Inoculation studies in some tree species with a Glomus sp. obtained from iron ore mines in Chhattisgarh.

Tanushree Chatterjee, Pradeep Kumar Sahu*, Shilpi Chatterjee

Department of Biotechnology, Raipur Institute of Technology, RITEE, Raipur Chhattisgarh (India)

Corresponding author E mail: sahupradeep47@gmail.com

ABSTRACT:

Vesicular- arbuscular mycorrhizal associations with plants are widely distributed and are geographically ubiquitous. Vesicular- arbuscular mycorrhiza (VAM) fungi a group of important soil micro-organism. VAM importance in iron ore mines for revegetation and reclamination. In these present work inoculation studies with VAM of existing five tree species are namely, Dulbergia sisso, Gmelina arborea, , Diospyros melanoxylon, Delonix regia, Dendrocalamus strictus, in nursery pot culture. Comparative studies in different parameter like percentage root infection, no. of VAM, root length, shoot length, fresh and dry biomass, root and shoot length ratio, fresh and dry biomass ratio within 90 days inoculation. In many parameters the VAM inoculated plants show better performance and encouraged results compare then non inoculated control plants.

KEYWORDS: vesicular arbuscular mycorrhiza, inoculation, revegetation

INTRODUCTION:

“Mycorrhizae are symbiotic associations essential for one or both partners, between a fungus (specialized for life in soils and plants) and a root (or other substrate-contacting organ) of a living plant, that is primarily responsible for nutrient transfer. Mycorrhiza occurs in a specialized plant organ where intimate contact results from synchronized plant-fungus development.” Mined land sites are generally known to be nutrient poor and contain soils that are in dire need of stabilization to prevent erosion. Marked by the beginning works of J. R. Schramm, mine reclamation practices have included the use of mycorrhizal inocula to establish successful plant communities on mined sites (Danielson 1985).

Chhattisgarh state has rich sources of mineral resource especially iron and coal. Open cost mining is the dominant form of mining. The immediate effect of open cost mining is the removal of soil and vegetation cover. The extent of damage depends on location of mining site, scale of operation, mining method, degree of mechanization etc.

The restoration of site is difficult if scientifically not attended well in time. The upper surface of the soil is very active and is full of micro organisms. They develop particular type of symbiotic relationship which is unique to the type of soil and environmental conditions prevailing in the area, and the stabilization of such symbiotic relationship takes long time to develop Mycorrhiza with plant roots. This association helps the plant to establish on sites such as saline, alkaline, eroded or distributed due to human activities. The present investigation is proposed with an objective to find out suitable host VAM associations in dominant plant species in selected mines of Chhattisgarh.

Vesicular- arbuscular mycorrhizal fungi, a group of important soil micro-organism, are known to improve plant growth through better up-take of nutrient and water resistance to drought and increased tolerance or resistance to root pathogens. However, no efforts seem to have been directed to-wards the isolation and the identification of VAM fungi inhabiting wastelands, soil of high salinity, and mining degraded soil. (S. Bhadauri and R. Yadav 1999).

These soils are deficient in organic matter, high pH and adversely both physical and nutritional properties of the soil and thus make these soils inhospitable for plant growth. Many experiments to raise tree plantation on degraded soil fail due to high mortality and poor establishment. Healthy and quality seedlings, through difficult to grow are a prerequisite to the successful establishment to hard wood plants, particularly for mines soil sites. Beside rhizobia, endomycorrhizae improve the quality of seedlings in the tree nurseries. (Kormanic1980).

The AM fungi were isolated from soil of the different area (untouched area, mines dumped area and active mining site) of different Iron ore mines of Chhattisgarh, like Rajhara iron ore mines, Mahamaya iron ore mines and Aridongari iron ore mines. The survey was conducted of mine site to known the existing plant present in the area. In these present work inoculation studies with VAM of existing five tree species are namely, Dulbergia sisso (Roxb), Gmelina arborea (Roxb), Diospyros melanoxylon (Roxb.), Delonix regia (Boj. Ex. Hook) Raf., Dendrocalamus strictus (Roxb.), in nursery pot culture.

MATERIALS AND METHOD:

The soil was collected in polythene bags from 10-30 cm depth of the different area (untouch area, mines dumped area and active mining site) of different Iron ore mines of Chhattisgarh, and brought for analysis. Isolation of AM fungi was carried out by following the method of (Gerdemann and Nicolson, 1963). AM spores obtained from the iron ore mines were purified following the funnel technique (Menge and Timmer 1982).

The spores were then observed for distinguishing morphological characters such as size, shape and wall characterstatics under a compound microscope (model: Labomed CXL PLUS). The spores were identified according to the manual of identification of vesicular-arbuscular mycorrhizal (VAM) fungi by Schenck and Perez (1990), Morton and Benny (1990) and Mukerjee (1994). The INVAM worksheet for spore characterization was used for diagnosing the spores. The pure culture of Glomus sp. was maintained and multiplied on maize roots using half strength Hogland solution for weekly irrigation (Menge and Timmer 1982).

Five tree species namely, Dulbergia sisso (Roxb), Gmelina arborea (Roxb), Diospyros melanoxylon (Roxb.), Delonix regia (Boj. Ex. Hook) Raf., Dendrocalamus strictus (Roxb.), were consider for the study. The seed were sterilized with 0.01% of sodium hypochloride, and checked the percentage germination of seed by Petri plate method. After germination, the 14-day-old seedlings were transplanted for the study. In the green house, pots of 4 kg capacity were filled with sterilized (autoclaved at 15 lb, 121° C, for three successive days), soil, sand and compost at the ratio of 2:1:1. Inoculation of different AM isolates was done by spreading soil inoculum (aprrox 100 g), containing about 300-350 spores in each pot separately for each treatment. Only sterilized soil was taken as control. Plants of individual tree species were transplanted in 5 replicates separately, and watering was done as per the requirement uninoculated pots (control) were supplied with 25 ml of half strength of Hogland solution every fortnight; inoculated pots were supplied with the same solution only once at the beginning of the experiment and with phosphate-free solution thereafter.

Plants (90 days old) were harvested and their roots were treated for clearing and staining following the method of Phillips and Hayman (1970) to determine AM infection. The number of VAM present in soil to known according to Gerdemann and Nicolson, (1963). Data obtained in triplicate were subjected to statistical analysis Sokal and Rolf (1973).

Table: 01 - Effect of AM inoculation on growth and AM root infection of perineal tree species (90 days).

S. No.	Parameter	Treatment	1	2	3	4	5
1	% Root infection	--	70±0.707	58.6±1.66	49.8±1.77	64.2±0.86	52.4±1.288
2	No. of mycorrhizal inoculum (100 gm of soil)	--	58.8±1.158	61±2.387	43±1.549	63.6±1.12	63.6±1.435
3	Shoot length (in cm)	NM	18.46±1.389	7.30 ±0.300	6.44± 0.236	7.8 ±0.29	6.56± 0.471
3	Shoot length (in cm)	M	27.74±0.817	14.98±0.32	9.80±0.239	3.66±0.20	2.08±0.166
4	Root length (in cm)	NM	12.68± 0.34	11.70± 1.32	6.92± 0.25	7.58± 0.22	0.687±0.898
4	Root length (in cm)	M	21.34±0.84	13.9±1.007	13.96±0.21	11.94±0.16	14.44±0.61
5	Biomass(fresh wt. in gm/plant)	NM	0.237±0.017	2.21±0.83	2.06±0.01	2.12±0.012	2.047±0.033
5	Biomass(fresh wt. in gm/plant)	M	0.672±0.029	3.347±0.12	3.472±0.062	3.82±0.037	3.390±0.047
6	Biomass(dry wt. in gm/plant)	NM	0.098±0.008	0.985±0.80	0.870±0.011	0.905±0.02	0.895±0.032
6	Biomass(dry wt. in gm/plant)	M	0.275±0.025	2.015±0.14	1.905±0.067	2.015±0.03	1.840±0.040

RESULTS AND DISCUSSION:

The results of AM inoculation conducted under green house conditions with AM fungi on five tree species are presented in Table 1. The isolated and purified AM culture was identified as Glomus sp.

All species of forest trees were found to be mycorrhizal. The maximum percentage root infection was recorded in the root of Dendrocalamus strictus (70±0.707), followed by the Dulbergia sisso (64.2±0.86), Gmelina arborea( 58.6±1.66), Delonix regia (52.4±1.288) and Diospyros melanoxylon (49.8±1.77). In this connection, no. of VAM spore present in 100 gm of soil from each pot also varied the more number are found Dulbergia Sisso and Delonix regia almost the same, then Gmelina arborea and Diospyros melanoxylon and the least value in Dendrocalamus strictus. Due to those plant shows more no. of VAM present in soil means the percentage infection are low. The results indicated a wide spectrum infection potential of this fungi. Since, low percentage of infection in forest tree species has been observed; it cannot be correlated as an efficient parameter for determination of AM infectivity and affectivity. Mycorrhizal association provides additional support to plants for nutrient uptake in deficient soil. Incidence of fairly low percentage of root infection was apparent in all five species.

The present study showed the relative effects of Glomus sp. on the growth of host plant. As for as symbiotic benefits were concerned, most of the host plants, achieved increment in all parameters like shoot length, root length, fresh weight and dry weight (see table no.01). Inoculation resulted in enhancement of plant height as compared to uninoculated (control). Similar differential responses due to AM inoculations were observed in plant root length. Fresh and dry biomass of plant enhanced to the maximum over to the control. All inoculated plants showed increased in fresh shoot length over the control. The maximum percentage of root infection was recorded in AM- infected plants. Results also attributed towards the suitability of this Glomus sp. is the best inoculants for forest trees species being used as a revegetation and reclamination of destroyed land soils after mining.

Another parameter studies shoot-root and biomass ratio in different tree species under non-mycorrhizal and mycorrhizal conditions (see table no-02). Through, AM inoculations enhanced growth of forest tree species, the differences in plant height and biomass among the control and inoculated plants were not very significant in Dendrocalamus stictus and Delonix regia. However, the overall performance of Gmelina arborea, Diospyros melanoxylon and Dul bergia sisso was encouraged through mycorrhization.

Table: 02 - Shoot-root and biomass ratio in different tree species under non-mycorrhizal and mycorrhizal conditions. (90 days)

S. No.	Tree species	Non-Mycorrhizal		Mycorrhizal
S. No.	Tree species	S/R lenght	F/D Biomass wt.	S/R length	F/D Biomass wt.
1	Dendrocalamus stictus	0.667	2.41	1.065	2.44
2	Gmelina arborea	0.623	2.242	1.077	1.66
3	Diospyros melanoxylon	0.931	2.367	0.702	1.822
4	Dulbergia sisso	1.02	2.34	1.14	1.895
5	Delonix regia	0.677	2.28	0.836	1.842

ACKNOWLEDGEMENTS:

The authors are thankful to the Department of Science and Technology, for giving INSPIRE FELLOWSHIP, Govt. of India, for financial support.

REFERENCES:

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Website

www .google.com;mycorrhizal association; the web resource.

www.google.com/ Rajhara mines wikipedia.

www.google.com/ Ari-dongari mines wikipedia.

http://invam.caf.wvu.edu/myc_taxonamy/species htm.

Received on 10.11.2011

Modified on 12.12.2011

Accepted on 24.02.2012

Research J. Science and Tech. 4(1): Jan.-Feb. 2012: 45-47