ISOLATION AND MORPHOLOGICAL

CHARACTERIZATION OF MYCORRHIZHA FROM

PINEAPPLE (Ananas comosus)

Ana Maria D. Carino1 and Daniel Marvin B. Ramel2

1
Undergraduate Student, 2Faculty Member, Institute of Natural Sciences
School of Arts and Sciences, Saint Mary’s University
Bayombong, Nueva Vizcaya, Philippines

ABSTRACT

A mycorrhiza is a symbiotic relationship between a fungus and a root of a

plant. Pineapple (Ananas comosus) is one of the major products of Bagabag, Nueva
Vizcaya. The dramatic increase in farm inputs to this plant is now a major concern of the
farmers of the said area. This study aimed to isolate and characterize mycorrhizal fungi
found in pineapple plants. There were 4 successfully isolated fungi. Their mycelial
growth configurations, margins, color and elevations are varied.

Introduction fumigated or receiving high rates of

A mycorrhiza (plural mycorrhizae
or mycorrhizas) is a symbiotic
relationship between a fungus and a
root of a plant. This mutualistic
association provides the fungus with
carbohydrates produced by plants. In
return, the plant uses the mycelium’s
very large surface area to absorb
mineral nutrients from the soil, thus
improving the mineral absorption
capabilities of the plant’s roots.
Mycorrhizae are beneficial for the plant
partner in nutrient poor soils
(http://science.jrank.org/pages/4537/My
corrhiza-Importance-mycorrhizae.htm).
Increase mobilization and transfer of
nutrients; better development of
Phosphorus solubilizing bacteria;
modification of plant-pathogen relations
as well as soil-plant-water relations; and
increased production of plant hormones,
are just some of its effects. To name
some of its applications to agriculture,
mycorrhizas are practically used 1) in
soil/substrate that are constantly
fungicides to eliminate/reduce soil borne
pathogens; 2) in arid and semi-arid
regions; 3) and in vegetation of eroded
or mined areas (R.M.
Muchovej2,_____). The dependence of
roots to mycorrhiza depends on plant
species, soil and climate conditions.
Mycorrhizal plants, ranging from 92%-
95% of the plant species, are more
resistant to diseases and to the effects
of drought. Mycorrhizas are classified
into Endomycorrhiza, and
Ectomycorrhiza. This mutualism can be
obligate or facultative.
Pineapple (Ananas comosus) is
an edible tropical plant native to the
southern part of Brazil, and Paraguay. It
belongs to the family Bromelieceae,
order Bromeliales. In Nueva Vizcaya
particularly in Bagabag, pineapple is
one of the major products. People in this
town adapted pineapple production
since 1990’s and used inorganic
fertilizers to nourish their plants for good
growth and better yield. Pineapple
production improves the economic

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status of the people in such ways that it
gives income and provides labor. Over
the years, the cost of farm inputs has
increased dramatically, therefore more
capital is needed. This created the
major problem of the farmers nowadays.
In addition, the continuous usage of
inorganic fertilizers and pesticides
caused poor soil condition, resulting to
lesser harvest and smaller the yield
(Farmer Production Guide on Pineapple).
Studies regarding Ananas
comosus revealed that mycorrhiza helps
the plant in absorbing more minerals
thus increases yield (S. Thamsurakul et Fig.2. Close up view of gathered Ananas comosus
al,_______). With the application of
Mycorrhiza, the pineapple yield has
increased and the efficacy of mineral The present study aimed to
fertilizer improved (S. Thamsurakul, et isolate and characterize them according
al., _____).The use of mycorrhizal to their mycelial growth rate and colony
symbioses to promote plant growth characteristics.
while reducing the inputs of fertilizers
and pesticides is one of the focuses of
most researches today (source?) Materials and Methods

Figure 3 shows the general

procedure employed in the study.

Collection of Plant Material

Isolation of Mycorrhiza
from Pineapple Roots

Pure Culture Production

of Mycorrhiza

Fig.! Ananas comosus in its habitat

Characterization

Figure 3. Schematic diagram of the experimental

procedure

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Collection of Plant Materials Distinct fungal colonies that have
grown from the PDA plates were
Five samples of Ananas carefully isolated using an improvised
comosus were gathered at Sitio scalpel. The isolated mycelia were
Kurasay, Baretbet, Bagabag, Nueva transferred in another PDA plate so as
Vizcaya on August 28, 2009. Abiotic to come up with a pure culture of the
factors such as soil and air fungus. The cultures were periodically
temperatures were measured. Soil inspected to check their purity.
nutrients such as nitrogen, phosphorus
and potassium were also measured to Fungi Characterization
ascertain potentially available nutrients
for the plant. Isolates were characterized
according to mycelial growth of colony,
Isolation of Mycorrhiza colony margin and configuration, color,
and colony elevation. Diameter of the
The pineapples were dug from mycelial growth was determined
the soil using shovel. Its roots were
dipped in sterile distilled water to DMG = DV+DH
remove the fungal spores together with 2
some soil particles. The soil solutions Where:
were poured through the 3 layers of
DMG – diameter of the mycelial
sieve plates, which are no. 50, no.100,
growth
and no. 200 respectively (from widest
mesh to the finest). The filtrates were DV – measurement of the vertical
then collected in sterile beakers and diameter of the colony
transferred in sterile bottles with cover. DH - measurement of the horizontal
Samples of filtrate were taken from the diameter of the colony.
surface of the solutions using a sterile
pipette and were transferred in a petri Other colony characteristics were
dish with sterile distilled water determined through inspection of five-
afterwhich observation was done under day old cultures grown in selective PDA.
the stereomicroscope to see tadpole-like
bodies floating at the surface. When Results
tadpole-like bodies are present, these
were siphoned and inoculated in Fungal Characteristics
prepared PDA plates amended with
amoxicillin. An improvised procedure Mycelial Growth
was also employed wherein a sterile
loop was dipped at the upper surface of Table 1. Mycelial Growth of the
the filtrate after which the loop was Fungal Isolates
streaked on PDA plates. The plates
were incubated at a temperature of 30- Mean
350C for 24 hours. Day Isolate N Diameter
(mm)
1 A3 3 12.75
Pure Culture Production of Isolates A5 3 40.02
A1a 3 42.04

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 A1b 3 90.4
A3 3 22.66
A5 3 73.83
2 Table 2. Colony Characteristics of A3.
A1a 3 72.58
A1b 3 *
A3 3 30.5 Configuration Round with scalloped
margin
A5 3 *
3
A1a 3 *
Elevation Flat
A1b 3 *
A3 3 38.875
Margin Wooly
A5 3 *
4
A1a 3 *
Color Light brown
A1b 3 *
A3 3 51.70833
A5 3 *
5
A1a 3 *
A1b 3 *

* means the whole surface area has

been fully colonized already

Table 1 shows that the isolate

A1b has the fastest mycelial growth as
manifested by the total colonization of
the agar plate in 2 two days. A5 and
A1a were able to colonize totally the
agar surface during the second day. it Fig. 5a. Day 5 of A1a
only shows that the A5 and A1a were
slower in mycelial growth than A1b but
faster than A3.
Table 2. Colony characteristics of A1a.

Configuration Concentric

Elevation Hilly
Margin Wooly
Color White with dark green

Fig.4a. Day 5 of A3

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 Table 4. Colony characteristics of A52.

Configuration Concentric

Elevation Crateriform

Margin Branching

Color White with dark green

Conclusions and Recommendations

Sampled pineapples in Bagabag

Fig.6a. Day 5 of a A1b
are host to a number of mycorrizhae.
Each of the isolated mycorrhiza
exhibited varying mycelial growth rates.
Table 3. Colony characteristics of A1b.
Table 5 shows the summary of colony
characteristics of the isolates.
Configuration Irregular and
spreading Table 5. Summary of the Colony
Elevation Hilly
Characteristics of the different fungal
isolates
Margin Thread-like
Fungal Configuration Elevation Margin Color
Color Pomelo orange Isolate
A3 Round with Flat Wooly Light
scalloped brown
margin
A5 Concentric Crateriform Branching White
with
dark
green
A1a Concentric Hilly Wooly White
with
dark
green
A1b Irregular Hilly Thread- Pomelo
and like orange
spreading

It is recommended that further

studies be conducted on (a) the
taxonomic identity of the isolates; and
(b) field testing of the isolates on
pineapples.

Fig. 7a. Day 5 of A52

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 BIBLIOGRAPHY

 R. M. Muchovej2, Importance of
Mycorrhizae for Agricultural
Crops.
 _____________, Farmer
Production Guide on Pineapple.
 S. Thamsurakul,et al.., Increasing
Pineapple Yield Using VA
Mycorrhiza Fungi.available online
at
http://science.jrank.org/pages/45
37/Mycorrhiza-Importance-
mycorrhizae.htm. date retrieved:
August 14, 2009.
 Mycorrhiza.available online at
http://science.jrank.org/pages/45
37/Mycorrhiza-Importance-
mycorrhizae.htm.date retrived:
August 14, 2009.