-
X
Conservation Strategies for Native Plant Species
and their Sustainable Exploitation: Case of the
Balkan Botanic Garden of Kroussia, N. Greece
Eleni Maloupa1 Nikos Krigas1,2 Katerina Grigoriadou1 Diamanto
Lazari3
Georgios Tsoktouridis1*
1 Laboratory of Conservation and Evaluation of the Native and
Floricultural Species-Balkan Botanic Garden of Kroussia, National
Agricultural Research Foundation, P.O.
Box 60125, GR-570 01, Thermi, Thessaloniki, Greece 2 Laboratory
of Systematic Botany & Phytogeography, Department of Botany,
School of Biology, Aristotle University of Thessaloniki, GR-54124,
Thessaloniki, Greece 3 Department of Pharmacognosy - Pharmacology,
School of Pharmacy, Aristotle University of Thessaloniki, GR-54124,
Thessaloniki, Greece
Corresponding author: * [email protected]
Keywords: GIS, ITS1, ITS2, propagation, molecular markers,
native plants, Origanum dictamnus, sustainable cultivation,
taxonomy
ABSTRACT
This study outlines a pioneer initiative concerning the native
plant conservation that is considered as one of the most important
plant refuges in
Europe. The Balkan Botanic Garden of Kroussia has designed a
system built on people, infrastructure and a strategy of seven
hierarchical and
complementary policies. To formulate and implement this
strategy, we have created a flexible research team with scientists
from different
disciplines that collaborate interdisciplinary. In this team,
information flows horizontally, experience and expertise are joined
and used collectively
and know-how is delivered vertically to all interested or
involved parties. In the frame of this strategy, target plants are
being initially explored,
located and collected from the wild and Important Plant Species
are maintained, evaluated and studied. Explicitly documented living
plant
collections are maintained with classical and innovative methods
and species-specific propagation protocols for wild plants are
being developed.
Environmental awareness is promoted and several educational
activities on the native biodiversity are organized. Numerous plant
conservation
actions are undertaken; all attempt to integrate the ex-situ
with the in-situ plant conservation and contribute to the
implementation of the targets
of the Global Strategy for Plant Conservation at local, regional
and (inter-) national levels. Furthermore, evaluation of the
medicinal, cosmetic,
flavouring, floricultural and ornamental value of selected
native plants is performed, aiming at their sustainable
exploitation in collaboration with
the state, the stakeholders and the market. This approach has
the ultimate goal to deliver promising and unique new crops that
are carefully
selected and designed, sustainably produced and managed,
successfully launched and fair traded internationally.
1. TARGET AREA: NATURAL FEATURES AND THE RICHNESS OF THE GREEK
FLORA
Greece has an exceptionally rich flora with more than 5,700
native taxa (species and subspecies); about 15-20% are unique,
found nowhere
else in the world (Greek endemic taxa), presenting the highest
degree of endemism for any comparable territory in Europe and
the
Mediterranean region (Strid and Tan 1997).
This figure is all the more striking considering the relatively
small area (132,000 km2) and the complex topography of the country.
Much of
the land is wild, rugged, mountainous, with more than 40% of the
national area lying above 500 m of altitude (including 314
mountains and 1,674
individual peaks above 1,000 m; Strid and Tan 1997). Varied
climatic conditions may be found in Greece ranging from subtropical
to subalpine
environments. The geology of the area is diverse and dissected,
mainly with limestone massifs, serpentine regions, schistose and
granite
mountains. A mosaic-like forest cover is estimated at ca. 18%,
including forest, open woodland and scrub and at least 400 wetlands
of various
sizes can be found in Greece. The Greek coastline stretches
along more than 15,000 km, one of the longest in Europe, while
about 3,000
islands and islets dispersed in the Aegean, Ionian and Cretan
Seas, comprising ca. 19% of the land area of Greece.
The Greek territory can be divided into 13 different floristic
regions, the borders of which almost follow natural geographic
features such as
rivers and lowland areas between mountains (Fig. 1).
Abbreviations: ABS, Access and Benefit-Sharing; BBGK, Balkan
Botanic Garden of Kroussia; CBD, Convention on Biological
Diversity; GIS, Geographical
Information Systems; GSPC, Global Strategy for Plant
Conservation; IPS, Important Plant Species; IPEN, International
Plant Exchange Network; ITS1, Internal
Transcribed Sequence 1; MAT, Mutually Agreed Terms; MTAs,
Material Transfer Agreements; PIC, Prior Informed Consent
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Maloupa et al. Conservation strategies for native plants in
Greece
Generally, mountain summits and islands seem to host the
rarest endemic wild plants. Such plants, confined as they are
to
Greece, are a treasure of international importance, whose
conserva-
tion will benefit future generations not just in Greece but in
the whole
world; the rarity of many of the Greek endemic plants makes
them
vulnerable to extinction through human activity, a loss not only
to
Greece but to the world (IUCN Threatened Plants Committee
Secretariat 1982).
In total, four areas in Greece are considered as European
Centers of Plant Diversity and Endemism (CPDEu14-17; Akeroyd
and Heywood 1994), 10 areas have been declared as National
Parks, 10 wetlands are included in the Ramsar Convention,
575
areas are considered under protection and 270 sites have been
pro-
posed to be included in the EU NATURA 2000 Network.
Nevertheless, the rich and unique native flora of Greece is
being
threatened by global warming, fires, land reclamation,
over-grazing
and current urban and tourism expansion.
Though Greece is no doubt a hot-spot for global biodiversity
(Akeroyd and Heywood 1994) due to its exceptionally rich and
unique flora, it has no long tradition in botanic gardens
dedicated to
plant conservation.
2. THE ONLY NATIVE PLANTS POLICY
In-situ conservation of rare and endangered plant species is
no
doubt indispensable and it may be achieved basically with
habitat
and ecosystem conservation. Nevertheless, in-situ conservation
is
also a difficult task to achieve since effective protection
regimes are
not easily applied in the wild, they are costly and difficult to
control and evaluate. The ex-situ conservation of rare plant taxa
in botanic gardens is
an internationally acknowledged contribution to species
conservation (CBD 1992), able to prevent the extinction of
endangered species (e.g.
Diplotaxis siettiana, Lotus berthelotii, Lysimachia
minoricensis) and able to guaranty their possible re-introduction
in the wild. The majority of
botanic gardens feel proud of their botanic collections of rare
or endangered plant species from every corner of the world and
these displays are
often a major attraction for visitors.
The Balkan Botanic Garden of Kroussia, N. Greece (BBGK) is a
newly established botanic garden founded in 2000, as an initiative
of the
National Agricultural Research Foundation of Greece (NAGREF).
Today it covers an area of 31 ha, located in Mt. Kroussia (Northern
Greece), at
600 m of altitude, within a natural deciduous oak forest. Since
its establishment, the BBGK in order to focus exclusively to the
ex-situ
conservation of native plants of Greece and the Balkans has
decided to leave aside the exotic plants and ornamental species
commonly found in
other botanic gardens of Europe and the world.
All plants of the displays and the ex-situ conservation sections
in the BBGK are native plant species originating in the wild (Fig.
2).
Fig. 1 Collection areas of native Important Plant Species (IPS:
rare, endangered,
vulnerable, protected and endemic taxa of Greece and/or the
Balkans, plants with
potential ornamental and/or medicinal value) currently in
ex-situ conservation (white
circles) at the Balkan Botanic Garden of Kroussia (BBGK), N
Greece and their distri-
bution across the different phytogeographical regions of Greece
(Strid and Tan
1997). Every red dot in the map concerns at least one botanic
expedition and col-
lection of at least one accession number of IPS (Krigas et al.
2007).
Fig. 2 Schematic representation of the Balkan Botanic Garden of
Kroussia, N Greece.
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Maloupa et al. Conservation strategies for native plants in
Greece
3. THE IMPORTANT PLANT SPECIES POLICY Since 2001, BBGK has
launched a series of
botanic expeditions to every different phytogeo-
graphic region of Greece in order to obtain wild
propagation material of native plants (Fig. 1;
Krigas et al. 2007).
With a special permit provided by the Ministry
of Rural Development and Foods, renewed every
year, the scientific staff of the BBGK is enabled to
collect wild plant material even from NATURA
2000 sites, Nature Reserves, National Parks and
other protected areas of Greece. The BBGK,
combining conservation principles and sustainable
plant exploitation policies, has formulated priorities
for the collection of propagation material of wild
plants. Five target-plant categories have been pri-
oritized:
(i) Greek endemic plants (plant species found
exclusively in Greece and nowhere else in the
world); This category includes (a) single-island en-
demics e.g. Origanum dictamnus, Campanula hie-
rapetrae and Ebenus cretica (Crete island), Limo-
nium arcuatum (Corfu island), Anchusa samothra-
cica (Samothraki island), Viola cephalonica (Fig. 3A; Mt. Aenos,
Cephallonia island) etc, (b) single-mountain endemics e.g.
Helichrysum sib-
thorpii (Fig. 3B) and Anthemis sibthorpii (Mt. Athos), Thymus
plasonii (Mt. Chortiatis), Crocus hadriaticus subsp. parnassicus
(Mt. Parnassos),
Achillea occulta (Mt. Koulochera), Centaurea cithaeronea (Mt.
Kithaeronas) etc., (c) single-area narrow endemics e.g. Limonium
antipaxorum
(Paxi and Antipaxi islands), Limonium ithacense (Cephallonia and
Ithaca islands), Muscari cycladicum (Cyclades and Crete) etc, (d)
regional
endemics (restricted to few phytogeographical regions) e.g.
Crocus hadriaticus subsp. hadriaticus, Scaligeria moreana, Stachys
ionica (Fig. 3D),
Campanula incurva (Fig. 3C) etc. and (e) national endemics
(restricted to numerous phytogeographical regions of Greece) e.g.
Anchusella
variegata, Cerastium candissimum, Dianthus corymbosus, etc.
(ii) Narrow Balkan endemics; This category includes native
plants occurring around the boundaries of Greece with neighbouring
Balkan
countries e.g. Lilium rhodopaeum, Centaurea pawlowskii,
Marrubium thessalum, Stachys iva, etc.,
(iii) Other rare taxa found in Greece; This category includes
native plant species of wider distribution than (i), (ii) or (iii)
with one or only a few
scattered populations in Greece (e.g. Poa molinieri, Datisca
canabina, Galanthus nivalis, Diathus crinitus, etc.),
(iv) Balkan (sub-) endemics; This category includes taxa found
exclusively in the Balkan countries and/or extending to W Turkey
and/or
parts of Italy with scattered populations e.g. Dianthus
giganteus, Hypericum olympicum, Scabiosa crenata subsp.
dellaportae, Thymus thracicus,
etc.) and
(v) Potentially ornamental and/or medicinal plants native to
Greece and/or the Balkans (e.g. Geranium macrorrhizum,
Coridothymus
capitatus, Digitalis grandiflora, Salvia officinalis, Crataegus
monogyna, etc.).
All plant species of groups (i), (ii) and (iii) and numerous
species of group (iv) have been characterized as Other Important
Plant Species
(IPS) of the NATURA 2000 Network (in total 1,853 IPS for Greece,
Kokkini et al. 1996).
To date, at least 60 botanic expeditions have been organized
(Fig. 1) and more than 1,200 taxa (>20% of the Greek flora) have
been
collected from the wild (ca. 2,300 accession numbers). All
native taxa are currently cultivated and maintained ex-situ in
BBGK. In total, ca. 40%
of them belong to target-plant groups (i), (ii), (iii), (iv) and
another ca. 45% belong to group (v).
4. THE EXPLICIT PLANT DOCUMENTATION POLICY
4.1. Preparation of the botanic expeditions
A complete work about the flora of Greece is absent nowadays.
Floristic data are widely scattered to numerous scientific papers
and various
modern floras. The Flora Hellenica (Strid and Tan 1997, 2002),
the only complete work for the flora of Greece, has produced
hitherto only two
volumes (out of 10 expected). The Mountain Flora of Greece
(Strid 1986; Strid and Tan 1991) covers exclusively mountain
summits above ca.
1,500 m. The Flora Europaea (Tutin et al. 1968-1980) lack
significant amount of information concerning the East Aegean
Islands of Greece,
recently described taxa and updated distribution areas of plant
species, while the Flora of Turkey and the East Aegean Islands
(Davis 1965-
1985) covers only the flora of the East Aegean Islands of
Greece. As a result, numerous areas of Greece are considered as
floristically ill-
explored and therefore botanists may still encounter and
describe taxa new to science (e.g. Constantinidis and Calpoutzakis
2005; Snogerup et
al. 2006).
Consequently, the scientists of the BBGK, prior to every botanic
expedition scheduled to a specific area, have to:
(a) Review and study in advance all the published floristic
literature concerning the target-area,
(b) Prepare working lists of the known IPS cited from this
area,
Fig. 3 (A) Viola cephalonica, (B) Helichrysum sibthorpii, (C)
Campanula incurva, (D) Stachys ionica in
their collection sites.
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Maloupa et al. Conservation strategies for native plants in
Greece
(c) Group target-plant species, target-sites and target-routes
that have to be followed in the field,
(d) Assess the target-plant groups, sites and routes based on
estimated cost vs. on the expected benefit.
4.2. Collection of data
Field and in-situ collection data are indispensable in many
ways. Every botanic collection is considered unique by the
combination of the
collector(s)-area-date information. Documentation takes place by
using accession numbers (specific codes for different plants from
different
localities), when only explicit and precise information is
provided i.e. geographical coordinates and site description,
specific location, region,
prefecture, and country; otherwise, no accession number is given
to plant collections arriving with poor documentation and no
subsequent
propagation is planned.
Habitat information for each plant individual collected from the
wild (substrate, soil type, forest zone, habitat type, slope and
altitude) is
valuable and necessary for future ex-situ conservation actions
(Krigas et al. 2007). All these site and habitat characteristics
mentioned above,
are documented in special collection forms in-situ, which
accompany each accession number throughout the subsequent
propagation.
4.3. Taxonomic identification and nomenclature
Taxonomic identification of herbarium specimens and collected
propagation material from the wild is indispensable (Jarvis 2007).
A plant species
without a name does not exist in terms of science and
conservation (Taylor 2007). Each plant species has a unique
scientific name; this is the
tag that allows to be found, counted, researched, and monitored,
encoding and classify among other plants in the world; additionally
this is the
index key that retrieves everything we know about it from
various sources (Taylor 2007). The value of accessions in botanic
gardens depends
critically on the correct identification and nomenclature,
either they are utilized as conservation resources, either as
reference material; the
consequences of inaccurate taxonomy could be very serious or
even fatal (Heywood 2007).
Nomenclature of plant taxa conserved in the BBGK follows Strid
and Tan (1997, 2002), for the taxa covered by Flora Hellenica,
Strid (1986)
and Strid and Tan (1991), for the taxa included in the Mountain
flora of Greece, and Tutin et al. (1968-1980) for all the rest.
4.4. Evaluation of the Important Plant Species (IPS)
The mother stock plants maintained in the nursery of the BBGK,
are periodically evaluated according to the IUCN status (IUCN
2001), endemism
category, inclusion in Other Important Plant Species of EU
NATURA 2000 Network (Kokkini et al. 1996), inclusion in national
and/or international
catalogues and/or conventions (e.g. Greek Presidential Decrees,
EU Directives and Annexes, Red Lists, CITES, Bern Convention,
etc.). This is
exercised in order to identify the Important Plant Species (IPS)
in the BBGKs mother plantations, meriting special ex-situ
conservation
programmes as defined by the International Agenda of Botanic
Gardens in Conservation (Wyse-Jackson and Sutherland 2000). Under
this view,
four groups of IPS are identified as Priority Important Plant
Species (Priority IPS; Table 2):
As Priority 1 IPS are designated (a) all taxa included in
national and/or international catalogues and/or conventions (e.g.
Greek Presidential
Degrees, EU Directives and Annexes, Red Lists, CITES, Bern
Convention etc), regardless of endemism, as well as (b) narrow
Greek endemics
(single-island, single-mountain, single-area or regional
endemics), which are not included in national and/or international
catalogues and/or
conventions.
Table 1 Ex-situ cultivation guidelines in the Balkan Botanic
Garden of Kroussia regarding Important Plant Species (IPS) based on
links of IPS collection data with
geodatabases (Krigas et al. 2007).
Source Type of variable Examples of attributes Guidelines for
the ex-situ cultivation of IPS in BBGK
ESDB v.2 (EC 2004) Soil moisture Topsoil and subsoil available
water capacity Selection of watering regimes
ESDB v.2 (EC 2004)
CORINE Soil
Classification (EC 1985)
Soil classes and types Textural class
FAO 1985 soil class
World Reference Base soil class
Dominant parent material
Selection of growing medium (texture, pH, drainage)
ESDB v.2 (EC 2004) Soil nutrient Topsoil and subsoil base
saturation
Cation exchange capacity
Selection of growing medium and fertilization regime
ESDB v.2 (EC 2004) Soil limitations Depth to a gleyed
horizon
Depth to rock
Depth of an obstacle to roots
Volume of stones
Selection of growing medium (texture, drainage)
and potting volume
WorldClim Database
(Guerin et al. 2002;
Hijmans et al. 2005)
Climate Mean minimum or maximum temperatures of the
coldest or the warmest month, respectively
Annual mean temperature range
Temperature seasonality
Mean diurnal temperature range
Precipitation of the driest month or the wettest month
Mean monthly precipitation
Annual precipitation
Mean precipitation of driest, wettest, coldest or
warmest quarter
Selection of appropriate growing sites and conditions in
greenhouse as well as in ex-situ cultivation
Selection of temperatures for seed germination
Selection of shading and ventilation
Variation of watering regimes per month
Digital Terrain Model
created
Topography Aspect
Slope
Altitude (elevation)
Customizing microclimate in ex-situ cultivation sites and
in positioning of plants in the displays
Mavromatis (1980) Vegetation zones 11 vegetation zones
CORINE Land cover
EC & ETC/LC (1999)
Land cover classes
and types
45 land-use classes and types
Selection of the ex-situ conservation sites
Selection of shading
Selection of species assemblages and sites for specific
plant displays
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Maloupa et al. Conservation strategies for native plants in
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Priority 2 IPS include all taxa which they are not included in
national and/or international catalogues and/or conventions but
they (a) are
endemic to more than a single phytogeopraphical area of Greece,
(b) are endemics of any other Balkan country and (c) are narrow
endemics to
the boundary areas of neighbouring Balkan countries.
All taxa which are endemic to the Balkan Peninsula and they are
not included in national and/or international catalogues and/or
conventions
are designated as Priority 3 IPS.
All taxa that are found exclusively in the Balkan countries
and/or extending to W Turkey and/or parts of Italy with scattered
populations and
they are not included in national and/or international
catalogues and/or conventions are designated as Priority 4 IPS.
Other plants native to Greece and/or the Balkans with potential
ornamental and/or medicinal value, which are not included to the
previously
mentioned categories, are designated as Priority 5 Plant
Species.
4.5. Species-specific baseline cultivation guidelines for IPS
using GIS
All collection data of taxonomically iden-
tified and evaluated as IPS are being in-
corporated in a GIS (Geographical Infor-
mation Systems) environment (Krigas et
al. 2007). The collection data are conse-
quently combined and/or verified with
data from ecological databases (Fig. 4);
this includes several topographic and
habitat features (altitude, slope, aspect,
vegetation zones, habitat types), climatic
conditions (minimum, maximum and
mean temperature per month, accumu-
lated temperature, minimum, maximum
and mean monthly and annual precipita-
tion, isothermality, Emberger ombrother-
mic quotient) and soil properties (soil
water capacity, soil types and dominant
parent material).
This approach (Krigas et al. 2007) is
being used to: (i) consider the in-situ eco-
logical amplitude of different IPS popula-
tions, (ii) inform and guide the ex-situ
growing conditions preferred and/or toler-
ated by each IPS, (iii) minimise the risk of
cultivation failure, (iv) select the best pos-
sible different treatments, (v) help or-
ganize better ecological groupings of
mother plants in the nurseries of BBGK
and (vi) provide species-specific baseline
cultivation guidelines for each IPS (Table
1; Figs. 5-6; Mouflis et al. 2007). This
approach may additionally facilitate gap
analysis of the botanic expeditions orga-
nized so far, permits better schedule of
next expeditions of BBGK, analyses and
reveals gaps in representation of IPS
from different altitudes, phytogeographic
and climatic regions of Greece, and fin-
ally is aimed to permit assessment of the conservation strategy
and actions of BBGK.
4.6. Distribution of plant material and the International Plant
Exchange Network Policy
The BBGK produces yearly an Index Seminum distributed in other
botanic gardens worldwide. All available plant accession numbers
maintained
in ex-situ conservation in BBGK are numbered according to the
International Plant Exchange Network (IPEN) (www.bgci.org). The
IPEN policy
has been adopted by botanic gardens in Europe as well as by the
BBGK in order to meet the provisions of 15 of the Convention on
Biological
Diversity (CBD 1992) in receiving, storing, and supplying plant
material, including free exchange only for non-commercial purposes
and free
exchange only between botanic gardens. Otherwise, regarding
institutions not sharing this policy and their request to access
the phytogenetic
resources of Greece, an Access and Benefit-Sharing (ABS) policy
is adopted by BBGK; this requires a Code of Conduct endorsing
bilateral
agreements with Mutually Agreed Terms (MAT), Prior Informed
Consent (PIC) and Material Transfer Agreements (MTAs), before the
distribution
of the plant material.
Fig. 4 Schematic representation of the link created in a GIS
environment between the collection data of IPS and
the data from ecological databases. (Table 1; Krigas et al.
2007)
Viola_cephThym_hol
Limo_ithSileCepCep
TmeanTmax
Tmin
17,917,7
15,0
9,2
32,232,3
29,5
23,96,7
6,3
3,9
-1,6-5,0
5,0
15,0
25,0
35,0
Temperature profiles
TmeanTmaxTmin
oC
Fig. 5 Temperature profiles (Tmean, Tmax, Tmin) regarding four
selected IPS of Cephalonia island (Viola
cephalonica (Fig. 3A), Thymus holosericeus, Limonium ithacense,
Silene cephallenia subsp. cephallenia. (Mouflis et
al. 2007).
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Maloupa et al. Conservation strategies for native plants in
Greece
Table 2 Example of evaluation of 50 native plant taxa (species
or subspecies) of Greece that are currently maintained in ex-situ
conservation at the Balkan Botanic
Garden of Kroussia aiming at assigning their potential
commercial value according to conservation priorities, aromatic and
pharmaceutical properties, impressive
botanical features, ornamental and/or horticultural interest,
propagation methods and cultivation regimes.
Use Impressive botanical features Taxon (species or
subspecies)
Family
Co
nse
rvat
ion
pri
orit
y
Aro
mat
ic p
rop
ertie
s
Ph
arm
aceu
tica
l pro
per
ties
Gar
den
orn
amen
tal
Cu
t fl
ow
er
Po
tted
pla
nt
Pro
pag
atio
n
Irri
gat
ion
Typ
e o
f so
il
Lar
ge
flow
ers
Den
se in
flore
scen
ce
Oth
er
Lea
ves
Po
st h
arve
st
/ tra
nsp
ort
atio
n
char
acte
rist
ics
Acanthus balcanicus Acanthaceae 3 T M Acid/ Clay Large d $
Achillea ageratifolia
subsp. ageratifolia
Asteraceae 2 A1 P1 S/iv L Lime Everlasting flowers
Glaucous b
$ Achillea chrysocoma Asteraceae 2 A P S/iv L Lime Hairy c
Achillea occulta Asteraceae 1 A1 P1 iv L Lime Everlasting
flowers
e
Adonis cyllenea Ranunculaceae 1 P1 T L Lime Large b Anthemis
sibthorpii Asteraceae 1 A1 P1 S/C L Lime Everlasting
flowers
a
Biebersteinia orphanidis Biebersteiniaceae 1 A P1 T M Sandy
-Clay
Large b #Campanula hierapetrae Campanulaceae 1 P1 S/iv L Acid
Velutinous a #Campanula incurva Campanulaceae 1 P1 S/iv M Acid
Pubescent a #Campanula pelviformis Campanulaceae 1 P1 S/iv L Acid
Hispid b Centaurea cithaeronea Asteraceae 1 A1 P1 iv L Lime
Everlasting
flowers
Glaucous b
Centaurea pawlowskii Asteraceae 1 A1 P1 S M Lime Coloured bracts
c #Cerastium
candidissimum
Caryophyllaceae 2 A C L Lime Glaucous c #Crocus cartwrightianus
Iridaceae 1 A1 P T M Lime Long orange
stigmas
b
#Crocus hadriaticus
subsp. hadriaticus
Iridaceae 1 A1 P1 T M Lime Coloured corolla throat
b
$ Crocus veluchensis Iridaceae 3 A1 P1 T M Lime Orange stamens
and stigma
b
$ Dianthus crinitus Caryophyllaceae 4 A1 P1 S/C/iv L Lime
Dissected petals Glaucous a Dianthus fruticosus
subsp. occidentalis
Caryophyllaceae 1 A1 P1 S/C/iv L Lime Toothed petals Fleshy b $
Dianthus
haematocalyx
Caryophyllaceae 2 A1 P1 S/C/iv L Lime Cushion form b $ Digitalis
lanata Scrophulariaceae 4 P S H Acid Coloured stems Hairy b $
Digitalis viridiflora Scrophulariaceae 4 P S M Acid b # Ebenus
cretica Fabaceae 1 P C L Lime d Erodium hartvigianum Geraniaceae 1
A1 P1 T L Lime Long fruits Hairy c Erysimum naxense Brassicaceae 1
P1 S L Lime Densely arranged
leaves
d
Fritillaria pelinea Liliaceae 1 A1 P1 T L Lime Glaucous-green
stems
Glaucous-
green shiny
c
$ Gentiana verna subsp.
balcanica
Gentianaceae 4 A1 P1 S H Acid Coloured calyces, compact form
a
$ Haberlea rhodopensis Gesneriaceae 1 Iv M Acid b # Helichrysum
sibthorpii Asteraceae 1 A1 P1 T M Lime Everlasting
flowers
Glaucous b
Heptaptera
colladonioides
Apiaceae 1 S L Lime Large d Hypericum rumeliacum
subsp. rumeliacum
Clusiaceae 3 A P C M Lime Densely arranged leaves
b
$ Inula ensifolia Asteraceae 1 A1 P1 T M Sandy/ Clay
b $ Iris reichenbachii Iridaceae 3 A1 P1 T M Sandy/
Clay
Compact form b $ Jasione heldreichii Campanulaceae 4 T M Lime b
Lilium rhodopaeum Liliaceae 1 A1 P1 S M Lime Orange long
stamens
Long, Shiny c
$ Linaria peloponnesiaca Scrophulariaceae 2 P1 T M Lime e
Muscari cycladicum Liliaceae 2 P1 T L Lime d # Origanum dictamnus
Lamiaceae 1 A P C/iv L Lime Hairy a $ Origanum onites Lamiaceae 4 A
P C L Lime a $ Paeonia mascula
subsp. hellenica
Paeoniaceae 1 A P T M Lime Large b $ Pancratium maritimum
Amaryllidaceae 4 P S/C L Sandy Glaucous-green
stems
Glaucous-
green
b
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Maloupa et al. Conservation strategies for native plants in
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5. THE DNA BARCODING POLICY
5.1. Molecular markers for plants
Molecular markers have been developed to
characterize/distinguish closely related plant species, moreover to
investigate the genetic diversity
and find the genetic relationship of plants grown in different
geographical areas. Potential target genes, which are conserved in
all plants, play
significant role in plant species evolution. These genes/genomic
areas are used to define polymorphisms among species and to
develop
molecular markers to characterize and find the genetic
relationships among them.
Selection of the appropriate molecular markers for species
identity is very difficult for plants. This is due to the large
genome of plants and
not unique high plasticity genomic regions they contain.
Depending on the intimacy of species individuals, the molecular
marker selection in
BBGK focus on using the matK and ndhF genes at family level,
while the nrDNA ITS1 and ITS2 regions for genus and species level.
To
differentiate individual plants at subspecies level, or within a
population having undistinguishable phenotypes, the MADS-box gene
pistillata and
three plastid DNA regions of the rbcL gene will be attempted to
apply. There are also many other useful molecular markers described
in the
literature applied in particular species, which may potentially
be used in cases that we could not find genetic differences in some
taxa.
The most widely used molecular markers for plants focus on the
cpDNA rapidly evolving genes, such as matK and ndhF (Soltis and
Soltis
1998), the phytochrome genes PhyA and PhyC (Mathews and Donoghue
1999), the RPB2 gene (encoding the RNA Polymerase II gene)
(Denton et al. 1998), two loci, Adc1 and Adc2, of the arginine
decarboxylase gene Adc, (Galloway et al. 1998), the granule-bound
starch
synthase gene (waxy) (Mason-Gamer et al. 1998), a portion of the
exon 1 of the phytochrome B gene PhyB (Mathews et al. 2000), a low
copy
nuclear gene encoding 4-coumarate: coenzyme A ligase (4CL) in
the lignin biosynthesis pathway (Wang et al. 2000), the alcohol
dehydrogenase
genes Adh1 and Adh2 (Sang et al. 1997), the
chloroplast-expressed glutamine synthase gene ncpGS (Emshwiller and
Doyle 1999), a portion of
the intron of a MADS-box gene pistillata and the cpDNA trnL
intron (Bailey and Doyle 1999), the vicilin gene (Whitlock and Baum
1999), the
single-copy nuclear genes Glb1 (Hilton and Gaut 1999) and cl
(Hanson et al. 1996), three plastid DNA regions of the rbcL gene,
the trnL-F
intron/intergenic spacer and the rps16 intron (Goldblatt et al.
2003) and many other molecular markers were also used and found in
literature
with very little significance. Additionally, the nrDNA ITS1 and
ITS2 regions, which have been widely used at interspecific level in
plant
phylogenetic studies (Baldwin et al. 1995) and it is nowadays
applied in thousands of species.
5.2. DNA barcoding for plants through the nrDNA ITS1 and ITS2
regions
Nucleotide sequence information of multicopy genes provide
accurate evidence of divergence in closely related organisms and
have been widely
used for phylogenetic inference. This is the main reason that
internal transcribed sequence (ITS) information of the nrDNA ITS1
and ITS2
regions were selected in BBGK to apply to the different
accession numbers of native Greek plants. The dynamics of DNA
sequences constitute
worldwide attention and significance for species identification
and origin. Therefore these sequences can be used as DNA barcodes
and
accompany every individual plant or vegetative clone propagated
in the nursery or tissue culture laboratories of BBGK, stimulating
the creation
Table 2 (Cont.)
Use Impressive botanical features Taxon (species or
subspecies)
Family
Co
nse
rvat
ion
pri
orit
y
Aro
mat
ic p
rop
ertie
s
Ph
arm
aceu
tica
l pro
per
ties
Gar
den
orn
amen
tal
Cu
t fl
ow
er
Po
tted
pla
nt
Pro
pag
atio
n
Irri
gat
ion
Typ
e o
f so
il
Lar
ge
flow
ers
Den
se in
flore
scen
ce
Oth
er
Lea
ves
Po
st h
arve
st
/ tra
nsp
ort
atio
n
char
acte
rist
ics
Pterocephalus perennis
subsp. bellidifolius
Dipsacaceae 3 P S/C M Acid Compact form c Satureja cuneifolia
Lamiaceae 2 A P C L Lime c $ Saxifraga rotundifolia
subsp. chrysosplenifolia
Saxifragaceae 2 A1 P1 T M Clay Dotted petals Rounded b Silene
orphanidis Caryophyllaceae 1 A1 P1 S L Lime Compact form c Stachys
ionica Lamiaceae 1 A P1 C M Clay/
Alkaline
Glaucous-green,
hairy
c
Staehelina uniflosculosa Compositae 2 P? C L Lime Everlasting
flowers
Large b
$ Thymus sibthorpii Lamiaceae 4 A P1 C L Lime d Thymus thracicus
Lamiaceae 4 A P1 C L Lime c Verbascum
xanthophoeniceum
Scrophulariaceae 4 A1 P1 T M Acid Rosette d Viola cephalonica
Violaceae 1 A1 P1 S M Lime b Empty cells concern absence of
information. Taxa marked with (#) concern narrow Greek endemics
that according to the Royal Horticultural Society Plant Finder
have
been or are currently purchased in nurseries of Great Britain,
while taxa (sub-) endemic to the Balkans are marked with
($).Conservation priorities 1: Taxa included in
national and/or international catalogues and/or conventions
(e.g. Greek Presidential Degrees, EU Directives and Annexes, Red
Lists, CITES, Bern Convention etc),
regardless of their endemism and/or narrow Greek endemics
(single-island, single-mountain or single-phytogeographic area
endemics). 2: Taxa endemic to more than a
single phytogeopraphical area of Greece or of other Balkan
country or endemic to the boundary areas of neighbouring Balkan
countries. 3: Taxa endemic to the Balkan
Peninsula. 4: Taxa found mainly in the Balkan countries and/or
extending to W Turkey and/or parts of Italy or other taxa rare in
Greece. A: Species with aromatic
properties. A1: Known aromatic properties in other species of
the genus. P: Species with pharmaceutical properties. P?: under
investigation. P1: Known
pharmaceutical properties in other species of the genus.
Propagation S: seed, C: cuttings, Iv: in vitro, T: testing in
process. Irrigation L: low. M: medium, H: high. Post
harvest / transportation characteristics: refer to desirable for
the market characteristics based on observations during
maintenance. a: very good, b: good, c:
medium, d: not good, e: bad.
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Maloupa et al. Conservation strategies for native plants in
Greece
of a database of documented reference sequences in a universal
library worldwide, to which comparisons of unidentified taxa can be
made.
In 2003, researchers at the University of Guelph in Ontario,
Canada, proposed DNA barcoding as a way to identify species
(see
www.barcoding.si.edu). DNA barcoding follows the same principle
like a basic taxonomic practice of associating a name with an exact
reference
collection, in conjunction with a functional understanding of
species concepts (i.e., interpreting discontinuities in
interspecific variation) (Kress et
al. 2005). In other words, it is a standardized genetic approach
that uses short but specific DNA tags, bar codes, from a uniform
locality on the
genome presented in all living organisms, in order to
distinguish one species from another. The aim and the benefits are
subject to problems
related to biodiversity and the tree of life (Stoeckle et al.
2004). The most important ones which meet the objectives and
standards of BBGK
focus on: (i) distinguishing species that look alike, enabling a
more accurate view of biodiversity, (ii) reducing ambiguity because
a sequence of
four discrete nucleotides CATG along a uniform locality on
genomes, providing a digital identifying feature which supplements
the more
analogue gradations of words, shapes and colours, (iii) making
expertise go further by facilitating plant identification, and (iv)
contributing to a
more democratized access to a standardized library of barcodes,
that will empower many more people to call by name the species
worldwide,
making possible the identification of species whether abundant
or rare, native or invasive, engendering appreciation of
biodiversity locally and
globally.
M ean m onth ly m in im um tem perature
-1,4
0
2,3
6
9,3
12
9 ,6
6,3
3
0
21 ,2
1 8,7
15,6
12
8,8
-1,6
11,9
6,7
6 ,8
8,6
11 ,2
14,5
18,1
20 ,8
-5
0
5
10
15
20
25
Jan
Feb
Mar
Apr
May
Jun
Jul
Aug
S ep
O ct
Nov
Dec
C
Vio la_cep Thym _hol
Lim o_ ith
SileC epCep
M ean m on thly m ax im um t t
57,1
11
16,4
20,7
23,8
20
14,8
10,1
6
32,2
28,5
24,1
19
15
4,6
23,9
32,2
28,7
24,5
20
16,11 4,113,4
0
5
10
15
20
25
30
35
Jan
F eb
M ar
A pr
M ay
Jun
Jul
A ug
Sep
O ct
N ov
D ec
C
V iola_cep
T hym _hol
L im o_ith
S ileC epC ep
Fig. 6 Differences in monthly temperature profiles (mean minimum
and maximum temperature), mean precipitation of the wettest and
driest quarter of the year, Emberger
pluviothermic quotient and variation of the mean monthly
precipitation regarding four selected IPS of Cephalonia island
(Viola cephalonica (Fig. 3A), Thymus holosericeus,
Limonium ithacense, Silene cephallenia subsp. cephallenia,
Mouflis et al. 2007).
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Maloupa et al. Conservation strategies for native plants in
Greece
Generally, four individual steps are required in
order to barcode each plant individual. The first
step is the specimen under investigation, which all
of them are currently maintained in living collec-
tions in the BBGK. The second step is the experi-
ments in the laboratory, which are subject to DNA
extraction, PCR for amplification of target genes,
purification steps and sequencing the obtained
sequences for each specimen. There are a large
number of protocols in the literature to apply and
work with even difficult samples, but there is not
any universal protocol yet to apply for every spe-
cimen and therefore, it is necessary to be deve-
loped. The third step is to deposit these sequen-
ces in a Database with free worldwide access.
Such Databases are found available at: (i) The
International Nucleotide Sequence Database
Collaborative, (ii) the GenBank in the U.S.A., (iii)
the European Molecular Biology Lab in Germany,
(iv) the DNA Data Bank of Japan and (v) the
Barcode of Life Database (BOLD). The forth step
applies to data analysis, where first the sequences
of the specimens are annotated and then they are
identified or compared to all closest relatives that are located
in the database.
To improve species concepts, there is a need to develop a more
sophisticated approach to barcoding, which would ideally
include
sequences from multiple (perhaps six to eight) independent
markers, a multi-locus barcode, and specific inference tools that
could be used to
explore species limits and identify genetic gaps. This second
type of barcode would improve the information, which DNA barcodes
depend on
(Chase et al. 2005).
Currently the BBGK maintains more than 1,200 different species
and more than 2,300 accession numbers (>20% of the native flora
of
Greece). Identification of these species is many times difficult
and a time-consuming process. Furthermore, there is a need to
reveal the genetic
identity of different accession numbers for many reasons i.e.
uniqueness of specimens from different native plant populations
originating in
different phytogeographical regions, possible sustainable
exploitation of selected accessions, explicit plant documentation,
and copyright.
Therefore a molecular DNA-based procedure has been adopted in
the laboratories of BBGK.
Currently there are three projects subject to the phylogenetic
relationships by using the nrDNA ITS1 and ITS2 molecular markers.
The first
one concerns the wild carnations Dianthus crinitus (Fig. 7A), D.
corymbosus (Fig. 7B), D. gracilis subsp. gracilis (Fig. 7C), D.
gracilis subsp.
drenowskianus (Fig. 7D), D. giganteus, D. deltoides, D.
petraeus, D. haematocalyx, D. monadelphus subsp. pallens, and 23
more unidentified
accessions belonging to wild Dianthus spp. The second one is an
EU financed project for saffron and its wild allies, investigating
cultivated
clones of Crocus sativus from different phytogeographical
regions with different accession numbers of wild relatives of
Crocus sativus such as C.
cartwrightianus and C. hadriaticus, collected from different
areas of Greece (see www.crocusbank.org). The third project is
related to different
accessions of plants belonging to Satureja sensu stricto (35
accessions of species belonging to Satureja montana group) and
representatives of
other closely related genera like Micromeria, Calamintha and
Acinos all belonging to Satureja sensu lato. Preliminary results
from this project
have revealed high levels of interspecific divergence in ITS1
and ITS2 ribosomal regions (Tsoktouridis et al. 2007).
In the future the aim is to extend and cover all the IPS and
further to apply to the whole collection of taxa maintained in the
BBGK. Data will
be deposited in the NCBI GeneBank DNA sequence database and the
sequences will be compared with the existed ones in order to
possibly
confirm and/or identify the accession numbers examined or to
contribute new reference material to the database for future
comparisons by other
scientists. In addition it is aimed that such an action would
act as a unique documentation of the phytogenetic resources
evaluated and managed
by the BBGK.
6. THE PROPAGATION OF THE IPS FIRST POLICY
6.1. Maintenance of IPS mother plants in the BBGK
In the BBGK, information from the literature and particular
ecological databases (Krigas et al. 2007, Mouflis et al. 2007) are
used in order to
provide the cultivation guidelines for the IPS (Table 1), aiming
at decreasing the transplanting sock and increasing cultivation and
propagation
success (Table 1; Figs. 5-6). In addition, IPS mother plants
with similar ecological requirements are grouped accordingly, in
order to save time
and labour.
All IPS are maintained on farm or in pots in the full equipped
extensive mother plantations and nurseries of BBGK under ideal
conditions,
which are as similar as feasibly possible to the wild habitat of
each taxon studied (Maloupa et al. 2003b).
Although great consideration is constantly given to climatic
conditions and soil properties, similar to the natural habitat of
the plants,
sometimes looses of few mother plants are unavoidable especially
regarding the mountainous plants and the (sub)alpine species. In
this case,
the maintenance of problematic species has been overcome by in
vitro culture methods. For instance, the single-mountain Greek
endemic
Achillea occulta (Contstantinidis and Kalpoutzakis 2005), which
grows in semi-shade, hidden limestone rock hollows, could not adapt
in the
C
A
D
B
Fig. 7 (A) Dianthus crinitus, (B) D. corymbosus, (C) D. gracilis
subsp. gracilis, (D) D. gracilis subsp.
drenowskianus
-
Maloupa et al. Conservation strategies for native plants in
Greece
environments of the nursery (sea level). Therefore, apical
meristems from mother plants have been successfully established in
vitro in MS
(Murashige and Skoog 1962) medium supplemented with 4 M
6-benzyladenine (BA), 0.5 M indole-3-butyric acid (IBA), 2%
sucrose, 0.6% agar and pH 5.8. Cultures were kept at 222oC and 16-h
photoperiod under cool white fluorescent light (40 mol m-2 s-1).
Explants produced
adventitious shoots and they were divided in small shoot
clusters which formed roots in MS medium supplemented with 10 M
IBA. Rooted plantlets were planted in trays filled with a
peat-perlite 1:1(v/v) mixture and they were maintained in a
glasshouse under a 90% RH fog-system
and 50% shading for 10 days. In the following 10 days, RH was
reduced (5%/day), while light intensity was gradually increased.
Plants were
acclimatized and transferred from the nursery to BBGK at 600m
altitude where the climatic conditions were more suitable for their
development.
Plant material at the nursery is now maintained only in vitro
until the elimination of the problems described above (Fig. 8).
Emphasis is also given to the hygienic state of the stock mother
plants. In some taxa, a water stress is recommended in order to
eliminate
pathogen infections prior propagation (Hadidi 1998; Smith 2002,
2005). Viruses and virus-like diseases are very common in native
species
(Dovas et al. 2002; Oshima et al. 2004) and the danger of
spreading could always become catastrophic, especially in some
species like
Dianthus spp. (Smyrnioudis et al. 2001; Terzakis et al. 2002).
Thus, all the initial propagation material is obtained from mother
plants treated with
exceptional care followed the standards of BBGK.
6.2. Propagation of the IPS in the BBGK
The major priority of the BBGK is to support, promote and
contribute to the combined in-situ and ex-situ conservation and
management of the
IPS. Propagation and mainly the asexual reproduction of the IPS
constitute the most significant stage prior to their evaluation for
sustainable
exploitation in order to improve human well being (Maloupa et
al. 2003a, 2003b, 2003c, 2005).
The plants collected from nature comprise the initial source
provided for the production of hundred thousands of plants, in a
very short
period of time; therefore, an error in this procedure could be
very critical (Hartman et al. 2002). The production of certified
propagation material of
the IPS requires a broad-spectrum of research strategies and
methods that have been extensively described and successfully
applied for other
cultivated plants (George 1993, 1996; Pierik 1997; Hartman et
al. 2002).
At the Laboratory-BBGK, each mother plant collected from the
natural environment is treated as a clone; it is asexually
propagated and
Fig. 8 In vitro propagation of Achillea occulta (up), Aubrieta
erubescens (middle) and Dianthus crinitus (bottom) in the BBGK.
-
Maloupa et al. Conservation strategies for native plants in
Greece
considered as the fundamental propagation material for the
reproduction of the IPS. The selection of certain clones requires
specific criteria
according to the standards followed by the BBGK. This certified
propagation material provided for the reproduction, should be
derived from a
selection procedure in order to supply products of high quality.
These criteria strictly focus on:
a. The characteristics of the plants related to their
sustainable exploitation (high production and quality of essential
oils, potential
exploitation in floriculture or landscaping etc),
b. True to type, certified for the genetic identity and
stability, of the selected clone (by using molecular DNA markers,
Section 5),
c. Disease free, to ensure a tolerable limit (if not absence) of
pathogens (Smith et al. 2002),
d. Yield, necessary to evaluate the selected material in order
to verify that the unique characteristics are maintained after
cultivation
treatments, achieving the maximum economic benefit, and
e. Asexual reproduction of the selected clones.
The BBGK currently maintains 2,300 accession numbers belonging
to 1,200 native plant taxa (more than 20% of the native Greek
flora). In
total, 326-selected Priority 1, 2, 3 and 4 IPS are maintained
(176 taxa as stock mother plants and 150 taxa as seeds preserved
within adjusted
environments). All of them have been or are currently subject to
propagation research in the nursery and laboratories of BBGK (Table
2). Already
ca. 45% of them (139 taxa) have been successfully propagated and
species-specific mass multiplication protocols have been developed
(Table
2). Taking into account their ornamental characteristics and/or
aromatic-medicinal properties, these taxa could be possibly
considered as
commercially valuable plants for sustainable cultivations (Table
2). Additionally, ca. 100 propagation protocols and cultivation
protocols have
already been developed regarding Priority 5 IPS (ca. 45% of the
total mother plants maintained in the nurseries of BBGK).
6.3. In vitro production of elite pre-basic material and IPS
mother plants
In vitro culture of any tissue used as explant material
(meristem, bud, shoot, node, etc.) makes possible vegetative
cloning from axillary buds, by
the adventitious shoot multiplication method (Daunay et al.
2007). Propagation is carried out under aseptic conditions, free
from pathogens,
yielding millions of plants within a year. The plant material
derived from in vitro cultures become the elite stock plant
material, from which mother
plantations of IPS are established for massive reproduction.
Currently, in the tissue culture laboratory of BBGK are mainly
maintained Priority 1 IPS, which either could not adjust and grow
at the
nursery conditions or they were not even possible to be
propagated using conventional methods. Furthermore, the massive
reproduction of
socio-economically valuable Priority 5 IPS is studied.
Characteristic examples are Aubrieta erubescens, Astragalus
maniaticus, Campanula
incurva, Centaurea cithaeronea, C. subsericans, Crithmum
maritimum, Dianthus spp., Melissa officinalis, Origanum dictamnus
etc. (Fig. 8).
Further propagation of the selected clones of the IPS is carried
out asexually, using mainly softwood cuttings from the elite mother
plants
(Table 2). The propagation material is preserved by the BBGK
with the possibility to be provided for research in other research
institutes. These
elite IPS materials are being also characterized genetically by
using the ITS1 and ITS2 molecular markers in order to certify true
to type
clone/species identity. However, the elite certified IPS becomes
the best value initial explant material for sustainable commercial
utilization and
therefore, many competitive companies are interested to
establish collaboration with the BBGK.
7. THE COMBINED EX-SITU AND IN-SITU PLANT CONSERVATION
POLICY
The Global Strategy for Plant Conservation (GSPC 2002)
acknowledges 16 main targets in order to halt biodiversity loss by
2010 and, the
Convention on Biological Diversity prioritizes the in-situ
conservation of rare and endangered plant species and their back up
by ex-situ
conservation (CBD 1992). If the steady decline of plant
diversity is to be halted in Europe as well, a thorough
understanding of the flora is
needed, including listing and assessment of wild plant species,
their abundance, and monitoring of changes in their distribution
and status
(Council of Europe 2002).
During the last years, BBGK has undertaken efforts in order to
contribute for the implementation of the Global Strategy for
Plant
Conservation targets (GSPC) at local, national, regional and
(inter-) national levels (Table 3).
7.1. Local scale actions
At the local level, BBGKs major plant display sections are
dedicated to the ex-situ and the in-situ conservation of native
plants (15 ha and 16 ha,
respectively). All propagated material of IPS is hosted in the
ex-situ conservation sections of BBGK, where plants are arranged
thematically and
ecologically (Fig. 9). Additionally, attention has been given in
incorporating the issue of native biodiversity within the
environmental activities in
order to increase public awareness (Krigas et al. 2006; Maloupa
et al. 2006; Krigas et al. 2007). In order to reveal distribution
changes due to
construction works in the garden or trampling from visitors and
spreading of involuntarily introduced invasive species, population
monitoring of ca.
300 plant species is in process in-situ at the 16 ha of BBGKs
natural oak forest which hosts ca. 10% of the regional flora.
Furthermore,
numerous micro-reserves are being created in-situ for wild
orchids protected from (inter-) national legislation as well as for
regional endemic
species (Fig. 10; Maloupa et al. 2007). Labeling of wild plant
populations is made evident along the Path of Biodiversity within
the oak forest by
using plant identity tags (ca. 500 tags for populations of 126
plant taxa, Fig. 10). Additionally, regular transplanting of
orchids from trampled sites
into protected display areas is being exercised (Maloupa et al.
2007).
7.2. Regional scale actions
At the regional level, the in-situ population monitoring, backed
up by ex-situ conservation actions, is currently in process at
different
phytogeographic regions of Greece (Maloupa et al. 2007): (a) in
Mt Athos, NE Greece, concerning the single-mountain endemics
Silene
orphanidis, Helichrysum sibthorpii, Anthemis sibthorpii, and
Aubrieta erubescens, the regional endemic Fritillaria euboica, as
well as the rare in
Greece Galanthus nivalis subsp. nivalis, and (b) in Mt Aenos
National Park and Cephalonia, Ionian Islands, SW Greece, concerning
some
single-area and regional endemic plant species (e.g. Teucrium
halacsyanum, Dianthus fruticosus subsp. occidentalis, Stachys
ionica, S. parolinii,
Silene cephallenia subsp. cephallenia, Viola cephalonica etc).
Additionally, in-situ population monitoring, investigation of the
genetic variability
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Maloupa et al. Conservation strategies for native plants in
Greece
Table 3 In-situ and ex-situ conservation actions of the Balkan
Botanic Garden of Kroussia (BBGK) contributing to the
implementation of the principal targets of The
Global Strategy for Plant Conservation (GSPC) at local,
national, regional, and international levels. Local level (LL):
actions within and/or around the grounds of BBGK
(31 ha). National level (NL): actions of BBGK in collaboration
with other institutes and/or organizations of Greece or actions in
different administrative and/or
phytogeopraphic areas of Greece. Regional (RL): actions of BBGK
in collaboration with adjacent countries and/or targeted in
borderline areas of Greece. International
level (IL): actions of BBGK in collaboration with institutes of
other countries and/or international organizations. BBGKs principal
target is the documentation and
conservation of the Important Plant Species (IPS) of Greece and
the Balkans, including single-mountain endemics, single-area
endemics, Greek endemics, and local
Balkan endemics, other rare, threatened and/or endangered taxa,
and Balkan subendemics with scattered distribution (Maloupa and
Krigas 2007).
Target number (#) and short description in GSPC
(2002)
BBGKs
contribution level
In-situ /
Ex-situ
BBGKs activities realized and/or in process
# 1: A widely accessible working list of known plant
species as a step towards a complete world flora
LL, NL In-situ Synthesis of existing floristic knowledge
scattered in various sources
and compilation of working lists of known IPS and their
distribution
for 10 areas in different phytogeographical regions of
Greece
(including a national park, 14 Natura 2000 sites, 8 islands, and
3
mountain areas)
# 2: A preliminary assessment of the conservation
status of all known plant species at national, regional
and international levels
LL, NL, RL, IL In-situ GIS mapping, monitoring and preliminary
assessment of the size, the
exact location and the distribution of known wild populations
of
globally and/or nationally threatened and endangered plant
species in
Mt Athos, Mt Aenos National Park and the Ionian Islands
Fieldwork searching for new populations
Recording of ecological and habitat preferences
# 3: Development of models with protocols for plant
conservation and sustainable use
LL, NL, RL, IL Ex-situ Plant propagation protocols for 109 plant
taxa
Cultivation protocols for 101 plant taxa
In vitro cultivation protocols
Establishment of propagation and conservation priorities
# 4: At least 10% of each of the Worlds ecological
regions effectively conserved
LL, NL In-situ In -situ conservation section in BBGK
Guided in -situ conservation in Cephalonia Botanica and in
Chios
Botanic Garden of the East Aegean Islands
# 5: Protection of 50% of the most important areas for
plant diversity assured
LL, NL In-situ Identification of the most important sites for
IPS diversity in a
mountain, a national park, 2 islands and 3 botanic gardens
GIS mapping of IPS
Labelling of plant populations in the wild in 3 botanic
gardens
Delimitation and protection of populations of wild orchids and
endemic
species in 3 botanic gardens
# 6: At least 30% of production lands managed
consistent with the conservation of plant diversity
LL
In-situ Fine scale pilot application of different forest
management actions in
BBGKs natural oak forest
# 7: 60% of the worlds threatened species conserved
in -situ
LL, NL Ex-situ
In-situ
Propagation, hardening and cultivation protocols of rare and
threatened plant species, aiming to potential future
re-introduction in
the wild
# 8: 60% of threatened plant species in accessible ex -
situ collections preferably in the country of origin and
10% of them included in recovery and restoration
programmes
LL, NL, RL, IL Ex -situ >2,300 accession numbers of >1,200
taxa (ca.20%) of the Greek flora
in ex -situ conservation in BBGK
Official call and request for the repatriation of IPS and native
species
of Greece from other botanic gardens
Integration of all living collections of BBGK in BGCIs website
in
preparation
# 9: 70% of the genetic diversity of crops and other
major socio-economically valuable plant species
conserved and associated indigenous and local
knowledge maintained
LL, NL, RL, IL Ex-situ Documentation, characterization and
evaluation of Crosus genetic
resources, including saffron and its allies (Crocusbank, EU
Project)
Pilot cultivation of 22 accession numbers of the protected
vulnerable
Cretan endemic Origanum dictamnus for essential oils research
and
production of natural cosmetics
Pilot cultivation of Melissa officinalis and Crithmum maritimum
for
essential oils research and production of natural cosmetics
# 10: Management plans in place of at least 100 major
alien species that threaten plants, plant communities
and associated habitats and ecosystems
LL, NL In -situ Identification and assessment of invasive alien
species in numerous
Greek cities (in collaboration with the Laboratory of Systematic
Botany
& Phytogeography, School of Biology, Aristotle University
of
Thessaloniki)
# 11: No species of wild flora endangered by
international trade
LL, NL Ex-situ IPEN numbering in all accession numbers of
BBGK
# 12: 30% of plant-based products derived from
sources that are sustainably managed
- - -
# 13: The decline of plant resources, and associated
indigenous and local knowledge, innovations and
practices that support sustainable livelihoods, local
food security and health care, halted
LL In-situ Ethnobotanical survey in villages of Mt Kroussia (in
collaboration with
the Department of Pharmacognosy, School of Pharmacy,
Aristotle
University of Thessaloniki)
# 14: The importance of plant diversity and the need for
its conservation incorporated into communication,
educational and public-awareness programmes
LL, NL, RL, IL In-situ
Ex-situ
Bi-lingual thematic calendars of native plant species produced
yearly,
distributed in Europe
Leaflets, presentations, posters, fliers
Environmental games, Environmental Trail, Path of
Biodiversity,
Educational Field Work, Experiential Conservation of Wild
Orchids, for
primary, secondary and tertiary education target groups
Organization of official events open to public and
policy-makers,
yearly
Design and 3D modelling of the Botanic Garden of
Environmental
Awareness, in Thermi, Thessaloniki
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Maloupa et al. Conservation strategies for native plants in
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and ex-situ conservation of Greek endemic Crocus spp. (C.
cartwrightianus, C. hadriaticus) which are considered as
socio-economically valuable
wild relatives of saffron is in process, through a multilateral
EU funded project (see www.crocusbank.org).
A C
B
ED
C
G F
Fig. 9 Ex-situ conservation sections with different
micro-environments in the BBGK. (A, B) Aromatic-medicinal plants,
(C) Stone-dwellers, (D, E) Aquatic and
hydrophillus (hydrophilic?) plants, (F) Educational herb garden,
(G) Mediterranean stone garden.
Table 3 (Cont.)
Target number (#) and short description in GSPC
(2002)
BBGKs
contribution level
In-situ / Ex-
situ
BBGKs activities realized and/or in process
# 15: The number of trained people working with
appropriate facilities in plant conservation increased,
according to national needs, to achieve targets of this
strategy
LL, NL - Increasing number of researchers, agriculturalists,
horticulturalists and
technicians in BBGK
Seminars concerning capacity building for the staff of BBGK
# 16: Networks for plant conservation activities
established or strengthened at national, regional and
international levels
NL, RL, IL In-situ
Ex-situ
In collaboration with Botanic Garden Conservation
International
(BGCI), efforts to establish a National Network of Greek
Botanic
Gardens of small size in different phytogeographic areas of
Greece,
dedicated to the combined ex-situ and in-situ conservation.
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Maloupa et al. Conservation strategies for native plants in
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7.3. (Inter-)national scale actions At the international level,
BBGK being an active member of Botanic Garden Conser-
vation International (BGCI, see www.bgci.org) and is currently
undertaking efforts
towards the establishment of a National Network of Greek Botanic
Gardens dedi-
cated to the combined in-situ and ex-situ plant conservation of
the native Greek and
Balkan flora (Maloupa et al. 2007). These efforts are aiming to
deliver the message to
all institutions and stakeholders involved in plant
conservation. The concept of this
initiative is subject to the establishment of numerous new,
small-scale botanic gar-
dens across different phytogeographical regions of Greece, which
will dedicate
accordingly to the ex-situ conservation of the endemic flora of
different regions. In
addition, BBGKs conservation actions of IPS attempt to meet
targets 1, 2, 3, 7, 8, 9,
14, 16 of the Global Strategy for Plant Conservation (GSPC
2002), contributing to the
effective conservation of globally threatened plant species,
implemented at local and
regional levels (Maloupa et al. 2007).
8. THE EVALUATION FOR SUSTAINABLE EXPLOITATION POLICY
8.1. The importance of medicinal plants and natural products
Natural products have been investigated and utilized to
alleviate disease since early
human history. Natural products were considered as a valuable
source of drug leads,
and the testing of natural product extracts was widely practiced
in the pharmaceutical
industry. In more recent times, natural products have continued
to be significant
sources of drugs and leads. Their dominant role is evident since
approximately 60%
of anticancer compounds and 75% of drugs for infectious diseases
are either natural
products or derivatives of natural products (Shu 1998; Newman et
al. 2003). Plants
continue to provide us with new chemical entities (lead
molecules) for the develop-
ment of drugs against various pharmacological targets, including
cancer, HIV/AIDS,
malaria, Alzheimer's disease and pain. The fact that many
currently used drugs have being derived from natural sources
(including paclitaxel,
camptothecin-derived analogues, artemether, galanthamine,
tiotropium to name a few), suggests that there might be compounds
in the plant
kingdom with promising clinical use.
The most interesting agents that are identified as natural
products come directly from the biodiversity, i.e., the richness in
variety of
organisms and genomes in the ecosphere. There are literally
millions of natural chemical structure types resulting from natures
combinational
chemistry effort supplying almost unimaginable chemical
diversity, which yields stereochemically complex structures with
diverse functional
groups, molecules ideal for interacting spherically with
biological target molecules. There are undoubtedly many more unique
and potent
biologically active natural products waiting to be
discovered.
In addition to medicinal plants, many aromatic plants are used
for the preparation of herbal drinks, in cooking or for
phyto-cosmetic uses
(Sideritis spp., Origanum spp., Rosa spp., etc.). More
opportunities for natural products discovery and development
concern agrochemicals,
cosmetics, new chemicals and nutraceuticals. However, wild
plants have played an important role in complementing staple foods
to provide a
balanced diet by supplying trace elements, vitamins, and
minerals, and may do so again in the future. Their interest as a
source of nutra-
ceuticals has been highlighted in recent studies (Heinrich et
al. 2005).
In the case of native plants of Greece, Alkanna tinctoria has
been known from ancient times for its wound healing properties.
Native A.
tinctoria plants, with alkannin and shikonin, two enantiomeric
hydroxynaphthoquinone red pigments found in their roots, have been
the base for
the development of a famous ointment which is approved and
marketed in Greece and abroad as a drug of botanical origin, as
well as for other
pharmaceutical and cosmetic preparations. Although Greece has
been blessed with a diversity of flora, most of it remains
unexploited regarding
species chemical constituents and biological potentials.
The BBGK, in collaboration with the Department of
Pharmacognosy-Pharmacology, School of Pharmacy, Aristotle
University of Thessaloniki,
are directed towards the study of bioactive molecules found in
IPS. The prime concern is to discover or develop pharmacologically
active agents,
which might be interesting for clinical practice in the future.
Up to date results show that a significant number of IPS appear to
contain
compounds with interesting biological activities i.e.
antioxidant, antivirus, antibacterial etc. (Skaltsa et al. 2003;
Mata et al. 2006; Sylignaki et al.
2006; Charami et al. 2007; Karalaki et al. 2007; Kreziou et al.
2007; Mata et al. 2007a, 2007b; Papaioannou et al. 2007).
Furthermore, IPS with potential high economic value are under
special investigation. In this approach, two types of plant
material are used
and analyzed: (a) plant material collected in-situ from wild
populations and (b) plant material originating at the same
populations and sites,
accordingly, which has been asexually propagated and ex-situ
cultivated in the BBGK (at sea level and at 600m). This approach
concern several
aromatic-medicinal plants (e.g. Crithmum maritimum, Origanum
dictamnus, Melissa officinalis, Staehelina uniflosculosa, Geranium
macrorrhizum,
Thymus spp., Sideritis spp., Achillea spp., Scutellaria spp.)
and is adopted aiming to evaluate and/or compare their quantitative
and qualitative
composition (essential oils and biologically active secondary
metabolites) under different cultivation regimes.
In the last few years, Greek and foreign companies have
developed applied research projects, aiming at the exploitation of
the unique
characteristics of the native Greek aromatic-medicinal species,
many of which are confined to Greece only (Korres 2005; Grigoriadou
et al.
2006). These research projects have been extensively considered
by the BBGK and are carried out with the joint collaboration of
Research
Institutes, Universities, National Research Centers and industry
in applied research programs, financed by different sources. The
aim for all of
Fig. 10 In-situ conservation actions in the BBGK.
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Maloupa et al. Conservation strategies for native plants in
Greece
these research projects focus on contributing to the integrated
conservation and management of the aromatic-medicinal plants as
well as other
major socio-economically valuable native Greek species (Maloupa
et al. 2005).
The most representative examples of these research projects
concern Origanum dictamnus and Melissa officinalis (both Lamiaceae)
and
Crithmum maritimum (Apiaceae) (Maloupa et al. 2003c, 2005;
Grigoriadou et al. 2006), all plants with exceptional
characteristics and valuable
properties, which could potentially play an important role at
the local economy (see www.bgci.org/cultivate/article/400/).
In addition, this initiative seems perfectly harmonized with the
key role of Botanic Gardens, ensuring that plant recourses are not
only
conserved but also used sustainably for the benefit of all
people, in order to improve human well-being (Wyse-Jackson and
Sutherland 2000;
GSPC 2002).
8.2. Evaluation of native plants prior commercialization
There is a rich base of information from many developmental
projects and case studies that have been explored on various
aspects on new crop
development. This information has been assembled in a non-unique
improvised mode, using a range of methods at different levels,
focusing
mainly on different elements, which are mostly related to
product production, processing and marketing systems.
A universal standardized evaluation system is required to be
established worldwide, in order to document and compare cases
using
consistent terms and definitions for particular range of
variables, following a data matrix as the basis for the comparative
analysis (Armitage
1996).
The ultimate goal is to provide guidance for action-oriented
interventions based on new crop development, moreover to identify
conditions
and types of cases that are amenable to development
interventions, as well as to flag types of cases that may not be
good investments.
Exploratory data analysis will be used to outline patterns,
gradients of variability, clusters of cases and key variables
associated with them.
The objective is to create typologies of cases, identify
conditions associated with particular kinds of development and
conservation outcomes
and identify/test hypotheses about new crop development (Becker
2004).
This chapter provides some guidelines and describes the
categories of information and the necessary steps required to take
under
consideration in order to select promising plant taxa for
commercialization. Interventions, and outcomes of new crop
development provide
definitions and rationales for the various descriptors used. The
evaluation parameters are described within five individual
interconnected stages.
Stage I: Targeting the market
It is very important to initially characterize the plant
material collected from the wild and to make a brief estimation of
the sector that is
targeting in the market. Depending on the characteristics of
each taxon it is necessary to be associated with a market category
of promotion
according to the relative importance and use (Roh and Lawson
1996; Armitage 1996; Daly 2005; Johnston 2005). These major
categories, which
lead to corresponded industries include:
a. Ornamental horticulture and floriculture; this sector holds a
huge number of species, cultivars and hybrids, therefore, it is
very
difficult to establish a new flower crop in a relative short
period of time. There is too much competition among the species
promoted but it is
also necessary to enrich the market with new taxa in order to
maintain and increase the consumption. The related ornamental
groups
include cut flowers, potted plants (usually for indoor use),
dried flower compositions, amenity and landscaping. Many promising
examples of
such plants originating in the wild Greek flora can be found in
Table 2.
b. Cosmetology; this sector includes plants with aromatic
properties for the production of fragrances, as well as taxa that
provide
substances extracted and mixed with crmes, lotions and other
cosmetics. This is a profitable category, due to the high demand of
products
based on natural extracts, therefore, new valuable crops provide
high income to farmers in rural areas. Many promising examples of
such
plants originating in the wild Greek flora concern Origanum
dictamnus, Crithmum maritimum, Melissa officinalis, etc.
c. Pharmaceuticals; this sector focus on taxa with very
particular chemical substances, which mainly used for
therapeutic/medicinal
purposes. These taxa automatically become valuable and research
focus on the production of high quality plants used as the initial
raw
material for drugs. In this case, advanced technology and
systems can be used throughout the production process, because of
the
importance and value of the raw materials. Many promising
examples of such plants originating in the wild Greek flora can be
found in Table
2.
d. Herbs and spices; this sector is nowadays of great interest
because people prefer on consuming more fresh and natural
healthy
products in their diet. Interesting examples originating in the
wild Greek flora concern Origanum dictamnus, O. onites, Thymus
thracicus, Th.
sibthorpii, Satureja cuneifolia, Crocus spp., etc. Most of the
products are available in open markets as row material, which
usually is not
certified. The production of culinary herbs in diverse
agroforestry systems may provide small-scale farmers on
competitively priced products
for local and potential export markets (Runham 1996). The
increased demand and knowledge of people require an organized
system that
provides certified and branded name products.
e. Sustainability and conservation; this category concerns taxa
with very particular characteristics. Usually they are not
commercialized but maintained for research and conservation
reasons. Plant species used for sustainability include taxa that
are aimed to
be re-introduced in the wild and they are part of a countrys
biodiversity. These taxa are massively propagated for plantations
in nature
under particular circumstances such as fires, flooding problems,
highway constructions and sinking of the ground, where there
are
governmental programs to sustain or re-introduce the regional
flora (Runham 1996). Conservation of threatened species, which tend
to
extinct from the wild, is subject from most Botanic Gardens,
which are aiming to rescue and maintain this genetic material and
germplasm of
plants. Moreover, national and international research programs
support the conservation, propagation of these endangered species
and
further re-introduction to nature. Such actions of the BBGK are
described in short in Table 3 and many examples of such species
originating
in the wild Greek flora can be found in Table 2.
Stage II: Basic marketing information
Basic value chain issues and information in marketing is
required in order to select the potential taxa for
commercialization. It is very difficult
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Maloupa et al. Conservation strategies for native plants in
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to explore and analyze multiple marketing parameters that may
affect the introduction of new species in the market. The reason is
that many
scientists who are specialized in the marketing of agricultural
products should exclusively work on the establishment of new crops
in the market,
increasing the cost and final price of the new products.
However, there is a lot of research developed on a variety of
agricultural products from Universities. Research topics are
considered from
time to time depending on the market. In this case, a joint
collaboration program will help to promote the native IPS of Greece
and the Balkans in
the market. Moreover, postgraduate research dissertations at MSc
or PhD level would be potentially applied on native flora, aiming
to
dynamically lead the IPS into the market. As the size of the
Greek market and sales are limited, the native plants are often not
appreciated by
the local consumers and therefore, it would be much more
plausible to promote them into the foreign markets.
The size of the market in the European Union is promising to
support the native plants from Greece produced with sustainable
methods
since Greek native plants collected from the wild have already
been on sale. From the native species mentioned in Table 2, a total
of 54% are
being purchased in nurseries of Great Britain (18% are narrow
Greek endemics and 36% are Balkan endemics).
It is not astonishing that many rare plants, which are endemic
to Greece or the Balkans, can be found and are actually being sold
in several
markets. For instance, everyone could easily find in the Royal
Horticulture Societys Plant Finder
(www.rhs.org.uk/rhsplantfinder/plantfinder.asp)
and purchase at least 115 Greek endemic taxa (ca. 10% of the
endemic flora of Greece; Maloupa and Krigas 2007). Many among them
are
included in the Red Data Book of Rare and Threatened Plant of
Greece (Phitos et al. 1995), Annexes II and/or IV of the EU
Directive 92/43
(Dafis et al. 1996) and WCMC lists (1991). To name but a few:
the Cretan endemic Origanum dictamnus, the regional Greek
endemic
Campanula incurva and the single-mountain (Mt. Athos) local
Greek endemic Helichrysum sibthorpii (the latter is also included
in the Bern
Convention). This fact brings about some legal and authority
issues: Although clearly stated in 15 of the CBD (1992), no
official agreement has
been made with the country of origin, regarding access to its
phytogenetic resources, fair trade and benefit sharing.
Furthermore, nowadays it is
recognized by most authorities that the collection of rare
species is a significant problem in Greece and the Balkans (Polunin
1980). Either
advent of botanical interest tours or independently organized
tours, the threat from the naturalist, amateur botanist, gardener
or plant hobbyist
who uproots plants for his own collection, whether of live
plants in a garden or pressed specimens in a private herbarium, is
internationally
deplored (IUCN Threatened Plants Committee Secretariat
1982).
Apart from the legal and authority issues mentioned above, it is
necessary to consider additionally the basic information adapted to
regional
or national standards. The suggested topics for consideration
are subject to: (a) Market characteristics (location, size,
distance, etc), (b) Number
of participants in the market, (c) Role of participants in the
market, (d) Production group processes, (e) Number of people with
capability and
skills (production capacity and schedule) (Johnston 2005).
Stage III: Production
Important keys to success in the high value horticulture crop
production are the low cost of production, high quality and the
likely returns. A
chain of value issues takes part during the production process
aiming to yield the best for the market and the consumer. The vast
number of
different species commercialized requires special treatments for
each taxon, therefore, a lot of research and many people are needed
to
optimize every production line. This procedure (Roh and Lawson
1996; Armitage 1996; Daly 2005; Johnston 2005) focus on particular
stages
which are subject to (see Table 2 and Figs. 4-6 for examples
originating in the native flora of Greece):
(a) Propagation by seed or vegetative in vivo or in vitro, (b)
Ability to produce planting material, (c) Climatic requirements and
limitations, (d)
Soil and nutritional requirements (soil type, drainage, nutrient
availability), (e) Production systems and their relative costs, (f)
Qualitative growth
characteristics (flowers/inflorescences, roots, habit/sprouts,
leaves, fruits/seeds, inner substances, habitat, diversity,
hybridization, stability,
durability, special sensibilities, etc.), (g) Controlled
flowering, flowering percentage and photoperiod, (h) Plant health
(pathogens, pests and
weeds), (i) Total cropping time, yield and supply period, and
(j) Price per unit.
Stage IV: Postharvest and transportation
Postharvest requirements focus mainly on packaging, maintenance
and transportation (see Table 2 for examples originating in the
native
flora of Greece). Packaging is depended on the size/shape of
plants or flowers, delicacy and the market of sale. For auctions
and wholesale
markets, packaging is of decisive significance; alternatively
packing which presents the products require means and features like
transparent foil,
text with the name, origin and indications for use etc., that
increase the final cost of the products (von Hentig 1996).
Maintenance is the most diverse parameter to estimate. There are
different requirements for each type of product and it is very
difficult to
hold simultaneously different products within appropriate
conditions. The main factors that affect maintenance include
humidity, temperature,
chemical treatments and stress of the plants. The durability of
the plants/flowers is connected also with the transportation; in
many cases
controlled environments are necessary for fragile products that
also increase the durability. The way of transportation and
distance from the
producers is also very critical, affecting the final price and
competence of the products in the market. Postharvest manipulations
for herbs and
spices, are much easier and simple, due to most of them are sold
as dried materials.
Stage V: Economics
Economics is a critical factor in producers adoption of new
crops and technologies. Aspects of economic benefits include the
maintenance
or restoration of the productivity of land or the provision of
low cost alternatives to fertilizer and soil conditioners, direct
economic benefits to
farmers by the addition of products or the diversification of
the range of farm outputs and capital reserves in the form of new
crops that accrue
value over time and can be harvested as needed (Arnold
1983).
The cost-benefit analysis can be used to determine which of a
combination of systems is the most efficient (Becker 2004)