Somatic embryogenesis for grapevine improvement: The induction and maintenance of embryogenic cell-lines of wine grape and rootstock cultivars for use in transformation experiments

Project Number
US WIN 95-01

Project title
Somatic embryogenesis for grapevine improvement: The induction and maintenance of embryogenic cell-lines of wine grape and rootstock cultivars for use in transformation experiments

Project leader
Goussard, P G

Institution
University of Stellenbosch: Department of AgriSciences, Department of Viticulture and Oenology

Project description
Somatic embryogenesis has been proven to be a powerful and versatile tool with particularly useful applications in plant improvement programs based on genetic manipulation and biotechnology. Embryogenic cultures have been shown to be excellent target material for the introduction of manipulated and enhanced traits in various crops due to its inherent high proliferation rates and regeneration abilities. Similarly, a simple and fast regeneration protocol allowing switching from a differentiated phase to a dedifferentiated phase and vice versa, is of the utmost importance in the development of an efficient and reliable transformation system for grapevine cultivars, which usually result in complex protocols useful for only one cultivar. Other problems encountered include excessivley long culture periods to obtain synchronised embryogenic cultures and the spurious germination of the cultured embryos. In this study the effects of different culture media, groth regulators and explant sources on the induction of embryogenic culture were examined.

Over the past two years a successful and reproducible protocol for the establishment and maintenance of synchromised embryogenic cell lines was established within a wide range of important wine grape cultivars for use in transformation systems. In all cases immature anthers were used as explant material. With the aid of specific culture conditions and culture media supplemented with specific growth regulators, it was possible to start within a two month period with synchronisation procedures, i.e. the selection of embryogenic cell lines of the same developmental stage exhibiting regeneration potential. At various stages it was possible to manipulated embryogenic calli for the production of small production of embryogenic calli (<2 mm) embryos as well as to achieve recallusing from mature embryos resulting once again in the production of embryogenic calli. A high genotype dependency was experienced in approaches to establish reliable and reproducible techniques for the induction, maintenance and augmentation of embryogenic call cell lines of important rootstock cultivars from immature anthers as explant material. Thus far embryogenic calli was successfully initiated in two important rootstock cultivars. Special attention was given to culture conditions and culture media supplemented with specific growth regulators and combinations as well as the utilisation of synergistic effects while combining speciif factors which will increase the efficiency and broaden the gentoype range for the successful establishment and maintenance of stable embryogenic cultures.

Presentation(s)
Goussard, P G and Vivier, M A. 1998. The introduction and maintenance of embryogenic cell lines of Chardonnay and Merlot noir for use in transformation experiments. 23rd National Congress of the South African Society for Enology and Viticulture, November, Somerset West, South Africa.

Vivier, M A, Goussard, P G and Pretorius, I S. 1998. Genetic transformation of cultivars Chardonnay and Merlot of Vitis vinifera. 23rd National Congress of the South African Society for Enology and Viticulture, November, Somerset West, South Africa.

Goussard, P G. 1999. Somatiese embriogenese: Projekbevindinge oor die afgelope jaar en doelwitte vir die volgende jaar. Virus discussion group. ARC Infruitec-Nietvoorbij, Stellenbosch.

Goussard, P G. 2000. Somatiese embriogenese: Projekbevindinge oor die afgelope jaar en doelwitte vir die volgende jaar. Virus discussion group. ARC Infruitec-Nietvoorbij, Stellenbosch.

Goussard, P G. 2000. The induction and maintenance of embryogenic cell-lines of important wine grape and rootstock cultivars for use in transformation experiments. 2nd International Congress of the South African Society for Enology and Viticulture, November, Cape Town, South Africa.

Note
No Final Report

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