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Catalogue
Training
on in vitro culture of AMF organised by the CESAMM
Reference book :
In
Vitro Culture of Mycorrhizas
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Cultivation techniques
Classical methodology
The most commonly used method to
propagate AMF remains the pot cultivation. However, this method
:
-
is time and space consuming ;
-
cannot guarantee the absence
of undesirable contaminants ;
-
requires regular assessment of
viability and monospecificity of inoculum ;
-
complicates the acquisition of quality
cultures.
In vitro
culture
When grown on a suitable synthetic growth
medium in association with excised roots, AMF cultures can be maintained
pure and viable for long term periods.
The methodologies commonly used
to grow AMF in GINCO are fully described by Cranenbrouck et al.
(methodologies for in vitro cultivation of arbuscular mycorrhizal
fungi with root organs) In : Declerck S., Strullu D.G. and Fortin
J.A. (Eds), In
Vitro Culture of Mycorrhizas, to be published in Springer-Verlag,
end March 2005.
Advantages
of the in vitro culture of AMF |
Reliability of
cultures |
contaminant free cultures
throughout life cycle. |
Monospecificity of fungal
inoculum |
no cross contamination
with other Glomeromycota throughout life cycle. |
Non-destructive monitoring |
morphological observations
conducted without disturbance of the intimate coexistence
of host roots and AMF during whole life cycle. |
Efficient system for |
- physiological studies
: culture conditions are standardized thus facilitating studies
on the impact of environmental factors ;
- molecular and biochemical studies : optimal conditions due
to the absence of undesirable contaminants ;
- mass-inoculum production : feasible through regular subculture.
|
Lower space and time requirements |
produced under growth
chamber controlled conditions. |
Easy quality control and
fungal growth quantification |
important for high scale
production of both in vivo and in vitro
propagation. |
Inconvenients
related to the in vitro culture of AMF compared to
pot cultures |
Sterile environment |
high precaution (controlled
atmosphere, laminar flow hood, …) are needed to succeed. |
Technical training |
may be
necessary to acquire expertise.
Training on in
vitro culture of AMF is organised by the Mycorhizal Research
Team (UCL).
[More
information...] |
Low sporulation levels
for some species |
make them less distributable
or distributable at a prohibitive price. |
Number of available strains |
several strains are cultivated
but few distributed. |
Continuous cultivation |
difficulty to maintain
some strains under continuous growth. |
Physiological and genetic
characters |
potential loss of strains
characters due to successive transfers. |
Perspectives
With the availability for the scientific
and the industry communities of high quality and contaminant-free
AMF inocula, very innovative research and development possibilities,
up to now neglected, will be greatly facilitated :
-
by much easier procedures for the
maintenance of clean and reliable fungal culture ;
-
by a much better control of inoculum
quality and effectiveness ;
-
by access to an experimental system
that allows refined physiological and molecular investigations up
to large-scale inoculum production.
References
Cranenbrouck S., Voets L., Bivort
C., Strullu D. G. and Declerck S., Methodologies for in vitro
cultivation of arbuscular mycorrhizal fungi. In : Declerck
S., Strullu D.G. and Fortin A.(eds.), In
Vitro Culture of Mycorrhizas, to be published in Srpinger Verlag,
end March 2005.
Published Glomeromycota species cultivated in vitro on
root organs (Cranenbrouck et al.)
Gigasporaceae
(non producing vesicles) |
G. gigantea
(Nicolson & Gerd.) Gerd. & Trappe |
Gadkar et al., 1997
Mosse, 1988 |
G. margarita
Becker & Hall |
Miller-Wideman and Watrud, 1984
Gadkar et al., 1997
Karandashov et al., 1999 |
G. rosea
Nicolson & Schenck |
Bécard and Fortin, 1988, 1992
Diop et al., 1992 |
S. reticulata
(Koske, Miller, Walker) Walker & Sanders |
de Souza and Declerck, 2003 |
Acaulosporaceae
(producing vesicle) |
A. rehmii
Sieverding & Toro |
Declerck and Dalpé, 2002 |
Glomaceae
(producing vesicles) |
G. caledonium
(Nicolson & Gerd.) Trappe & Gerd. |
Karandashov et al., 1999, 2000 |
G. cerebriforme
McGee |
Samson et al., 2000 |
G. clarum
Nicolson & Schenck |
de Souza and Berbara, 1999 |
G. constrictum
Trappe |
Mathur and Vyas, 1999 |
G. deserticola
Trappe, Bloss & Menge |
Mathur and Vyas, 1995 |
G. etunicatum
Becker & Gerd. |
Pawlowska et al., 1999 |
G. fasciculatum
(Thaxter sensu Gerd.) Gerd.&Trappe emend. Walker & Koske |
Strullu an Romand, 1986
Declerck et al., 1998 |
*G. fistulosum
Skou & Jakobson |
Nuutila et al., 1995 |
G. intraradices
Schenck & Smith |
Strullu and Romand, 1987
Chabot et al., 1992
Diop et al., 1994a, b
St-Arnaud et al., 1996
Karandashov et al., 1999
Declerck et al., 1998
Boisson-Denier et al., 2001
Douds, 2002 |
G. macrocarpum
Tulasne &Tulasne |
Declerck et al., 1998 |
*G. mosseae
(Nicolson & Gerd.) Gerd. & Trappe |
Mosse and Hepper, 1975
Mugnier and Mosse, 1987
Karandashov et al., 1999
Douds, 1997
Raman et al., 2001 |
G. proliferum
Dalpé & Declerck |
Declerck et al., 2000 |
G. versiforme
(Karsten) Berch |
Diop et al., 1994
Declerck et al., 1998 |
Caption :
* : non-sporulating species
Additionnal unpublished AMF species cultivated
in vitro
Caption :
* : non-sporulating species
? : identification to be confirmed
Gigasporaceae
(non producing vesicles) |
Gigaspora
albida
Schenck & Smith |
Wu C-G (pers. comm.) |
*Scutellospora castanea
Walker |
Declerck S (pers. comm.) |
S. nigra
(Redhead) Walker & Sanders |
Wu C-G (pers. comm.) |
Acaulosporaceae
(producing vesicle) |
*Acaulospora
Laevis
Gerd. & Trappe |
Declerck S (pers. comm.) |
A. morrowae
Spain & Schenck |
Wu C-G (pers. Comm.) |
Glomaceae
(producing vesicles) |
Glomus aggregatum
Schenck &Smith emend. Koske |
Moutoglis P (pers.comm.) |
?G. lamellosum
Dalpé, Koske & Tews |
Dalpé Y (pers. comm.) |
| Glomus sp. |
Declerck S (pers. Comm.)
Dalpé Y (pers. comm.) |
Sclerocystis sinuosa
(Gerd.& Bakshi) Almeida & Schenck |
Wu C-G (pers.comm.) |
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