Brussels, 27 May 2022
 

COST 071/22

 

DECISION

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Subject: Memorandum of Understanding for the implementation of the COST Action “European
Network for Innovative Woody Plant Cloning” (COPYTREE) CA21157

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The COST Member Countries will find attached the Memorandum of Understanding for the COST Action European Network for Innovative Woody Plant Cloning approved by the Committee of Senior Officials through written procedure on 27 May 2022.
 

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MEMORANDUM OF UNDERSTANDING

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For the implementation of a COST Action designated as

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COST Action CA21157

EUROPEAN NETWORK FOR INNOVATIVE WOODY PLANT CLONING (COPYTREE)

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The COST Members through the present Memorandum of Understanding (MoU) wish to undertake joint activities of mutual interest and declare their common intention to participate in the COST Action, referred to above and described in the Technical Annex of this MoU.

 

The Action will be carried out in accordance with the set of COST Implementation Rules approved by the Committee of Senior Officials (CSO), or any document amending or replacing them.

 

The main aim and objective of the Action is to address the major research challenges of in vitro cloning of woody plants, its public acceptance, risk assessment and promotion of commercial applications, in order to achieve a well connected and informed scientific community and better informed policy makers, stakeholders and market. This will be achieved through the specific objectives detailed in the Technical Annex.

 

The present MoU enters into force on the date of the approval of the COST Action by the CSO.

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OVERVIEW
Summary
In vitro culture of woody plants is leaving the academic laboratories and is now being developed in a range
of commercial applications in horticulture and forestry that respond to the challenges of climate change and
changing global food and wood consumption habits. It is therefore urgent that the research challenges,
public acceptance, risk assessment and commercial application are confronted now in order to establish a
well informed scientific community, policy makers and market place. This proposal concerns the following
challenges, whose solution will have a significant scientific, social and economic impact: How can we
overcome recalcitrance in a lot of woody plants? What are the best tools for diagnosis, sanitation and
storing clean stocks? How can the production of elite clones be scaled up at a acceptable price? What are
the real risks of this technology and how can the public be informed so that they appreciate and accept the
applications ? How can foresters and landowners be persuaded to invest in planting poly-clonal
forests? Taking these aspects into account, it seems more than urgent to us to set up a European network
to connect the researchers involved from various domains, so that they can share innovations and develop
new research strategies, assess the risks of the technology and improve communication with stakeholders
and the general public.

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Areas of Expertise Relevant for the Action
● Agricultural biotechnology: Biotechnology (non-medical)
● Agriculture, Forestry, and Fisheries: Agriculture related to
crop production, soil biology and cultivation, applied plant
biology, crop protection
Keywords
● Woody
● shrub
● biotechnology
● somatic
● micropropagation

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Specific Objectives
To achieve the main objective described in this MoU, the following specific objectives shall be
accomplished:
Research Coordination
● Improving the understanding of all aspects of micropropagation of woody plants. The action aims to take
this technology to the next level to address the challenges of 'Recalcitrance', 'Diagnosis and sanitation' and
'Scaling up and automation', as well as the challenges 'Technological risk and public acceptance' and
'Commercialisation'.
● Comparative performance evaluation of emerging technologies through the exchange of research tools,
know-how and practical experience. This platform will benefit from the skills of the different groups to
integrate in vitro protocols, diagnostic methods and production experience to address the above
challenges.
● Encouraging open access publication of joined research results
● Vulgarisation of breakthroughs and their impact, tailored to stakeholders, including the general public,
and providing input on future market application for investors
● Providing information on and raising awareness of the potential risks of the technology

Capacity Building
● Bridging in vitro technology with other disciplines to achieve breakthroughs that require an interdisciplinary approach
● Promoting knowledge exchange and the development of joint projects and doctoral research themes
around new or emerging research areas.
● Building a stakeholder platform for exchanging information and stimulating interaction
● Stimulating investments in cost effective and innovative business models for in vitro production of woody
plants.

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TECHNICAL ANNEX

1. S&T EXCELLENCE

1.1. SOUNDNESS OF THE CHALLENGE

1.1.1. DESCRIPTION OF THE STATE OF THE ART
Trees and shrubs together are called woody plants. They differ from herbaceous plants because they
form wood as a structural tissue. This action focuses on woody plants that produce timber wood, edible
fruits, nuts, berries, saps, forage and plant-based medicines or have an ecological, ornamental or even
cultural value. Although historically, there has been a focus on herbaceous ornamentals, nowadays
micropropagation of trees and shrubs is gaining great importance. The past years have seen an

evergrowing need for superior planting material for fruit and nut orchards, commodity and biomass
plantations and timber forests due to changing food patterns and climate change. Sustainable re- and
afforestation and the restoration of degraded forests can increase absorption of CO2 while improving
the resilience of forests and promoting the circular bioeconomy. In Europe, researchers from
universities and institutes are anticipating this evolution and trying to solve a number of challenges,
which are listed below.

Challenge 1: Recalcitrance
In vitro recalcitrance is the inability of plant cells, tissues, and organs to respond to tissue culture
manipulations. With respect to plant propagation and regeneration, recalcitrance is a major limiting
factor for the biotechnological exploitation of economically important woody plant species, and it can
also im- pair the wider application of in vitro conservation techniques.
Gene regulation
In vitro explants derived from mature woody plants usually exhibit low morphogenic ability. In contrast
to juvenile seedlings, they often lack vigorous growth and root poorly. Only recently detailed studies are
highlighting the physiological and molecular events involved in plant rejuvenation. DNA methylation,
histone modifications, microRNAs and telomere length could play an important role. In the last decade,
successes in the field of -omics have stimulated scientists to elucidate the molecular mechanism
initiating and controlling in vitro growth and development. Nevertheless, insights obtained in the model
plant Arabidopsis are not readily applicable to woody plants due to their large size, long life cycle and
large genome size. To advance, there is a need to outline the major regulatory genes implicated in in
vitro rooting, shoot organogenesis and somatic embryogenesis in woody crops as well as to understand
the metabolic and signalling pathways involved.
Proteomics
The proteome refers to the set of proteins expressed by the genome of an organism at any given time
or condition. Proteomics allows the quantitative and qualitative evaluation of differentially expressed
proteins in different physiological events occurring in cells, tissues, and organs. It generated
remarkable results that improved our understanding of, for example, somatic embryogenesis in trees.
Recently, gel-free separation and tag-free quantitation considerably enhanced the sensitivity of protein
identification and the recently introduced protein networks could help to find solutions to the irregular
or even non-existent germination of somatic embryos of so many species.
Epigenetics 

Plant developmental processes, as differentiation and proliferation of shoots, roots and somatic
embryos are accompanied by chromatin remodelling and epigenetic reprogramming. Methylation
constitutes a prominent epigenetic modification that can lock the DNA in a transcriptionally inaccessible
con- formation. Analysing DNA methylation distribution patterns informs on the regulatory mechanisms
of these processes, helping in the design of efficient protocols in different species. in situ localization
approaches using modern bioimaging technology have become potent tools to analyse spatial and
temporal patterns of methylation.
A new toolbox to solve old problems
New in vitro chemicals are regularly developed, but their application to woody crops is delayed due to
a lack of knowledge transfer. Examples are the so-called topolin cytokinins. Many research groups have
shown that these compounds can make the difference between success or failure in the micropropagation system of a plant species. Moreover, derivatives of phenylurea compounds have recently
been developed that act as pure cytokinin oxidation inhibitors and hold promise as a new tool in the in
vitro toolbox. Furthermore, the "chemical genetic approach" has become a popular method for
identifying interesting plant growth regulators and compounds that interfere with signal formation. Using
pipette robots and image recognition, chemical libraries are screened with literally tens of thousands of
small molecules. Almost always, the model systems used are based on a particular developmental
stage of young Arabidopsis seedlings in multiwell plates. Compounds have been identified that affect
cell elongation (brassinosteroid signalling), cell division and lateral root induction (auxin signalling),
shoot organogenesis (cytokinin signalling) and protoplast regeneration, apical hook opening (ethylene
signalling). Together, these studies have resulted in an impressive collection of unique molecules. The
application of these molecules in cloning woody plants is still a virtually unexplored area. Other tools
include affordable LED lamps that allow morphology and development to be altered in vitro through
manipulation of phytochromes and cryptochromes. New bioimaging technologies also offer great
opportunities. Worth mentioning are the huge range of cameras and imaging software that can track
bio- luminescent genetic reporters, estimate stress factors using epifluorescence and enable 3D microscanning.

 

Challenge 2: Diagnosis, sanitation and germplasm conservation
Virus/bacteria diagnosis by Next Generation Sequencing
Vegetatively propagated woody plants can be systemically infected with one or more plant pathogens,
not only in the field, but also in nurseries. Traditionally, selected viruses have been detected by ELISA
and RT-PCR and bacteria have been characterized by using culture-based methods followed by further
confirmation. These methods will not detect other potential viruses that are not included in the test with
specific antiserum or primers and the bacteria that are not able to grow on standard artificial media.
Biological indexing has been applied to screen potential pathogens in relative long indexing period with
several indexing cycles. However, high-throughput sequencing (HTS), also called next-generation
sequencing (NGS) or deep sequencing, are powerful tools for both targeted and non-targeted pathogen
analysis for certification of nuclear stocks and propagation materials. HTS has the potential to re- place
the traditional testing procedures, shortening the indexing cycles and increasing pace flow of novel
varieties to the market. In addition, microbial profiling using HTS can be applied to identify the
community of non-culturable microorganisms that often exist in in vitro mother plants. They often
present a real burden for reliable micropropagation protocols because they produce plant hormones
and toxins that influence growth and development. We are at a turning point in this regard because
routine pathogen diagnosis and cleaning in woody plants will be needed to set new international
standards, which are essential for research, production, and international trade.
Sanitation technology
In addition to the development of sensitive techniques for detection, identification and characterization
of viruses and bacteria, substantial progress must be realized in plant sanitation. For viruses, these
methods are essentially based on meristem culture, preceded by thermotherapy, and followed by shoot
tip culture. In order to improve the efficiency of those methods, additional tools such as chemotherapy, 

electrotherapy and cryotherapy have been developed. For many species, making an in vitro mother
plant completely bacteria-free is an enormous challenge.
Medium and long-term storage
Today, conservation of plant species has become a high priority to ensure sustainable use of
biological resources and prevent further loss of plant diversity. Efficient conservation of plant
germplasm is essential not only to prevent further loss of interesting genotypes, but also to preserve
the certified stocks obtained with such great effort.
Cryopreservation, i.e. the storage of plant material in liquid nitrogen has become the only safe and costefficient option for the long-term conservation of various categories of plants, including non-ortho- dox
seed species, vegetatively propagated plants and rare and endangered species. The utilization of new
vitrification-based techniques including vitrification, encapsulation-dehydration, encapsulationvitrification, droplet-vitrification, and V-cryo-plates has extended the applicability of cryopreservation to a
broad range of plant species. However, the effectiveness of these protocols is highly genotype
dependent and as the regeneration capacity of cryopreserved tissues is affected by the physiological
stage of plant material, its tolerance to osmotic, chemical, desiccation and cold stress etc. Hence,
additional research is needed and should focus on additional genotypes and species and validation of
improved cryopreservation protocols in different laboratories.

 

Challenge 3: Lack of scaling up and automation
Somatic embryogenesis
Somatic embryogenesis (SE) is the developmental process by which somatic cells, under suitable

induction conditions, undergo restructuring and go through a series of morphological and biochemical
changes, resulting in the formation of a somatic embryo. The regulation of SE development, maturation
and subsequent germination requires the concerted action of several signalling pathways that integrate
genetic and epigenetic programs, as well as hormonal and metabolic signals. SE has been observed in
a wide range of tissue culture systems but in Gymnosperms it is the only available micro- propagation
method (Park 2013). For many commercial conifer species, many research efforts have already been
made to render SE industrially applicable. However, despite these efforts, large scale industrial
application has so far been problematic due to a limited number of species that react well. A lot of effort
is needed, especially to improve the frequency of conversion of embryos into viable plants. Somatic
embryogenesis has a lot of advantages in comparison with other propagation techniques, but the main
advantage is that embryogenic tissue can be cryopreserved while field testing is still in progress. Once
field tests show which genotypes perform best, the corresponding embryonic tissue can be thawed and
used for mass multiplication of elite plants for commercial reforestation.
Temporary Immersion Bioreactors
To compete with plants coming from countries at low labour costs, new approaches in micropropagation

have recently been explored. Experiences with temporary immersion systems are
promising in this respect and deserve to be shared. The technology to produce plants in bioreactors
has caught a lot of attention of commercial in vitro laboratories. The main reasons are reduction of space
and cutting of labour costs. To avoid problems such as hyperhydricity, other variants were explored.
Among those, Temporary Immersion Systems (TIS) become popular as they allow short-term contact
of shoot clusters and somatic embryos with the liquid medium. These short immersions are appropriate
to take up the nutrients by the complete plant surface, permitting rapid growth under a head space that
allows gas-exchange. In the last decades, several TIS bioreactors have been developed, based on
single or double containers. They allow automation, as the media can easily be renewed without
changing the container. Moreover, the atmosphere can be renewed, providing adequate gas transfer to
the cultures. The use of forced ventilation may promote photoautotrophic behaviour, improving plant
propagation and acclimation. As rooting and acclimation are still important challenges for woody plant
culture, the use of bioreactors for tree propagation may represent a real improvement in this matter.
However, there are still some points that should be addressed in order to extend the use of this system,
such as the occurrence of hyperhydricity and the risk of loss of explants by fungal or bacterial

contamination. The bioreactor model, immersion frequency and duration, phytohormone type and
concentration, explant type and the use of support material are some of the parameters that merit
careful evaluation.
Robotics
The micropropagation process is tedious and labour intensive; thus, limiting the application of this
technology due to the high labour cost, which still remains one of the major costs in tissue culture
industry in the developed countries. From the 1990s onwards, there was worldwide interest in the
development of automated technologies for micropropagation. In addition to reducing labour costs,
robotised systems have other advantages: guaranteed uniformity and quality of plants, reduced levels
of contamination, elimination of human error and improved control of production. This is why several
companies tried to include automation systems in their tissue cultures. Although an automated
micropropagation process has been in progress step by step for decades, the technology is only now
becoming sufficiently efficient and affordable to implement. The problem is that plant morphology differs
a lot, which makes it technically challenging to develop a generally automated process that can be used
for multiple species. The flexibility achieved with the human eye-brain combination can only be matched
by the current level of artificial intelligence (A.I.) and Machine Learning. In Germany, Robotec's Robocut
won many awards. By means of Machine Learning, this robot is taught where to cut the laser cutter.

 

Challenge 4: Pro-active management of technological risk, public acceptance and legislation
Somaclonal variation
Somaclonal variation has more to do with epigenetic changes (methylation) than with DNA mutations.
DNA methylation can be quantified with (i) sodium bisulfite conversion and sequencing, (ii) differential
enzymatic cleavage of DNA, and (iii) affinity capture of methylated DNA.
Public concern and acceptance
While the production of clonal plants in fruits (rootstocks and cultivars) and ornamentals is today largely
accepted and considered economically-important, public acceptance could become very significant if the
use of clonal forestry does not adequately address the long-term impact on genetic variation. The main
public concern about the use of vegetatively propagated (forest) trees stems from an aversion of man’s
“manipulation of nature”, specifically the concern that vegetatively propagated mate- rials could
potentially result in a loss of genetic diversity which could lead to a catastrophic decline of forests. There
is indeed a scientific consensus that monocultures with their limited genetic variation are at risk. But it is
entirely possible to produce a vegetatively propagated plantation that has a greater genetic diversity than
is available even in natural forests by including material from a wide range of populations. An increase
in productivity balanced with environmental sustainability will decrease the pressure to allocate more
land for plantations, thus mitigating risks as the replacement of native species and potential long-term
loss of diversity at the landscape level. A process for addressing environ- mental concerns basically
depends on sharing the experience with testing of poly-clonal forestry to demonstrate that public
concerns are being addressed. Continued communication is essential in this process. Failure to address
these concerns could lead to increased legislation.
Legislation
The Council Directive 1999/105/EC on the marketing of forest reproductive material contains definitions
of how clones and clonal mixtures are defined. But is the individual member state that can regulate the
number and proportion of clones as well as how long clones are to remain in production in forestry. In
Germany and France only tested material can be vegetatively propagated and must be planted in clonal
mixtures. In Finland, “qualified” mother plants can be vegetatively propagated up to one million copies
(two million for birch). More than 11 clones must be used to plant clonal mixtures. In Sweden up to 5 %
of a site (up to 20 ha) can be planted with one or more clones. In Norway, a mini- mum of 30 clones
from 10 unrelated families are required with a maximum of 50 plants per clone per site. In Denmark only
the use of clones of poplar are regulated for planting in Denmark. As for fruit species, the Italian
legislation on the production of certified plants imposes a maximum of 12 subcultures before the line 

must be renewed, starting from genetically- and sanitary-controlled stock plants. In most member states
no specific regulations exist. However, this could change due to public pressure (Lelu-Walter et al.,
2013).
Challenge 5: Insufficient commercialization
Convincing stakeholders and investors
Pome, stone and berry fruits (cultivars and rootstocks) are largely produced today by micropropagation.
European commercial laboratories offer rootstocks of peach, plum, cherry, apricot and walnut, and
cultivars of kiwi, blueberry, raspberry, fig, olive, hazelnut, pear and date palm and their
commercialization is still growing. But the situation is different for conifers and broadleaved forest trees.
Micropropagation of selected well growing and stress resistant phenotypes would speed up
reforestation. It gives an opportunity to rapidly deploy selected varieties and to design diversity directly
in the field. But In Europe there is no large, well-developed market for genetically improved forestry tree
planting stock, especially when price becomes the major factor in deciding what type of planting stock
to use.
Unlike tropical forestry, European forestry is generally a very conservative business, where profits seem
to be limited because costs have to be borne for many years before a return on investment is achieved.
An important problem is that the advantages of planting micropropagated elite material have not been
fully and clearly demonstrated, proven and communicated to foresters and landowners. Further efforts
are needed here. “Lack of interest in improved material” is a serious bottleneck for the in- creased use
of micropropagated trees. (Lelu-Walter et al., 2013).

1.1.2. DESCRIPTION OF THE CHALLENGE (MAIN AIM)
In vitro culture of woody plants is leaving the academic laboratories and is now being developed in a
range of commercial applications in horticulture and forestry that respond to the challenges of climate
change and changing global food consumption patterns. It is therefore urgent that the research
challenges, public acceptance, risk assessment and commercial application are confronted now in order
to establish a well-informed scientific community, policy makers and marketplace. This proposal
concerns the following questions/challenges, the answers to which will have a significant scientific,
social and economic impact:
How can we overcome recalcitrance in a lot of woody plants?
Our knowledge of plant development, genetics, physiology, biochemistry and molecular biology has
expanded exponentially, often through work on mutants of Arabidopsis, and benefited from the
breakthroughs in genomics and proteomics. It opened up many new avenues for the plant propagator
to explore. However, Arabidopsis is not a tree, and so translation to woody plants just started. There is
a need for outlining and sharing knowledge about the key regulatory genes and proteomics concerning
in vitro shoot, root and somatic embryo induction, growth and development. New compounds, LEDs
and imaging systems should be introduced to the toolbox of every academic and commercial laboratory.
What are the best tools for diagnosis and sanitation?
Detecting and eliminating viruses and bacteria has enormous potential as a therapeutic tool. The field
is currently at a tipping point and, with prices falling, it can easily be integrated into woody plant research

and production,
How can the production of elite clones be scaled up at an acceptable price?
Since the 1980s, micropropagation of plants has grown into a multi-billion-dollar industry, mainly
focused on ornamental plants, a market that now seems saturated. In recent years, however, the
demand for superior planting material for fruit and nut orchards, commodity and biomass plantations
and timber wood has steadily increased. Therefore, in Europe, companies and researchers in the field
of micropropagation translate the new insights and technologies into practical applications. The

dissemination of the results of this action is vital for efficient, reliable and competitive in vitro production
of woody plants.
What are the real risks of this technology and how can the public be informed so that they appreciate
and accept the applications?
There is already a large public acceptance for the clonal production of fruit and ornamental woody
species. Nevertheless, the situation is different for forest species. Lack of public acceptance of clonal
forestry can become a very important issue if we do not communicate clearly about the risk to genetic
variation and how we want to reduce it.
How can foresters and landowners be persuaded to invest in planting poly-clonal forests?
Not only the sharing of scientific results and technical progress is important to make investment
decisions in forestry, but the sharing of the results of long-term, multisite growth performance field trials
is at least as important.

 

1.2. PROGRESS BEYOND THE STATE OF THE ART

1.2.1. APPROACH TO THE CHALLENGE AND PROGRESS BEYOND THE
STATE OF THE ART

We will address the five challenges by integrating our different skills and know-how and by sharing the
expertise, opinions and experiences of all stakeholders, from scientists to the general public and foresters.

We believe the following innovations are very important to meet the challenges:

Recalcitrance
• New molecular insights regarding the initiation and development of shoot and root meristems
and somatic embryos
• Recently gel-free separation and tag-free quantitation technology that will considerably
enhanced insight in protein networks
• DNA methylation quantification with (i) the very promising sodium bisulfite conversion and
sequencing, (ii) differential enzymatic cleavage of DNA, and (iii) affinity capture of methylated
DNA
• A chemical toolbox recently expanded to include new in vitro chemicals: new classes of cytokinins,

auxin-like compounds and signal amplifying drugs
• Phytochrome and cryptochrome manipulation by means of monochromatic light (blue, red and
far-red LEDs)
• Advanced bioimaging technologies and imaging software (Artificial Intelligence)

Diagnosis, sanitation and conservation
• High-throughput new generation sequencing (HTS) has become affordable and is extremely
powerful in combination with robust bioinformatics for diagnosing plant viruses, viroids and
bacteria.
• Additional tools to remove microorganisms, such as thermotherapy, chemotherapy,
electrotherapy and cryotherapy were developed in combination with tissue culture
techniques but have yet to prove their validity.
• New encapsulation-, vitrification-, and droplet-based techniques for the long-term storage of
mother plant stocks that were sanitized with great efforts, as well as ancient and endangered
fruit germplasm

Scaling up and automation

• Methods are being developed to enable cryopreservation and subsequent thawing of embryonic
tissue for an increasing number of tree species. They allow 'on demand production' when, for
example, the results of field trials are finished.
• Various temporary immersion systems are industrially produced and compete for the interest
of commercial enterprises
• Fully automatic micropropagation systems are currently for sale for ornamental plants. They
combine mini-robots, laser cutting, machine learning and artificial intelligence. It is obvious
that these robotized systems will also be usable for woody crops.

Technological risk assessment and public acceptance
• Vulgarization of the technology of micropropagation and somatic embryogenesis
• translating the results of long-term, multisite growth performance field trials
• modern communication tools (appealing website, facebook, twitter, youtube, blogs and podcasts)

Commercialization
• Creating a platform to share valuable results of long-term, multisite growth performance field
trials with the forester community
• Networking events with elevator pitches by Ph.D. students to convince venture capitalists and
business angels to invest in cost effective and innovative business models for in vitro
production
• Introducing the concept of crowdfunding projects by scientists and foresters for polyclonal
forestry, nut plantations, cloning special historical and cultural heritage individual trees for the
public, etc.

 

1.2.2. OBJECTIVES
We propose this Action to collect, share and disseminate the available information and experience in
all aspects of the micropropagation of woody plants. We want to move to the next level of this technology
in order to address the challenges ‘Recalcitrance’, ‘Diagnosis and sanitation’ and ‘Scaling up and
automation’. Concerning challenges ‘Technological risk and public acceptance’ and ‘Commercialization’
we want to summarize and communicate the relevant field trial data and experience so that legitimate
steps can be taken to minimize risks and to convince all stakeholders of the economic benefits of
planting cloned elite trees and shrubs. In this way, in-vitro technology can achieve its full eco nomic
potential for supporting European forestry and horticulture. In summary, our main objectives and results
of this Action:

1.2.2.1 Research Coordination Objectives
A. Improving the comprehension of the already defined challenges and the possible strategies to
address them
B. Comparison and performance assessment of emerging technology and tools
C. Encouraging open access publication of joined research results
D. Vulgarization of breakthroughs and their impact, tailored to stakeholders including the general
public and input on future market application for investors
E. Informing and raising awareness of the possible risks of the technology 

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1.2.2.2 Capacity-building Objectives
F. Bridging in vitro technology with other disciplines to achieve breakthroughs that require an

interdisciplinary approach
G. Promoting knowledge exchange and the development of joint projects and doctoral research
themes around new or emerging research areas
H. Building a stakeholder platform for exchanging information and stimulating interaction
I. Stimulating investments in cost effective and innovative business models for in vitro production of
woody plants.

 

2. NETWORKING EXCELLENCE

2.1. ADDED VALUE OF NETWORKING IN S&T EXCELLENCE
2.1.1. ADDED VALUE IN RELATION TO EXISTING EFFORTS AT EUROPEAN
AND/OR INTERNATIONAL LEVEL
In the last 20 years, there have been three relevant COST Actions dedicated to Plant Tissue Culture,
but they were never specifically dedicated to woody plants:
• 822 - Development of integrated systems for large scale propagation of elite plants using in
vitro techniques (23/5/1995-23/5/1999)
• 843 – ‘Quality Enhancement of Plant Production through Tissue Culture’ (7/12/00-7/12/04)
• 871 – ‘Cryoplanet, Cryopreservation of crop species in Europe’ (12/12/06-11/12/10)
As the research community of researchers is both limited and fragmented, a major objective of this
COST Action is therefore to strengthen collaboration in the field of biotechnology applied to woody
plants at the European level. This will be achieved by sharing research tools (e.g. embryogenic cell
lines, in vitro model plants, compounds, innovative micropropagation technologies) and know-how
(experimental protocols, procedures and practical experience) to tackle common challenges. This

platform will integrate in vitro protocols, diagnostic needs (identification of virus, viroids, phytoplasma and
bacteria) and production needs (automatisation, temporary immersion bioreactors) by integrating the
skills of the different groups.
This is not the first networking initiative at international level since a large number of researchers in
this field are already federated within the IUFRO 2.09.02 Unit entitled 'Somatic embryogenesis and
other vegetative propagation technologies' (686 researchers and stakeholders from 67 countries).
Every two years an international conference is organized and dissemination of results is done by its
proceedings/books. We will try to combine the activities of IUFRO and COST to create a synergy for
the European in vitro community of woody plants. This will boost the dissemination of results, notably
through the temporal coordination of meetings or the organization of side events whenever possible.

 

2.2. ADDED VALUE OF NETWORKING IN IMPACT
2.2.1. SECURING THE CRITICAL MASS AND EXPERTISE
The challenges in woody plant cloning in terms of techniques and know-how can only be tackled on a
European scale, as it is impossible to find all expertise in one country. Although the European re- search
in this area can be considered being of high level, it nevertheless suffers from fragmentation in small
research groups and a lack of coordination. This implies a poor integration of interdisciplinary research
and spreading of results towards forestry and horticulture. This project is designed to over- come the

shattering of the European research field, stimulate mobility, and improve links with less

research intensive countries across Europe. Thus, it will be possible to maintain its level on a

world-leading stage.
All participants have extensive experience in the in vitro culture of woody species, but while some of the
members lead projects related to the fundamental questions of plant morphogenesis, others focus
mainly on more applied aspects, such as the sanitization of the starting material, cryopreservation and
the use of bioreactors for large-scale propagation. The interaction and feedback between fundamental
and applied strategies will ultimately allow cutting-edge research to reach companies and society as a
whole. The complementation based on expertise extends to the type of species with which each group
works and their main uses (hardwoods, conifers, currently productive species, endangered
germplasm...), as well as the type of challenges their native forests and plantations suffer in the

current climatic change scenario. The creation of a diverse and interactive group that detects the problems
that are currently affecting forests and plantations at the local level will allow scientific solutions to be
sought before they become global.

2.2.2. INVOLVEMENT OF STAKEHOLDERS
Following stakeholders should be made aware of the progress of this technology and the new
opportunities it offers to contribute to the bioeconomy:
• Scientific community
• Commercial laboratories: In Europe, many large commercial laboratories are producing highquality

woody plants
• Centers for woody plant germplasm conservation, elimination of pathogens, and genetic
engineering
• Nurseries and producers of fruit, ornamental and forest tree planting material
• Suppliers of chemicals and equipment for In vitro laboratories
• Educational institutions and training in the sector of in vitro technologies
• Land owners and associated foresters,
• Policymakers and risk assessors (EU or national, regional institutions and government
authorities)
• NGO’s: IUCN
• Organisations for biodiversity: BIOVERSITY INTERNATIONAL, CGIAR, FORESTS FOR
FUTURE, EVOLTREE
• Media: journalists
These main stakeholders will benefit greatly from the innovations produced by this COST Action. Their
involvement will take place through active participation in meeting initiatives organized within the Action
(workshops, one-day meetings on specific topics, “traditional” seminars and webinars), as well as a
specific website supported platform of the Action, through which operators will be able to interact with
the participants in the COST Action, propose their problems, get in touch with operators from other
Countries and be informed about the potential benefits and risks so as to enable multi-perspective
discussions on the sociological and ethical aspects of the technology.

2.2.3. MUTUAL BENEFITS OF THE INVOLVEMENT OF SECONDARY
PROPOSERS FROM NEAR NEIGHBOUR OR INTERNATIONAL PARTNER
COUNTRIES OR INTERNATIONAL ORGANISATIONS
There is a tendency for research groups to work in networks dedicated to temperate or tropical fruit
rootstock, berry shrubs, ornamental tree, conifers or forest tree rather than tool or process specific

networks. As such, isolated networks have to “reinvent the wheel” and cannot benefit from the learning of
others working on other plants. Given the size of the problems, and the current dynamics of the field, a
concerted and aligned effort is required, also involving research from tropical or arid regions groups, to
learn from each other’s new fundamental insights and experiences with new technical tools. 

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3. IMPACT

3.1. IMPACT TO SCIENCE, SOCIETY AND COMPETITIVENESS,
AND POTENTIAL FOR INNOVATION/BREAKTHROUGHS
3.1.1. SCIENTIFIC, TECHNOLOGICAL, AND/OR SOCIOECONOMIC IMPACTS
(INCLUDING POTENTIAL INNOVATIONS AND/OR BREAKTHROUGHS)
By joining complementary forces, COPYTREE will serve as a network of excellence in innovation
based woody plant production for food, timber, feed, fuel, fiber, ornamental and medicinal use and thus
provides a long term and sustainable service to society. The impact of this network on the academic
quality of the participants will be highly positive. During the annual meetings, not only the COST Action
members, but also external specialists in other scientific fields will be invited to talk about their expertise
which will stimulate new connections, creativity and inspire innovation. Indeed, meetings, staff
exchange, shared PhD supervision and training programs will allow cross fertilization of ideas and
methods and building a critical mass. Improved access to each other’s state-of-the-art facilities and
technologies and multidisciplinary expertise will further stimulate the academic quality and career of all
involved participants.
The immediate results of this action will be scientific and technological advances, including a better
understanding of the initiation and development of shoots, roots and somatic embryos, as well as the
genes, proteins and signalling pathways involved. In the short to medium term, it is expected that the
European plant production industries will rapidly absorb and benefit from the practical results of the
action, given the potential economic advantage over traditional plant production methods.

3.2. MEASURES TO MAXIMISE IMPACT
3.2.1. KNOWLEDGE CREATION, TRANSFER OF KNOWLEDGE AND CAREER
DEVELOPMENT
COPYTREE intends to become a platform that centralizes the skills and resources of researchers and
companies to tackle the challenges in mass in vitro cloning of woody plants by knowledge transfer.
Through concerted pursuit for extra funding, the network will further improve its impact. The members
of the network will make use of each other’s expertise during conferences, workshops, summer schools
and Short-Term Scientific Missions (STSM). They will be preferred partners in research proposals, PhD
and master thesis student exchange and assessment and jointly work on publication. This will help in
scientific and educational career development of the participants. The frequent meetings and
communications will further shape a true sense of community that will survive the COST Action for a
long time ahead.

3.2.2. PLAN FOR DISSEMINATION AND/OR EXPLOITATION AND DIALOGUE
WITH THE GENERAL PUBLIC OR POLICY
We foresee a specific, focused, and structured plan for dissemination, based on clever online
strategies, life presence at stakeholders’ events and complementary activities. Furthermore, we are
interested in education of the broad public on innovative plant biotechnologies. In order to carry out
this task, the following activities will be coordinated by a dedicated workgroup: WG5.
Online strategies
Website: Conceived as a key element for the communication strategy, a dedicated and independent
project website with a distinctive URL will be developed, published, updated, and maintained for the
duration of the Action and running even after its end. The website will be organized in the following
sections: General description of the action, consortium composition, progress update, news and events,

documents, news, links and contact. It will be comprehensive and attractive, enhanced with
photography, video, graphs and audio presentations. The Action website will serve throughout the
duration of the project as a constant point of reference for the partners as well as for current and
potential stakeholders. The homepage will be clearly and visibly marked with the COST and
COPYTREE’s logo, together with all the partner´s logos and will acknowledge the European Community
contribution. The web site will also host links or sub-pages where databases, datasets, e-manuals. Link
with others COST Actions and European Projects with similar aims will be provided in the Action web
page to mutually increase the audience of interested parties. Scientists involved in the project will
continue to peri- odically update the site and respond to the questions arising. The visitors of the website
will have the possibility to submit questions and clarifications directly to the scientists/technicians who
have been involved in the COST Action.

Facebook, Twitter, YouTube, blogs and podcasts: In order to improve the dissemination to a wide
range of stakeholders and eventually, to maintain a two-way and interactive communication, the project
will offer linking widgets to social media tools such as Facebook page and Twitter account. In particular,
an Action´s Facebook profile will be created to target the most dynamic audience and facilitate the
rapid information diffusion and to stimulate discussions. A YouTube channel will be dedicated to video
related to the COPYTREE development. YouTube movies will be recorded at the laboratories with
interviews and attractive images of in vitro culture practices and cloned trees. This will be placed on
the website, available for online visitors for many years. Twitter will also be effectively used to create
a hashtag for the Action. Each article posted on the website will have a share button below to promote

a regular sharing and promotion of the news. The URL will also be promoted in other relevant
forestry stakeholders’ webs (forestry research institutions, Universities, nurseries, wood processing
industry, forestry associations, etc.). Podcasts will be recorded in interview format. A blog about the
conference will appear on the website.
TV: Actions will be taken to allow scientists/technicians, participating in the COST Actions, to be
hosted by local/national radio/tv channels to give information, details and, essentially, to raise the
awareness of the public on the Action topics.
Video: A video/documentary will be produced in English, explaining the most relevant characteristics
of the COPYTREE Action in a simple and direct language. The video length won´t exceed the 15
minute duration and will also be uploaded on the Action´s website and on YouTube. It will be
broadcasted on screens during events (Conferences, Congresses, Symposia) related to plant
propagation and in vitro culture that will accept to offer a space to the COST Action.
Scientific E-community: To maintain informed the scientific community, the scientists involved in the
COST action will provide information about the project also in their Researchgate profiles.

Information stands
A demonstration and information stand will be installed at selected events where stakeholders are

present, such as important professional forestry and horticultural trade expositions in Europe. Panels and
posters will present the Action, in vitro material will trigger the passing visitors to start an informative
talk. This will also contribute to advertise and promote the website and social media activities.

Annual reports
A scientific report will be made before the annual meeting to explain all the activities carried out.

Promotional materials
Production and distribution of digital brochures in pdf format will be made.

Scientific dissemination of the project
• Specialized papers in peer-reviewed open access journals.
• Presentation of the Action by means of talks and a stand at International
Conferences/Symposia such us IUFRO Unit 2.09.02 (biotechnology section of the
International Forestry Research Organization), IAPB (International Association for Plant
Biotechnology) meeting, ISHS Symposia on plant biotechnology and tissue culture 

(International Symposium on Production and Establishment of Micropropagated Plants, IS on
In Vitro Culture and Horticultural Breeding, IS on Micropropagation and In Vitro techniques)
SLTB (Society for Low Temperature Biology) Annual Conferences.
• Furthermore, participants will inform about initiatives and outcomes resulting from the
COPYTREE Action also in National events (Conferences, Congresses).

Complementary actions
• Exchange of technology between participants: all questions regarding IPR, material
exchanges, responsibilities and further scientific disseminations shall be regulated in a
detailed specific agreement document according to COST policies.
• Establishing links with International Associations (IUFRO, EVOLTREE, EFI, SLTB, etc…).
• Invitation of different stakeholders, especially of the host country of the annual conference, to
participate in the event.

 

4. IMPLEMENTATION

4.1. COHERENCE AND EFFECTIVENESS OF THE WORKPLAN
4.1.1. DESCRIPTION OF WORKING GROUPS, TASKS AND ACTIVITIES
Taking into account the aforementioned challenges, we are planning four working groups (WG).

WG1: Recalcitrance
Tasks: Outlining knowledge, sharing and communicating insights in genetics, epigenetics and protein
networks. Re- viewing new chemical and physical tools that might break recalcitrance.
Demonstrating the power of bio-imaging systems.

WG2: Diagnosis, sanitation and conservation
Tasks: Updating, sharing and communicating the latest technology for the diagnosis of viruses, viroids
and bacteria, sanitation protocols for clean stock production and identifying new breakthroughs in mid-and long-term storage of clean stocks and germplasm.

WG3: Scaling up and automation
Tasks: Sharing and communicating breakthroughs in the initiation, multiplication, preservation,
germination and conversion of somatic embryos of new species. Critical comparison of the results of
micropropagation in different models of temporary immersion systems. Performance assessment of
advanced fully automatic micro- propagation systems combining robotics, laser cutting, machine
learning and artificial intelligence. To provide a forum for industrial suppliers of this technology.

WG4: Technological risk assessment, public acceptance, legislation and commercialization (transversal
working group)
Tasks: Fostering knowledge ex- change dealing with risk assessment, developing a strategy to
stimulating technology awareness and acceptance of stakeholders, stimulating investment and
commercialization. 

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WG5: Communication, dissemination and technology transfer (transversal working group)
Tasks: Coordination of the reports, organizing a stakeholder platform for communication and
feedback, dissemination and technology transfer by website, social media, information stands,
promotional materials.
Working groups interact at the yearly conferences
The yearly conferences represent the heydays of the Action. The four working groups will have strong
ties and interactions that will bring mutual benefits. To promote new ideas and exchange of information

among different WGs, ensure the specific needs of each WG and to guarantee dissemination
towards the stakeholders, the yearly conference will be organized as follows. During Plenary sessions,
each WG provides a key-note speaker who gives an overview of the state of the art and the challenge.
However, there will also be a forum for external scientists in new evolutionary domains such HTS for
virus and bacterial load detection, large throughput robotic screening of chemicals, big data algorithms,
artificial intelligence, and image analysis. During parallel sessions, the working groups organize
specific lectures, including an in-house poster session. The working groups come together to set up
joint experiments in which each lab examines the same tool (connection, technique, apparatus,
service) with specific model plants (ring test). There will also be an opportunity to visit stakeholders in
the vicinity of the conference: woody plant propagators, commercial laboratories, etc.

4.1.2. DESCRIPTION OF DELIVERABLES AND TIMEFRAME 

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