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Life Science Research and Sustainable Development ISBN: 978-98-84663-33-9

2.2 Root and rhizospheric soil sample collection
The roots of the plants was collected along with the rhizospheric soil sample of Heteropogon
contortus L. widely growing in area of Mauli Mahavidyalaya, Wadala , N. Solapur , without any
harm to the root system. The collected material brought into laboratory, washed the roots
thoroughly with tap water until the removal of adhering soil. The washed roots were cut into 1cm
length each. The arbuscular mycorrhizal root colonization was studied by using the method
suggested by Phillips and Haymann (1970). The root segments were kept in 10% KOH and
autoclaved it for 15 minutes at proper pressure. After that KOH was removed and dipped these
root segments into 1N HCL for 5 minutes. After that root segments were stained in cotton blue
with lactophenol overnight. In next day such stained root segments were screened under
microscope, for its arbuscular mycorrhizal root colonization.
The whole mount of root segments showed presence of arbuscles, vesicles and mycelium
as the roots were positively mycorrhizal with colonization. The percentage of root colonization
was calculated by using following formula (Giovanneti M and Moss B, 1980).

Percentage of root colonization= X100

Along with calculation of root colonization, the air dried rhizospheric soil sample was preserved
for study of spore population. Isolation of spore was done by using method of Gerdmann and
Nicolson, (1963). 100 gm of air dried rhizospheric soil dissolved in water with few drops of tween-
20. The above solution was continually stirred with glass rod and finally allowed to settle down
for about 20 to 25 minutes.
Such settled solution then sieved through different sized sieves as 710mµ, 210mµ, 150mµ,
75mµ, 45mµ, 25mµ respectively. The content of each sieves now poured into separate Petri dishes
for examination of spores. The identification of spores were done by using the manual given by
Schenck and Perez, (1990).

3 Results and Discussion
The roots of Heteropogon contortus L. Showed the presence of all type of arbuscular mycorrhizal
root colonization as vesicular, arbuscular and mycelial with about 90%. The present vesicles were
with globular, rounded and elongated while hyphae were with branched structure. The hyphae
and vesicles were predominately observed.
The analysis of rhizospheric soil sample showed presence of 350 spores/100gm of soil. In
rhizospheric soil sample different AMF spores were observed in which Glomus was the genera
found predominately present as compared to other generas. Glomus was with G. microcarpum, G.
aggregatum, and G. mossae.
The roots of Vetiveria zizanioides L.c ollected from the botanical garden of Swami
Ramanand Teerth Marathwada University, Nanded showed the presence of 90%arbuscular
mycorrhizal fungal root colonization while the soil analysis showed presence of 240 spores in
100gm of soil (R.M. Mulani and S.B. Wankhede, 2015). The grasses show fibrous root system and
are efficient in nutrient absorption. However some grasses are heavily mycorrhizal (de la Pena,
E. Echeverria et al, 2006).Muthukumar and Udaiya (2000) noticed that 53 grasses were collected
from Western Ghats of South India only 18 species showed arbuscular mycorrhizal
colonization.Javaidet al (1995) also worked on the mycorrhizal root colonization in grass species.

https://jesjalna.org/Zoology-Publications/index.html 28 Department of Zoology, J. E. S. College, Jalna