You are on page 1of 9

Symbiosis (2012) 58:2937

DOI 10.1007/s13199-012-0208-9

Development of arbuscular mycorrhizal biotechnology

and industry: current achievements and bottlenecks
Miroslav Vostka & Ale Ltr & Silvio Gianinazzi & Jana Albrechtov

Received: 2 October 2012 / Accepted: 30 November 2012 / Published online: 8 January 2013
# Springer Science+Business Media Dordrecht 2013

Abstract Advanced scientific knowledge on arbuscular

mycorrhizal symbioses recently enhanced potential for implementation of mycorrhizal biotechnology in horticulture
and agriculture plant production, landscaping, phytoremediation and other segments of the plant market. The advances consist in significant findings regarding:new
molecular detection tools for tracing inoculated fungi in
the field;the coexistence mechanisms of various fungi in
the single root system;new knowledge on in vitro physiology of the AM fungi grown in root organ cultures;
mechanisms of synergistic interactions with other microbes
like PGPR or saprotrophic fungi; discovery of mycorrhiza
supportive compounds such as strigolactones. Scientific
knowledge has been followed by technological developments like novel formulations for liquid applications or seed
coating, mycorrhiza stimulating compounds or new application modes. Still the missing components of biotechnology are appropriate, cheap, highly reproducible and effective
methods for inocula purity testing and quality control. Also
there is a weak traceability of the origin of the mycorrhizal
fungi strains used in commercial inocula. Numerous poor
quality products can still be found on the markets claiming
effective formation mycorrhiza which have very low capacity to do so. These products usually rely in their effects on
plant growth not on support of host plants via formation of
M. Vostka (*) : J. Albrechtov
Institute of Botany, Academy of Sciences,
Pruhonice, Czech Republic
A. Ltr
Symbiom Ltd., Lanskroun, Czech Republic
S. Gianinazzi
InoculumPlus Ltd., Dijon, France
J. Albrechtov
Faculty of Science, Charles University, Prague, Czech Republic

effective mycorrhizal symbiosis but on fertilizing compounds added to products. There is growing number of
enterprises producing mycorrhiza based inocula recently
not only in developed world but increasingly in emerging
markets. Also collaboration between private sector and scientific community has an improving trend as the development of private sector can fuel further research activities.
Last but not least there is apparent growing pull of the
market and increasing tendency of reduction of agrochemical inputs and employment of alternative strategies in planting and plant production. These circumstances support
further developments of mycorrhizal inocula production
and applications and maturation of the industry.
Keywords Arbuscular mycorrhizal fungi . Sustainable
agriculture . Inoculum quality . Inoculum tuning .
Large-scale trials/verification . Mycorrhizal technology

1 Introduction
For the majority of plant species including agricultural and
horticultural crops, uptake of water and mineral nutritions,
particularly phosphorus, is mediated by the mycorrhizal
fungi. As applies to nitrogen (N), the contribution of mycorrhizal uptake and the costs to the plant are still not fully
clear (Smith and Smith 2011). Mycorrhizal fungi have also a
lot of non-nutritional effects on plant physiology often
alleviating plant stress caused by biotic and abiotic factors,
acting as biocontrol agent, stabilising soil aggregates, preventing erosion and influencing plant fittness and sustainability of the whole plant-soil system (Smith and Read
2008). Many lines of scientific evidence prove not only
improved crop yield and resistance of mycorrhizal plants
to environmental factors but improvement of many foodquality properties, such as increased contents of desirable


antioxidants, vitamins and mineral elements (Gianinazzi et

al. 2010; Albrechtova et al. 2012). Mycotrophic agricultural
crops are usually forming symbiosis with mycorrhizal fungi
and arbuscular mycorrhiza (AM) is the most common type
occurring in more than 80 % of plant families. Sustainable
management of agricultural ecosystems includes efficient
management of soil microorganisms (Jeffries et al. 2003;
Selosse et al. 2004; Bunemann et al. 2006; Barrios 2007;
Vosatka and Albrechtova 2009; Gianinazzi et al. 2010).
Responding to the need of fast-growing world population, in the middle of the previous century the Green
Revolution brought development of dwarf and semidwarf genotypes of high-yielding varieties of wheat and rice
responsive to higher-fertilisation inputs (Khush 2001; Lynch
2007) while improving the yield by high dosage of pesticides with multiple environmental haphazards (Pimentel
1996). The focus of new green revolution is to improve
the yield of crops grown in infertile soils (Lynch 2007)
and attention has to be focused to below-ground crop traits
such as root architecture including root branching (De Smet
et al. 2012), nutrient uptake and nitrogen fixation (Den
Herder et al. 2010) and management of beneficial soil
microorganisms in general and to arbuscular mycorrhiza
(AM) in particular (Gianinazzi et al. 2010). Abundance
and diversity of mycorrhizal fungi populations in soils
decreases with soil degradation, pollution, agricultural practices (e.g. tillage, crop station mode) and agrochemical
applications (Plenchette et al. 2005; Bunemann et al. 2006;
Vostka and Albrechtov 2008). Comparison of AM fungi
abundance and diversity indicates a potentially severe loss
of ecosystem function under conventional high-input farming compared to more ecologically-friendly low-input agriculture (Oehl et al. 2004). Particularly, high phosphate
availability in soils has negative effects on mycorrhiza development and usually no mycorrhizal effects on yield are,
thus, recorded (e.g. Janos 2007; Douds et al. 2012). Degree
of mycotrophy plays an evident role particularly when cultivation conditions involve either soil-less or fumigated
substrates used often in horticulture where indigenous mycorrhizal fungal populations are either lacking or greatly
reduced. Fields in conventional agriculture can have significantly reduced AM populations. In organic agriculture
where low-input cropping systems are maintained, the role
of mycorrhiza has increased significance in production of
mycotrophic crops in comparison to conventional agriculture where mycorrhiza function and importance has been
marginalised by high inputs of agrochemicals (Harrier and
Watson 2003; Plenchette et al. 2005; Gosling et al. 2006).
Scientific knowledge on mycorrhiza effects on various
crop plant parameters are recently increasing emphasising
potential use of mycorrhiza application in production systems (Vostka and Albrechtov 2008, 2009; Gianinazzi et
al. 2010). Mycorrhizal fungi are widely recognized as a

M. Vostka et al.

natural plant health insurance (Gianinazzi and GianinazziPearson 1988). During the last decade, mycorrhizal inoculations are becoming recognized as a modern feasible biotechnology in plant production and the whole industry on its
own with its specifics has emerged (Vostka et al. 2008a,
2008c); there is an increasing number of examples of the
positive impact of mycorrhiza in crop production and in
particular AM. Mycorrhizal technology is currently reaching an industrial stage supported by extensive applied research and commercial applications emphasizing an
ecological, sustainable aspects of the use of mycorrhiza
(Vostka and Dodd 2002; Vosatka and Albrechtova 2009;
Gianinazzi et al. 2010).

2 Current mycorrhiza industry achievements

The production systems of arbuscular mycorrhizal fungi
have evolved significantly during the last years from relatively trivial technologies of in vivo cultivation on the roots
of plants grown in the field in non sterile open nursery or
farm cultures (e.g. Douds et al. 2012) or pot production (e.g.
Feldmann and Schneider 2008; Vostka et al. 2008b). The
AM fungal inoculum production in association with the host
in non axenic conditions is simple and relatively cheap
method, however, it has a disadvantage of limited application, and it might be easily contaminated and it is not well
adapted for the development of an industrial activity
(Gianinazzi and Vosatka 2003). Due to simple set up and
easy control, the most abundant AM fungi production
occurs in greenhouses with different inert substrates with
plants inoculated with pure fungal cultures or their mixes in
containers or pots (Feldmann and Schneider 2008; Vostka
and Albrechtov 2008; Vostka et al. 2008b). However, the
most important issue about commercial production in in situ
systems remains the microbial quality and purity of AMF
strains contained inocula and presence of other contaminating microbes. The purity amd quality of inoculum can be
checked by new-generation molecular sequencing methods
(454 pyrosequencing) recently being used for AMF diversity studies (Opik et al. 2009). More sophisticated production
methods in vitro have been recently devloped e.g. production on transformed hair root cultures (ROC - root organ
cultures) in axenic conditions (Declerck et al. 2005, 2011).
Currently, there are about 3 small enterprises producing in
vitro in Europe, two large ones outside Europe, however, the
proportion of both types of production modes is difficult to
estimate globally, since the real commercial data are not
always publically available.
During the last decade a number of companies manufacturing mycorrhiza based inocula has increased significantly.
By our estimation, there are over 15 small medium enterprises just in Europe producing mycorrhizal fungi, however,

Development of arbuscular mycorrhizal biotechnology and industry

it is often difficult to distinguish the primary producers of

inoculum from distributors of re-branded products. Previously each company has established its own local market,
mainly in their own territories but recently markets have
started to overlap and several distributors chains have been
formed. Few companies have been bought out by multinational corporations and have started to market mycorrhiza
world wide.
Nevertheless, the majority of the companies are small
scale and have focused primarily on retail hobby markets.
The major product lines they have launched are small users
packaging for the garden centres. Partly the companies
suceed to engage also professional horticulture market and
proved economic feasibility of inoculation for specific market segments like tree planting at landscaping, high value
fruit trees and shrubs production. Also an important target
segments for inoculations are nursery production of forest
and ornamental trees (Vostka and Albrechtov 2008) and
phytoremediation projects where plants encounter severe
stress imposed by high pollutants contents such as heavy
metals or organic pollutants. In general, inoculation outcome is more apparent and thus economically feasible in
low-fertile and arid soils where nutrients and water resources are scarce. Applications of mycorrhizal inoculations in
convential production systems for crops in extensive agriculture are still disputable for uncertain effects since inoculation can be effective but economically unfeasible.
There is a significant move of some companies to develop new emerging markets in developing countries. There is
little information available on commercial implementation
of mycorrhizal technology and particular projects but it is
clear that there has been a lot of applications in several
countries of Africa, significant development of markets in
India (Adholeya 2012) and other countries (Kasuya and
Costa 2009).

3 New potentials for mycorrhizal research and

Until now the most emphasis has been given on nutritional
effects of mycorrhiza for plantsparticularly uptake of
phosphorus (P). It is generally known that AM develops
and functions more effectively for crop yield, growth and
plant resistence under conditions of low availability of
phosphates in the soil for plant uptake. This is particularly
important in these days when sources of rock phosphate are
going to be exhausted in decades from now on and mineral
fertiliser production is going to be hampered severely
(Hinsinger et al. 2011). There are also many nonnutritional effects on plants and their functioning in ecosystems, which might be important and support ecosystem
services provided by mycorrhizal fungi (Barrios 2007;


Gianinazzi et al. 2010). One of the most important ecosystem services are enhancement of soil erosion control by soil
binding capacity via mycorrhizal extraradical mycelium
(ERM) (Caravaca et al. 2002; Rillig 2004; Piotrowski et
al. 2004), and significant allocation of plant carbon products
of photosynthesis into mycorrhizal structures being a significant carbon sink on global level (Gianinazzi et al. 2010).
There are also new findings on AM role in mediation of
allelopathy through easier transport mediated common mycorrhizal networks (Cipollini et al. 2012).
Currently, soil microbe management leading to sustainable management practices belongs to possible applications
of mycorrhizal or combined-microbial inoculations with
ever increasing importance. During time of ongoing climate
change desertification of agricutlrural land increases, however, microbe management still plays a secondary role in
conventional agricultural practices. One of the reasons is an
ongoing lack of legislative policy focusing explicitly on soil
ecosystems and degradation processes, scarce international
policy framework to guide sustainable soil management
(Thomas et al. 2012). According to the authors, the United
Nations Convention to Combat Desertification (UNCCD)
remains the only Rio Convention that is not supported by
involvement of scientific community and lacks the equivalent of an IPCC (Intergovernmental Panel on Climate
Change) or the proposed IPBES (Intergovernmental SciencePolicy Platform on Biodiversity and Ecosystem
Implementation of mycorrhizal inoculum into practice of
phytoremediation (phytoextraction, phytostabilisation) has
some potential (Miransari 2011a; Bhargava et al. 2012;
Meier et al. 2012). There are several evidences that certain
selected AM fungi have capacity to reduce translocation of
heavy metals into above-ground plant tissues and therefore
improve safety and quality of edible parts of the food crop
(Rivera-Becerril et al. 2002).
On the other side, some fungi can increase phytoextraction and then help in decontamination of polluted soils.
Nevertheless careful selection of inoculants in this aspect
should be performed prior large scale applications because
some fungi can have an opposite effects on heavy metal
translocations (Janoukov et al. 2012).
Newly exploited potential of AM became enhancement
of food quality in crop plants. There has been recently
reports on increase of sugar content in plant, increase of
essential elements (Zn, Mg etc.) and mainly antioxidants
and beneficial mineral elements (Perner et al. 2008;
Albrechtova et al. 2012; Gianinazzi et al. 2010).
Currently, a lot of attention has been devoted to the
research on the impact on interactions between mycorrhizal
plants and pathogens, herbivores, and parasitic plants (Jung
et al. 2012). Modulation of plant defense responses as a
consequence of mycorrhiza establishment results in a mild,


but effective activation of the plant immune responses not

only local but also systemical (Jung et al. 2012; Hao et al.
2012). An interesting issue in mycorrhizal plant-parasite
interaction is a recently reported potential of mycorrhiza to
provide partly protection and control of Striga (FernandezAparicio et al. 2011).
Recently defined phytohormones strigolactones induce
suicidal germination of Striga seeds, control shoot branching and seems to be part of host recognition signals for
mycorrhizal fungi (Fernandez-Aparicio et al. 2011; Kohlen
et al. 2011). However, this phenomenon should be further
investigated and eventually exploited since a role of strigolactones in plant signalling and how strigolactones were
co-opted as an allelochemical signal by parasitic plants still
has not been fully elucidated (Tsuchiya and McCourt
2012). Recently other molecules active in signalling and
development of AM symbiosis have been discovered, e.g.
sulphated and non-sulphated simple lipochitooligosaccharides (LCOs), and they hold promising potential of AM
symbiotic efficacy enhancement in agriculture (Maillet et
al. 2011).
Important issue for development is also combinations
with other beneficial microbes. Synergistic inoculations
bring benefits to plant production, so they are already sought
after. This, for example, applies to dual inoculations consisting of AM fungi and rhizobacteria, which promote plantgrowth together (Vazquez et al. 2000; Gryndler et al. 2002;
Miransari 2011b). Dual inoculations with AM fungi and
saprotrophic fungi decomposing supplied organic plant matter can exhibit a beneficial effect on the crop growth and
food quality, as we recently proved on mycorrhizal onions
(Albrechtova et al. 2012). Members of the saprobic genus
Trichoderma have emerged as an especially promising
group of microbial inoculants when Trichoderma genotypes
supporting plant growth were found (Camprubi et al. 1995;
Tchameni et al. 2011). Consistency of given interactions is
still to be assured before safe implementation of the microbe
As examples of newly emerged knowledge based on
recent findings, we can emphasize ecosystem services of
mycorrhiza in agriculture (Gianinazzi et al. 2010), crucial
role of AM in phytoremediation (Bhargava et al. 2012;
Meier et al. 2012), new findings on AM role in mediation
of allelopathy via common mycorrhizal networks (Cipollini
et al. 2012).

4 Problems and bottlenecks

In commercial AM fungi production, the main issue remains
quality assurance. The only way is to produce pure, noncontaminated AM inoculum is currently in production systems on transformed roots in axenic conditions. However,

M. Vostka et al.

still up-scaled mass in vivo production has some limitations,

for example not all fungal species can be cultivated successfully in axenic conditions. In non-axenic production systems
this issue remains a crucial point in the production and has
been dealt with since many years ago (Von Alten et al. 2002;
Vostka and Dodd 2002; Gianinazzi and Vosatka 2003;
Feldmann and Schneider 2008).
During the in situ host-plant mediated inoculum production, root samples of host plants used for AM fungi inoculum production should be microscopically checked for the
presence of potential pathogenic fungi. Additional test based
on trap plants susceptible to soil-borne pathogens can be
also be used, like cress, because it is very susceptible to root
rots. However, none of these methods is actually satisfactory
and there is an urgent need for molecular tests, for detecting
fungal pathogens in AM fungi inocula. Mycorrhizal industry is taking the necessary measures in order to ensure that
the inoculum producers will respect the defined criteria of
quality. For example, the Federation of European Mycorrhizal Fungi Producers (FEMFiP; has been
founded (Vostka et al. 2003) with the aims to achieve and
maintain the highest standards of inoculum quality. Also,
the internationally recognised IMS (International Mycorrhizal Society, is becoming very important,
which among others also has an aim to help corporations
producing inocula in promoting their products name and
quality. Nevertheless it still remains responsibility (at least
moral and selfdefending ones) of scientific community to
serve as watchdogs of the mycorrhizal products fairness and
Following mass production, fungal propagules must be
formulated in such a way that they can be stored and
distributed under a wide range of temperatures and without
loosing viability for quaranteed period. Formulation should
be simple, economic, and the inocula should be easy to
transport and apply. Some companies have adopted the
approach of single formulations for all markets, while others
produce a range of products for their target buyers. However, inoculum tuning seems to be necessary in particular
when targeted to more extreme conditions like arid, saline
soils, polluted soils or climate zones with distinct features
(Vostka and Dodd 2002).
The form of final product implies also the consequent
mode of application. Dry formulation of inocula can be
homogenised into a cultivation substrate or placed as a layer
below the seeds. It can also be applied as a gel formulation
on bare root seedlings or root balls or even cuttings. There
are several products on the market in a liquid form, however, viability of the AM fungal propagules in non-sterile
liquid media is generally very short, usually only few days,
since bacterial and mold contamination builds up in few
days (Vostka M, Ltr A, Kolom P, unpublished results
of experiments conducted in the scientific division of the

Development of arbuscular mycorrhizal biotechnology and industry

Symbiom company.). For example, commercial micropropagated plants can be inoculated post-vitro (Vestberg et
al. 2002; Gryndler et al. 2002) straight at the transplantation
stage or bare roots plants can be dipped into gel formulations of mycorrhizal inocula before transplantation, or the
dry formulations of inocula can be spread into the planting
hole. For large-scale application, machinery is needed, i.e.
mixing tanks for application in substrates or sowing
machines for field applications.
One of the main issues will always be economical feasibility of mycorrhizal applications especially in low value
and extensive crops. Cost benefit ratio of different applications is crucial for any large scale use of mycorrhiza. Savings on fertilizers, water, pesticides should be compared
with increased yield of target crops and cost of inoculum
and inoculum application should be taken into account.
Cost/benefit calculations for commercial rose production
(production facilities size about 100 ha) showed that by
reducing NPK to 50 % and application of the AM fungi at
planting time, a total 267 Euro is saved per hectare per
month (cost of AM fungi is calculated 2 Euro per L) and
resulting saving is about 1.5 M Euro for commercial production of roses in Kenya (Vostka M, Maimba F, unpublished results on rose production in Kenya.). The inoculation
is, however, too costly for extensive crops like cereals.
Main problem of inoculations is that it is not always working and there might be lack of consistent positive results in
field applications. Sometimes present native fungi are abundant in target cultivation systems and they can be sufficiently
effective and costs of inoculation is not outbalanced by the
outcomes. Advances in molecular techniques enable to prove
effectiveness of AM fungal inoculation expressed by improved
yield even in high presence of indigenous populations of AM
fungi in the field soil (Hernadi et al. 2012; Pellegrino et al.
2012; Sykorova et al. 2012). Future research should focused
more on the interaction of introduced and native mycorrhizal
populations and on tracing of inoculated strains to determine
their persistence of inocula in field soils and their effectiveness
in forming colonization with either annual or perennial crops.
Another problem often encountered in mycorrhiza application is that in conventional practice of plant production there are numerous agricultural practices (e.g.
agrochemical applications, tillage, cropping systems),
which inhibit mycorrhiza development, particularly in top
soil layers (Oehl et al. 2004, 2005). There are concerns
particularly on the use of pesticides. Insecticides and fungicides (particularly systemic, copper ones) have been
reported to exhibit many negative effects on soil organisms
whereas few significant effects of herbicides have been
documented (Bunemann et al. 2006). Another concern
arose based on new findings on the role of mycorrhiza in
allelopathy via common mycorrhizal networks (Cipollini
et al. 2012). Authors suggest, that implications of


microbial reintroduction (or controlled inoculations) in

agricultural systems on allelochemical modification should
be explored, since fungal network seem to serve as highways for allelochemical movement directly from donor to
target plants. But it is still unclear how important this
mechanism can be in field conditions.
Other aspect of different mycorrhizal responsiveness of
common crops currently under consideration is orientation
of selection and breeding programs during the last century
towards maximum plant performance in high-input production agroecosystems and concern of selection of modern
lines towards phosphate uptake without AM fungi (Sawers
et al. 2008). As an example is often given wheat (Triticum
aestivum) varieties developed before and after 1900, more
and less responsive to AM colonization, respectively
(Hetrick et al. 1992). However, recent study based on a
meta-analysis on 39 studies from 1981 to 2010 (Lehmann
et al. 2012) stated that new crop plant genotypes did not lose
their ability to respond to mycorrhiza due to agricultural and
breeding practices.
The development of an industrial activity producing microbial inocula is a complex procedure that involves not
only the achievement of the necessary biotechnological
know-how, but also the ability to respond to the specifically
related legal, ethical, educational and commercial requirements (Gianinazzi and Vosatka 2003). There are numerous
snake oils (unreliable products) on the market and not all
mycorrhizal products are capable to enhance effective mycorrhizal symbiotic associations (Tarbell and Koske 2007).
They are often labelled as mycorrhizal inoculum and list
numerous AM fungi (and other mycorrhizal fungi, bacteria
etc.), however, true content is different and presence of
viable microorganisms is very low or none. In the 10- week
test of 5 different commercial inocula performed repeatedly
on maize it was shown that some inocula used in dosage
recommended was capable to enhance maize growth in inert
substrate (autoclaved zeolite) fertilized only by Hoagland
solution once a week (Fig. 1a), however, only two out of 5
products were capable to form any mycorrhizal colonization
(Fig. 1b). It shows that the effects was rather accountable
to content of nutrients, aminoacids, humic acids and similar components rather than development of beneficial
mycorrhizal association. Similar failures in testing commercial inocula have been reported previously (Rowe et
al. 2007). Obviously the end users of the inocula i.e.
farmers or horticulturalists have little chance to estimate
mechanisms of the effect eventhough it is visible and
therefore with these inferior quality products that just
buy packs of very expensive organic fertilizers which
can do the job of making plant greener and larger in a
short term but would never provide formation of effective
mycorrhizal symbiosis desirable for longer term and sustainable mycorrhizal effects.


M. Vostka et al.

5 Solutions and ways forward in mycorrhizal

Mycorrhizal applications in agriculture are dependent on
scientific evidence supplied by fundamental and applied
research in collaboration with end usersthis is the main
direction of the way forward: deepening of collaboration of
mycorrhizal scientists, mycorrhizal industrial producers and
mycorrhiza end users. Challenge for areas with welldeveloped conventional agricultural cultivation systems
remains selection of appropriate inocula formulations and
adjustment to more-mycorrhiza friendly cultivation practices in order to ensure inoculation efficacy.
Furthermore, crop breeding programs could include aspect of mycorrhizal responsiveness to help to attain maximum benefits following mycorrhizal symbiosis in crop
production. Recent advances in molecular mechanisms of
AM symbioses (Smith and Smith 2011) and breeding
approaches are helping to optimize the use of AM fungi
for improvement of crop productivity. Recent study based
on meta-analysis indicates that in general, new crop plant
genotypes have not lost their ability to form mycorrhiza and
that plant breeders can include new cultivars in their germplasms (Lehmann et al. 2012).
To improve consistency in inocula applications there is
essential need to ascertain that inocula will be standardized
and will have always the same quality. The development of
internationally recognized methods of quality assurance is
an important step to be taken on the way to achieve standardization of mycorrhizal products.
Based on scientific evidence, to meet various demands of
soils and climate it is better to adhere to inoculum tuning and to
use native fungal strains in inocula formulations (Vostka and
Dodd 2002). To reduce costs of products the inocula should be
optimally produced locally especially for overseas regions.
There are also important concerns about shifting foreign unautochtonous inocula between continents (Schwartz et al. 2006)
and local production with native strains of AM fungi can help
to solve this problem. There certainly are ubiquitous AM

dry shoot weight in g




Inoc 1

Inoc 2

Inoc 3

Inoc 4

Inoc 5

Fig. 1 Test of five commercial AM fungi based products regularly

sold on EU and USA markets (Inoc 1 to Inoc 5, their real names
are not disclosed): a Dry shoot weight (g) and b the root mycorrhizal colonization (%) evaluated on maize after 10-week-

mycorrhizal colonization in %

fungal species found in variable soil conditions of several

continents, however, there seems sometimes be evidenced
level of the AM fungal species specificity towards pH, plant
species, pollutants level or degree of soil disturbance and
inoculum tuning often offers suitable solution (Vosatka and
Albrechtova 2009), what applies also to ectomycorrhizal fungal inoculum (Vostka et al. 2008b). The main question is
whether for perrenial crops the resilience of introduced AM
fungal strain is sufficient so the inoculated fungal species can
still be found after few years persisting in the root system. This
has already been proven possible in the field conditions
(Sykorova et al. 2012).
An important issue is also to adjust application modes
suitable for a given cultivation system. For example, development of functioning seed coating is essential for large scale
application on extensive crops as cereals. Innovation of products leading to use of highly concentrated, high quality and
efficacy of biological component can substantially enhance use
of mycorrhiza in plant production. More intensive application
modes via liquid formulation applicable in irrigation system
can increase substantially feasibility of mycorrhization in hydroponic cultivation systems. Also retrofitting of mycorrhizal
products into soil around mature perrenial plants like trees and
shrubs via injection can be supporting tool for further increase
in mycorrhiza use. It has been proven by molecular tools that
injected AM fungal strain into root system of mature plant is
capable to colonise root already colonized by other AM fungal
species (Vostka M, Ltr A, Kolom P, unpublished results of
experiments conducted in the scientific division of the Symbiom company) or that two fungal species can coexist and
interact in single plant root system (Janoukov et al. 2012).
There are several supporting aspects for further development of mycorrhizal technology and that is certainly growing demands for sustainability and organic products. Also
constantly increasing cost of fertilizers and future scarcity of
some fertilizing components like phosphates support development of biological alternatives.
One of the most important aspects for the development of
mycorrhizal industry is to increase awareness and education




Inoc 1

Inoc 2

Inoc 3

Inoc 4

Inoc 5

cultivation in zeolite. Bars above columns represent SD, the different letters above the bars indicate significant differences betwen
means (n06) P<0.05, ANOVA

Development of arbuscular mycorrhizal biotechnology and industry

leading to organic approach as until recent times there was a

little understanding among farmers and potential users. In
developing countries, many farms are not using agrochemicals but only due to the fact that they are either not available
or are too expensive. Intensive agriculture practices are
accepted without adherence to the scientific principles and
ecological aspects once economic situation allows it to
enhance productivity. For professional growers consistent
and convincing data are crucial for their decision making
whether to use or not to use biological alternatives. Therefore good scientific large-scale trials in real practice and
education of farmers are paramount. Currently each company in mycorrhizal industry is and should in fact be not only a
producer and marketer but also to certain extent a research
and educational body collaborating with scientists, because
of rather low awareness of mycorrhizal products.

6 Conclusions
The main way forward in mycorrhizal applications in agriculture depends on the scientific knowledge derived from
fundamental and applied research and collaboration of both
mycorrhizal science and mycorrhizal industry. That is driving force for advances in safe and effective applications of
mycorrhizal products in agricultural practices.
Population growth together with climate change, ongoing soil degradation and increasing costs of chemical
fertiliser is making the need for the envisaged New
Green Revolution which comprises better exploitation
of existing soil resources including proper soil-microbe
management (Lynch 2007).
There is growing number of enterprises producing inocula based on mycorrhizal fungi and recently not only in
developed world but increasingly in emerging markets. Also
collaboration between private sector and scientific community has an improving trend as the development of private
sector can fuel further research activities. Last but not least
there is apparent growing pull of the market and increasing
tendency of reduction of agrochemical inputs and employment of alternative strategies in planting and plant production. These circumstances support further developments of
mycorrhizal inocula production and applications and maturation of the industry.
Still the missing components of biotechnology are appropriate, cheap, highly reproducible and effective methods for
inocula purity testing and quality control. Also there is a
weak traceability of the origin of the mycorrhizal fungi
strains used in commercial inocula. Appropriate and accessible methods for quality and products certification are
paramount as there are numerous low quality inocula occurr
on the markets. We should be aware that mycorrhiza inoculation is not a Panacea, which is going to sort out all the


problems of plant production. Nevertheless, mycorrhizal

inoculation can be a part of a good practice in
ecologically-friendly plant cultivation, in particular in
specific conditions of degraded, arid and nutrient-poor
soils or horticultural substrates lacking mycorrhiza. The
crucial point is, however, that protection and proper
management of native AMF populations in soils is a
primary tool to exploit positive effects of mycorrhizal
symbiosis phenomena.
In summary the achievements of mycorrhizal industry are
very promising, challenges though remain and there are
numerous bottlenecks to solve. Further research is necessary
to facilitate commercial exploitation of mycorrhizal fungi.
In overcoming these drawbacks the collaboration of science
and industry is essential. Appeal is very clear and consist in
working together to support achievements and overcome
bottlenecks of the mycorrhizal industry. Main solution is
to join efforts of fundamental and applied research, farmers
and industry to provide consisting evidence for benefits of
mycorrhiza in the real conditions of plant production. Only
then it will be possible that mycorrhiza will be considered as
a part of good practice in sustainable horticulture and agriculture and it will also help to select reliable products from
the market from an inferior ones and help mycorrhizal
industry (and research) to grow.
Acknowledgments The authors are thankful to the manuscript editor
and two anonymous referees for all their suggestions and comments,
which helped to improve manuscript. The authors acknowledge funding of R&D from TACR TAO2020544, projects funded by the Ministry of Education, Youth and Sports of the Czech Republic in the EUcoordinated scheme Eurostars MycoDripSeed E!4366, and the project
of the Ministry of Industry and Trade of the Czech Republic FR-T11/
299 and the R&D project 9210AAO003S3427 funded by the Conseil
Rgional de Bourgogne (France). Two co-authors Ph.D. Ale Ltr and
Prof. Silvio Gianinazzi are employed by mycorrhizal inoculum producer and inoculum distributor, respectively. They declare that their
contribution to the manuscript is based on purely scientific knowledge
and, thus, does not constitute any conflict of interest.

Adholeya A (2012) Development and testing of mycorrhiza in multilocation field trials for improved crop yield under different cultivation systems and soils. In: Book of Abstracts, 7th International
Symbiosis Society Congress (The earths vast symbiosphere, July
2228, 2012), Krakw, Poland, p 254.
Albrechtova J, Latr A, Nedorost L, Pokluda R, Posta K, Vosatka M
(2012) Dual Inoculation with Mycorrhizal and Saprotrophic Fungi Applicable in Sustainable Cultivation Improves the Yield and
Nutritive Value of Onion. Sci World J. doi:10.1100/2012/374091
Barrios E (2007) Soil biota, ecosystem services and land productivity.
Ecol Econ 64:269285. doi:10.1016/j.ecolecon.2007.03.004
Bhargava A, Carmona FF, Bhargava M, Srivastava S (2012)
Approaches for enhanced phytoextraction of heavy metals. J
Environ Manage 105:103120. doi:10.1016/j.jenvman.2012.04.002

Bunemann EK, Schwenke GD, Van Zwieten L (2006) Impact of
agricultural inputs on soil organismsa review. Aust J Soil Res
44:379406. doi:10.1071/SR05125
Camprubi A, Calvet C, Estaun V (1995) Growth enhancement of
Citrus reshni after inoculation with Glomus intraradices and Trichoderma aureoviride and associated effects on microbial populations and enzyme activity in potting mixes. Plant Soil 173:233
238, View at Scopus
Caravaca F, Hernandez T, Garcia C, Roldan A (2002) Improvement
of rhizosphere aggregate stability of afforested semiarid plant
species subjected to mycorrhizal inoculation and compost addition. Geoderma 108:133144. doi:10.1016/S0016-7061
Cipollini D, Rigsby CM, Barto EK (2012) Microbes as targets and
mediators of allelopathy in plants. J Chem Ecol 38:714727.
De Smet I, White PJ, Bengough AG, Dupuy L, Parizot B, Casimiro I,
Heidstra R, Laskowski M, Lepetit M, Hochholdinger F, Draye X,
Zhang HM, Broadley MR, Peret B, Hammond JP, Fukaki H,
Mooney S, Lynch JP, Nacry P, Schurr U, Laplaze L, Benfey P,
Beeckman T, Bennett M (2012) Analyzing lateral root development: how to move forward. Plant Cell 24:1520. doi:10.1105/
Declerck S, Strullu DG, Fortin JA (eds) (2005) In vitro culture of
mycorrhizas. Springer, Heidelberg, p 388. ISBN 9783540240273
Declerck S, Cranenbrouck S, Ijdo M (2011) Methods for large-scale
production of AM fungi: past, present, and future. Mycorrhiza
21:116. doi:10.1007/s00572-010-0337-z
Den Herder G, Van Isterdael G, Beeckman T, De Smet I (2010) The
roots of a new green revolution. Trends Plant Sci 15:600607.
Douds DD, Lee J, Rogers L, Lohman ME, Pinzon N, Ganser S (2012)
Utilization of inoculum of AM fungi produced on-farm for the
production of Capsicum annuum: a summary of seven years of
field trials on a conventional vegetable farm. Biol Agric Hortic
28:129145. doi:10.1080/01448765.2012.693362
Feldmann F, Schneider C (2008) How to produce arbuscular mycorrhizal inoculum with desired characteristics. In: Feldmann F,
Kapulnik Y, Baar J (eds) Mycorrhiza Works. Deutsche Phytomedizinische Gesellschaft, Braunschweig, pp 292310. ISBN 978-3941261-01-3
Fernandez-Aparicio M, Westwood JH, Rubiales D (2011) Agronomic,
breeding, and biotechnological approaches to parasitic plant management through manipulation of germination stimulant levels in
agricultural soils. Botany 89:813826. doi:10.1139/B11-075
Gianinazzi S, Gianinazzi-Pearson V (1988) Mycorrhizae: a plants
healt insurance. Chimica Oggi, Octobre, 5658
Gianinazzi S, Vosatka M (2003) Inoculum of arbuscular mycorrhizal
fungi for production systems: science meets business. Can J Bot
82:12641271. doi:10.1139/B04-072
Gianinazzi S, Gollotte A, Binet MN, van Tuinen D, Redecker D, Wipf
D (2010) Agroecology: the key role of arbuscular mycorrhizas in
ecosystem services. Mycorrhiza 20:519530. doi:10.1007/
Gosling P, Hodge A, Goodlass G, Bending GD (2006) Arbuscular
mycorrhizal fungi and organic farming. Agric Ecosyst Environ
113:1735. doi:10.1016/j.agee.2005.09.009
Gryndler M, Vostka M, Hrelov H, Catsk V, Chvtalov I, Jansa J
(2002) Effect of dual inoculation with arbuscular mycorrhizal
fungi and bacteria on growth and mineral nutrition of strawberry.
J Plant Nutr 25:13411358. doi:10.1081/PLN-120004393
Hao Z, Fayolle L, van Tuinen D, Chatagnier O, Gianinazzi S, Li X,
Gianinazzi-Pearson V (2012) Local and systemic mycorrhizainduced protection against the ectoparasitic nematode Xiphinema
index involves priming of defence gene responses in grapevine. J
Exp Bot 63:36573672. doi:10.1093/jxb/ers046

M. Vostka et al.
Harrier LA, Watson CA (2003) The role of arbuscular mycorrhizal
fungi in sustainable cropping systems. Adv Agron 79:185225.
Hernadi I, Sasvari Z, Albrechtova J, Vosatka M, Posta K (2012)
Arbuscular mycorrhizal inoculant increases yield of spice pepper
and affects the indigenous fungal community in the field. Hortscience 47:603606
Hetrick B, Wilson GWT, Cox TS (1992) Mycorrhizal dependence of
modern wheat-varieties, landraces, and ancestors. Can J Bot
Hinsinger P, Brauman A, Devau N, Gerard F, Jourdan C, Laclau JP, Le
Cadre E, Jaillard B, Plassard C (2011) Acquisition of phosphorus
and other poorly mobile nutrients by roots. Where do plant
nutrition models fail? Plant Soil 348:2961. doi:10.1007/
Janos DP (2007) Plant responsiveness to mycorrhizas differs from
dependence upon mycorrhizas. Mycorrhiza 17:7591.
Janoukov M, Krak K, Caklov P, Vostka M, torchov H (2012)
Intraradical dynamics of two coexisting isolates of the arbuscular
mycorrhizal fungus Glomus intraradices sensu lato as estimated
by real-time PCR of Mitochondrial DNA. Appl Environ Microbiol 78:36303637. doi:10.1128/AEM.00035-12
Jeffries P, Gianinazzi S, Perotto S, Turnau K, Barea JM (2003) The
contribution of arbuscular mycorrhizal fungi in sustainable maintenance of plant health and soil fertility. Biol Fertil Soils 37:116.
Jung SC, Martinez-Medina A, Lopez-Raez JA, Pozo MJ (2012)
Mycorrhiza-induced resistance and priming of plant defenses. J
Chem Ecol 38:651664. doi:10.1007/s10886-012-0134-6
Kasuya MCM, Costa MD (2009) Abstracts, ICOM6, 6th International
Conference on Mycorrhiza Beyond the Roots, 914 August
2009, Bell Horizonte, Brazil.
Khush GS (2001) Green revolution: the way forward. Nat Rev Genet
2:815822. doi:10.1038/35093585
Kohlen W, Ruyter-Spira C, Bouwmeester HJ (2011) Strigolactones: a
new musician in the orchestra of plant hormones. Botany 89:827
840. doi:10.1139/B11-063
Lehmann A, Barto EK, Powell JR, Rillig MC (2012) Mycorrhizal
responsiveness trends in annual crop plants and their wild
relatives-a meta-analysis on studies from 1981 to 2010. Plant Soil
355:231250. doi:10.1007/s11104-011-1095-1
Lynch JP (2007) Roots of the second green revolution. Aust J Bot
55:493512. doi:10.1071/BT06118
Maillet F, Poinsot V, Andr O, Puech-Pags V, Haouy A, Gueunier M,
Cromer L, Giraudet D, Formey D, Niebel A, Martinez EA,
Driguez H, Bcard G, Dnari J (2011) Fungal lipochitooligosaccharide symbiotic signals in arbuscular mycorrhiza. Nature
469:58U1501. doi:10.1038/nature09622
Meier S, Borie F, Bolan N, Cornejo P (2012) Phytoremediation of
metal-polluted soils by arbuscular mycorrhizal fungi. Crit Rev
Environ Sci Technol 42:741775. doi:10.1080/10643389.
Miransari M (2011a) Hyperaccumulators, arbuscular mycorrhizal fungi
and stress of heavy metals. Biotechnol Adv 29:645653.
Miransari M (2011b) Soil microbes and plant fertilization. Appl Microbiol Biotechnol 92:875885. doi:10.1007/s00253-011-3521-y
Oehl F, Sieverding E, Mader P, Sieverding E, Mader P, Dubois D,
Ineichen K, Boller T, Wiemken A (2004) Impact of long-term
conventional and organic farming on the diversity of arbuscular
mycorrhizal fungi. Oecologia 138:574583. doi:10.1007/s00442003-1458-2
Oehl F, Sieverding E, Ineichen K, Ris EA, Boller T, Wiemken A
(2005) Community structure of arbuscular mycorrhizal fungi at
different soil depths in extensively and intensively managed

Development of arbuscular mycorrhizal biotechnology and industry

agroecosystems. New Phytol 165:273283. doi:10.1111/j.14698137.2004.01235.x
Opik M, Metsis M, Daniell TJ, Zobel M, Moora M (2009) Large-scale
parallel 454 sequencing reveals host ecological group specificity
of arbuscular mycorrhizal fungi in a boreonemoral forest. New
Phytol 184:424437. doi:10.1111/j.1469-8137.2009.02920.x
Pellegrino E, Turrini A, Gamper HA, Cafa G, Bonari E, Young JPW,
Giovannetti M (2012) Establishment, persistence and effectiveness of arbuscular mycorrhizal fungal inoculants in the field
revealed using molecular genetic tracing and measurement of
yield components. New Phytol 194:810822. doi:10.1111/
Perner H, Rohn S, Driemel G, Batt N, Schwarz D, Kroh LW, George E
(2008) Effect of nitrogen species supply and mycorrhizal colonization on organosulfur and phenolic compounds in onions. J
Agric Food Chem 56:35383545. doi:10.1021/jf073337u
Pimentel D (1996) Green revolution agriculture and chemical hazards.
Sci Total Environ 188(Suppl 1):S86S98. doi:10.1016/0048-9697
Piotrowski JS, Denich T, Klironomos JN, Graham JM, Rillig MC
(2004) The effects of arbuscular mycorrhizas on soil aggregation depend on the interaction between plant and fungal species. New Phytol 164:365373. doi:10.1111/j.14698137.2004.01181.x
Plenchette C, Clermont-Dauphin C, Meynard JM, Fortin JA (2005)
Managing arbuscular mycorrhizal fungi in cropping systems. Can
J Plant Sci 85:3140
Rillig MC (2004) Arbuscular mycorrhizae, glomalin, and soil aggregation. Can J Soil Sci 84:355363
Rivera-Becerril F, Calantzis C, Turnau K, Caussanel JP, Belimov AA,
Gianinazzi S, Strasse RJ, Gianinazzi-Pearson V (2002) Cadmium
accumulation and buffering of cadmium-induced stress by arbuscular mycorrhiza in three Pisum sativum L. genotypes. J Exp Bot
Rowe HI, Brown CS, Claassen VP (2007) Comparisons of mycorrhizal
responsiveness with field soil and commercial inoculum for six
native montane species and Bromus tectorum. Restor Ecol 15:44
52. doi:10.1111/j.1526-100X.2006.00188.x
Sawers RJH, Gutjahr C, Paszkowski U (2008) Cereal mycorrhiza: an
Ancient symbiosis in Modern Agriculture. Trends Plant Sci
13:9397. doi:10.1016/j.tplants.2007.11.006
Schwartz MW, Hoeksema JD, Gehring CA, Johnson NC, Klironomos JN, Abbott LK, Pringle A (2006) The promise and
the potential consequences of the global transport of mycorrhizal fungal inoculum. Ecol Lett 9:501515. doi:10.1111/
Selosse MA, Baudoin E, Vandenkoornhuyse P (2004) Symbiotic
microorganisms, a key for eco-logical success and protection of
plants. C R Biol 327:639648. doi:10.1016/j.crvi.2003.12.008
Smith SE, Read DJ (2008) Mycorrhizal symbiosis, 3rd edn. Academic,
Smith SE, Smith FA (2011) Roles of arbuscular mycorrhizas in plant
nutrition and growth: new paradigms from cellular to ecosystem
scales. Annu Rev Plant Biol 62:227250. doi:10.1146/annurevarplant-042110-103846
Sykorova Z, Borstler B, Zvolenska S, Fehrer J, Gryndler M,
Vosatka M, Redecker D (2012) Long-term tracing of Rhizophagus irregularis isolate BEG140 inoculated on Phalaris
arundinacea in a coal mine spoil bank, using mitochondrial
large subunit rDNA markers. Mycorrhiza 22:6980.

Tarbell TJ, Koske RE (2007) Evaluation of commercial arbuscular
mycorrhizal inocula in a sand/peat medium. Mycorrhiza 18:51
56. doi:10.1007/s00572-007-0152-3
Tchameni SN, Ngonkeu MEL, Begoude BAD et al (2011) Effect of
Trichoderma asperellum and arbuscular mycorrhizal fungi on
cacao growth and resistance against black pod disease. Crop Prot
30:13211327. doi:10.1016/j.cropro.2011.05.003
Thomas RJ, Akhtar-Schuster M, Stringer LC, Marques MJ, Escadafal
R, Abraham E, Enne G (2012) Fertile ground? Options for a
science-policy platform for land. Environ Sci Pol 16:122135.
Tsuchiya Y, McCourt P (2012) Strigolactones as small molecule communicators. Mol Biosyst 8:464469. doi:10.1039/c1mb05195d
Vazquez MM, Cesar S, Azcon R, Barea JM (2000) Interactions between arbuscular mycorrhizal fungi and other microbial inoculants (Azospirillum, Pseudomonas, Trichoderma) and their effects
on microbial population and enzyme activities in the rhizosphere
of maize plants. Appl Soil Ecol 15:261272
Vestberg M, Cassells AC, Schubert A, Cordier C, Gianinazzi S (2002)
Arbuscular mycorrhizal fungi and micropropagation of high value
crops. In: Giananazzi S, Schepp H, Barea JM, Hasselwandter K
(eds) Mycorrhizal technology in agriculture. Birkhuser Verlag
Basel, Switzerland, pp 223233
Von Alten H, Blal B, Dodd JC, Feldmann F, Vosatka M (2002) Quality
control of arbuscular mycorrhizal fungi inoculum in Europe. In:
Gianinazzi S, Schuepp H, Barea JM, Haselwandter K (eds) Mycorrhizal technology and agriculture. Birkhuser Verlag, Basel, pp
Vostka M, Albrechtov J (2008) Theoretical aspects and practical uses
of mycorrhizal technology in floriculture and horticulture. In:
Teixeira da Silva JA (ed) Floriculture, ornamental and plant
biotechnology: advances and topical issues 5, 1st edn. Global
Science Books, Isleworth, pp 466479
Vosatka M, Albrechtova J (2009) Microbial strategies for crop improvement. In: Khan MS, Zaidi A, Musarrat J (eds) Benefits of
arbuscular mycorrhizal fungi to sustainable crop production.
Springer, Dordrecht, pp 205225
Vostka M, Dodd JC (2002) Ecological considerations for successful
application of arbuscular mycorrhizal fungi inoculum. In: Gianinazzi
S, Schuepp H, Barea JM, Haselwandter K (eds) Mycorrhizal technology in agriculture. Birkhauser Verlag, Basel, pp 235248
Vostka M, Dodd JC, Patten R, von Alten H, Hutter I, Blal B, Dorrego
A, Estaun V, Kapulnik Y, Giovannetti G, Cassels A, Gianinazzi S
(2003) A joint initiative for the use of mycorrhizal fungi in plant
production (the establishment of the Federation of European
Mycorrhizal Fungi Producers FEMFiP). Folia Geobot 38:235
237. doi:10.1007/BF02803155
Vostka M, Albrechtova J, Patten R (2008a) The international market
development for mycorrhizal technology. In: Varma A (ed) Mycorrhiza. Springer, Dordrecht, pp 419438
Vostka M, Gajdo J, Kolom P, Kavkov M, Oliviera RS, Franco AR,
Sousa NR, Carvalho MF, Castro PML, Albrechtov J (2008b)
Applications of ectomycorrhizal inocula in nursery and field plantings: the importance of inoculum tuning to target conditions. In:
Feldmann F, Kapulnik Y, Baar J (eds) Mycorrhiza works. Desutsche
Phytomedizinische Gesellschaft, Braunschweig, pp 112124
Vostka M, Ltr A, Albrechtov J (2008c) How to apply mycorrhizal
inocula in a large-scale and what outcome can be expected in
respect to plant growth and cultivation costs. In: Feldmann F,
Kapulnik Y, Baar J (eds) Mycorrhiza works. Desutsche Phytomedizinische Gesellschaft, Braunschweig, pp 323339