Functional Analysis of Central Metabolic Pathways with Regard to Roles in Stress Tolerance, Colour Development or Sugar Metabolism


Project Number
IWBT 05-09B

Project title
Functional analysis of central metabolic pathways with regard to roles in stress tolerance, colour development or sugar metabolism

Project leader
Vivier, M A

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

Project description
Two central metabolic pathways are targeted in this project to gain insight and develop technologies to manipulate them with regard to their prominent roles in stress-tolerance, colour development as well as sugar metabolism. The following has been achieved:

Functional analysis of carotenoid pathway members
Isoprenoid precursor- and carotenoid biosynthetic encoding genes from grapevine have been isolated (refer to IWBT 05-09A), cloned into plant expression vectors and transformed into the model plants, Arabidopsis thaliana and Nicotiana benthamiana. The resulting transgenic plants will be evaluated to elucidate their role in photosynthesis, photoprotection and drought tolerance. The transgenic plant lines expressing carotenoid biosynthetic genes should provide valuable insights into plant responses to stress conditions (especially drought, salinity, temperature and light) that will serve as potential targets for subsequent grapevine manipulations.

Unravelling sugar transport mechanisms in grapevine. The aim of this part of the study is to examine sugar transport in grapevine. To this end heterologous expression-cassettes containing a yeast-derived invertase targeted to either the symplast, vacuole or apoplast have been constructed and their functionality verified through the transformation and analysis of transgenic tobacco plants. These constructs are currently utilised in the transformation of grapevine embryogenic calli. The regenerated transgenic grapevine plants will be analysed to elucidate sugar loading and unloading mechanisms as well as to examine hexose and sucrose transporter expression through collaboration with the Institute for Plant Biotechnology, Stellenbosch University.

Poster(s)
Champanis, R, Vivier, M A, Botha, F C and Pretorius, I S. 1999. Manipulation of grapevine sugar metabolism. Poster presented at the 24th National Congress of the South African Society for Enology and Viticulture. 18-19 November, Somerset West, South Africa.

Champanis, R, Vivier, M A, Botha, F C and Pretorius, I S. 2000. Transformation of grapevine with a yeast-derived invertase (SUC2) targeted to different cellular location. Poster presented at the International BioY2K Combined Millennium Meeting, Grahamstown, South Africa.

Taylor, K L, Young, P R, Brackenridge, A E and Vivier, M A. 2004. Overexpression of grapevine carotenoid biosynthetic genes in Arabidopsis thaliana. Poster presented at the South African Microbiology Society and South African Genetics Congress, Stellenbosch, South Africa.

Taylor, K L, Young, P R, Brackenridge, A E and Vivier, M A. 2004. Overexpression of grapevine carotenoid biosynthetic genes in Arabidopsis thaliana. Poster presented at the South African Microbiology Society and South African Genetics Congress , Stellenbosch, South Africa.

Presentation(s)
Champanis, R, Vivier, M A, Botha, F C and Pretorius, I S. 1998. Sweetening up to Vitis. Presentation to Experimental Biology Group of South Africa. Cape Town, South Africa.

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 International Congress of the Federation of African Societies of Biochemistry and Molecular Biology. 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 24th National Congress South African Society for Enology and Viticulture. 18-19 November, Cape Town, South Africa.

Champanis, R, Vivier, M A, Botha F C and Pretorius, I S. 1999. Manipulation of grapevine sugar metabolism. Paper presented at the 24th Congress of the South African Society for Enology and Viticulture. Cape Town. South Africa.

Young, P R, Vivier, M A and Pretorius, I S. 2000. The structural analysis of a ß carotene hydroxylase-encoding gene from Vitis vinifera. BioY2K Combined Millennium Meeting, Grahamstown. South Africa.

Champanis, R, Vivier, M A, Botha F C and Pretorius, I S. 2000. Transformation of grapevine with a yeast-derived invertase (SUC2) targeted to different cellular locations. International BioY2K Combined Millenium Meeting, 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. 26th World Wine and Vine Congress, 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. 19th International Congress of Biochemistry and Molecular Biology, 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. 7th International Symposium for Grapevine Physiology and Biotechology, Davis, California. United States.

Vivier, M A, Joubert, D A, Becker, JvW, Wentzel, L, Taylor, K and Young, P R. 2005. Functional analysis of grapevine genes, International Grape Genomics Synposium, St Louis, United States.

Thomas, M R, Sreekantan, L, Iocca, P, Davies, C, Franks, T, Boss, P K, Tarr, C, Clingeleffer, P R, Vivier, M A, Young, P R and Torregrosa, L. 2006. Grapevine functional genomics. 9th International Conference on Grape Genetics and Breeding, Udine, Italy.

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
Champanis, R. 2004. Aspects of sucrose metabolism in transgenic tobacco. PhD. University of Stellenbosch, Stellenbosch, South Africa.

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. The generation and subsequent analyses of transgeneric Arabidopsis plant lines that over express grapevine carotenoid genes. MSc. University of Stellenbosch, Stellenbosch.

FinalReport.pdf

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