Methods for Mass Production of AM Inoculants
M
ASS
PRODUCTION
OF
AM
INOCULANTS
M
ASS
P
R
ODUCTION
:
P
R
OBLEMS
AND
PROSPECTS
•
Being
obligate
symbionts
AM
fungi
could
be
mass
produced
only
in the
presence
of
living
roots.
•
Since
AM
fungal
associations
are
universal
and
have
been
reported
in
almost
all
terrestrial plants,
these
can
be
reproduced
on
a
wide
range
of
host plants.
•
There are
several
techniques
reported
for
mass
production
of
AM
inoculum.
A
.
I
N
VIVO
CULTURE
•
AM
fungi
are grown
on
roots
of green
house
plants
and
chopped
mycorrhizal
roots,
often
mixed
with
growth
media
containing
hyphae
and
spores,
are
used
as source
of
inoculum.
•
Soil
could
be
replaced
by
inert
substances
such as
vermiculite,
perlite,
sand or
a
mixture
of these
for crude
inoculum
production.
M
ASS
PRODUCTION
Spores
+
antibiotic
solution
(
streptomycin
of
220
ppm
concentration
for
15
min
)
Wash
spores
with mercuric
chloride
Wash
with
distilled
water
Inoculate
the
plant
pots ( Guinea
grass
or Bahiya
grass
)
Keep
in
green
house
for
3
-
4
weeks
Uproot
the
plants
Co
n
t….
C
ONT
….
Check
for
colonization
Again
keep for
field
growth
( 1 – 1½
months
)
Macerate
the
root
Check
for
moisture
content
(
only
5 %
should
be
there)
Use
as
biofertilizer
1.
Tank
for
mass
multiplication
of
AM
2.
Sprinkling
of
water
in
tank
with
vermiculite
3.
Making
of
furrows
to
sow maize
seeds
M
ETHOD
OF
PRODUCTION
OF VAM
4
.
S
o
w
i
n
g
t
h
e
s
e
e
d
s
i
n
f
u
r
r
o
w
s
5
.
V
i
e
w
o
f
t
h
e
m
a
i
z
e
s
o
w
n
A
M
p
i
t
6
.
V
e
r
m
i
c
u
l
i
t
e
c
o
n
t
a
i
n
e
d
r
a
i
s
e
d
A
M
i
n
f
e
c
t
e
d
m
a
i
z
e
p
l
a
n
t
s
•
M
ASS
PRODUCTION
OF
VAM
B
.
I
N
VITRO
/
AXENIC
CULTURE
TECHNIQUES
i)
Solution
culture
ii)
Ae
r
oponic
cultu
r
e
iii)
Root
organ
culture
I
)
S
OLUTION
CULTURE
•
Involves
growing infected
roots
in aqueous medium enriched
with
mineral
nutrients
required for the growth
of the roots
under
controlled
biotic
and
abiotic
conditions.
ii)
AEROPONIC
CULTURE
•
Involves
applying a
fine
mist
of nutrient
solutions
to colonized
roots
for
AM
fungal
inoculum
production.
III
)
R
OOT
ORGAN
CULTURE
•
Use of a modified agar medium (MS
rooting
medium)/
liquid
medium
for
creation of increased amount of
roots
from callus
tissue
and these
roots
are infected by
AM
spores
or
by
surface
sterilized
root
bits
obtained
from
mycorrhizal plant.
C
ROP
Mycorrhiza
species
Barley,
maize,
wheat
Glomus
spp.
Bean
Asaulospora
morrowiae,
Glomus,
Gigaspora
Pean
u
t
Glomus
fasciculatum,
Sclerocystis
dussi
Pea
Glomus
intraradices
C
o
t
t
on
Glomus
sp.,
Sclerocystis
sinuosa
Gigaspora
margarita,
Glomus
spp.,
Acaulospora
sp.
Black
pepper
Entrophosphora
colombiana,
Scutellospora
sp.
Carda
m
om
Glomus
fasciculautm
Cit
r
us
Glomus
faciculatum,
G.
mosseae
Mar
i
g
o
ld
Glomus
mosseae
Tomato,
potato
THANK YOU
Techniques for mass production of arbuscular mycorrhizal (AM) inoculants involve in vivo culture on green house plants, spore production with antibiotic treatment, field growth monitoring, and in vitro/axenic culture techniques. Solutions include using living roots, inert substances like vermiculite, and root organ culture. These methods aim to efficiently produce AM inoculants for enhancing plant growth and soil health.
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Presentation Transcript
MASS PRODUCTION OF AM INOCULANTS
MASS PRODUCTION: PROBLEMS ANDPROSPECTS Being obligate symbiontsAM fungi could be mass produced only in the presence of living roots. SinceAM fungal associations are universal and have been reported in almost all terrestrial plants, these can be reproduced on a wide range of host plants. There are several techniques reported for mass production ofAM inoculum.
A. IN VIVO CULTURE AM fungi are grown on roots of green house plants and chopped mycorrhizal roots, often mixed with growth media containing hyphae and spores, are used as source of inoculum. Soil could be replaced by inert substances such as vermiculite, perlite, sand or a mixture of these for crude inoculum production.
MASS PRODUCTION Spores + antibiotic solution ( streptomycin of 220 ppm concentration for15 min ) Wash spores with mercuric chloride Wash with distilled water Inoculate the plant pots ( Guinea grass or Bahiya grass ) Keep in green house for3 - 4 weeks Uprootthe plants Cont . CONT .
Checkforcolonization Again keep for field growth ( 1 1 months ) Macerate the root Check formoisture content ( only5 % shouldbe there) Use as biofertilizer
METHODOF PRODUCTIONOF VAM 2. Sprinkling of water in tank with vermiculite 3. Making of furrows to sow maize seeds 1.Tank for mass multiplication of AM
6. Vermiculite contained raised AM infected maize plants 4. Sowing the seeds in furrows 5. View of the maize sownAM pit
B. IN VITRO/ AXENIC CULTURE TECHNIQUES i) Solution culture ii)Aeroponic culture iii) Root organ culture
I) SOLUTION CULTURE Involves growing infected roots in aqueous medium enriched with mineral nutrients required for the growth of the roots under controlled biotic and abiotic conditions. ii) AEROPONIC CULTURE Involves applying a fine mist of nutrient solutions to colonized roots forAM fungal inoculum production. III) ROOTORGANCULTURE Use of a modified agar medium (MS rooting medium)/ liquid medium for creation of increased amount of roots from callus tissue and these roots are infected by AM spores or by surface sterilized root bits obtained from mycorrhizal plant.
Mycorrhiza species CROP Barley, maize, wheat Glomus spp. Bean Asaulospora morrowiae, Glomus, Gigaspora Peanut Glomus fasciculatum, Sclerocystis dussi Pea Glomus intraradices Cotton Glomus sp., Sclerocystis sinuosa Tomato, potato Gigaspora margarita, Glomus spp., Acaulospora sp. Entrophosphora colombiana, Scutellospora sp. Glomus fasciculautm Glomus faciculatum, G. mosseae Black pepper Cardamom Citrus Glomus mosseae Marigold
