Grapevine constitutes one of the most important fruit species globally,
thanks to its remarkable propagation characteristics. Evolving from a bushy
plant to a trailing climber, this ancient species has been domesticated with
ease, and its fruit is today the basis for major worldwide industries.
Desirable traits in vines have long been identified as critical to wine
quality. Unfortunately the complex network of genes that control these
desirable traits also complicate traditional breeding programmes. It is currently
believed that the efforts of the grapevine industries to preserve and encourage
desirable characteristics can accentuate the shortcomings of practices that
have traditionally allowed for sustainable yields.
From an economic viewpoint, sustainable, high-quality production is a
direct consequence of the fitness of vines in the context of their available
environmental resources. It is widely believed that there is immense potential for enhanced cost-effective production of
grapes and wine with minimised resource inputs, improved quality and low
environmental impact. Among a host of factors at play in enhancement
efforts include improved nutrient capture from soils and better adaptation to
adverse conditions. Cultivation practices could benefit from enhanced
phytosanitary features of cultivars, resulting in the reduced use of
agrochemicals and fungicides. But the availability of excellent starting
material is fundamental to establishing new or replacement vineyards; germplasm
maintenance and propagation is key.
The fundamental principles of grapevine cultivation and winemaking have
remained the same for millennia, while the role of wine has changed
dramatically, from a practical, storable beverage, to a cultural luxury
product, placing wine between market-pulling forces and technology-pushing
developments, between tradition and innovation. This metamorphosis of the
global wine sector has resulted in a fundamental change in the focus of
research and development activities of the leading wine-producing countries.
New initiatives are being seized from recombinant DNA technology;
extensive efforts are underway to characterise the genomes of agriculturally
important species. Grapevine biotechnology is currently practised in all major
viticultural research centres worldwide, several technical factors dictating
the speed of the progress being made. The success rate has increased
exponentially over the last few years, focussing research activities on some of
the most interesting traits of genetically modified grapes and creating huge
potential for the wine industry.
Improvement
of methods for the transformation of grapevines
The grapevine genome is huge and complex, and its accessibility is
currently relatively restricted. This will soon be overcome by the intense study
being performed on it and the number of initiatives working on molecular
markers for the Vitis genome.1
New molecular techniques, such as bombardment technologies, are
enabling the addition of interesting genes into plant genomes, and are opening
up new possibilities for plant improvement in grapevine. The first significant
progress was made when embryonic cell lines were used as target tissue for
transformations, leading to routine production of transgenic grapevines,
involving a few commercially important grapevine scion and rootstock cultivars.2,3
Figure 1 depicts a summary of the processes involved in grapevine
transformation, as well as a time scale of the individual components of the
process.
Targets
for genetic improvement
Despite the limited successes achieved in early attempts to introduce
genes with known functions into plant species to express a desired phenotype,
the use of molecular biology to study fundamental processes and to combine
process knowledge with application has produced a growing list of genes and
their regulatory sequences from economically important species, including
grapevine.
Disease resistance, and other factors impacting cultivation and quality
are the main targets for genetic improvement of grapevine cultivars and
rootstocks, as indicated in Table 1. Several approaches have been used to
enhance disease tolerance in plants, almost all of them making use of some part
of the natural interaction between host and pathogen. Most transformation
strategies involve the introduction into the host, at high copies or in an
inducible manner, of a gene that produces anti-pathogenic activity, to optimise
the plant defence response. Also, disease tolerance can be improved by
expressing a pathogen–derived gene at an inappropriate time or in an
inappropriate form or amount during the infection cycle, thus preventing the
pathogen from maintaining infection. This is the case in most antiviral
strategies applied in grapevine genetic improvement research.
Transgenic approaches have also speeded up the development of plant
lines able to adapt to adverse climatic conditions, among them drought and salt
stress, photo-damage and freezing tolerance (Table 1). These involve
complex pathways of interacting proteins elicited by signals attenuated or amplified
by equally complex processes. Manipulation with single or multiple gene
additions is difficult and greater knowledge of the complex control mechanisms
is needed.
Gene technology offers at least as much promise to the product as to
the vine. Basic quality factors, such as suitable colour and sugar development,
are of generic importance to all segments of the wine industry and are
currently targeted in grapevine molecular biology. Grapevine biotechnology,
however, is in its infancy in this regard; significant efforts are underway to
shed light on the processes required prior to targetting genes and manipulating
biochemical pathways to produce novel and desirable products.
Obstacles
to the commercialisation of genetically improved grapevine cultivars
Despite the interesting possibilities of the use of gene technology in
the improvement of grapes and wine, there has yet to be a single transgenic
grapevine variety used on a commercial scale. Producers, consumers and
regulatory authorities all have their concerns, and much more knowledge of the
complex peculiarities of the grapevine genome and improvements offered by
genetic transformation technology is required to answer questions.
In most countries, the approval of genetically modified (GM) products
and the release of genetically modified organisms (GMOs) requires a number of
guarantees: (i) a complete definition of the DNA sequence introduced;
(ii) the elimination of any sequence that is not indispensable for
expression of the desired property; (iii) the absence of any selective
advantage conferred on the transgenic organism that could allow it to become
dominant in natural habitats; (iv) no danger to human health and/or the
environment from the transformed DNA; and (v) a clear advantage to both
the producer and the consumer.4
There is a growing consensus that risk is primarily a function of the
characteristics of a product, rather than the use of genetic modification per
se. Thus «substantial equivalence», equal safety compared to analogous
conventional food and beverage products,4 should be demonstrated.
This safety consideration is often considered sufficient.
Issues of intellectual property might also threaten the release of
genetic improvements. Patents covering many of the tools and methods commonly
used in genetic engineering may require formal agreements that could cause
ownership disputes and serious impediment to the commercialisation process.
The claim is also made that patents on genetically engineered organisms
confer an unfair advantage on certain producers;4 thus there is
pressure to justify trade bans and technical barriers to free trade on the
basis of «social interest».
The marketing of wine currently relies to a great extent on label
integrity and product identity. In the most profitable market segments, the
varietal name, the origin of production and the vintage are the key bits of
information presented on the label. Genetically improved grapevines should not
interfere with established varietal names and predictable wine styles, and an
industry relying on a few select cultivars would be very hesitant to introduce
new varietal names.2,5 The description and naming of transgenic
grapevines will determine, to a large extent, their acceptance by grape
growers, winemakers and consumers.5
Given the immense marketing value contained in some varietal names,
there is an urgent need for consensus that GM grapevines are little different
to grapevine clonal selections, which have been selected on the basis of
beneficial, spontaneous genetic variations. In fact, when clonal selections are
used, the identity might be known to the grapegrower but the wine is still
marketed under the varietal, and not the clonal name. It is not yet clear,
though, whether improved transgenic grapevines will be assigned a new varietal
name or just a new clonal number.
These uncertainties complicate the wine industry’s evaluation of
transgenic grapevines. And as a consequence of its strong identities and deep
cultural roots, this industry is less receptive than others to technologies
that promise revolutionary changes. There is a consequent fear that gene
technology could accelerate the tendency to standardise wines, leading to loss
of local identity, variety and uniqueness.
The successful application of recombinant DNA technology in the wine
industry will depend on assuring the commercial users of transgenic grapevines
that existing, desirable characteristics have not been damaged, that the
requirements of legislation are met and that the engineered cultivar will be
stable in practice, with suitable procedures for monitoring. If convinced,
these producers would be in a strong position to develop a new niche market for
«GM wine». Wine consumers attracted to such niche markets are usually well
informed and very curious; therefore, GM wines produced by a limited number of
interested producers would certainly attract widespread attention.
This technology holds a significant array of benefits, but it is
essential that the consumer be educated if the fear of the unknown is to be
removed. Scientists must consistently inform the public and remain open about
experiments, research and products. Consumers must be reassured of first-class,
transparent regulatory systems and should be persuaded by clear demonstrations
of safety. Only then will they feel empowered to make informed decisions.
References
1 Sefc, K.M.:
«Microsatellite markers for grapevine: a state of the art». In: Molecular
Biology and Biotechnology of the Grapevine 2001 (Roubelakis – Angelakis,
K.A:, ed.), Kluwer Academic Publishers; 433-464.
2 Colova-Tsolova,
V. et al.: «Genetically engineered grape for disease and stress
tolerance». In: Molecular Biology and Biotechnology of the Grapevine
2001 (Roubelakis – Angelakis, K.A:, ed.) Kluwer Academic Publishers; 411-432.
3 Kikkert, J.R. et
al.: «Grapevine genetic engineering». In: Molecular Biology and
Biotechnology of the Grapevine 2001 (Roubelakis – Angelakis, K.A:, ed.) Kluwer
Academic Publishers; 393-410.
4 Pretorius, I.S.:
«Tailoring wine yeasts for the new millenium: novel approaches to the ancient
art of winemaking», Yeast 2000; 16: 675-729.
5 Vivier, M.A. i
Pretorius, I.S.: «Genetic improvement of grapevine: tailoring grape varieties
for the third millenium», S Afr J Enol Vitic 2000; 21: 5-26.
Table 1
Targets for genetic improvement of grapevine cultivars and rootstocks
Target traits
|
Target processes
|
Target genes and proteins
|
|
Disease resistance
|
|
|
|
Tolerance to fungal diseases
|
Defence signalling in response to fungal pathogens
Pathology of fungal pathogens
Innate resistance towards fungal pathogens
|
Glucanase and chitinase from fungi, yeast and plants
Ribosome inactivating proteins
Thaumatin-like protein (antifungal peptides from plants and insects)
Polygalacturonase- inhibiting proteins from plant species and stilbene
phytoalexins
Phenylalanine ammonia lyase
CuZn superoxide dismutase
Detoxification enzymes
|
|
Tolerance to bacterial diseases
|
Defence signalling in response to bacterial pathogens
Pathology of bacterial pathogens
Innate resistance towards bacterial pathogens
|
Anti-microbial peptides
Disfunctional import and integration protein from Agrobacterium
|
|
Tolerance to viral diseases
|
Epidemiology and molecular biology of virus infections and vectors
Pathogen-derived resistance strategies
|
Virus coat proteins
Virus movement proteins
Replicase
Proteinases
Oligoadenylate synthase
|
Stress tolerance
|
|
|
|
Water stress
|
Aquaporins
Isolation of root-specific promoters
|
Tonoplast integral proteins
Plasma membrane integral proteins
|
|
Oxidative damage
|
Carotenoid biosynthesis and control
Anaerobiosis
|
Carotenoid biosynthesis factors
Alcohol dehydrogenase
Chloroplast and
mitochondrial CuZn superoxide dismutase
|
|
Osmotic stress
Other abiotic stresses
|
Proline accumulation
Polyamines in stress
Freezing tolerance
|
Ornithine aminotransferase
Glycine betaine and antifreeze peptides from Antarctic fishes
|
Quality factors
|
|
|
|
Colour development
|
Ripening related signals and anthocyanin biosynthesis and control
Isolation of berry-specific promoters
|
Glucose:flavanoid glucosyltransferase and regulatory sequences
Production of pelargonidin-based anthocyanins
Anthocyanin methyl-transferases
|
|
Sugar accumulation and transport
|
Phloem loading and unloading
Sugar transport
Isolation of berry-specific promoters
|
Plant and yeast invertases
Hexose transporters
|
|
Reduced browning (table and dried grapes)
|
Oxidation reactions
|
Polyphenol oxidase (silencing)
|
|
Seedlessness (table grapes)
|
Seed formation
|
Baranase
|