louis, Missouri, United States.

Article
Taylor, K L, Brackenridge, A E, Vivier, M A, Oberholster, A. 2006. High performance liquid chromatography profiling of the major carotenoids in Arabidopsis thaliana leaf tissue, Journal of Chromatography A, v. 1121 (1) (p. 83-91)

Dissertation
Young, P R. 2004. Molecular analyses of candidate carotenoid biosynthetic genes in Vitis vinifera L. PhD. University of Stellenbosch, Stellenbosch, South Africa.

Taylor, K L. 2006. Isolation and characterisation of carotenoid biosynthetic genes from Vitis vinifera. PhD. University of Stellenbosch, Stellenbosch, South Africa.

Thesis
Brackenridge, A E. 2006. Over-expression of two Vitis vinifera carotenoid biosynthetic genes in transgenic Arabidopsis. MSc. University of Stellenbosch, Stellenbosch.

FinalReport.pdf

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