John Ruane and Andrea Sonnino
15.1 INTRODUCTION
On the occasion of World Food Day 2004, the United Nations Secretary-General Kofi Annan urged “individuals and institutions alike to give greater attention to biodiversity as a key theme in our efforts to fight the twin scourges of hunger and poverty and achieve the Millennium Development Goals.” He also noted that the unprecedented loss of biodiversity over the past century was a major cause for alarm, where “many freshwater fish species, which can provide crucial dietary diversity to the poorest households, have become extinct, and many of the world's most important marine fisheries have been decimated. Food supplies have also been made more vulnerable by our reliance on a very small number of species: just 30 crop species dominate food production and 90 percent of our animal food supply comes from just 14 mammal and bird species - species which themselves rely on biodiversity for their productivity and survival. There has been a substantial reduction in crop genetic diversity in the field and many livestock breeds are threatened with extinction (www.un.org/News/Press/docs/2004/sgsm9539.doc.htm)”.
On the same occasion, FAO's Director-General Jacques Diouf also underlined that although forests are among the world's most important repositories of biological diversity, the world forest cover is decreasing at an alarming rate (www.fao.org/wfd/2004/dgmessage_2004_en.asp).
It is in this context of declining agricultural biodiversity that the FAO Biotechnology Forum (www.fao.org/biotech/forum.asp) hosted an e-mail conference from 6 June to 3 July 2005 to consider the role that biotechnology can play in the characterization and conservation of crop, forest, animal and fishery genetic resources in developing countries. Biotechnology is a broad collection of tools that can be applied for a range of different purposes (e.g. genetic improvement of populations; disease diagnosis and vaccine development; and improvement of feeds). This conference focused on biotechnology tools such as molecular markers, cryopreservation and reproductive technologies that can be used directly for the characterization and/or conservation of genetic resources for food and agriculture. Genetic modification and GMOs will not be considered here.
On 5–7 March 2005, as part of the preparations for this e-mail conference, an international workshop was held in Turin, Italy, entitled “The Role of Biotechnology for the Characterisation and Conservation of Crop, Forestry, Animal and Fishery Genetic Resources”, organized by the FAO Working Group on Biotechnology, the Fondazione per le Biotecnologie, the Econogene project and the Società Italiana di Genetica Agraria. Proceedings of the workshop (available at www.fao.org/biotech/torino05.htm) include 20 papers and 37 poster abstracts covering applications of molecular markers, cryopreservation and reproductive technologies and can be consulted by anyone looking for more detailed technical information in this area. In addition to the poster session, the workshop was split into Session I on the status of the world's agricultural biodiversity, Session II on the use of biotechnology for conservation of genetic resources, and Session IV on genetic characterization of populations and its use in conservation decision-making. Session III presented results from Econogene, a European Union-funded project combining a molecular analysis of biodiversity, socio-economics and geostatistics to address the conservation of sheep and goat genetic resources and rural development in marginal agrosystems in Europe.
This chapter provides background information for the e-mail conference. First, a brief overview of genetic resources for food and agriculture is provided (Section 15.2), followed by more specific information regarding the current status of the genetic resources in the different food and agricultural sectors (Section 15.3). A brief description of the relevant biotechnologies is then given (Section 15.4), followed by a discussion of some key issues and some questions that might be addressed in the email conference (Section 15.5).
15.2 A BRIEF BACKGROUND TO GENETIC RESOURCES FOR FOOD AND AGRICULTURE
As described in Diamond (2002), domestication of crops and livestock, where species selected from the wild adapt to a special habitat created for them by humans, was a momentous development with major consequences for human societies, since “it provides most of our food today, it was prerequisite to the rise of civilization, and it transformed global demography.”
Domestication seems to have arisen independently on at least five different occasions in different parts of the world where, for example, hunter-gatherers in the Fertile Crescent (an area in Asia, including parts of Iran, Iraq, Israel, Jordan, Lebanon, Syria and Turkey) began domesticating barley, peas, wheat, cows, goats, pigs and sheep about 10 000 years ago. Occasionally the same species (e.g. cow, sheep) was domesticated independently in different places. The process of domestication has continued ever since, for example, in the Middle Ages with rabbit and strawberry.
Domestication had major impacts on human societies, to enable the transition from a hunter-gatherer society to a settled farming society. It also led to major changes in the species that were domesticated. Since their domestication, crops and livestock have been used in almost all environments of the world and many have been domesticated in particular environments for very long periods of time. Soay sheep on Hirta, off the west coast of Scotland, for example, are thought to be the survivors of the sheep first brought to Britain by the first farmers 5 000 years ago. Each environment has a certain set of stressors that impacts the production environment. If we consider livestock, these might include climatic features (heat/cold, amount of rain, humidity), feeding conditions (quality/quantity vegetation available, access to water) or diseases (parasitic infections, etc.), and over time, populations adapted to the environment and its specific stressors. Farmers have also exercised a major influence on the domesticated species by preferentially selecting for specific morphological or production traits.
Unlike crops and livestock, domestication is generally a very recent phenomenon in the fishery and forestry sectors. Only a small number of forest tree species have been domesticated to some degree, and only over the past half century. Such tree species, used in plantation forestry, are usually no more than one to two generations removed from their “wild relatives” (see, for example, Sigaud, 2005). Concerning fish species, with the exception of a few species such as common carp domesticated about 2 000 years ago, aquaculture (fish farming) is a relatively new development (Bartley, 2005) and most of the fish consumed today still come from wild populations, although it is predicted that by 2030 consumption from the ever-growing aquaculture industry will exceed capture fisheries.
In order to consider the agricultural genetic resources in this e-mail conference, a wide range of species has therefore been included, from those domesticated 10 000 years ago to those domesticated in recent times, as well as those that have not been domesticated but that are nevertheless of importance for mankind's food and agriculture.
As will be described in more detail in Section 15.3, many of these genetic resources are endangered. The need to conserve them is now widely accepted and generally justified for one or more of a variety of reasons: their importance in ensuring against future changes in market needs and production conditions; as a source of material for scientific research and future germplasm development; and as a cultural and historical part of our heritage, passed down from previous generations (e.g. Ruane, 2000). The issue has received considerable attention at the international policy-making level. For example, it is addressed under the Convention on Biological Diversity (CBD), a legally-binding agreement with 188 parties, through its agricultural biodiversity work programme (www.biodiv.org/programmes/areas/agro/default.asp), and under the International Treaty on Plant Genetic Resources for Food and Agriculture, a legally-binding treaty with 69 parties that entered into force in June 2004 (www.fao.org/ag/cgrfa/itpgr.htm).
Two major strategies for conservation can be distinguished: in situ conservation, where a population is maintained in its natural or agricultural habitat and ex situ conservation, where it is maintained outside of this habitat. Ex situ conservation can involve living individuals (e.g. animals in zoos, fish in aquaria, crops or trees in botanical gardens and arboreta), or tissues/genetic material from individuals (such as seeds, pollen, sperm, embryos and DNA). Ex situ conservation is important for crops, where an estimated 6 million plant, mainly crop, accessions (i.e. samples of a variety collected at a specific location and time) are stored in national, regional and international collections (FAO, 1997, Chapter 3). Of the 6 million accessions, over 80 percent are in national collections and around 90 percent are stored in seed gene banks which, depending on the seed storage facilities, may allow short-, medium- or long-term storage. A gene bank is a storage facility where germplasm of plant or animal origin is stored in forms such as seeds, pollen, semen or embryos; in in vitro culture (germplasm kept as sterile plant tissues or plantlets on nutrient gels); in cryogenic storage; or in a field gene bank, i.e. as plants growing in the field. In situ conservation is currently more important than ex situ conservation for the livestock, forestry and fisheries sectors.
Characterization of genetic resources goes hand in hand with their conservation since it is fundamental both for understanding what is being conserved and for choosing the genetic resources that should be conserved. Characterization enables us to identify the key features, both the strengths and weaknesses, of the available genetic resources and this knowledge can also be used to develop breeding programmes for sustainable use of the genetic resource to harness and/or disseminate the positive attributes identified in the population. Characterization can also play an important role regarding issues of access to and benefit-sharing of agricultural genetic resources, as well the promotion and control of “bioprospecting” (i.e. biodiversity prospecting, the search for commercially valuable compounds, substances or genetic material in nature). If a country has characterized its genetic resources, it should be appropriately positioned to develop and implement conservation strategies for targeted species. It should also be able to promote and oversee any potential commercial or other enterprises arising from the knowledge generated to protect the resources (or any of their products) from inappropriate bioprospecting by foreign companies or countries. It should also be able to negotiate appropriate compensation for use of these resources by third parties, be they national or foreign.
The kinds of features that could be characterized for the different agricultural populations include their morphological and physiological features; their phenotypic and economic performance in different environments; their interaction with and relation to the environment; their value as social or cultural objects; their role in scientific research; their diversity at the gene and DNA sequence level; their population size and their degree of endangerment.
15.3 STATUS OF GENETIC RESOURCES IN THE DIFFERENT FOOD AND AGRICULTURAL SECTORS
This section aims at providing an overview of the current situation regarding genetic resources in each of the four sectors.
15.3.1 Crop genetic resources
Compared to the large numbers of wild plant species available, only very few were actually domesticated (Diamond, 2002). Although this number is small, just a subset of them, again, are of special importance globally. For example, considering plant-derived energy intake worldwide, just three crops (wheat, rice and maize) provide over 50 percent of the intake and 30 crops provide 95 percent (FAO, 1997, Chapter 1). Splitting the world into 17 subregions, there are only 12 major crops that supply more than 5 percent of the plant-derived energy intake in one or more subregions, i.e. wheat, rice, maize, millet, sorghum, potato, sugar cane, soybean, sweet potato, cassava, the common bean and related species (Phaseolus), and banana/plantain. It should be kept in mind, however, that although some specific crops may not have any global/regional significance, they may nevertheless be important at the individual country level (e.g. tef in Ethiopia).
Although the number of major crop species is quite small, as seen above, the diversity within each species can be very substantial. Within a species, cultivated crop varieties (cultivars) can be broadly categorized as farmer's varieties (also known as landraces or traditional varieties, i.e. populations that are the product of breeding or selection carried out by farmers, either deliberately or not, continuously over many generations) and modern varieties (also known as high-yielding or high-response varieties, which are the products from professional breeders working in publicly-funded research institutes or private companies). Farmer's varieties tend to maintain high levels of genetic diversity and to be the focus of conservation efforts (FAO, 1997).
The first comprehensive worldwide report on the status and use of crop genetic resources was published by FAO in 1997 (www.fao.org/ag/AGP/AGPS/Pgrfa/pdf/swrfull.pdf). This State of the World's Plant Genetic Resources for Food and Agriculture (SoW-PGRFA) was developed through a participatory country-driven process and the primary data sources for the publication were detailed country reports from a total of 154 countries. Annex 2 of the report (FAO, 1997) documents the state of diversity within each of the 12 major crops. It shows, for example, that the global ex situ collection of millet contains roughly 90 000 accessions, of which 2 percent are wild relatives, 33 percent landraces and old cultivars, and 5 percent advanced cultivars and breeding lines, while 60 percent are unknown and mixed. Although incomplete data makes it difficult to give a definitive picture of the kind of material stored in the world's ex situ collections, it is estimated that roughly half the accessions for which the type of material is known are advanced cultivars and breeding lines, while a third are landraces and around 10 percent are wild species (FAO, 1997, Chapter 3).
In their country reports nearly all countries stated that genetic erosion (i.e. the loss of genetic diversity, generally as a result of social, economic and agricultural changes) was a serious problem. The main cause cited was the replacement of local varieties or landraces by improved and/or exotic varieties and species, and numerous examples were provided (e.g. displacement of traditional barley and durum wheat varieties by introduced varieties in Ethiopia). Other important causes cited were: overexploitation of plant genetic resources, including overgrazing and reduced fallow periods in shifting cultivation; deforestation and land clearance; population pressure and urbanization; environmental degradation (e.g. desertification, flooding); and changes in agricultural systems (FAO, 1997, Chapter 1). Preparations are underway for the second Report on the SoW-PGRFA, which should be finalized in 2008.
15.3.2 Forest genetic resources
Most forest tree species are characterized by inherently high levels of variation and extensive natural ranges. They are long-lived, frequently outcrossing organisms with generally large distribution ranges. A high level of intra-specific genetic variation is needed to ensure present-day and future adaptability to changing environmental conditions. It is also needed to maintain options and potential for improvement to meet changing end-use requirements. Forests provide a wide range of goods and services such as timber, fibre, fuelwood, food, fodder, gum, resins, medicines, pharmaceutical products and environmental stabilization. Similar goods and services are often provided by a wide range of genera and tree species. Despite the availability of a large number of forest tree species, less than 500 have been systematically tested for their present-day utility for human beings and less than 40 species are included in intensive selection and breeding programmes (FAO, 2000a).
Estimates for 2000 the year indicated that the world's forest cover was about 3.9 billion ha, of which 47 percent was in the tropics, 33 percent in the boreal zone (i.e. the northernmost forest zone), 11 percent in temperate areas and 9 percent in the subtropics (FAO, 2001). About 95 percent of the forest cover was in natural forest and 5 percent in forest plantations (defined as forest stands established by planting or/and seeding in the process of afforestation or reforestation). Of the roughly 190 million ha of forest plantation, 48 percent had an industrial end-use (e.g. pulpwood for paper), 26 percent a non-industrial purpose (e.g. fuelwood, soil and water conservation), and for 26 percent it was not specified.
Most of the forest genetic resources worldwide are therefore found in natural, largely unmanaged forests which, however, are frequently intensively used for the provision of both wood and non-wood products for local, national and international use. Conversion of forests to other land uses represents the greatest threat to forests and their diversity. According to FAO (2001), “increasing pressure from human populations and aspirations for higher standards of living, without due concern to the sustainability of the resources underpinning such developments, heighten these concerns. While some land use changes are inevitable, it is important that such changes be planned and managed to address complementary goals. Concerns for biological and genetic conservation should be major components of land use planning and forest management strategies”. Unlike the situation in crops, however, no global overview of the status of forest tree genetic resources has yet been carried out, although the idea of developing a first global assessment of forest genetic resources is currently being discussed (Sigaud, 2005).
15.3.3 Animal genetic resources
Of the 50 000 known bird and mammalian species, around 30 have been used extensively for agricultural purposes, with fewer than 14 accounting for over 90 percent of global livestock production (FAO, 1999). In the millennia following their domestication, animal species were influenced by both natural and artificial selection in a wide range of geographic and climatic areas and production systems, leading to the development of large numbers of genetically diverse domestic animal breeds.
Many of these breeds are currently endangered. In developing countries, the introduction of exotic breeds and their spread through indiscriminate crossbreeding is a major reason for loss of local, indigenous breeds. Other factors include changes in breeders' preferences due to short-term socio-economic influences, degradation of the ecosystem in which the breeds were developed, natural disasters (e.g. drought and diseases), wars and other forms of political unrest and instability (FAO, 1998).
In 1993, FAO published the first World Watch List for Domestic Animal Diversity, which provides a basic global overview of the state of the genetic resources of seven mammalian species (buffalo, cattle, donkey, goat, horse, pig and sheep). The third edition, expanded to cover 30 mammalian and bird species, showed that of the over 4 000 breeds with population data, roughly 30 percent could be classified at a high risk of loss and that chicken and cattle have large numbers of breeds at risk while horses and geese have the highest percentages of breeds at risk of loss (FAO, 2000b). In addition, with the exception of the wild boar and wild red jungle fowl (ancestors of the domestic pig and chicken respectively), the putative wild ancestors of the major livestock species are in danger of extinction or are already extinct (Hanotte and Jianlin, 2005).
In a similar process to that described for the crop sector, FAO is preparing the first State of the World's Animal Genetic Resources, which is expected to be completed by 2006 and for which 151 countries have accepted to submit country reports to date. The aim of the country and global assessments is to provide a comprehensive analysis of the status and trends of the world's farm animal biodiversity and of their underlying causes, as well as of local knowledge regarding its management (Cardellino, 2005).
15.3.4 Fishery genetic resources
Like forestry but unlike crops and livestock, wild populations currently play a greater role than domesticated populations in the fishery sector. When considering the status of the world's fishery genetic resources, both the aquatic species that are fished (capture fishing) and those that are farmed (aquaculture) can be included. Bartley (2005) gives an overview of the status of aquatic species that are fished or farmed worldwide.
For capture fisheries in 2002, FAO Members reported that 974 taxa (i.e. taxonomic units such as a family, genus or species) of fin-fish, 143 taxa of crustacea, 114 taxa of molluscs, 26 taxa of plants and 73 taxa of miscellaneous animals (such as sea urchins, sea cucumbers and marine mammals) were taken from the world's capture fisheries. These numbers are likely to be underestimates, however, because reporting is often incomplete. Although over 1 000 taxa are represented, about ten species make up about one-third of capture fisheries production. Unlike the crop and animal sectors, there is no systematic effort to describe the state of the world's fishery genetic resources below the species level. Nevertheless, description of the fish genetic stocks at the within-species level is important for fisheries management and is an active research area (Bartley, 2005; Primmer, 2005).
For aquaculture in 2002, FAO Members reported that apart from a small number of aquatic plants and animals, a total of 153 species of fish, 60 species of molluscs and 44 species of crustaceans were farmed. The relative novelty of aquaculture can be shown by the fact that its contribution to global supplies of fish, crustaceans and molluscs has grown from just 4 percent in 1970 to 30 percent in 2002 (Bartley, 2005).
Aquatic science in general lags behind terrestrial sciences in identifying species, understanding ecosystem relationships and assessing potential uses for genetic resources. Communities of life on the ocean floor are poorly understood, and every year some aquatic species become extinct before they have even been identified. Fish genetic resources worldwide are currently threatened by overfishing, but also by habitat destruction (e.g. dam building) and pollution from human activities (Greer and Harvey, 2004).
Every two years FAO publishes the State of World Fisheries and Aquaculture, which, inter alia, provides a global overview of capture fisheries (marine and inland) and aquaculture. Regarding exploitation of marine fish stocks (estimated production at 84.5 million tonnes in 2002), the most recent edition indicates that there has been a consistent downward trend since 1974 in the proportion of stocks offering potential for expansion, together with an increase in the proportion of overexploited and depleted stocks, rising from about 10 percent in the mid-1970s to close to 25 percent in the early 2000s. The percentage of stocks exploited at or beyond their maximum sustainable levels varies widely among fishing regions. The report says that the information available continues to confirm that despite local differences, the global potential for marine capture fisheries has been reached, and more rigorous plans are needed to rebuild depleted stocks and prevent the decline of those being exploited at or close to their maximum potential (FAO, 2004a).
Inland fish stocks (estimated production was 8.7 million tonnes in 2002), unlike the major marine fish stocks, are less well defined and occur over much smaller geographical areas such as individual lakes, rice fields or rivers, or over vast areas such as transboundary watersheds that are often situated in areas that are difficult to access. These factors make it costly to monitor the exploitation and status of fish stocks and, in fact, very few countries can afford to do so. As a result, most countries report only a small fraction of their catch of inland fisheries by species, further compounding the problem of accurate assessment. There are indications, however, that these resources are undervalued and threatened by habitat alteration, degradation and unsustainable fishing activities (FAO, 2004a).
15.4 OVERVIEW OF RELEVANT BIOTECHNOLOGIES
As seen in Section 15.3, a wide range of genetic resources are currently being used by humankind for food and agricultural purposes and many of them are in danger of being lost for reasons such as over-exploitation (fish), replacement of local with international germplasm (crops/livestock), habitat change and destruction (crop/fish/forest/livestock). A brief overview will be presented of molecular markers and cryopreservation, in vitro culture and reproductive technologies and how they can be used for the characterization and/or conservation of these genetic resources.
15.4.1 Molecular markers
The importance of characterizing genetic resources was described in Section 15.2 as well as the wide range of features that can be characterized in each population, such as morphology, phenotypic performance and degree of endangerment. Whereas phenotypes (e.g. yield, growth rate) or morphological traits (coat colour, seed shape) are influenced by both genetic and environmental factors, the use of molecular markers reveals differences at the DNA level that are not influenced by the environment (de Vicente et al., 2005). Molecular markers usually do not have any biological effect and are normally assumed to represent neutral loci unaffected by selection. They are identifiable DNA sequences found at specific chromosomal locations on the genome and transmitted by the standard laws of inheritance from one generation to the next. A number of types of molecular markers exist, such as RFLP markers, random amplified polymorphic DNA (RAPD) markers, amplified fragment length polymorphism (AFLP) markers, and microsatellites and single nucleotide polymorphism (SNP) markers, which can differ in a variety of ways, such as the amount of time, money and labour needed or the amount of genetic variation found at each marker in a given population. Microsatellites, for example, are simple DNA sequences, usually two or three DNA bases long, repeated a variable number of times in tandem. (For more background information on molecular markers, see FAO, 2003).
Molecular markers are used in a variety of approaches to characterize and conserve genetic resources, the most important of which will be described below. Some of the approaches are applied in each of the crop, forestry, animal and fishery sectors (e.g. estimating the genetic relationships between populations within a species), while others are more sector-specific (e.g. identifying duplicate accessions in crop gene banks or monitoring effective population sizes in capture fish populations).
Estimating the genetic relationships between populations within a species
This is one of the major uses of molecular markers in agricultural research. Molecular markers are typed in populations of the same species (for instance, those located in different places) and about which inferences can be made on how genetically related they are to each other by comparing the frequencies and presence/absence of marker loci in the different populations. The more differences in markers found between the populations, the greater the genetic distance between them, and the populations are inferred to be less genetically related to each other than to populations that have more similar marker loci frequencies. Taking a simple example for illustrative purposes (in practice many markers would be used simultaneously), if individuals in populations A, B and C are typed for a molecular marker M (with two alternative forms or alleles called M1 and M2) and all individuals typed in populations A and B have allele M1 while those in population C have M2, this suggests that A and B are genetically more similar to each other than to population C. In using molecular markers to estimate genetic distances between populations, it is assumed that they represent neutral loci (i.e. do not reflect genetic differences between populations at genes that are or have been under natural or artificial [farmer] selection) and that the use of a relatively small number of independently segregating marker loci will be a good predictor of the overall genomic diversity of a population (Hanotte and Jianlin, 2005).
Typically, a genetic diversity study involves sampling biological tissue (e.g. blood, leaves, fish scales) from a certain number of individuals from a number of populations of interest and typing them in the laboratory for a number of molecular marker loci. A statistical analysis of the marker data is then carried out to estimate the genetic relationships between the populations. For example, Lanteri and Barcaccia (2005) sampled 30 individuals each from seven populations of wild cardoon identified in Sardinia and Sicily, and each individual was genotyped for 32 microsatellite markers as well as seven AFLP primer combinations. They used a statistical analysis method called principal coordinate analysis to show that the Sardinian and Sicilian populations were clearly differentiated, representing two distinct gene pools.
Such studies have been routinely applied in each of the agricultural sectors. For example, in developing a management strategy for commercially exploited fish, molecular markers are used to characterize the within-species genetic structure of the populations being harvested to determine the units between which limited gene flow occurs, i.e. to identify units that should not be overfished (Primmer, 2005). A recent questionnaire-based survey of animal diversity studies carried out during the last ten years provided information on a total of 86 projects involving 13 livestock species from 93 countries (FAO, 2004b). Most of the projects were for ruminants, especially cattle. Blood was the most frequently used biological sample and microsatellites were the most commonly used molecular markers.
As pointed out by Lenstra et al. (2005), these kinds of studies are not only relevant for the purposes of characterization of genetic resources, but they can also provide useful insights into the history of domestication. Evidence from molecular markers has been used often to elucidate whether individual crop or livestock species were domesticated just once at a single site or independently several times in different parts of the world (Diamond, 2002).
Establishing and managing gene banks
As described in Section 15.2, gene banks are an important tool for conservation of crop genetic resources, although currently of more limited importance for livestock, forestry and fish. Molecular marker information can be used in a number of gene bank-related activities, such as sampling of material, management of the collections and stimulating use of germplasm stored therein (Lanteri and Barcaccia, 2005).
When sampling germplasm to create a gene bank, information from the use of molecular markers to assess the within- and between-population genetic diversity (as described in the section above) can be used in conjunction with data on other characteristics of the populations to assist decision-making regarding which material should be sampled (e.g. Simianer, 2005). Since the amount of germplasm to be stored may be limited, for example, due to financial reasons, it would be important to sample material from genetically diverse individuals.
Once a gene bank has been established, molecular markers can also be used to assist in a number of gene bank management activities. As a first example, regeneration of seeds or other reproductive plant material in storage is an important task in a gene bank, and the regeneration requirements (how frequently accessions need to be regenerated) depend on factors such as the species concerned, the storage conditions and the quality of the individual accessions (FAO, 1997, Chapter 3). Markers can be used to check whether changes in alleles or in allele frequencies are taking place over time as a result of the adopted conservation strategies, for instance of frequent regenerations (de Vicente et al., 2005). Second, while safety duplication of unique accessions acts as an insurance against possible accidents, unintentional duplication or overduplication of accessions is wasteful, however (FAO, 1997), and markers can be used to detect them (de Vicente et al., 2005). Third, to reduce costs and/or encourage greater and more efficient use by farmers and breeders, core collections (i.e. a subset of accessions selected to contain the maximum available variation in a small number of accessions) have been established for many crop species. Again, molecular markers can assist in this task.
Finally, it is argued (e.g. Ferreira, 2005) that molecular markers offer opportunities to increase the use of crop genetic resources from gene banks and will therefore stimulate increased use of gene bank material for research purposes, thereby playing an important role in the sustainable conservation of these genetic resources. Since a major justification for the existence of germplasm collections is use of the conserved accessions, de Vicente et al. (2005) underline the importance of genetically characterizing them so that valuable genes can be identified for use in developing new plant varieties by conventional and advanced breeding methodologies.
Gene flow from domesticated populations to wild relatives
Wild relatives of domesticated species are of conservation interest because they represent one of the components of agricultural biodiversity. They have also proven useful for genetic improvement of a range of cultivated varieties (FAO, 1997, Chapter 1). As described in FAO (2002), crossing of domesticated populations with their wild relatives has been well documented in some crop, forest tree, animal and fish species. For example, in crops, gene flow has been observed between rice and perennial rice, between maize and teosinte, and between sugar beet and wild beet, while in animals there is evidence for crossing of domestic cattle with wild North American bison, and of domestic pigs with European wild boars. In forestry, gene flow from intensively bred forest trees to natural populations of the same or closely related species (“wild relatives”) has been a cause for concern in many species, for example, the European black poplar (Populus nigra). For fish, crossing of escaped farmed Atlantic salmon with wild Atlantic salmon is also a much-discussed problem.
Gene flow from a domesticated population will cause changes in the genetic diversity of wild populations. In cases where diversity is systematically reduced over time by such gene flow, it may potentially lead to genetic extinction of the wild population in its original genetic state (see, for example, Papa, 2005). Molecular markers can assist in conservation of the wild relatives through their use to distinguish hybrids from non-hybrid wild relatives and to monitor gene flow from domesticated populations to their wild relatives.
Estimating and monitoring the effective population size
When developing conservation strategies for wild species, accurate estimates of the effective population size (Ne) are important for predicting a number of parameters, such as the rate of inbreeding (Primmer, 2005). Knowing the rate of inbreeding is vital because increases in inbreeding (due to mating of genetically related individuals) can lead to the reduced abilities of individuals to survive and reproduce (a phenomenon called inbreeding depression). Ne can be estimated using pedigree information and/or knowledge about the population breeding structure (male/female mating ratio, family sizes, etc.). However, for wild populations, especially of aquatic species, it can be difficult to get this information. Molecular markers can be used to estimate Ne, which has been done, for example, in several commercial fish populations (Primmer, 2005). Similarly, molecular markers can also be used as a tool to monitor fish population sizes over time and to detect any major recent declines in Ne.
15.4.2 Cryopreservation, in vitro culture and reproductive technologies
As described in Section 15.2, there are two major strategies for conservation, in situ and ex situ, the latter involving, for instance, storage of genetic material in gene banks, keeping live animals in zoos, or keeping plants or botanical gardens. Biotechnology, through cryopreservation (freezing) technologies and in vitro culture, provides additional tools for ex situ conservation.
Cryopreservation involves the preservation of germplasm in a dormant state by storage at ultra-low temperatures, usually in liquid nitrogen (-196 °C), and can be used to preserve biological material (e.g. seeds, sperm, embryos) of crop, livestock, forest or fish populations indefinitely in gene banks. It allows large quantities of genetic material to be stored in a limited amount of space. If the living populations later become extinct, they can in many instances be regenerated using the frozen genetic material. In addition, if the living populations are declining in numbers and develop problems due to inbreeding, these problems can be alleviated by introducing unrelated genetic material from the gene bank. In animals, collection of germplasm for cryopreservation and production of live offspring from cryopreserved genetic material depends on a number of associated reproductive technologies, such as embryo transfer. In vitro culture is a plant biotechnology that involves application of slow growth procedures to germplasm accessions kept as sterile plant tissues or plantlets on nutrient gels.
Here below, we will provide a brief overview of the current status of these biotechnologies, focusing on cryopreservation since it is applicable in all four sectors. Information provided on cryopreservation in livestock/fish and crops/forest trees is based primarily on Hiemstra et al. (2005) and Panis and Lambardi (2005), and the references therein.
Livestock and fish
Before considering the main types of genetic material that have been cryopreserved, a brief overview of relevant terminology may be useful. In sexual reproduction, through fertilization there is a fusion of two haploid gametes to form the diploid zygote, a cell that begins dividing and develops to become an embryo. (Note: haploid and diploid cells have one or two sets of chromosomes, respectively.) The female gamete is called the ovum or egg, and is derived from a cell called an oocyte that has undergone two meiotic divisions. The male gamete is called the sperm, contained in fluid called semen.
For cryopreservation purposes, semen is currently the biological material of greatest importance, although embryos and, to a lesser degree, oocytes and somatic cells, have also received considerable research attention. Success in freezing these materials varies considerably from species to species, which is not surprising given the wide diversity in reproductive systems (e.g. fertilization is usually external in fish and internal in birds where the female stores the sperm before fertilization). Collection of biological materials for the gene bank and production of live offspring following thawing of formerly cryopreserved material is made possible through a variety of reproductive technologies, such as artificial insemination (AI) using semen, multiple ovulation and embryo transfer using embryos.
Freezing of semen and subsequent successful AI with thawed semen was first achieved over 50 years ago and semen of most livestock species can now be frozen adequately. There are large differences between species in insemination techniques and pregnancy rates using fresh or frozen semen, where AI with frozen semen is most successful in cattle. For domesticated birds, although AI is less widely used than in mammals, reasonable success rates with frozen semen, albeit at times highly variable, have been reported for the major bird species. For fish, sperm cryopreservation has been tested in over 200 fish species with external fertilization and the present state of the art for many species of fish seems to be adequate for the purpose of gene banking (Hiemstra et al., 2005).
For embryos, cryopreservation in cattle is now a routine procedure. It has also been reported for other mammalian species, such as horse, pig, rabbit and sheep (ERFP, 2003). In order for embryos to be cryopreserved, they can be collected using non-surgical (cattle and horses) or surgical methods (other mammalian species). Inducing multiple ovulation using hormones can increase the efficiency of the process. Alternatively, embryos can be collected through in vitro maturation and in vitro fertilization of oocytes, a more technically demanding approach. Production of live offspring following cryopreservation involves non-surgical (only in cattle and horses and, to a lesser extent, pigs) or surgical embryo transfer to recipient animals. For bird and fish species, however, cryopreservation of embryos has not been successful, mainly because of the large size, the high lipid content and the polar organization of the ova and early embryos.
For mammalian oocytes, live offspring have been reported in cattle and horse from embryos produced using cryopreserved oocytes, although the efficiency and reliability of using oocytes for generating offspring is still much lower today than with cryopreserved embryos. Oocytes for cryopreservation in the gene bank can be collected from ovaries of dead animals or by ovum pick-up (recovery of oocytes using an ultrasound probe) on live animals. Production of live offspring following thawing of cryopreserved oocytes involves in vitro maturation and fertilization, and eventual embryo transfer to recipient animals. Cryopreservation of bird and fish oocytes has not yet been successful, mainly for the same reasons given earlier for embryos.
For somatic cells (i.e. cells not involved in sexual reproduction), a number of cell types (e.g. skin fibroblasts, mammary cells) can now be cryopreserved. Collection of material for the gene bank is easy and cheap. Production of live offspring (which, with the exception of mitochondrial DNA, are genetic clones of the individual that donated the somatic cells) is complicated, however, and has a low rate of success. It involves culturing the thawed cells and then transferring their nuclei (i.e. the organelle that contains the genetic material) to, or fusion of the somatic cells with enucleated oocytes (i.e. without a nucleus) from another individual. The resulting embryos are then cultured and eventually transferred to recipient animals. In mammals, live offspring have been obtained from embryos generated from somatic cells in a number of mammalian species (cattle, goats, horse, pigs, rabbits and sheep) and in fish (zebrafish), but not yet in poultry. The low success rates, as well as the many developmental problems encountered, are current major limitations to application of the technology.
Crops and forest trees
As described in Section 15.2, about 90 percent of the six million plant accessions in gene banks, mainly crops, are stored in seed gene banks. However, storage of seeds is not an option for crops or trees that do not produce seed, such as bananas, or that produce recalcitrant or non-orthodox seed (i.e. seed that does not survive under cold storage and/or the drying conditions used in conventional ex situ conservation), such as mango, coffee, oak and several tropical forest tree species. The same applies to plant species that are propagated vegetatively to preserve the unique genomic constitution of cultivars, such as fruit and several timber and ornamental trees. It also presents practical problems in most forest tree species in regard to the necessary periodic regeneration of seedlots, because tree species usually have a long vegetative period prior to producing flowers and seed (lasting from several years to several decades), and because they require large areas to carry out such an operation due to outcrossing and their size.
Genetic resources of these species are normally conserved either in vivo, i.e. living collections, or by in vitro culture, where slow growth procedures allow the plant material to be held for 1–15 years under tissue culture conditions with periodic regeneration (subculturing), depending on the species. Normally, growth is limited using low temperatures often in combination with low light intensity or even darkness. Temperatures in the range of 0–5 °C are employed for cold-tolerant species and 15–20 °C for tropical species. Growth can also be limited by modifying the culture medium and reducing oxygen levels available to the cultures (Rao, 2004). The former option (in vivo) is the cheapest and thus the most widely adopted, but exposes the material to the harms of the external environment, e.g. contamination with pathogens or parasites. In vitro culture, on the other hand, ensures that the collections are kept free from pest and diseases and protected from adverse conditions. Maintenance of in vitro collections is labour-intensive, however, and there is still a risk of losing accessions due to contamination, human error or somaclonal variation, i.e. mutations that occur spontaneously in tissue culture, with a frequency that increases with repeated subculturing (Panis and Lambardi, 2005).
In all these situations, as well as for long-term storage of seeds from orthodox species, cryopreservation offers an alternative tool or complementary strategy for ex situ conservation. Following plant cell, tissue or organ storage at low temperatures, plants can then be regenerated and there seems to be no clear evidence so far of morphological, cytological or genetic alterations arising as a result of cryopreservation (Panis and Lambardi, 2005). They conclude that, however, despite the fact that cryogenic procedures are now being developed for an increasing number of recalcitrant seeds and a wide range of tissues and organs, the routine utilization of cryopreservation for the preservation of plant biodiversity is still limited. They note that the main drawback for a wider application of plant cryopreservation is the unavailability of efficient cryopreservation protocols for many plant species.
Panis and Lambardi (2005) nevertheless report that for various herbaceous (i.e. non-woody plants), hardwood (i.e. broadleaf, deciduous trees) and softwood species (i.e. coniferous trees), cryopreservation has been achieved of a wide range of tissues and organs such as cell suspensions, embryogenic cultures, pollen, meristematic tissues and seeds. Cell suspensions (cells in culture in moving or shaking liquid medium) have been cryopreserved from banana and grape while cryopreservation of embryogenic cultures (i.e. callus or suspension cultures with potential to differentiate somatic embryos) is an advanced technology in conifers and is already being successfully applied to numerous commercial species, including pines. There are methods for the cryopreservation of pollen from many crops, but its application is still rather limited and restricted to a few research centres. Meristematic tissues (also called meristems, i.e. plant tissues such as shoot tips, in which the cells are capable of active division and differentiation into specialized tissues such as shoots and roots) are the most common explants (i.e. a portion of a plant aseptically excised and prepared for culture in a nutrient medium) for the cryopreservation of vegetatively propagated species, such as fruit trees and many root and tuber crops. There is largescale application of shoot tip cryopreservation in fruit crop germplasm collections, such as in plum and apple, while the number of cryopreserved herbaceous meristem samples is significantly smaller, although continuously growing. Seeds of most common agricultural and horticultural species can be cryopreserved, and for orthodox seeds this can be considered as an alternative since it offers long-term storage, to traditional storage (e.g. in celery).
15.5 SOME ISSUES AND QUESTIONS RELEVANT TO THE DEBATE
As with each conference hosted in the FAO Biotechnology Forum, the focus is on application of biotechnology in developing countries. In the debate on the role of biotechnology for the characterization/conservation of genetic resources for food and agriculture in developing countries, some of the potential factors that should be considered are briefly described below as they may influence applications of biotechnology for these purposes. In addition, a selection of some of the specific questions that participants might wish to address in the e-mail conference are also given.
15.5.1 Capacity issues
Some of the biotechnologies described in Section 15.4 require a considerable amount of infrastructure and capacity. For example, application of molecular markers to marine capture fishery populations requires appropriate sampling of individuals from the populations of interest, accurate laboratory typing of the samples and good statistical analysis of the data, inter alia. Some technologies that are and have been routinely applied in developed countries are rarely used in many developing countries. For example, AI is a commonplace tool and is used to breed most of dairy cows in the Nordic countries, but is little used in many developing countries.
A rough idea of the status of developing countries' capacities in some of the biotechnologies discussed in this chapter can be derived from a first analysis of about 2 000 crop sector entries from 71 countries in FAO-BioDeC (www.fao.org/biotech/inventory_admin/dep/default.asp), a database providing information on biotechnology products/techniques in use or in the pipeline in developing countries (FAO, 2005). It reports over 400 molecular marker activities, most at the research level, in 43 developing countries. Considering the different regions, most activities were reported in Latin America, with 93 trials and 165 molecular marker projects at the research phase in nine countries. Species reported to be included in molecular marker programmes in the Latin American countries are sugar cane, rice, cocoa, banana, bean, maize and Andean local roots and tubers. The survey also indicates that most countries in Asia are undertaking a wide spectrum of crop research using molecular markers. Molecular marker-related research activities in Africa are reported to be underway in only a few of the countries, such as Ethiopia, Nigeria, South Africa and Zimbabwe; the range of African crops under study with molecular markers is very wide, however, from traditional commodities to tropical fruits. Of the different marker systems, RAPDs, microsatellites, RFLPs and AFLPs were commonly used. While the analysis does not distinguish between use of molecular markers for genetic improvement (marker-assisted selection) or for genetic characterization/conservation, it does indicate that, in crops at least, there is a wide range of marker-based research activities ongoing in developing countries, although some countries and regions seem to have far greater capacity than others.
The database also includes information on in vitro germplasm conservation and cryopreservation, indicating that there are only few activities in developing countries, although there may be substantial under-reporting. The report argues that low uptake of micropropagation techniques for in vitro germplasm conservation may also be due to the existence of established whole plant germplasm collections for species where in vitro conservation is appropriate, leading to a reluctance to provide funding for in vitro facilities. The balance between in vitro and whole plant collections may change as facilities for and capabilities in in vitro conservation increase and existing plant collections need rejuvenation (FAO, 2005).
Some of the specific questions on this issue that participants might wish to address in the e-mail conference might include:
How important is this issue of capacity in different countries or regions?
What are the current limiting factors to capacity building in developing countries? How can these limiting factors be overcome?
What role should international organizations, such as FAO, the World Bank, or the Consultative Group on International Agricultural Research (CGIAR) research centres have in this area?
What role should public-private partnerships play in this area?
How important should regional capacity-building projects be with respect to national initiatives?
15.5.2 Economic issues
While obviously dependent on a range of factors such as the species or the size of the initiative, the use of biotechnology for characterization/conservation of genetic resources can be expensive.
For example, in a survey of livestock molecular marker biodiversity projects, respondents also provided information on costs of 41 projects (FAO, 2004b). They ranged from US$500 to 14 million, averaging US$ 130 000. For projects providing a breakdown of costs, it was estimated that, on average, 65 percent of the total project expenses were spent on genotyping, about 20 percent on sampling animals, 10 percent on the statistical analysis and 5 percent on project coordination. There was also considerable variation between projects, with genotyping costs ranging from 20–90 percent of the total costs in individual projects.
Cryopreservation technologies also can bear a considerable cost. FAO (1997, Annex 1–2) concludes that “the techniques required for successful cryopreservation of non-orthodox seeds are sophisticated and implementing a cryopreservation programme requires trained technical staff, advanced plant tissue culture facilities and increased transportation/handling costs to assure fresh materials. These constraints may present a barrier to effective technology transfer to many developing countries”.
Some of the specific questions on this issue that participants might wish to address in the e-mail conference could include:
In a situation where genetic resources for food and agriculture are being lost rapidly and where financial and human resources to support conservation activities are often limited, how much emphasis should be given to the application of biotechnology in this area?
Which applications should be prioritized in this situation and in which sectors/species?
15.5.3 Prioritization issues
While in an ideal world, all available conservation options should be used, for financial reasons reality often dictates that tough choices have to be made. Although alternative tools or strategies can complement each other, priorities often have to be made. This is also valid for the biotechnologies and conservation strategies described in this chapter.
Molecular markers
Given the large numbers of animal breeds and crop varieties in danger of becoming extinct and limited resources, governments may need to identify the breeds and varieties they consider to be most important for conservation purposes in terms of financial support and storage in gene banks, inter alia. A range of factors may then be considered for the different populations, including their degree of endangerment, their economic value, their impact on the environment, their scientific value, their social and cultural importance and their genetic diversity (can be studied using DNA molecular markers). Characterizing populations for any one of these factors can be time-consuming and costly. Results from FAO (2004b) suggest that many studies, at least in livestock, are characterizing populations for more than one factor at a time. In 56 of the 86 livestock genetic diversity projects surveyed (i.e. about two-thirds), additional breed characteristics such as production performance, morphological traits, disease resistance, behavioural traits and cultural values were also recorded.
If information on these different factors is available, the question is then raised on how best to prioritize the different sources of information for conservation purposes. This is a valid question for each of the agricultural sectors where, for example, efforts have been made to develop methodologies for prioritizing between brown trout populations for conservation using factors such as the relative risk of extinction, the socio-economic and scientific value and the genetic/evolutionary and ecological legacy of the different fish populations (Primmer, 2005).
Some of the specific questions on this issue that participants might wish to address in the e-mail conference could include:
How should genetic differences be weighted with respect to non-genetic differences (cultural importance, current market value, degree of endangerment, etc.) when wishing to prioritize between populations for conservation purposes?
How should differences between populations in genetic diversity at neutral loci (measured using molecular markers) be weighted with respect to differences in genetic differences for traits that are or have been under selection (estimated using production/adaptation data)?
Which kinds of molecular markers or statistical analysis methods are most appropriate for the different kinds of population of interest?
Cryopreservation, in vitro culture and reproductive technologies
As outlined earlier, in situ and ex situ conservation represent two major strategies. Both have their disadvantages (e.g. in situ populations are vulnerable to disease, drought and human interference, etc. while germplasm storage in ex situ collections may be very technology-intense and thus require relatively sophisticated equipment and skilled labour) and advantages (in situ populations can adapt to environmental changes while ex situ collections can be stored indefinitely). Economic merits of one over the other may depend on the species involved and the kind of ex situ collection, among other considerations.
Some of the specific questions on this issue that participants might wish to address in the e-mail conference might include:
When is it more appropriate to support biotechnology as a tool for ex situ conservation in preference to supporting in situ conservation?
What kind of biological material should be cryopreserved, considering that the goal may be to eventually regenerate a living population? For example, in animals, should the focus be on semen, embryo, oocytes, or a combination thereof?
The birth of the Scottish sheep Dolly in 1996, produced through somatic cell cloning using udder cells and nuclear transfer, was a major scientific breakthrough, and since then, individuals from a number of other species have been cloned. How important is this biotechnology for conservation of animal genetic resources?
15.6 REFERENCES
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Cardellino, R.A. 2005. Status of the world's livestock genetic resources. Preparation of the first Report on the State of the World's Animal Genetic Resources. In Proceedings of the International Workshop on the Role of Biotechnology for the Characterisation and Conservation of Crop, Forestry, Animal and Fishery Genetic Resources. (available at www.fao.org/biotech/docs/cardellino.pdf).
de Vicente, M.C., Guzmán, F.A., Engels, J. & Ramanatha Rao, V. 2005. Genetic characterization and its use in decision-making for the conservation of crop germplasm. In Proceedings of the International Workshop on the Role of Biotechnology for the Characterisation and Conservation of Crop, Forestry, Animal and Fishery Genetic Resources. (available at www.fao.org/biotech/docs/vicente.pdf).
Diamond, J. 2002. Evolution, consequences and future of plant and animal domestication. Nature, 418, 700–707. (also available at www.nature.com/cgi-taf/DynaPage.taf?file=/nature/journal/v418/n6898/full/nature01019_fs.html&content_ filetype=pdf).
ERFP. 2003. Guidelines for the constitution of national cryopreservation programmes for farm animals. In S.J. Hiemstra, ed. The European regional focal point on animal genetic resources. Publication no. 1. (also available at www.zum.lt/agroweb/Tekstai/Guidelinest.pdf).
FAO. 1997. The state of the world's plant genetic resources for food and agriculture. (also available at www.fao.org/ag/AGP/AGPS/PGRFA/pdf/swrfull.pdf).
FAO. 1998. Secondary guidelines for development of national farm animal genetic resources management plans: management of small populations at risk. Rome. (also available at dad.fao.org/en/refer/library/guidelin/sml-popn.pdf).
FAO. 1999. The global strategy for the management of farm animal genetic resources: executive brief. Rome. (also available at dad.fao.org/en/refer/library/idad/ex-brf.pdf).
FAO. 2000a. How appropriate are currently available biotechnologies for the forestry sector in developing countries? Background Document to Conference 2 of the FAO Biotechnology Forum (25 April to 30 June 2000). Rome. (available at www.fao.org/biotech/C2doc.htm).
FAO. 2000b. World watch list for domestic animal diversity. 3rd edition. Rome. (also available at dad.fao.org/en/refer/library/wwl/wwl3.pdf).
FAO. 2001. Global forest resources assessment 2000. Rome. (also available at www.fao.org/forestry/site/fra2000report/en).
FAO. 2002. Gene flow from GM to non-GM populations in the crop, forestry, animal and fishery sectors. Background Document to Conference 7 of the FAO Biotechnology Forum (31 May to 5 July 2002). Rome. (available at www.fao.org/biotech/C7doc.htm).
FAO. 2003. Molecular marker assisted selection as a potential tool for genetic improvement of crops, forest trees, livestock and fish in developing countries. Background document to Conference 10 of the FAO Biotechnology Forum (17 November to 14 December 2003). Rome. (available at www.fao.org/biotech/C10doc.htm).
FAO. 2004a. The state of world fisheries and aquaculture. Rome. (also available at www.fao.org/DOCREP/007/y5600e/y5600e00.htm).
FAO. 2004b. Measurement of domestic animal diversity - a review of recent diversity studies. Information Document prepared for the Meeting of the Intergovernmental Technical Working Group on Animal Genetic Resources for Food and Agriculture, 31 March to 2 April 2004, Rome, Italy. (available at dad.fao.org/en/refer/library/reports2/itwg/CGRFA_WG_AnGR_3_04_Inf3.pdf).
FAO. 2005. Status of research and application of crop biotechnologies in developing countries: preliminary assessment by Z. Dhlamini, C. Spillane, J.P. Moss, J. Ruane, N. Urquia & A. Sonnino. Rome. (also available at ftp.fao.org/docrep/fao/008/y5800e/y5800e00.pdf).
Ferreira, M.E. 2005. Molecular analysis of genebanks for sustainable conservation and increased use of crop genetic resources. In Proceedings of the International Workshop on the Role of Biotechnology for the Characterisation and Conservation of Crop, Forestry, Animal and Fishery Genetic Resources. (available at www.fao.org/biotech/docs/ferreira.pdf).
Greer, D. & Harvey, B. 2004. Blue genes: sharing and conserving the world's aquatic biodiversity. Ottawa, Canada, International Development Research Centre. (also available at web.idrc.ca/openebooks/157-4/).
Hanotte, O. & Jianlin, H. 2005. Genetic characterisation of livestock populations and its use in conservation decision-making. In Proceedings of the International Workshop on the Role of Biotechnology for the Characterisation and Conservation of Crop, Forestry, Animal and Fishery Genetic Resources. (available at www.fao.org/biotech/docs/hanotte.pdf).
Hiemstra, S.J., van der Lende, T. & Woelders, H. 2005. The potential of cryopreservation and reproductive technologies for animal genetic resources conservation strategies. In Proceedings of the International Workshop on the Role of Biotechnology for the Characterisation and Conservation of Crop, Forestry, Animal and Fishery Genetic Resources. (available at www.fao.org/biotech/docs/hiemstra.pdf).
Lanteri, S. & Barcaccia, G. 2005. Molecular markers based analysis for crop germplasm preservation. In Proceedings of the International Workshop on the Role of Biotechnology for the Characterisation and Conservation of Crop, Forestry, Animal and Fishery Genetic Resources. (available at www.fao.org/biotech/docs/lanteri.pdf).
Lenstra, J.A. & Econogene Consortium. 2005. Evolutionary and demographic history of sheep and goats suggested by nuclear, mtDNA and Y-chromosome markers. In Proceedings of the International Workshop on the Role of Biotechnology for the Characterisation and Conservation of Crop, Forestry, Animal and Fishery Genetic Resources. (available at www.fao.org/biotech/docs/lenstra.pdf).
Panis, B. & Lambardi, M. 2005. Status of cryopreservation technologies in plants (crops and forest trees). In Proceedings of the International Workshop on the Role of Biotechnology for the Characterisation and Conservation of Crop, Forestry, Animal and Fishery Genetic Resources. (available at www.fao.org/biotech/docs/panis.pdf).
Papa, R. 2005. Gene flow and introgression between domesticated crops and their wild relatives. In Proceedings of the International Workshop on the Role of Biotechnology for the Characterisation and Conservation of Crop, Forestry, Animal and Fishery Genetic Resources. (available at www.fao.org/biotech/docs/papa.pdf).
Primmer, C. 2005. Genetic characterisation of populations and its use in conservation decision-making in fish. In Proceedings of the International Workshop on the Role of Biotechnology for the Characterisation and Conservation of Crop, Forestry, Animal and Fishery Genetic Resources. (available at www.fao.org/biotech/docs/primmer.pdf).
Rao, N.K. 2004. Plant genetic resources: advancing conservation and use through biotechnology. African Journal of Biotechnology, 3: 136–145. (also available at www.academicjournals.org/AJB/PDF/Pdf2004/Feb/Rao.pdf).
Ruane, J. 2000. A framework for prioritizing domestic animal breeds for conservation purposes at the national level: a Norwegian case study. Conservation Biology, 14: 1385–1393. (also available at www.nordgen.org/ngh/download/bokartikkel-ruane.doc).
Sigaud, P. 2005. Efforts towards assessing the global status of forest genetic resources. In Proceedings of the International Workshop on the Role of Biotechnology for the Characterisation and Conservation of Crop, Forestry, Animal and Fishery Genetic Resources. (available at www.fao.org/biotech/docs/sigaud.pdf).
Simianer, H. 2005. Use of molecular markers and other information for sampling germplasm to create an animal genebank. In Proceedings of the International Workshop on the Role of Biotechnology for the Characterisation and Conservation of Crop, Forestry, Animal and Fishery Genetic Resources. (available at www.fao.org/biotech/docs/simianer.pdf).
John Ruane and Jonathan Robinson
16.1 SUMMARY
Characterization and conservation of genetic resources of crops, forest trees, livestock and aquatic species are important for all countries, but particularly for developing countries whose economies depend heavily on these sectors, and where genetic resources are often threatened. A number of biotechnology tools are available that can help in characterization and conservation of such genetic resources, ranging from relatively cheap and uncomplicated technologies to sophisticated, resource-demanding ones. In each of the crop, forestry, animal and fishery sectors, albeit to different degrees, biotechnology tools are currently being applied in developing countries for these purposes and numerous examples of the wide range of applications were provided during this FAO e-mail conference. Of the different biotechnologies, most discussions were about molecular markers, in particular their use for characterization of genetic resources, where issues such as the advantages or disadvantages of different marker systems and the proposal to develop a universal molecular marker database were debated. in situ ations involving potential use of marker and non-marker information, such as development of a core collection of plant gene bank accessions or prioritization of animal breeds for conservation purposes, there was general consensus that decisions should not be based on marker information alone and that other factors, such as morphology and agronomic performance, should also be considered. The merits of several in vitro techniques, including tissue culture, cryopreservation and DNA storage, were considered with a view to conservation of genetic resources, where e.g. DNA banks for plants were seen as potentially complementing but not replacing seed banks, at least in the near future. The ability to apply these biotechnologies in developing countries is currently limited by the lack of sufficient funds, human capacity and adequate infrastructure. The importance of human resource capacity building was highlighted. There was a general call for greater collaboration among researchers and practitioners, particularly at the regional level, to reduce costs and pool limited resources, and between developed and developing country institutions. A role was seen for international organizations, including FAO, and the centres of the Consultative Group on International Agricultural Research (CGIAR), in coordinating these collaborative efforts and in supporting these capacity-building activities.
16.2 INTRODUCTION
The theme of this e-mail conference, which was hosted by the FAO Biotechnology Forum between 6 June and 3 July 2005, was the role of biotechnology for the characterization and conservation of crop, forest, animal and fishery genetic resources in developing countries. This chapter provides a summary of the principal issues and opinions received from contributors to the conference. Specific messages are referenced in the document using participants' surnames and the message number. All messages can be read at http://www.fao.org/biotech/logs/c13logs.htm. About 650 people subscribed to the conference and 127 e-mail messages were posted from people living in 38 different countries; over 60 percent of messages were from developing countries. Most of the messages came from people working in research organizations, including CGIAR centres, and universities.
Most participants directed their messages to issues in one of the crop, forestry, animal or fishery sectors, with greatest attention given to crops and animals. Some also addressed cross-sectoral issues such as resource availability and constraints and international collaboration. In Section 16.3 of this chapter, the main issues discussed during the conference are summarized. Section 16.4 provides information on participation and Section 16.5 represents a list of names and countries of those who sent referenced messages.
16.3 MAIN THEMES DISCUSSED
16.3.1 Current applications and potential of biotechnology in the different sectors
Most of the messages addressed the current situation and potential of using biotechnology tools for the characterization and conservation of genetic resources in one of the individual sectors (crop, forestry, animal or fishery). Although each sector has its specificities, some of the discussions were also very relevant for other sectors.
Crop genetic resources
The potential importance of biotechnology for locally important crop genetic resources was raised right at the beginning of the conference. Nkhoma (1), describing activities at the Southern African Development Community Plant Genetic Resources Centre (SPGRC) in Zambia, noted that while their programme had succeeded fairly well with common cereals and legumes, they were in some cases unable to work on indigenous crops - for example when difficult to cultivate - which were locally useful and on which nobody else was working. He said that biotechnology facilities would help them to tackle these problems. He also reported that the SPGRC collections were only characterized using agronomic and morphological data, although molecular markers would be helpful to allow removal of duplicate accessions. The usefulness of applying biotechnology to genetic diversity studies of less common crops was reported by Infante (8), who described their results in Venezuela where they found genetic variability in clonally reproduced henequen and cocuy following analysis with molecular markers. This had implications for conservation because clonally reproduced species were assumed to be of uniform genetic constitution. Morphological data were also gathered and analysed which supported the molecular data.
Kisha (6) noted that molecular markers are now an accepted and widely used tool for measuring genetic diversity, where “molecular marker technology can be used to characterize the extent of diversity within a collection and for the development of collection management strategies, which may include establishment of core collections, identification of redundancies or contamination, guidance for future collection efforts, and identification of gaps of ancestral crop relatives. Additionally, analysis of world-wide genetic diversity can identify areas suited for the establishment of in-situ conservation sites.”
Ndjiondjop (41) reported that recent studies with molecular markers based on repetitive DNA sequences had provided useful information on genetic diversity present in rice, Oryza glaberrima. Previous studies using isozyme and restriction fragment length polymorphism (RFLP) markers had failed to identify as many polymorphisms, although considerable variation for many morphological and agronomic traits had been recorded. The microsatellite marker information will be used to develop the core collection of O. glaberrima accessions in the Africa Rice Center (WARDA) gene bank. Also regarding core collections, Huaman (38) said that “molecular markers are the most valuable source to get data on genetic diversity of a given crop or plant species”, but added that when selecting a core collection, other factors, such as eco-geographical data, disease and pest reaction and morphological diversity, had to be taken into account. Ghamkhar (28) suggested that molecular markers should be “definitely employed as the best technique for screening of genetic diversity but they must also be double checked by morphological data to make certain there is no major loss in our breeding and/or core collection development programs.”
Molecular marker data were considered to be only one aspect of characterization (Huaman, 38), but one that would be of greater use were a universal molecular marker database to exist (Kisha, 6). He noted that in the current situation, few, if any, plant genetic diversity studies can be directly compared or compiled; that marker data can be lost or forgotten after publication; and that studies of genetic diversity are usually limited to a few accessions or accessions from a limited area of interest. The development of such a database was supported by several participants, including Vijay (18), who urged, however, that consideration should be given to selection of a set of universally reliable and reproducible markers; to consensus on the outcome from different markers; and to standardization of methodologies, including the mode of analysis. In this context, both de Vicente (26) and Ford-Lloyd (30) mentioned an initiative by the International Plant Genetic Resources Institute (IPGRI) to define community standards for documenting information about genetic markers so that researchers can generate and exchange genetic marker data that are standardized and replicable. Barker (24) noted that a large number of molecular markers were already available in the public domain, but to assemble a universal database would require considerable effort and it was hard to see any single country willing to invest in such a project. He did envisage, however, that it would be easier to establish databases on a single species basis. Kisha (51, 103) expanded on his original proposal, describing how the database would need to be curated and why a core set of primers would be useful to make studies comparable. Sales (19) noted that some microsatellite databases already exist and Ghamkhar (28) mentioned some Webbased databases, mostly for cereals. R. Jones (54) hoped that a global marker database might also include data for key fish and crustacean species.
Kisha (51) also commented on the relative usefulness of different types of molecular markers, arguing that amplification fragment length polymorphism (AFLP) markers can cover a large area of the genome with less cost than simple sequence repeat (SSR) markers, also known as microsatellites. Ghamkhar (28, 80) proposed that inter-simple sequence repeat (ISSR) techniques were also cheap and efficient, and Varshney (43) supported using single nucleotide polymorphism (SNP) markers for characterization, while acknowledging their high cost and the relative paucity of species for which markers currently existed. In a similar vein, Ghamkhar (53) suggested that SNPs might be useful in well-studied crops such as wheat and maize but not for less common crops because of the sequencing work required to get the markers. Warburton (42) described experiences in using SSR markers to study molecular diversity in maize among several laboratories and highlighted problems of comparison and reproducibility among laboratories even when using the same protocols and platforms for genotyping. She said that the possibility of combining datasets was “virtually non-existent”if laboratories used different techniques and that there were problems of repeatability with some other kinds of markers as well. Varshney (43) had had similar experiences and suggested that expressed sequence tag (EST)-derived SSR markers showed higher reproducibility than genomic SSR markers. Krishna (88), supported by Buso (93), felt that all molecular marker data were nevertheless useful, and even though random amplified polymorphic DNA (RAPD) and AFLP markers might have some problems with reproducibility (highlighted by Vijay [79]), they also had advantages (highlighted by Muchugi [77]).
Dulieu (95) also compared different markers systems with respect to the reproducibility of their results, noting that some markers, such as microsatellites, were more reliable than others but required more preliminary research, and suggested that biotechnology companies should be encouraged to produce kits for the most important species, following the example of human DNA fingerprinting kits which are used universally. De Vicente (26) noted that as part of the CGIAR Generation Challenge Program, microsatellite kits were being put together which they hoped to have available in the near future. Dulieu (95) also pointed out that many of the molecular markers revealed differences between populations that were not highly correlated with performance or phenotypic characters. Gupta (44) suggested that if molecular markers were to be used for genetic diversity analysis, they should be functional markers rather than random genomic markers. He (44, 87) also reported that they got different results from genetic distance analyses of bread wheat when different kinds of markers (SSR, AFLP or selective amplification of microsatellite polymorphic loci [SAMPL]) were used, suggesting that the best estimate of diversity might be obtained from data on large numbers of morphological traits. Kisha (88) wrote that it would be useful to compare at least two different marker systems for agreement in the resulting relationships. Ghamkhar (118) agreed that there can be inconsistencies between results obtained with different sets of molecular and morphological data, concluding, “more molecular techniques/data, more resolution or better results.”
The merits of tissue culture as a means of genetic resource conservation were discussed by several participants (e.g. Muchugi, 68). Lin (2) suggested that tissue culture and other forms of micropropagation were useful tools for the conservation and multiplication of plant species, noting that low-cost options were also available. Cummins (9), however, felt that because of somaclonal variation (i.e. mutations that occur spontaneously in tissue culture), tissue culture was not a good way of conserving local genetic material. Muralidharan (22) agreed and favoured use of slow-growing shoot culture. He suggested also that molecular markers could be used to study the extent of somaclonal variation in slow-growing or cryopreserved cultures. Lin (21) reported that improved protocols for in vitro conservation, developed for a range of species, could overcome the potential problems of somaclonal variation and that in vitro conservation was useful, particularly for plants that do not produce seeds or that produce seeds of limited viability. Ford-Lloyd (30) pointed out that in vitro conservation was being used to support genetic conservation, despite genetic instability, as it represented a better option than the currently available alternatives.
There were several responses to a question from Muralidharan (22) regarding the potential of DNA as a means of long-term conservation of genetic material. Wang (32) pointed out that germplasm conservation as pure DNA was already a reality in some countries, which was supplemented by concrete examples in later messages (Ghamkhar, 38, 48; Widjaja, 40; Vijay, 47). De Vicente (46) reported results from a 2004 worldwide survey on plant genetic resources DNA banking activities showing that 20 percent of the 243 institutions that replied to the questionnaire kept DNA as a genetic resource. Although noting that both DNA banks and seed banks have advantages and disadvantages, Ghamkhar (34) suggested that seed banks were currently preferable to DNA banks since, for example, contamination was more immediately apparent and morphological screening and maintenance were easier. Vijay (47) agreed with Ghamkhar (34) and described the major limitations he saw to the use of DNA banks, suggesting they could complement, but not replace, seed banks, at least in the near future.
Dulieu (96) preferred phage genomic libraries over DNA banks for genetic resource conservation because they are easier to prepare and maintain. However, he warned against using more technology to counter effects of misuses of technology (mainly the destruction of traditional agricultural systems), a point echoed in a different context by Magalhães (72), supported by Kante (105) and Adediran (112), who argued that the use of biotechnology to conserve or characterize biodiversity could not be considered a solution but only a palliative when biodiversity in developing countries was being destroyed by an economic model based on the economic exploitation of developing countries and their natural resources.
Nassar (4) suggested that apomixis (i.e. where seeds are produced through asexual processes so that the genetic make-up of the seeds is identical to that of the mother plant) could contribute to conservation of certain crop genetic resources and reported that they had produced apomictic cassava clones in Brazil, confirmed using molecular markers. Vijay (11) agreed and felt that identification and transfer of apomixis to other cultivated species would have important consequences for conservation and hybrid seed production.
During the conference, Gupta (23, 87), supported by Vijay (73), also mentioned the new possibilities of using DNA barcoding, allowing different plant species to be identified and discriminated. Ghamkhar (118) felt it could help taxonomists to classify, re-classify or identify taxa or new species, when used together with traditional taxonomy methodologies based on morphological data. Gupta (120) also pointed out the potential of DNA microarrays for the study of genetic diversity.
Forest genetic resources
There were relatively few contributions specifically addressing the use of biotechnology in characterization and conservation of forest genetic resources. Oluawsegun (101) reported on the loss of forest species in Nigeria due to factors such as poverty and low education levels among the rural people, concluding “for any meaningful and lasting conservation programme to be effectively carried out there must be a conscious effort in involving the local people in maintaining and managing their environment since the needs of these dwellers must be respected.” Muralidharan (67) bemoaned that there were insufficient effort and funding put into conservation of forest genetic resources compared to the crop and livestock sectors and wondered whether tropical forest genetic resources could be successfully conserved in DNA banks as they “are in danger of mass erosion due to degradation of the habitat.” He emphasized, however, that wherever feasible, the more conventional conservation methods should be used. Ghamkhar (76) shared his concerns and supported the use of ex situ conservation methods (seed banks, storing tissues, or DNA banks) as there did not seem to be other options.
Muchugi (68) was also concerned about conservation of indigenous forest genetic resources, which are threatened by factors such as increasing population sizes (requiring land for settlement and farming). Although biotechnology could be of value, through e.g. use of molecular markers to investigate gene flow or to assist in establishment of ex situ conservation programmes, she noted that little work had been carried out on tropical tree species compared to temperate species. She (68, 77) described the advantages, in terms of relative costs and speed, of using RAPD markers and isozymes for molecular characterization of tropical tree species, but was saddened that renowned molecular genetics journals refused to publish results using these simpler techniques, concluding: “this is placing scientists in the developing world with simple labs in a tricky position; we would love to employ the modern state of art sequencers but financial limitations will not allow it. What is the way out then considering the need to study these taxa before we lose them on the earth's surface.”The importance of molecular markers for management of endangered natural forest tree species was also underlined by Dulieu (96).
Animal genetic resources
Aziz (7), supported by Silva (20), noted that livestock production in developing countries is characterized by several major constraints, such as the absence of national recording systems, paucity of breeding programmes, small herd sizes, lack of awareness of the importance of animal genetic resources, national policies to replace local breeds with exotic ones, scarce resources and weak infrastructure. He (82) proposed that in developing countries a systematic approach be taken to the documentation, evaluation, conservation and utilization of their animal genetic resources. Babar (107, 115) emphasized the importance of conserving local breeds in developing countries, especially in countries such as Pakistan that are home to several important breeds. He (107) encouraged use of cryopreservation technologies, including embryo and semen storage, and establishment of DNA banks (as they had done in Lahore). Sales (84) said that developing countries had been “swamped” with new livestock breeds and little had been done to conserve their native breeds. All these issues represented obstacles to the application of biotechnology, as communicated by Silva (20) and Tantia (65).
Hassan (61) described the limitations to livestock production in sub-Saharan Africa, which were similar to those previously outlined by Aziz (7). He suggested that although molecular genetics might have a great role to play in revolutionizing livestock production in developing countries, “the stage is not yet set”for them to do so and that developing countries should continue the ongoing phenotypic characterization. Maddul (89) mentioned their work on phenotypic characterization and conservation of pigs and chickens in the northern Luzon highlands of the Philippines, where molecular characterization had not been carried out due to the lack of a laboratory. Tan (12) described results from molecular characterization studies of buffaloes in Southeast Asia carried out in the 1990s. Silva (20) reported that in Sri Lanka, applications of biotechnology are limited, involving artificial insemination (AI) and, in isolated cases, genetic characterization of local animals. Osakwe (104) suggested that although some progress had been reported in plants, there “is no meaningful characterisation and conservation of genetic resources using biotechnology in animals in most developing countries.”
Köhler-Rollefson (31) felt that although scientifically interesting, the relevance of molecular characterization of livestock breeds for livestock keepers and for poverty alleviation had yet to be proven. She also asked whether, when studying livestock domestication and dispersal, molecular data were superior to data from archaeological and ethno-historical investigations, or if they merely confirmed what was already known. Lenstra (35) said that there were several examples where DNA studies had provided additional insights into these issues. For Hanotte (36), molecular characterization had provided an additional source of information that, together with data from archaeology, linguistics and indigenous knowledge, was making it possible to “complete a puzzle about origin and history of agriculture.” P. Jones (59) reported that recent molecular work had substantiated the long-held belief that most of today's donkeys have their origins in northern Africa and that those in southern Africa have different origins, concluding “such a history has implications for management practices and technology transfer, and thus indirectly on poverty alleviation.”
Köhler-Rollefson (31) also asked whether genetic uniqueness should be the key criterion for deciding which breeds to prioritize for conservation purposes. Lenstra (35) argued that such a decision should not be made on the basis of marker data alone and that other factors, such as the breeds' relevance to regional tradition, were also important. Tantia (65) thought that molecular characterization with microsatellites was an excellent tool in this context. Toro (71) argued that the important parameters to consider were phenotypic ones, adaptation to specific environments, possession of economically important traits and cultural/historical value, etc., and that “to use molecular markers is not harmful as long as we recognize its subordinate relevance in this context.” Hanotte (36) suggested that molecular criteria should be considered very important when selecting animals for ex situ conservation, but caution should be advised for in situ conservation at the farmer community level. Nimbkar (45) argued that areas directly related to poverty alleviation, such as characterizing the performance of indigenous breeds (leading to genetic improvement programmes based on simple principles and methods of animal breeding), were being neglected in developing countries such as India in favour of biotechnologies, such as molecular characterization or embryo transfer, blaming this trend on the “glamour” of these biotechnologies. This view was supported by Steane (55), who suggested that one of the problems is that in most countries the funding bodies emphasize research rather than application, concluding “certainly biotech has an important role but it will never be the sole criterion for decisions about conservation and characterisation of breeds.” On the general issue of weighting genetic versus non-genetic differences, Rakotonjanahary (92) felt the former should be ranked at a lower level because “socio-economic traits such as culture, market value, degree of endangerment are more related to humankind, which is to be considered the top priority.”
Galal (60, 66) suggested that it would be useful for conservation purposes to use marker data to verify if phenotypically similar breeds were actually the same or not, although Toro (63, 71) did not see how this could be verified since molecular markers refer to neutral or non-coding genetic variation and genetic distances ignore within-breed variation. Lenstra (70), based on results from two recent European Union research projects, suggested that phenotypic distinctness and molecular diversity could both represent valid reasons for conservation, but they were often negatively correlated. Toro (99) argued that genetic distances based on molecular markers were only partially informative, and that there had been too much emphasis on molecular markers, with very few papers published on phylogeny based on phenotypic information compared to “hundreds” on genetic distances using molecular markers. Ghamkhar (100), in response, listed several advantages of molecular markers over morphological data for conservation purposes, including their relative abundance, stability under changing environmental conditions, consistency among laboratories, reduced time requirement and ease of data analysis. Chagunda (113) saw a need for decision support tools to assist in animal genetic resource conservation strategies, where information from different sources (molecular, phenotypic, genotypic, production system, indigenous knowledge, socio-economics, etc.) could be combined in a meaningful way to support decisions for conservation strategies.
Fishery genetic resources
R. Jones (14, 54) felt that “fisheries resources are undervalued and underrepresented in discussions on the conservation and sustainable use of germplasm” and that “the application of molecular tools in the relatively unknown world of fisheries and aquatic biodiversity will continue to play roles in determining stock structures of multi-species fisheries (meta-population dynamics) and important taxonomic classification work”. Following the message of Oluawsegun (101), describing the pressures on natural habitats in developing countries, R. Jones (102) urged that those working on conservation of genetic resources in developing countries should think seriously about how to prioritize their efforts. He (64) noted that because of the high costs involved, most of the molecular work in fisheries was done on commercial or potentially commercial species and wondered how these results might be applied to alleviation of hunger and poverty.
Tan (12) wrote that microsatellites were being used to study three commercial aquatic species in Malaysia - the giant freshwater prawn, the green-lipped mussel and the Asian river catfish. Chinsembu (37) described recent research on the mitochondrial DNA analysis of cichlid fish from five southern African rivers. Results of this work had helped to explain details of the evolution and radiation of the species. Bhassu (62) was enthusiastic about the impacts of biotechnology in stock improvement and characterization of fish, citing recent work on characterization of red tilapia stocks with microsatellites; on the phylogeny of freshwater prawns (where mitochondrial DNA analysis confirmed previous findings from morphological and protein data); and on the use of AFLP markers for sex determination in freshwater prawns, noting, however, that little funding was available for such research. Ablan (75) felt that molecular analysis was not as meaningful for cultivated populations as for wild populations, for which they can define spatial distribution of stocks and provide guidance on where to obtain brood stock for restocking programmes. She noted that “we've had cases where molecular genetics provide answers, others where it simply validates observation of the phenotype, local knowledge, value judgements, or even gut feel. And yes, sometimes its usefulness can be forced.”
Tan (69) reported that protein and molecular marker analysis had indicated low levels of genetic diversity in many cultured stocks of prawn and sea bass, which were associated with abnormalities and reduced fertility. He argued that such results had important economic implications, justifying investments in genetic marker studies, and that stock deterioration could therefore be halted based on this knowledge. Toro (71), arguing that the phenomenon of inbreeding depression was already well known, was not convinced that molecular markers would provide useful additional information here. Tan (81), however, noted that aquaculturists in Malaysia usually did not maintain breeding records, paid little heed to inbreeding and so were often surprised when molecular studies revealed that their problem stocks were inbred. Only after seeing the molecular typing results would they begin outcrossing. Kalamujic (110) reported that in Bosnia and Herzegovina, molecular marker characterization of salmonid species had led to a new freshwater fisheries law, including a provision on obligatory genetic control of material for stocking. R. Jones (14) described how DNA fingerprinting of sturgeon had helped to provide information on stock make-up, potential loss of genetic diversity in Russian brood stocks, the caviar trade and use of ex situ conservation.
16.3.2 Priorities and resource constraints in developing countries
Some participants discussed the priority that developing countries give or should give to applying biotechnology for the characterization and conservation of their genetic resources. For example, Qureshi (3) wrote that conservation of animal genetic resources was neglected in developing countries as the state had failed to support it and it was also not a priority for the farmers. He saw, however, that in this situation, universities could play an important role in applying biotechnology in this area. Huque (52) suggested that policy-makers in developing countries did not support animal biotechnology as they preferred to invest in areas promising short-term rather than long-term benefits. Rakotonjanahary (92) said the weak capacity to use biotechnology for characterization/conservation in developing countries was understandable given that, in general, national policy prioritized poverty alleviation, food self-sufficiency and increased agricultural productivity over research activities. Komwihangilo (57) argued that farmers in, for example, sub-Saharan Africa were not concerned about characterization but about their livelihoods and their animals being able to produce. Therefore, molecular characterization and other new technologies will only be of value to the farmers “if they deliver them from the present trials of life and assure them sustainable futures.”
As most of the world's poor live in the rural areas of developing countries, Djoulde (16) was skeptical about the merits of using advanced technologies, such as molecular markers or cryopreservation and reproductive technologies, in these areas. De Vicente (27) wondered whether work should therefore cease on solving such problems if the rural people did not care about the solutions and whether effort should be concentrated on ensuring that the research findings reach the rural people. Djoulde (29) supported this latter point and emphasized that the complicated technologies needed to be adapted for easier application in developing countries. Ghamkhar (33) suggested that farmers in rural areas were not expected to employ the new methodologies, but should benefit from the better adapted germplasm resulting from their application. Considering the instability and threats that many gene banks faced in developing countries (due to human or environmental factors, such as war, hurricanes or famine), Murphy (124) questioned the merits of progressing to advanced methods such as molecular markers or tissue culture, concluding, “in places where seed banks are being destroyed by looters who are just after the plastic bottles, or where stocks are dying from want of electricity to refrigerate them, we need to question our priorities”.
Many participants (e.g. Nkhoma, 1; Krishna, 74) commented on the lack of financial, human and infrastructural resources in developing countries for applications of biotechnology for the characterisation and conservation of genetic resources. As a typical example, Huaman (38) highlighted the difficulties facing researchers in developing countries wishing to use molecular markers, due to lack of laboratory equipment or materials. Aziz (7) and Galal (60) noted that the high costs of biotechnology, along with training, equipment and infrastructure, directed at livestock, represented a constraint. Komwihangilo (57) pointed out that the problem was not biotechnology-specific as there were, in general, low levels of funding for agricultural research. Oluawsegun (101) said economic problems meant that his government was “unable to allocate enough resources for conservation, for research and monitoring of conservation programmes, for the creation of gene banks and education of the public concerning the importance of preserving the biosphere.”Urriola (50) reported that despite insufficient funding, human capacity, equipment and infrastructure, researchers in Panama were nevertheless trying to take advantage of the biotechnology resources available. Murphy (124) indicated that there were also serious resource shortages in the flagship centres of the CGIAR.
Despite the constraints, some participants urged that developing countries should not be left behind. For example, Vijay (73) considered that, despite the large amount of resources needed, it was important to keep up with the advancing technology: “Developing countries cannot stay aside from the mainstream knowledge as most of the diversity and its end users belong to them.” Similarly, Kapoor-Vijay (117) urged that developing countries should not become “technologically excluded” and that “information, knowledge, and expertise associated with biotechnology which is relevant and needed to conserve unique plant, animal and microbial species thriving in diverse and especially extreme environments should be strengthened.” For Edema (123), although developing countries did not, on their own, have enough resources to advance in biotechnology, they could not afford to be left behind either. R. Jones (15), on the other hand, warned of the dangers of being seduced by technology in a developing country context, “where the capacities to understand, absorb and if necessary fix and upgrade may be limited or non-existent.” He emphasized, however, that no country should be deprived of the knowledge of the technologies available and that it was their choice if or how to apply them.
Not all biotechnologies are, however, equally resource-demanding. Lin (2) commented on the high costs of establishing and operating tissue culture and micropropagation facilities in developing countries, but pointed out that low-cost options had been developed. Thro (106) described one such low-cost initiative for propagating Andean root crops in rural areas. R. Jones (15) commended work being done on development of low-cost, portable cryopreservation technology for fish gametes. Kisha (98), commenting on the application of molecular markers, suggested that inexpensive, high throughput technology should be a primary consideration. Muchugi (68, 77) proposed the use of cheaper marker systems, such as RAPDs, in preliminary studies to form the basis of conservation strategies. Vijay (79), however, disagreed because of problems of reproducibility of RAPDs, concluding that although funding was the main problem for scientists in developing countries, the “use of outdated technology because of its low cost is not the answer.” He (49) emphasized the importance of adoption, moving from “lab to land”, to make technology useful for the common good, and that further research could allow the technologies developed to be made user-friendly. Similarly, Prana (58) urged a down-to-earth approach, focusing on appropriate technology adjusted for the real-life resources, local needs and socio-economic factors
The importance of human capacity was highlighted with, for example, Prana (56) urging that human resource development be placed highest on the priority list. Uzochukwu (83), supported by Krishna (86) and Osakwe (104), argued that the large funding bodies “are interested in providing financial support for biotechnology research but not for training and updating local scientists in the developing countries. Krishna (74), supported by Uzochukwu (83), called for a massive capacity-building effort at the national and international level. Caesar (126) outlined the key features of a potential global biotechnology capacity-building project, based on regional and subregional groupings of developing countries and including a comprehensive scholarship/fellowship programme for developing countries. Sales (84) identified a need for developing country scientists to be regularly updated on the current trends and innovations relevant in biotechnology. She cited the Asian Maize Biotechnology Network, established to strengthen the biotechnology capacity of national maize research programmes in Asia, as a good model that, for example, livestock scientists might follow. Ghamkhar (85) also pointed out that the CGIAR Generation Challenge Program, previously described by de Vicente (26), provided relevant training programmes. He argued, however, that developing countries could not expect international organizations to cover all their training needs and that the countries should prepare a strategic plan and national training programme to transfer the knowledge accumulated by their already-trained senior scientists to the national research centres and universities.
16.3.3 Cooperative approaches
A recurring theme throughout the conference was that biotechnology research and application of results to characterization and conservation of germplasm can benefit from collaborative efforts, particularly at the regional level (e.g. Silva, 20; Ghamkhar, 78). In livestock, Galal (60), noting that the costs of equipment and materials for molecular characterization were too high for most local institutions, argued that “some regional coordination, possibly with international input, is required to carry out such work.” In a similar vein, Muchugi (68), arguing that molecular characterization of tree species was best approached from an ecological/geographical perspective as their distributions cut across political boundaries, called for “greater collaboration among scientists within the regions in exchange of plant materials and knowledge gathered.” The importance of collaboration between developed and developing country institutions was also highlighted (e.g. Prana, 56; Babar, 107, 115), with Vijay (121) arguing that “with the advancement of technology (like use of molecular markers for conservation) there should be a proper collaboration between these two parts of the world.” Rakotonjanahary (92) thought that public-private partnerships in developing countries could play an important role, but “only on the condition that characterization and conservation have an evident economic impact, which is not the case for the moment.”
Suggestions for increased cooperation were generally made in the interests of pooling scarce resources and reducing costs (e.g. Galal, 60; Muchugi, 94; Chinsembu, 108). De Vicente (25), arguing that it was not realistic for all countries to have facilities to carry out their own work, proposed that institutions consider the possibility of having a hub centre in their region where they could either send their samples for analysis or go there to do the work. Muchugi (94) cited the Biosciences Eastern and Central Africa (BECA) facilities in Nairobi as a good example to show how pooling of resources could help in the advancement of biotechnology. A universal molecular marker database was suggested by Kisha (6), based on collaboration among germplasm conservation centres and other interested parties. This idea was supported by several participants (e.g. Sales, 19), with Barker (24) emphasizing the requirement for “international collaboration and for the data to be maintained in the public domain.”The role of international organizations, such as FAO, and the CGIAR centres, in coordinating these collaborative efforts, providing funds and contributing to capacity building was emphasized in several messages (e.g. Ghamkhar, 53, 78; Muchugi, 68; Vijay, 73, 79; Rakotonjanahary, 92; Babar, 107; Caesar, 126).
16.4 PARTICIPATION
The conference ran for four weeks, from 6 June to 3 July 2005. There were 645 subscribers to the conference, of whom 64 (10 percent) submitted at least one message. There were 127 messages in total, of which 61 percent came from people living in developing countries. Contributions to the conference came from all major regions of the world, with 28 percent of messages from Asia, 20 percent from Africa, 17 percent from Europe, 13 percent from Latin America and the Caribbean, 13 percent from North America and 10 percent from Oceania. Contributors represented 38 countries, the greatest numbers of messages coming from India, Australia, Canada, Brazil, United States, France, Kenya, Malaysia and Nigeria respectively. Most of the messages came from people working in research organisations (45 percent), including centres of the CGIAR and universities (43 percent). The remainder were from independent consultants or from people working for an inter-governmental institute, non-governmental organization, national development agency or private company.
16.5 NAME AND COUNTRY OF PARTICIPANTS WITH REFERENCED MESSAGES
Ablan, Menchie. Malaysia
Adediran, Samuel Adeniyi.
Gambia Aziz, Mahmoud Abdel. Egypt
Babar, Masroor Ellahi. Canada
Barker, Guy. United Kingdom
Bhassu, Subha. Malaysia
Buso, Glaucia Salles Cortopassi. Brazil
Caesar, John. Guyana
Chagunda Mizeck. Denmark
Chinsembu, Kazhila Croffat. Namibia
Cummins, Joe. Canada
de Vicente, Carmen. Colombia
Djoulde, Darman Roger. Cameroon
Dulieu, Hubert. France
Edema, Olayinka. Nigeria
Ford-Lloyd, Brian. United Kingdom
Galal, Salah. Egypt
Ghamkhar, Kioumars. Australia
Gupta, P.K. India
Hanotte, Olivier. Kenya
Hassan, W. Akin. Nigeria
Huaman, Zosimo. Peru
Huque, Quazi M. Emdadul. Bangladesh
Infante, Diogenes. Venezuela
Jones, Peta. South Africa
Jones, Ron. Canada
Kalamujic, Belma. Bosnia and Herzegovina
Kante, Bocar. Italy
Kapoor-Vijay, Promila. Switzerland
Kisha, Theodore. United States of America
Komwihangilo, Daniel. United Republic of Tanzania
Köhler-Rollefson, Ilse. Germany
Krishna, Janaki. India
Lenstra, Hans. Netherlands
Lin, Edo. France
Maddul, Sonwright. Philippines
Magalhães, Vladimir. Brazil
Muchugi, Alice. Kenya
Muralidharan, E.M. India
Murphy, Denis. United Kingdom
Nassar, Nagib. Brazil
Ndjiondjop, Marie Noelle.
Benin Nimbkar, Chanda. Australia
Nkhoma, Charles. Zambia
Oluawsegun, Adegoke Adedayo. Nigeria
Osakwe, Isaac. Nigeria
Prana, Made Sri. Indonesia
Qureshi, Muhammad Subhan. Pakistan
Rakotonjanahary, Xavier. Madagascar
Sales, Emma. Philippines
Silva, Pradeepa. Sri Lanka
Steane, David. Thailand
Tan, S.G. Malaysia
Tantia, M.S. India
Thro, Ann Marie. United States of America
Toro, Miguel. Spain
Urriola, Jazmina. Panama
Uzochukwu, Sylvia. Nigeria
Varshney, Rajeev. Germany
Vijay, D. India
Wang, Richard. United States of America
Warburton, Marilyn. Mexico
Widjaja, Elizabeth. Indonesia
