Plant Genetic Resources and Conservation

INTRODUCTION
Global concern about loss of valuable genetic resources prompted international action. The
reasons for this loss are many and include deforestation, developmental activities such as
hydroelectric projects, road laying, urbanization and changes in agricultural practices, and
finally modern agriculture and introduction of new and uniform varieties. Programs for
conservation of plant genetic resources for food security and agriculture were thus initiated
and genebanks established in many countries. The main objective was to collect and
maintain the genetic diversity in order to ensure its continued availability to meet the needs
of different users. The concept of germplasm conservation demands that collection methods
initially capture maximum variation and subsequently, conservation and regeneration
techniques minimize losses through time. To this effect, plant genetic resources (PGR)
conservation activities comprise of collecting, conservation and management, identification
of potentially valuable material by characterization, and evaluation for subsequent use.
Advances in biotechnology, especially in the area of in vitro culture techniques and
molecular biology provide some important tools for improved conservation and
management of plant genetic resources.

Types of Germplasm

Advanced (or elite) germplasm includes (a) "cultivars," or cultivated

varieties, which are suitable for planting by farmers, either recently
developed cultivars or "obsolete" cultivars that are no longer grown, and
(b) advanced breeding material that breeders combine to produce new
cultivars (sometimes referred to as "breeding materials").

Improved germplasm is any plant material containing one or more traits of interest that have
been incorporated by scientific selection or planned crossing.

Landraces are varieties of crops improved by farmers over many generations without the
use of modern breeding techniques. Within a modern breeding program, landraces are
sometimes used for resistance traits, and extensive efforts are generally required before
their genes can be used in a final variety.

Wild or weedy relatives are plants that share a common ancestry with a
crop species but have not been domesticated. These plants can serve as
another source of resistance traits, but these traits can be very difficult to
incorporate in final varieties.

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 Plant Genetic Resources and Conservation

Genetic stocks are mutants or other germplasm with genetic

abnormalities that may be used by plant breeders for specific purposes.
Genetic stocks are often used for highly sophisticated breeding and basic
research.

Why Is Germplasm Important?

The relationship between access to genetic resources and agricultural production is often
overlooked. The plant breeding process is complex and continual, and diverse genetic
resources are a critical input. Advances in yield potential, pest resistance, quality, and other
desirable traits in modern varieties have resulted from professional breeders crossing
diverse parental genetic material.

Farmers who rely on their crop output for seed or consumption and professional plant
breeders both depend on crop genetic resources. In turn, the efforts of farmers and plant
breeders can generate new genetic resources. About 10,000 years ago, people in parts of
Asia, the Near East, and Mesoamerica (modern-day Mexico and Central America) began to
deliberately cultivate specific species. Over the generations, farmers selected and improved
particular crops. In many parts of the world, this process continues today with farmer-
developed varieties known as landraces. Landraces have been adapted to specific
environments, and the areas in which they grow host many diverse varieties.

GERMPLASM COLLECTING AND TECHNIQUE USED FOR PREPARATION AND COLLECTION

Collecting involves gathering samples of a species from populations in the field or natural
habitats for conservation and subsequent use. Collecting may be easy in species producing
small botanic seeds in abundance.

However, it becomes problematic when seeds are unavailable or non-viable due to (a)
damage of plants by grazing or diseases (b) large and fleshy seeds that are difficult to
transport; or (c) where samples are not likely to remain viable during transportation due to
remoteness of the collecting site from the genebank.

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 Plant Genetic Resources and Conservation

Advances in biotechnology provide useful solutions for collecting such problem species:

Plant Difficulty Solution

in vitro techniques have
been developed that allow
collecting of the relatively
small zygotic embryos in
coconut the field and transporting
large size of the seeds
Cocos nucifera them back in sterile
conditions to the
laboratory to inoculate and
germinate them on a
culture medium.
simple in vitro method that
involved collecting shoot
rapid deterioration of
nodal cuttings, followed by
cacao samples during transit as the
sterilization and inoculation
Theobroma cocoa seeds do not withstand
of tissue into prepared
desiccation.
culture vials containing
semi-solid medium.

In vitro collecting methods were also developed for a range of other species including oil
palm, forage grasses, banana, coffee, grape, Prunus and Citrus spp.

CONSERVATION – MODERN BIOTECHNOLOGY METHODS

There are two approaches for conservation of plant genetic resources, namely in situ and ex
situ. In situ conservation involves maintaining genetic resources in the natural habitats
where they occur, whether as wild and uncultivated plant communities or crop cultivars in
farmers’ fields as components of the traditional agricultural systems. Ex situ conservation on
the other hand, involves conservation outside the native habitat and is generally used to
safeguard populations in danger of destruction, replacement or deterioration. Approaches
to ex situ conservation include methods like seed storage, field genebanks and botanical
gardens.

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 Plant Genetic Resources and Conservation

Conventional ex situ conservation

seed storage
Methods Problems Solution
• most convenient for • tropical and sub-tropical clonal propagation
long-term conservation. tree produce recalcitrant preferred to conserve elite
• involves desiccation of seeds that quickly lose genotypes.
seeds to low moisture viability and do not
contents and storage at survive desiccation.
low temperatures. • also a number of other
important crop species
that are sterile or do not
easily produce seeds, or
seed is highly
heterozygous

field genebanks
Methods Problems Examples
provide easy access to run the risk of destruction by banana, sweet potato,
conserved material for use natural calamities, pests and sugarcane, cassava, yam,
diseases potato, and taro.

Biotechnology contributed significantly by providing complementary in vitro conservation

options through tissue culture techniques. In vitro conservation also offers other distinct
advantages. For example, the material can be maintained in a pathogen-tested state,
thereby facilitating safer distribution. Further, the cultures are not subjected to
environmental disturbances. In general, they fall under two categories: (i) slow growth
procedures and (ii) cryopreservation.

Slow growth
allow clonal plant material to be held for 1-15 years under tissue culture conditions with
periodic sub-culturing, depending on species
Methods
• a low temperature with low light intensity is used to limit growth.
• temperatures in the range of 0-5ºC are employed with cold tolerant species, but for
tropical species, temperatures between 15º and 20ºC are used.
• limit growth by modifying the culture medium, mainly by reducing the sugar and/or
mineral elements concentration and reduction of oxygen level available to cultures.
Protocols
• Protocols for clonal multiplication are well established for several species.
• Generally, organized cultures such as shoots are used for slow growth storage since
undifferentiated tissues such as callus are more vulnerable to somaclonal variation.
Although slow growth procedures have been developed for a wide range of species,
they are routinely used for conservation of genetic resources of only a few species
including Musa spp., potato, sweet potato, cassava, yam, Allium spp. and temperate
tree species.

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 Plant Genetic Resources and Conservation

Cryopreservation
storage of plant material at ultra-low temperatures in liquid nitrogen (-196°C)
• At -196°C, cell division and metabolic activities remain suspended and the material
can be stored without changes for long periods of time.
• only available method for long-term conservation of vegetatively propagated plant
germplasm.
Choice of materials
cells, protoplasts, shoot apices, somatic embryos, seed or excised zygotic embryos
Advantages
• limited space.
• protects material from contamination.
• involves very little maintenance.
• cost-effective.
Technique
• older classical techniques based on freeze-induced dehydration of cells.
• newer techniques based on vitrification
Classical technique
• tissues are cooled slowly at a controlled rate (usually 0.1-4°C/min) down to about –
40ºC, followed by rapid immersion of samples in liquid nitrogen using a
programmable freezing apparatus.
• cryoprotectants are added to the freezing mixtures to maintain membrane integrity
and increase osmotic potential of the external medium.
• have been successfully applied to undifferentiated culture systems such as cell
suspensions and calluses

Vitrification
• involve removal of most or all freezable water by physical or osmotic dehydration of
explants, followed by ultra-rapid freezing which results in vitrification of intracellular
solutes.
• more appropriate for complex organs like embryos and shoot apices; they are also
less complex and do not require a programmable freezer, hence are suited for use in
any laboratory with basic facilities for tissue culture.
• there are seven vitrification-based procedures in use for cryopreservation:
(1) encapsulation-dehydration, (2) vitrification, (3) encapsulation-vitrification,
(4) desiccation, (5) pregrowth, (6) pregrowth desiccation, and (7) droplet freezing.

In general, cryopreservation is well established for vegetatively propagated species.

However, it is much less advanced for recalcitrant seed species due to some of their
characteristics, including their very high sensitivity to desiccation, structural complexity and
heterogeneity in terms of developmental stage and water content at maturity. The new
cryopreservation techniques have been successfully applied for more than 80 species (some
examples: rice, wheat, barley, mustard and coconut) and they are under development or
vigorous testing for several other species. However, examples of their routine use for long-
term conservation are still limited only to oil palm and potato.

CHARACTERIZATION OF DIVERSITY
The ability to identify genetic variation is indispensable to effective management and use of
genetic resources.

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 Plant Genetic Resources and Conservation

Traditional method
• measuring variation in phenotypic traits such as flower colour, growth habit or
quantitative agronomic traits like yield potential, stress tolerance etc.
• has certain limitations: genetic information provided by morphological characters is
often limited and expression of quantitative traits is subjected to strong environmental
influence.
Biochemical method
• based on seed protein and enzyme electrophoresis; introduced in 1960s.
• useful in analysis of genetic diversity as they reveal differences between seed storage
proteins or enzymes encoded by different alleles at one or more gene loci.
• eliminates the environmental influence.
• limited due to their inability to detect low levels of variation.
Molecular method
• used as complementary strategies to traditional approaches; increasingly playing an
important role in conservation and use of plant genetic resources.
• can be performed at any growth stage using any plant part and it requires only small
amounts of material.
• methods differ with respect to technical requirements, level of polymorphism
detected, reproducibility and cost.
• generally based on the use of restriction enzymes that recognize and cut specific short
sequences of DNA (e.g., Restriction Fragment Length Polymorphism, RFLP) or
polymerase chain reaction (PCR), which involves amplification of target DNA sequences
using short oligonucleotide primers.
• Specific areas in which molecular marker techniques have been used are: developing
sampling strategies and identification of gaps in the collections to plan for future
collecting and acquisition, and managing conserved germplasm – including
identification of redundancies, development of core collections, fingerprinting,
identification of genetic contamination and quantification of genetic drifts/shifts.

Nevertheless, it is difficult to compare the different techniques and determine which one is
best and for what purpose because each method has its advantages and disadvantages. The
appropriateness of individual marker systems also varies depending on the objective of
study and the properties of the species.

Sampling strategies

Molecular markers have been applied to study genetic diversity from natural populations
and formulate efficient sampling strategies to capture maximum variation for genetic
resources conservation. For example, the substantially higher level of RFLP variation
observed in self-incompatible, as compared with self-compatible species of Lycopersicon was
used to recommend predominant sampling of self-incompatible species for germplasm
acquisition. Studies of distribution of genetic diversity using AFLP markers in Sri Lankan
coconut populations showed that emphasis should be placed on collecting relatively large
numbers of plants from few populations since most of the diversity is within populations
rather than between populations. Genetic variation within and between natural populations
of Digitalis obscura was quantified using RAPDs and the results were used for optimizing
sampling strategies for conservation of genetic resources of the species. Recommendations
to focus on the sampling of marginal pawpaw (Asimina triloba) populations in future
collection missions were derived from the genetic structure across natural distribution areas,
established by RAPD analysis.

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 Plant Genetic Resources and Conservation

Managing genetic resources

Molecular techniques proved useful in a number of ways to improve the conservation and
management of PGR. In particular, genetic diversity data provides information on gaps in
terms of coverage in gene pools as well as redundancies, i.e., material with similar
characteristics that wastes resources through increased cost of management. For example,
RAPD analysis in Brassica oleracea revealed that 14 phenotypically uniform accessions could
be reduced to 4 groups with minimal loss of genetic variation. Intraaccession variation was
determined by AFLP markers in ex situ conserved barley, and results were used to evaluate
the efficiency of splitting heterogeneous accessions into distinct lines in order to avoid the
negative effects of selection and genetic drift during regeneration. Recent examples of use
of molecular markers to identify redundancies in collections include perennial kales, wheat,
grapevine, sorghum, cassava, flax, and barley.

Molecular markers have been employed for fingerprinting, verification of accession identity
and genetic contamination. For example, microsatellites were used to distinguish different
cultivars of grapevine, and to compare landraces and develop unique DNA profiles of
soybean genotypes. Variation within species has also been studied to explore geographic or
ecological patterns of distribution of diversity in many different crops and their wild relatives
that include wild bean (Phaseolus vulgaris), banana, mango, bambara groundnut, vetch,
Cicer spp., sorghum, sweet potato, tea, and chicory.

Molecular markers are being increasingly used to resolve problems of taxonomy and
phylogenetic relationships, as a good knowledge of genomic homologies helps in devising
suitable breeding strategies for appropriate conservation as well as transfer of genes from
one species to another.

In situ conservation

Diversity studies using molecular markers have also assisted in developing in situ
conservation strategies. The advent of molecular genetic markers made it easy to
discriminate between wanted and unwanted agronomic genes in segregating populations. If
linkages are established between a heritable agronomic trait and a genetic marker, markers
can be used to identify the location of genes. Such linkages allow direct selection for the trait
using marker assisted selection in a backcrossing programme. Molecular markers, which
densely cover an entire crop genome, can be applied to develop a molecular map for a crop,
which could be used to determine linkage between a specific molecular marker and a
strongly heritable trait. This holds great promise for breeding programmes, as many traits
are difficult to select for directly from breeding populations.

CONCLUSION
Biotechnology has made significant contributions to improved conservation and use of plant
genetic resources. The rapid progress made in in vitro culture technology has helped in
improving the conservation of genetic resources especially of problem species. Some of the
most important contributions have been in the areas of in vitro collecting, slow growth and
cryopreservation. Slow-growth techniques are in a more advanced state of development
than cryopreservation techniques, which still require improvement before they can be used
on a routine basis in a number of species. By facilitating better understanding of diversity,
both in extent and structure, molecular marker techniques are proving extremely useful in
identification of redundancies in collections, in testing accession stability and integrity, and
in supporting the development of effective management strategies both for ex situ and in

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 Plant Genetic Resources and Conservation

situ conservation. Molecular genetic studies are also being increasingly used to support
improved use of plant genetic resources. The sequence data that are becoming available for
increasing numbers of genes as a result of the rapid advances in DNA technology have
stimulated the development of novel molecular technologies which allow the screening of
germplasm for functional diversity and identify variation at an early developmental stage
without the need for performing the time-consuming evaluation tests.

International Issues and Agreements

Historically, plant genetic material was generally freely collected and shared. Today’s
developing countries-with a wealth of biological diversity in situ (in the wild and on fields)-
were often the source of raw genetic material collected by public gene banks worldwide.

Now, however, critics argue that unrestricted access to germplasm unaccompanied by

benefit sharing results in an inequitable system of exchange. For example, freely shared crop
traits from donor countries could be incorporated into varieties by researchers in developed
countries and then sold back to donor country farmers by private seed companies. The lack
of direct compensation is seen as giving donor countries little incentive to conserve genetic
resources, some of which are now at risk of extinction. Proponents counter that a system of
“free exchange” indirectly compensates lower income countries for donations of raw genetic
materials in two ways. First, these countries have had free access to public gene banks,
whose holdings include improved varieties. Second, many lower income countries are net
importers of food, and consumers in those countries benefit from lower world food prices
made possible by genetic improvements, regardless of where the improvements were made.

Several international agreements have sought to further the preservation of genetic

resources and to balance the sharing of benefits generated by their use. In 1983, the
Commission on Plant Genetic Resources (now the Commission on Genetic Resources for
Food and Agriculture) was established under the auspices of the Food and Agricultural
Organization (FAO) of the United Nations. The Commission developed the International
Undertaking, a nonbinding treaty to govern the exchange of genetic resources, but some
developing and developed countries (including the U.S.) did not commit to its
implementation. In 1992, the U.N. Convention on Biological Diversity (CBD) was established,
with a focus on the preservation of biodiversity, especially those genetic resources with
pharmaceutical and industrial rather than agricultural uses. In an attempt to ensure
equitable returns to donor countries for the use of native resources (and to spur
conservation), the CBD granted nations sovereign rights to genetic resources within their
borders, which in practice meant both nonagricultural and agricultural germplasm.

International agreements on intellectual property rights also have implications for genetic
resource conservation. Stronger intellectual property rights provide incentives for private
research and development (R&D) investment, and, in theory, also enhance incentives for
conserving genetic resources. However, intellectual property law varies from country to
country and may not cover unimproved germplasm and farmer-developed varieties. The
World Trade Organization’s (WTO) agreement on Trade-Related Aspects of Intellectual
Property Rights has provisions that can affect the exchange of germplasm. WTO member
countries must commit to implementing a system protecting intellectual property for plant
genetic resources, and noncompliance can result in sanctions.

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 Plant Genetic Resources and Conservation

The New Treaty

The new International Treaty on Plant Genetic Resources for Food and Agriculture was
intended to bring the International Undertaking into conformity with the CBD. After lengthy
negotiations, delegates from 116 countries adopted the text of the treaty in November
2001, with the American and Japanese delegates abstaining.

The new treaty has several objectives. First, it mandates the conservation and sustainable
use of plant genetic resources for food and agriculture. Second, it seeks fair and equitable
sharing of benefits arising out of the use of these resources. Finally, it establishes a
multilateral system to facilitate access to all crops listed in Annexes I and II of the treaty and
to share the benefits derived from such facilitated access under the terms of a standard
Material Transfer Agreement (MTA). The treaty specifies that the Governing Body at its first
meeting will establish the terms of the standard MTA after the treaty enters into force.

Much remains to be resolved. Application of intellectual property rights to plant genetic

resources remains a contentious issue. Precisely how benefits will be shared has yet to be
determined and is complicated by: (a) A lack of consensus regarding what “equitable”
benefit sharing means (b) Disagreement over how to estimate the magnitude of benefits
derived from use of shared Germplasm (c) Substantial variability in benefit estimates derived
from similar assessment methods.

Unlike the CBD, which provides for bilateral negotiations to establish the terms of access and
benefit sharing for each specific exchange of materials, all germplasm exchanges under the
multilateral system will be subject to the standard MTA. Monetary benefits will be paid to a
fund established by the Governing Body. This fund will be used primarily to support farmers
who conserve and sustainably use plant genetic resources for food and agriculture,
especially such farmers in developing countries or in countries with economies in transition.

The new treaty addresses the financing of germplasm conservation only in general terms,
making this aspect of the treaty potentially difficult to implement. The overall impact of the
treaty is also limited by its omission of soybeans, peanuts, and other major world crops from
the list of 35 crops covered.

Crops covered under the International Treaty on

Plant Genetic Resources for Food and Agriculture
Apple Breadfruit Pea
Major aroids: includes Carrot Pearl millet
taro, cocoyam, dasheen, Cassava Pigeon pea
and tannia Chickpea Potato
Brassica complex: Citrus Rice
includes Coconut Rye
cabbage, rapeseed, Cowpea Sorghum
mustard, cress, Eggplant Strawberry
rocket, radish, and turnip Faba bean /Vetch Sunflower
Asparagus Finger millet Sweet potato
Banana/Plantain Grass pea Triticale
Barley Lentil Wheat
Bean Maize (corn) Yam
Beet Oat

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 Plant Genetic Resources and Conservation

Despite the progress made, there remain many unresolved questions, the most important
being that of determining the most appropriate molecular markers for the required
understanding of the patterns of diversity in specific studies. While there is a pressing need
to ensure that available technologies are made accessible to a wider range of users through
improved training and other capacity building initiatives, the existing technologies are also
expensive and given that most of the crop diversity is to be found in developing countries,
the issue of resources assumes importance. Hence, there is real need to maximize synergy
through appropriate collaboration between various national, sub-regional and international
levels, including sharing burdens and responsibilities, in order to use these techniques for
effective conservation and use of plant genetic resources.

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