The mass production or commercial production of VAM fungi (Vesicular Arbascular Mycorrhiza) or AM Fungi (Arbuscular Mycorrhiza Fungi) is given here.

1. Professor Jayashankar Telangana State Agricultural University
College of Agriculture, Rajendranagar, Hyderabad-30
Submitted By :
Arunodaya Maji
CARA-2018-110
Batch - D
Under the guidance of,
Course In-charge :
Dr. S. Triveni

2. CONTENTS :
•Introduction
•Types of mycorrhizae
•Mycorrhizal biofertilizer
•Applications of mycorrhizae
•Conclusion
•References

3. Introduction :
1. Mycorrhizae are mutualistic symbiotic associations formed between the roots of
higher plants and fungi.
2. It is an Greek word, mykes: mushroom or fungi; rhiza: root.
3. Fungal roots were discovered by the German botanist A B Frank in the last
century (1855) in forest trees such as pine.
4. In nature approximately 90% of plants are infected with mycorrhizae. 83%
Dicots,79% Monocots and 100% Gymnosperms.
5. Convert insoluble form of phosphorous in soil into soluble form.

4. An arbuscule of Glomus
versiforme in a root cortex cell
with branch hyphae densely
packed in the cortex cell of the
host
Arbuscule of Gigaspora margarita
with an elongated trunk hypha (T)
and tufts of fine branch hyphae
(arrows)

5. Developing arbuscule of
Glomus mosseae in a root
cell with fine branch hyphae
(arrows). The trunk (T) of
this arbuscule branched
from an intercellular
hyphae.

7. Mycorrhiza
• The term “mycorrhiza” was coined by A. B. Frank,
1885.
• Mutualistic symbiosis (non-pathogenic
association) between soil-borne fungi and roots of
plants.
• Key components of ecosystems
• Link plants within a habitat
• Labelled CO2 fed to tree can be
found in seedlings growing nearby
• Retain and conserve mineral
nutrients

8. Types of mycorrhizae :
• On the basis of morphological and anatomical features,
mycorrhizae are divided into the three types.
• 1. Endomycorrhizae
• 2. Ectomycorrhizae
• 3. Ectendomycorrhizae

9. Endomycorrhizae further classified in to
five types.
• 1. AM fungi (Arbuscular mycorrhizae)
• 2. Orchidoid mycorrhizae
• 3. Monotropoid mycorrhizae
• 4. Ericoid Mycorrhizae
• 5. Arbutoid mycorrhizae

10. Acaulospora dan Entrophosphora
Glomus, Gigaspora, Scutellospora
Spore formation and spore-based taxonomy

11. Vesicular-Arbuscular Mycorrhiza (VAM)
• Also called as Arbuscular mycorrhiza as vesicles are not formed in all
cases.
• Develop arbuscules and vesicles within root cortical cells.
Vesicles Arbuscles
• Intercellular hyphae form large
swellings (vesicles) at ends of hyphae
or intercalary
• Typically rich in lipids & used as
storage organs
• Surrounded by plant cell membrane
• Typically disintegrate after 2 weeks in
plant cell and release nutrients
• sites where carbohydrates and
nutrients are exchanged between cell
and VAM hyphae.

12. AM fungi (Arbuscule mycorrhizae) :
• Fungi formed AM association with plants may belongs to ascomycetes ,
basidiomycetes and zygomycetes.
• All AM fungi are obligate biotrophic, as they are completely dependent on
plants for their survival.
• Many microorganism form symbioses with plants that range from parasitic to
mutualistic. Among this the most widespread mutualistic symbiosis is the
arbuscular mycorrhizal association. Arbuscular mycorrhizal (AM) symbiosis
occurs between the fungi of the Glomeromycota (Schubler et al., 2001) and
majority of terrestrial plants. The phycobiont correspond to 80% of plant
species and this association involves an intimate relationship between plant
roots and fungal hyphae. This mutualism is manifested in bidirectional nutrient
exchange: the fungus is nourished by plant photosynthates and plant mineral
nutrition particularly phosphate is enhanced by the fungus (Smith and Read,
1997). AM fungi are obligate biotrophs, depending on living root tissue for
carbohydrate supply to complete their asexual life cycle.

13. Spores: These are the "fruiting bodies" of the fungi and are
formed both inside the roots and externally in the soil.
• Spores of diameters
ranging from 50 to 400
μm
• Depending upon the
season and conditions,
spores can make up a
significant amount of
mycorrhizal biomass.

14. Mycorrhiza as root extension
• The fungus also produces hyphae outside the roots
that serve as root extension; thereby increasing the
plant’s potential to absorb water and nutrients from
the soil.

16. Need to use mycorrhiza
Intensive management practices limit viability and
infectivity of native mycorrhizal fungi
Excessive applications of chemical
fertilizers and pesticides
Severe soil disturbances like
erosion, tillage, compaction
Non-native transplants grown in soil and
climactic conditions different from the
areas where they are planted
Areas under heavy environmental stresses

17. Different functions played by the arbuscular mycorrhizal (AM) fungi in the
physiology and ecology of their host plants.
Mycorrhizal hyphae interconnect roots with soil particles (3), provide direct connections of root
systems of different plant individuals (2), and interact with a number of soil microbes (4).

18. Mechanism of Improved plant growth
due to VA Mycorrhiza
 Increased nutrient uptake
 Synergistic beneficial interaction with other soil
micro-organisms
 Resistance to plant pathogens
 Better drought tolerance
 Production of growth promoting substances
 Stimulate the growth of beneficial microorganisms
 Improve soil structure – hyphal polysaccharides
bind and aggregate soil particles

19. Farm -level VAM root inoculants production
A. Materials Required:
• Sterilized soil
• Clay / plastic pots
• Corn / sorghum /raagi / Rice seeds
• VAM starter inoculants

20. Culturing of AMF
• The obligate symbiotic nature of the fungus makes axenic
cultivation an important challenge for both scientific and
practical point of view. Inability to culture AM in the laboratory is
the major limiting factor in their application in agriculture.
Though AMF has very broad specificity towards plants including
various agricultural horticultural and forestry plant species, but
the ability to produce AM in bulk quantities is a major bottleneck.
AM biofertilizer is currently recommended only for transplanted
and nursery raised crops because of the difficulty in inoculum
production as well as the bulk requirement of the inoculum.
• Various methods were developed for mass production of AM fungi
world wide.

21. Soil sample + sterile water
Hot water
Filter and sieve
( 719μm → 250μm → 50μm → 45μm)
Spores separated from soil particles
Mix with carrier material
Use when required as biofertilizer
i) Isolation
A ) Sieving method :

22. B ) Floatation method : Soil sample + sterile water
Separate the soil particles using
membrane filter
Centrifuge
( Density gradient centrifuge = at 3000rpm
for 30 min )
Spores separated from soil particles
Mix with carrier material
Use when required as biofertilizer

23. Mass Production :
• 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.

24. 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.

25. Mass production of VAM

26. 1. Tank for mass
multiplication of
AM
2. Sprinkling of water in
tank with vermiculite
3. Making of furrows to
sow maize seeds
Method of production

27. Method of production
4. Sowing the
seeds in furrows
5. View of the maize
sown AM pit
6. Vermiculite
contained raised AM
infected maize plants
•

28. In-Vitro Culture/Axenic Culture
Techniques
• i) Solution culture
• ii) Aeroponic culture
• iii) Root organ culture

29. 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.

30. ii)Aeroponic culture
• Involves applying a fine mist of nutrient solutions to colonized
roots for AM fungal inoculum production.

31. iii)Root 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.

32. On Farm Mass Production of Vessicular Arbuscular Mycorhiza
Method
Sterilize soil by heating for 2-4 hours using a big pan or talyasi or by drying under intense heat of
the sun for 2-3 days.
Place the sterilized soil in thoroughly cleaned and dry clay pots.
After cooling the soil, place a pinch of root starter inoculants then cover with a thin layer of soil.
Sow 3-5 seeds in each pot.
Grow the plants for three months under normal conditions. Protect the plants from pest and
diseases. Stop watering the plants after 3 months.
Cut the plants or stalks when they are completely dried. Allow the soil in the pot to dry further.
Remove soil adhering to the roots. Cut the roots finely and save some rot inoculants for future
use. Mix the finely cut roots with the soil/ vermi compost from the pot to produce VAM soil
inoculants.
Store the root and soil inoculants in sealed plastic bags in a dry and cold place.

33. VAM application
• Mycorrhizal application goal is to create physical contact between
the mycorrhizal inoculant and the plant root.
• Mycorrhizal inoculant can be sprinkled onto roots during
transplanting,
• worked into seed beds, blended into potting soil, “watered in” via
existing irrigation systems,
• applied as a root dip gel or probed into the root zone of existing
plants.
• Mycorrhizae inoculant can be applied by broadcast thinly on prepared
seedbed. Mix into the soil or in case of nursery, cover VAM soil
inoculant with a thin layer of soil.
The type of application depends upon the conditions and needs of the
user. Sow seeds on the seedbed and cover seeds with a thin layer of
soil. If necessary, apply fertilizer 15 days after sowing.
Do not use chemical fertilizer / fungicide on seedbed
before sowing if VAM is to be applied.

34. Application of AM fungi
• Nursery application
• 100 g bulk inoculum is sufficient for one m2. The inoculum should be applied a
2-3 cm below the soil at the time of sowing. The seeds/cuttings should be
sown/planted above the AM inoculum to cause infection.
• For polythene bag raised crops
• 5 to 10 g bulk inoculum is sufficient for each packet. Mix 10 kg of inoculum with
1000 kg of sand potting mixture and pack the potting mixture in polythene bag
before sowing.
• For out-planting
• 20 g of AM inoculum is required per seedling. Apply inoculum at the time of
planting.
• For existing trees
• 200 g AM inoculum is required for inoculating one tree. Apply inoculum near the
root surface at the time of fertilizer application.

35. Micorrhiza Application Rates
• Row Crop Application : side dress seed furrows or transplants @
6 to 9 kg/ha. Incorporate at 2-6 inches (5 to 15cm) below soil
surface.
• Seed Drilling: incorporate into the soil at a depth below the
seed.
• Broadcast and Till: Evenly distribute across seedbed after
seeding. Cover the exposed seed and inoculum by harrowing ,
chain dragging or applying an organic topdressing .
• Nursery Medium: Evenly blend @ 3 kg/ m3.
• Do not leave inoculum exposed to sunlight.

36. Application :
• Increase nutrient uptake of plant from soil.
• P Nutrition and other elements: N, K, Ca, Mg, Zn, Cu, S, B, Mo, Fe, Mn, Cl
• Increase diversity of plant. Produce uniform seedling.
• Significant role in nutrient recycling.
• More tolerant to adverse soil chemical constraints which limit crop production.
• Increase plant resistance to diseases and drought. Stimulate the growth of
beneficial microorganisms. Improve soil structure.
• Stable soil aggregate – hyphal polysaccharides bind and aggregate soil
particles.
• Increases absorption of phosphate by crops. uptake of zinc also increases.
• Increases uptake of water from soil. Increases uptake of sulphur from the soil
• Increases the concentration of cytokinins and chloroplast in plants.
• They protect plants during stress condition.

37. Benefits :
• Produce more vigorous and healthy plants.
• Increase plant establishment and survival at seedling or transplanting.
• Enhance flowering and fruiting.
• Increase yields and crop quality.
• Improve drought tolerance, allowing watering reduction.
• Optimize fertilizers use, especially Phosphorus.
• Increase tolerance to soil salinity.
• Reduce disease occurrence.
• Contribute to maintain soil quality and nutrient cycling.
• Contribute to control soil erosion.

39. No mycorrhizal treatment Mycorrhizal treatment No mycorrhizal treatment Mycorrhizal treatment

41. Few mycorrhizal products available
commercially

42. Conclusion :
• Mycorrhizal association is very essential for the plants because it
has several benefits like absorption of nutrients, increases drought
resistance, enhance plant efficiency in absorbing water and
nutrients from soil. Especially, AM fungi are very useful in the
agriculture because it serves as biofertilizers as it helps in the
absorption of phosphorus, and other nutrient uptake.

43. References :
• N S subba Rao, Soil microorganisms and plant growth, 3rd edition.
Oxford & IBH publishing co. Pvt. Ltd, New Delhi, page no 287-296.
• Tanuja Sing, S S Purohit and Pradeep Pairhar, Soil
• microbiology. Agrobios (India), page no 383-405.
• Himadri Panda and Dharamvir Hota, Biofertilizers and organic
farming, 2007 gene- Tech books New Delhi, page no 147-187.
• Rangaswamy G and Bhagyaraj. Agricultural microbiology and 2nd
edition. Prentice- hall of Indian private limited.
• Dr. H A Modi, Biofertilizers & organic forming, Aaviahkar
publishers, Distrubutros jaipur 302003(Raj) India.