The word "mycorrhizae" literally means "fungus roots" and

defines the close mutually beneficial relationship between

specialized soil fungi "mycorrhizal fungi" and plant roots. Duclos
et al., (1983) concluded that inoculation with mycorrhizae
stimulated the growth and flowering of Erica carnea plants. The
mycorrhizae formed during the first month remained localized
and after 6 months, the roots were usually no longer colonized by
the fungus. Macroscopic and microscopic characters of the
mycorrhizae were studied using light and scanning (SEM) and
transmission (TEM) electron microscopy. The hyphae contained
simple septa and woronin bodies (ascomycytes). The abundance
of lipid globules and the presence of electron-dense granules in
the vacuoles characterized the intracellular hyphae. These hyphae
are enclosed in a network of fibrils, particularly at the penetration
points but also on external hyphae. These mycorrhizae seem to
survive for approximately 1 month only. The importance of the
inoculum as nutritional source must not be neglected since similar
stimulation of the plants was also produced by inoculation with
autoclaved slurry of mycelium.

Fusconi (1983) reported that the short root system plants

(Cistue incanus) examined at different times after inoculation
with mycorrhizae exhibited short lateral roots which could be
categorized as follows: (i) uninfected; (ii) with hyphae in the dead
outer root cap cells; (iii) with devolved sheaths. In the last case
the inner hyphae of the sheath penetrated through remains of dead
root cap cells which were trapped in the inner sheath. Remains of
root cells were also found in the outer sheath. Growth of hyphae
into the dead layers of root cap thus leads to formation and
development of the sheath.
 Formation of mycorrhizae on plant roots could be obtained
by inoculation of mycelium from in vitro cultures (Ceruti et al.,
1987) or from mycelial slurries papered from agar plants
(Danielson et al., 1984). Ceruti et al., 1987 compared the
mycorrhizae structure with sterile roots of the same age. They
found that the mycorrhizae were thin and the heritage net often
penetrated deeply into the periblem, with cells losing their nuclei
and cytoplasm. In the mycorrhizae there was a decrease in tannin
content. There was no evidence of hyperplasia or hypertrophy.
There were occasionally pores in the heritage net, in which case
periblem cells were alive.

Boudaraga and Dexheimer (1988) revealed that the

mycorrhizae observed in the cortical cells of some plant roots, 6
months after inoculation with glomus Fasciculaatum, had the
same basic structure as other known examples of mycorrhizae. In
the cells invaded by the endophyte, there was a large increase in
the amount of cytoplasm, which was rich in mitochondria. The
endoplasmic reticulum was well developed and its element
showed frequent contacts with host plasmalemma running along
the interface. Those may correspond to the transfer of the
precursors of the isolating layer into the matrix.

Mafia et al., (1993) examined the roots of 27 epiphytic and

terrestrial species of piperaceae. Terrestrial roots of 2 peperomia
spp. , 2 piper spp. And 2 pothomorephe umellatum contained
internal vesicles and/or arbuscular. No internal vesicles and/or
arbuscular were found in 3024 cm of fine roots of epiphytic
piperaceae, even through 15% of these root segments had
associated external typical glomalean hyphae. Glomus and
Acaulospora spores, and Gigaspora auxiliary cells occurred in
both canopy and terrestrial habitats.
 Nasim and Zahoor (1994) mentioned that both vesicles
and arbuscular were recorded in roots of 7 Arisaema spp.
examined , while only thick-walled vesicles and beaded mycelia
were found in scales and epidermis of the corms. VA-my
corrhizal hyphae and vesicles were observed in and along the
vascular tissue of the roots. Vesicles attained the shape of the
infected corm epidermal cells.

Untch and Weber (1995) indicated that the fungi

Ceroprgia spp., produced intercellular running hyphae and all
other structures common for VAM fungi in flowering plants. In
Periploca laevigate, colonization of the hyphae within the root
cortex changed from the intercellular type to an intercellular
growth, characteristic for VAM fungi in Gentianaceae.

Symbiotic germination tests between Gastrodia elata and

Mycena dendrobii were demonstrated by Fan et al., (1999). M.
dendrobii stimulated seed germination. Many seeds germinated
and formed protocorms, which were colonized by fungal hyphae.
The hyphae were commonly distributed in stipe cell, sub
epidermal parenchyma cell (SEP) and inner cortical parenchyma
cells (ICP), and were separated from protocorm cell cytoplasm by
electron lucent materials and the protocorm cell plasma
membrane.

Gao et al., (1999) pointed that in the first stages of

development, the spores and hyphae outside the roots served as
inoculum and infected the 0.5-1.0 cm zone behind the growth
point on the root tip. The hyphae of G.mosseae began to infect
and form arbuscules in the cortex of the host 2 weeks after
inoculation. The vesicles appeared betwwen the host cells in 4
weeks. The arbuscules became closer in 6 weeks and started to
disintegrate, but many besicles occurred in the cortex in 8 weeks.
The spores formed and gradually increased in 10 weeks. The
mycorrhizal infection rate, the vesicle number and the hyphae
infection rate on roots increased with time. Arbuscular
mycorrhizal fungi were mainly distributed in the upper and
middle levels of the soil and the infection rate and amount
decreased with increase in soil depth.

In pot experiments, Hemalatha and Selvaraj (2003)

determined the association of indian borage with VAM.
Extensive (92%) colonization of the roots of Indian borage in the
field by VAM was observed. Based on morphological characters,
the VAM isolate was identified as belonging to the genus
Glomus. Glomus aggregatum. Glomus ambisporum were
predominant, whereas Gigaspora, Acaulospora, Sclerocystic and
Scutellospore were rarely isolated.

Panwar and Tarafder (2006) pointed that filed study of 12

districts of arid zones of rajasthan wan undertaken to evaluate the
ooccurrence of three selected endangered medicinal plant species
(Leptadenia reticuata, Mitragyna parvifolia, Withania coagulans),
and arbuscular mycorrhizal fungal (AMF) associations with
them. Five genera were identified in the rhizosphere of these
selected plant species. A high diversity of AMF was observed
which varied between different host plant species. Among the
five genera, Glomus occurred most frequently, with ten species,
Acaulospora and scutellospora were found with three species,
respectively. While Gigaspora and paraglomus were detected
with one species each. Glomus constrictum, Glomus
fasciculatum, Glomus geosporum, Glomus intraadices, Glomus
mosseae and Glomus rubiforme were the most dominant species.
The AMF spore density was not clearly affected by the host plant
suggesting that biotic factors may be relatively less important
than abiotic/edaphic factors for establishing population pattern.
The spore density of AMF had a strong positive correlation with
soil pH and organic carbon content and a negative correlation
with Olsen's P content of the soil. The association with AMF of
these plant species native to the harsh environmental conditions
of the Indian Thar Desert may play a significant role in the re-
establishment and conservation of these multipurpose halophytic
plants.

Schalamuk et al., (2006) investigated the influence of

tilling, N fertilization and crop stage on arbuscular mycorrhizae
(AM) fungal species diversity in a wheat monoculture in the
pampa region of Argentina. Glomalean spores were isolated by
wet sieving and decanting from conventionally tilled and
nontilled soils cropped with wheat with or without N fertilization,
at three phenological stages of the crop (tilling, flowering and
grain filling) and fallow. Morphological characterization yielded
at least 24 AM fungi taxa in the field samples, belonging to six
genera of AMF: Acaulospora Archaeospora, Entrophospora,
Gigaspora, Glomus and Scutellospora. Tilling and fertilization
treatments did not result in decreased spore biodiversity. Wheat
phenology influenced AM communities, with highest spore
biodiversity during grain filling.

2-2- Types of mycorrhizae:

The two common types of mycorrhizae are the ectomycorrhizas
and endomycorrhizae the (more commonly known as arbuscular
mycorrhizas). The two groups are differentiated by the fact that
the hyphae of ectomycorrhizal fungi do not penetrate the cell wall
of the plant's root cells, while the hyphae of arbuscular
mycorrhizal fungi penetrate the cell wall.
2-2-1- Endomycorrhizae
Arbuscular mycorrhizas, or AM (formerly known as vesicular-
arbuscular mycorrhizas), are mycorrhiza whose hyphae enter into
the plant cell walls, producing structures that are either balloon-
like (vesicles) or dichotomously-branching invaginations
(arbuscules). The fungal hyphae do not in fact penetrate the
protoplast (i.e. the interior of the cell), but invaginate the cell
membrane. The structure of the arbuscules greatly increases the
contact surface area between the hypha and the cell cytoplasm to
facilitate the transfer of nutrients between them.

Arbuscular mycorrhizas are formed only by fungi in the

division Glomeromycota, which are typically associated with the
roots of herbaceous plants, but may also be associated with
woody plants. Fossil evidence and DNA sequence analysis
suggest that this mutualism appeared 400-460 million years ago,
when the first plants were colonizing land. Arbuscular
mycorrhizae were likely to have been very helpful at that time,
protecting plants from adverse conditions such as lack of water
nutrients.

Arbuscular mycorrhizae fungi are quite extraordinary

organisms. They have been asexual for many million years.
Unusually, individuals can contain many genetically different
nuclei (a phenomenon called heterokarosis).

(Kosuta et al., 2003).

This type of association is found in 85% of all plant families in
the wild, including many crop species such as the grains.
2-2-2 Ectomycorrhizae
Ectomycorrhizas, or EcM, typically form between the
roots of woody plants and fungi belonging to the divisions
Basidiomycota, Ascomycota, or Zygomycota. They are external
mycorrhizae that form a cover on root surfaces and between the
root's cortical cells.

Besides the mantle formed by the mycorrhizae, most of the

biomass of the fungus is found branching into the soil, with
some extending to the apoplast, stopping short of the
endodermis.

Ectomycorrhizas are found in 10% of plant families, mostly

the woody species, including the oak, pine, eucalyptus,
dipterocarp, and olive families.

2-2-3-Other forms of mycorrhizae:

Arbuscular and ecto- mycorrhiza associating with plants

belonging to the order Ericales form ericoid mycorrhiza, while
some Ericales form arbutoid and monotropoid mycorrhiza. All
orchids are mycoheterotrophic at some stage during their
lifecycle and form orchid mycorrhiza with a range of
basidiomycete fungi.

Research by Klironomos and Hart at the University of

Guelph, Ontario has found that the mycorrhizal fungus Laccaria
bicolor can lure springtails and kill them (probably with a toxin);
the fungus utilises the nitrogen from the decaying springtails, and
some of this nitrogen also becomes available to the plant that the
fungus forms mycorrhizae on (in this study, the Eastern White
Pine). Klironomos found that up to 25% of the plant nitrogen
came from springtails or insects. (Kabir and Koide 2000 and
Marschner and Timonen, 2004)

2-3- Benefits of mycorrhizae:

Brejda et al., (1993) pointed that mycorrhizae inoculated
plants had a greater number of tillers, greater shoot weight, root
weight, tissue P concentration and percentage P recovered, and a
lower root/shoot ratio than non-inoculated plants.

Mastsubara et al., (1994) concluded that VAM inoculation

caused both leaf sheaths and leaf blades to thicken in welsh
onions and enhanced shoot and crown development in asparagus.
Shot and root fresh weight (FW) increased when the plants were
inoculated with VAM. in most of the vegetables, the increase in
root FW was caused by an increase of the number of roots, but in
welsh onions and asparagus it was caused by an increase in the
number of roots and a thickening of the main roots. VAM
infection was recognized in most vegetables except Persia
(Brassica campestris) cv. Hamrumaki gokuwase. A high infection
level was obtained mainly in plants with marked growth response
to the fungus. In most vegetables (except Welsh onions), VAM
infection was observed only in lateral roots; the root caps were
uninfected.

Khanizadeh et al., (1995) found that VAM inoculation and

phosphorous (P) supply had no effect on inflorescence or flower
number, total yield, fruit weight, or crown number. Increasing
rates of P did not affect total dry shoot weight (DW), fresh weight
(FW) leaf area, total root DW or leaf number in the presence of
VAM. The cultivars responded differently to the VAM species.
VAM inoculation in combination with P at all rates increase total
shoot DW and FW, leaf area and leaf number compared with
application of P alone.

Torres et al., (1996) indicated that inoculation of onion,

delayed white rote epidemics by 2 weeks and provided significant
protection against the disease for 11 weeks after transplanting.
Mycorrhizal plants showed an increase of 22% in yield.

Vinita and Singh (1996) evaluated that inoculation of some

forest trees with mycorrhizae. Response of the inoculated plants
was significant when compared with controls regarding plant
height, root weight, shoot weight and total biomass. Inoculation
of Acacia nilotica plants with Glomus fasciculatum resulted in the
highest increases in root and shoot weight. Root: shoot ratio was
low seedings inoculation with in mycorrhizae compared to the
control.

Neelima and Janardhanan (1996) found that inoculation

with the VAM fungus, Glomus aggregatum increased the growth,
yield and oil content of palmarosa (Cymobopogan martini var.
motia) significantly over un-inoculated control.

El-Sawy et al., (1998) showed that inoculation with a mixture of

Azotabacter and Azospirilla, with a full dose of rock phosphate
and inorganic N-fertilizer, in combination with VAM remarkably
increased plant growth, and N and P content. the highest plant
content of Khellin (1.35%), an important medicinal active
constituent, was detected 180 days from cultivation in plants
inoculated with a symbiotic diazotrophs and VAM, and provided
with the full dose of inorganic N and rock phosphate.
 Saleh et al., (1998) showed that dual inoculation with
asymbiotic N2-fixers and VAM, in treatments supplemented with
full dose of rock-phosphate and inorganic N-fertilizer,
significantly increased the dry matter, nitrogen content and
alkaloids in the plants. This increase was accompanied by a high
percentage of mycorrhizae root infection.

Wani and Konde (1998) indicated that vesicular arbuscular

mycorrhizae (VAM) (Glomus mosseae, Gigaspora maragarita
and Acaulospora calospora) significantly increased the plant
height, number of functional leaves, dry matter, fresh and dry
weight of bulbs, and N and P uptake by garlic cv. Godavari plants
and available P2O5 in soil.

Banerjee et al., (1999) revealed that VAM, generally

enhance uptake and translocation of P and encourages plants and
development in nutrient deficient soils. Contradictory reports
about the role of VAM in sulphure (S) uptake and translocation
may result from insufficient consideration of soil S-status. In this
study, using soils of low S-status, VAM inoculation increased the
content of radioactively labeled S(35s) in shoots of maize plants.

Kothari et al., (1999) inoculated a pasteurized sandy loam

soil either with rhizosphere microorganisms VAM fungi (non-
mycorrhizal) or with the VAM fungus, Glomus intraradices and
supplied with 0,50 and 100 mg P /kg soil. G.sintraadices
subsatantially increased root and shoot ratio, specific and total
root length, nutrient (Pm Zn and Cu) concentrations in root and
shoot of the plants, total nutrient uptake and nutrient acquisition
per unit root length, compared to soil inoculation with
rhizosphere microorganisms excluding VAM fungi when the soil
was not amended with P. little or no response to the VAM fungus
was observed when the soil was amended with 50 or 100 mg P/Kg
soil.

Liu and Zhang (1999) showed that inoculation with VA

mycorrhizae increased the uptake of phosphorous and calcium
and growth of wheat and maize, especially in soils with low
phosphorous content and reduced the gab in phosphorus content
between rhizospheres and bulk soil.

Senthilkumar et al., (1999) found that, nitrate reductase

activity and protein content were higher in all the mycorrhizae
colonized maize (Zea mays) seedlings. Zinc content was
estimated in leaves and roots of infected and non-infected
seedlings. About 31% of heavy metal (zinc) accumulation was
found in the roots and about 69% in the leaves in the absence of
vesicular arbuscular mycorrhizae. In the VMA colonized
seedlings about 40% was accumulated in the leaves and 59% in
roots which point out to the retention of the observed heavy metal
in the roots.

Shirani and Alizadeh (2000) showed that application of

VMA in addition to nitrogen fixer Bradyhizobium japonicum to
soybean as inoculum caused a significant increase in potassium
uptake efficiency. A high phosphorus uptake efficiency (517
g/ka) resulted when 6 g phosphorus/m2 was applied to soybean
inoculated with both bacterium or fungus. Treatment with 12 g
phosphorus/m2 without using bacterium or fungus, decreased the
phosphorus uptake (160.6 g/Ka). The highest potassium uptake
efficiency (95.5 g/Ka) was obtained when soybean was
inoculated with both bacterium and fungus + 6 g phosphorus/m2
the lowest potassium uptake efficiency was obtained when none
of the treatment (bacterium, fungus) was applied.
 Abdel-Ati (2000) showed that inoculation of cowpea plants
with VMA, increased plant height, number of branches per plant,
number of fresh pods per plant, dry matter percentage, weight of
100 seeds and total dry seed yield per feddan in two successive
seasons.

Ratti et al., (2000) determined the effect of vesicular

arbuscular mycorrhizae (VAM) fungus, Glomus mosseae on the
growth and yield of menthol mint (menthe arvensis subsp.
Haplocalyx). The result showed that shoot height, shoot and root
fresh and dry weight, and oil yield were significantly high in
plants inoculated with VMA compared to the control.

Rupam et al., (2001) studied the effect of inoculation with

two species of VMA fungi (Glomus macrocarpum and G.
fasciculatum) in the presence or absence of fertilizer on the
growth and yield of essential oil in the seeds of Anthum
graveolens. was investigated. Singnificant increase was observed
in vegetative growth of mycorrhizal treated plants. The essential
oil concentration increased by 19.18% and 41.3% with G.
fascicualatum and G. macrocarpum inoculation, respectively in
plants grown with the recommended level of phosphorus in soil.

Liu et al., (2001) studied the effect of inoculation with VA

mycorrhizae on phosphorous transformation in the rhizosphere of
wheat plants. The result showed that VAM inoculation
significantly increased the uptake of the fractions (Olsen -P, Ca2
-P1, Al -P and Fe -P) by the wheat plants and improved their
growth, especially in the low-P soil.
 Gupta et al., (2002) mentioned that the VAM inoculation
significantly increased mint root colonization, plant height, fresh
herbage and dry matter yield, oil content and oil yield as
compared to non-inoculated cultivars. The effect of VAM
inoculation on the root colonization, growth and yield of mint was
more pronounced with the cv Shivalik than the cvs Kalka and
Gomati, indicating Shivalik as a highly mycorrhizal dependent
genotype. VAM inoculation significantly increased the uptake of
N, P and K by shoot tissues of mint, but most markedly increased
the uptake of P. The VAM-inoculated mint plants depleted the
available N, P and K in the rhizosphere soil as compared to non-
inoculated control plants, however the extent of nutrient depletion
was greater for P than N and K.

El-Sayed et al., (2002) indicated that the growth,

nodulation, nutrients content and nitrogenase activity of
mycorrhiza-treated Vicia faba plants were significantly increased
compared to untreated ones. Such increases were related to the
increase in mycorrhizal root infection.

Elwan et al., (2002) revelated that inoculation with VAM

along with P and Zn application in pot experiment using natural
field soil and soil infected with fusarium solani, significantly
reduced the disease incidence of cotton seedlings. Furthermore,
these treatments significantly increased the survival of plants and
gave the heaviest dry weights of shoots and roots compared to
non-mycorrhizae application, in either infected soil or natural
field soil. The highest percentage of mycorrhizal root
colonization was found in the plants grown in infected soil at low
P level. Mycorrhizal plants had a higher content of total
chlorophyll and total carbohydrates than non- mycorrhizal plants
in either infested or natural field soil. In infested soil, zinc
treatment increased K-concentration, regardless of mycorrhizal
inoculation at two P-levels. The same trend was observed with P
concentration. Mycorrhizal inoculation decreased Fe, Mn, Zn and
Cu contents in shoot of cotton plantsgrown in natural field soils.
On the other hand, mycorrhizal inoculation increased thier
concentrations in shoots of cotton plants grown on infested soils,
regardless of P and Zn levels.

Sharma et al., (2002) studied the effect of VAM and P

application at rates of 25, 50, 100, 150, 200 and 250 mg P/Kg soil
on maize plants. The VAM fungi tested were Glomus
fasciculatum, G. mosseae, G. macrocarpum and G. versiforme.
Regardless of P added to the soil, VAM inoculation increased the
concentration of P, Zn, Cu, Mn, and Fe in leaves. There was no
indication of nutrient interaction between P and Zn with respect
to VAM application. Plants grown with supplemental P had
greater P tissue concentration than those not receiving P. The
maximum nutrient concentration was observed when the plants
were fertilized with 50 ppm P and inoculated with Glomus
macrocarpum.

Galal et al., (2003) found that maximum wheat grain yield

was obtained with single inoculum of AM fungi while N and P
uptake increased to their maximum with dual inoculation
(AM+Rhizobium). Inoculation with rhizobium either alone or in
combination with AM had an enhancing effect on wheat growth
and N and P uptake. Except for the soil treated with the lowest
level of fertilizer, there was an increase in microbial biomass N
following biofertilizer application.

Ryan and Angus (2003) studied the role of arbuscular

mycorrhizae fungi (AMF) in nutrition and yield of dry land
autumn-sown wheat and field pea through 2-years crop sequence
experiment on a red loam in (SE) Australia. Hosts of AMF (wheat
and filed pea) and non-mycorrhizal canola were grown with 20
kg ha1 of P as superphosphate. These treatments led to 23 fold
increase in colonization by AMF for wheat and field pea. High
colonization however did not correspond with greater crop
growth in some cases.

Hemalatha and Selvaraj (2003) determine the symbiotic

response of Indian borage (Plectranthus amboinicus) to 8
vesicular arbuscular mycorrhizas (VAM) (Acaulospora
morrowae, G. aggregatum, Gigasporea maragarita, G.
ambisporum, G. deserticola, G. geosporum, Sclerocystis sinuosa
and Scuellospora calospora) during transplanting of the crop in
pot experiments. Inoculated plants generally had higher values of
plant height, biomass production, and nutrient uptake and content
compared to the uninoculated controls. Plants inoculated with
Glomus aggregatum recorded the highest values for the
parameters measured. Mycorrhizal inoculation also resulted in an
increase in the number of spores in the rhizosphere. No significant
increase in the essential oil content of Indian borage duo to VAM
inoculation was recorded.

Regvar et al., (2003) indicated that mycorrhizae

inoculation significantly increased the plant biomass parameters
of pepper and parsley, and the root biomass of carrots.

Ding et al., (2004) observed that, the survival rate of

Chamaenerion angustifolium seedlings was enhanced after
inoculation, with AM fungi. The indigenous AM fungi
significantly increased the leaf area and plant height of the
seedlings compared with that of the uninoculated controls.
 Grandcourt et al., (2004) reported that mycorrhizal
colonization, growth, phosphorus content, net photosynthesis and
root respiration of Dicorynia guianesis and Eperua falcate were
determined during 9-months growth period grown at three soil
phosphorus concentrations, with or without inoculation with
arbuscular mycorrhizas. * Seedlings of both species were unable
to absorb phosphorus in the absence of mycorrhizal association.
Both species benefited from the mycorrhizal symbiosis in terms
of phosphorus acquisition but the growth of E. falcata seedlings
was unresponsive to this mycorrhizal improvement of
phosphorus status, probably because of the combination of high
seed mass and P reserves, with low growth rate.