Research Articles

2017  |  Vol: 3(4)  |  Issue: 4 (July- August)
Diversity of Vesicular Arbuscular mycorrhizal fungi in Parthenium hysterophorus from different sites of Jabalpur M.P. India

Jitendra Nagpure1*, Sajad Mir2, A.K. Pandey3, Jamaluddin4

1,2Research scholars, Mycological Research lab, Rani Durgavati Vishwavidyalaya, Jabalpur, M.P. 482001 India

3Professor at Mycological Research lab Rani Durgavati Vishwavidyalaya, Jabalpur, M.P. 482001 India

4Emeritus scientist at University Grant Commission, New Delhi India

*Corresponding author

Jitendra Nagpure

Mycological Research Lab, Rani Durgavati Vishwavidyalaya (RDVV), Jabalpur (M.P.) 482001 India


Abstract

Background: Arbuscular mycorrhizal fungi are found in a wide range of habitats usually in the roots of angiosperms, gymnosperms and pterydophytes, whose diversity can be estimated by studying rhizopheric soil and also by colonization patterns in roots of the host plants. Objective: To find diversity of Vesicular Arbuscular mycorrhizal (VAM) fungi associated with Parthenium hystophorusMaterials and methods: Thus, an investigation was carried out on a weed plant ‘Parthenium hysterophorus’ to determine the mycorrhizal association potential of different fungi in Jabalpur district of Madhya Pradesh, India. Ten species of arbuscular mycorhiza fungi of the genus Glomus aggregatum, Glomus geosporum, Glomus fasciculatum, Glomusmosseae, Sclerocystiscorymeoides, Scetullospora jabalpurensis, Acaulospora appendiculata, Acaulospora rehmii, Gigaspora margarita and Archeospora were studied. Results: The highest VAM spore density was measured from RDVV Campus of approximately 119 density/50 gram soils and least was noticed from site Gauri ghat area. Species richness was maximum from RDVV Campus and from site Dumna park while site Sadar, Gorabazar, Adhartal contains species richness of 3 each. Site Gorabazar area has species richness of 4 each. The result suggested that AM fungi are well distributed in Parthenium plant species in central Jabalpur area.

KeywordsParthenium hysterophorus, Vesicular Arbuscular mycorrhizal fungi, Jabalpur district and frequently occurring Glomus species


Introduction

Increased efficiency of mycorrhizal roots and non efficiency in non-mycorrhizal roots is due to active uptake of following nutrients like, Phosphorous, Zinc and Copper (Phiri et al., 2003, Jamel et al., 2002). Khubato et al. (2005) described that the morphology of A.M. type association depend upon the interaction between plant and fungal species. Frank (1885) first gives this term ‘Mycorrhiza’ to define it as the essential structure and functioning of the peculiar connection between the roots and ectomycorrhizal fungi. They are known to enhance plant tolerance to biotic and abiotic stresses like nutrients, drought, metal toxicity, salinity and pathogens, which may affect successful establishment. Glomus was found to be most dominant as trace metal tolerant, as Arbuscular mycorhiza fungi isolated from contaminated part (Joner, 2003; Malcova, 2003). The AM fungi can increase the survival rate of plants, reduce susceptibility to environmental stresses, and increase nutrient aquision in plants, and increase carbon sequestration and nitrogen fixation in the soil (Almas et al., 2004). Spain et al., (2006) classified AM fungi into different groups based on the structure of their soil-borne spores and DNA sequences. The AM fungi, belonging to Glomeromycota  phylum, was classified into four orders, namely Archaeosporales with two families (Ambisporaceae and Archaeosporaceae), Diversisporales with four families (Gigasporaceae, Entrophosporaceae, Diversisporaceae and Acaulosporaceae), Paraglomerales with only one family of Paraglomeraceae and Glomerales with only one family of Glomeraceae. Glomus is the largest genus of AM fungi, belonging to Glomeraceae family, currently defined as non- monophyletic. Mycorrhizal associations may be initiated by spore germination. Most of the plant species can be colonized by AM fungi under natural stressed rangeland conditions (Neeraj et al., 1991). Miller (1979) showed that when soil from rhizosphere is removed, it resulted in decrease of mycorrhizal propagules. Local variant of AM fungi are more beneficent compared to foreign AM fungi used in that region (Requenta et al., 1997). A number of factors for successful colonization of AM fungi are pH, soil nutrients, organic matter, moisture, temperature, and the age of disturbed sites, which have shown correlation with AM root colonization and diversity (Mukhopadhyay and Mati, 2009). AM fungi colonise the plant roots to derive carbon for their survival. AM fungi give both intra and extra radical structures. It is fact that rate of translocation of nutrients becomes high during plant-microbe interaction. Microbes present in plant root rhizosphere are treated as plant growth promoting rhizo bacteria (Kumar et al., 2014-2015). Mycorrhizal diversity is mostly in forests compared to other area (Chaturvedi et al., 2009).

Materials and methods

Site and location

The location of Jabalpur is 23°10' North latitude and 79° 59' East longitude. Jabalpur is situated on Varanasi-Nagpur NH-7. Nestled in the 'Mahakaushal' region in the central part of India, it is has a peaceful ambiance and a tranquil environment, 
Jabalpur enjoys a prime location. It is located at the centre of Madhya Pradesh.

Figure 1. Location of Jabalpur

 

 

Collection of soils and root samples

Roots and soil samples were collected from the rhizosphere of plants growing in that area. From each site, 3-4 healthy plants of Parthenium were selected. The roots of plant and rhizosphere soil was dug out with a trowel to a depth of 0-15 cm after scrapping away the top 1 cm layer of soil. Samples were collected randomly from different zone in each site, pooled and homogenized. Before processing, all the samples were sieved (< 2 mm mesh size) to remove stones, coarse roots and other litter, and fine roots were collected from each sample. Soil samples were air dried and stored at 4°C for further experiments.

Isolation of Arbuscular mycorrhizal Species from plant roots

After sample collection, next step was isolation of Arbuscular mycorrhizal species collected from root samples. The following steps were made:

Fine root samples were collected and then washed with running tap water and fixed in FAA (Formalin Acetic acid). Roots were segmented into 1cm bits. Three replicates of 100 root bits each, selected at random were processed separately for determining the mycorrhizal intensity in the roots. Root bits were treated with 10% KOH solution for 30 min. at 40 ºC temperatures. The concentration of KOH and time of incubation of roots depend upon the age and softness of the roots.  Pour off the KOH solution and rinse the roots well in a beaker using at least three complete changes of tap-water or until no brown colour appears in the rinse water. After thorough washing, root bits were stained with trypan blue (0.01% trypan blue) for 24 hours at room temperature.  Stained root pieces were mounted in lactoglycerol and examined under microscope for the mycorrhizal colonization and its spore’s structures study.

Isolation of VAM spores from rhizosphere Soil mixtures

 Spores were isolated from field-collected root-rhizosphere soil mixtures. Spores of AM fungi were isolated by using the ‘wet sieving and decanting method’ described by (Gerdemann and Nicolson, 1963). In soil remove the coarse materials like straw, debris and rocks should be removed with a 2-mm sieve.100 g of air-dried root-rhizosphere soil mixture were put into a glass container with 1000 ml of tap water. The root-soil mixture was vigorously mixed with a glass rod for 30 seconds. A 10-second pause enabled to settle heavier particles and organic material, the remaining soil-water suspension were slowly poured through a set of two sieves. The sieves used are those with pores of diameters of 0.5mm (the top one) and 0.045 mm (lowest one). Most spores retain on the 0.045 mm sieve. The extracts were washed away and spores collected from the sieves in to Petri dishes. Using a microscope, spores and aggregates were picked by means of dropper and needle. Selected spores were separated with a needle. A drop or two of (poly vinyl lacto glycerol) was spread on the centre of a clean and dry slide so as to hold cover slip. Spores were placed on the mount and the cover slip was placed gently by avoiding air bubbles. Such prepared slides were allowed to dry in a dust free chamber for 3-5 days. The edge of the cover slip was sealed with clear nail polish to prevent the desiccation and entry of air bubbles. Spores were examined under microscope and photographs were taken.

Analysis of Soil Samples

Soils constitute the weathered surface of the earth’s crusts, which is mixed with organic material and microorganisms live and plants grow. Soil testing is one of the most important tools to determine the status of plant nutrients in a field. The air dried and sieved soil samples were analyzed for pH, Organic carbon, macro nutrients and micro nutrients.

Results and discussion

The highest VAM spore density was measured from RDVV Campus of  approximately 119 density/50 gram soils and least was noticed from site 3rd. Species richness was maximum from RDVV Campus and from site 5thDumna park, while site 2nd , 3rd, 4th and site 9th contains species richness of 3 each. Site 7th and 10th has species richness of 4 each. Lowest species richness was to be seen in site 6th. Organic matter was highest located from site 6th while lowest located from site 8th.

Table 1. Analysis of soil samples by different parameters

Soil sites

Water holding capacity

 

     pH

Organic         matter %

Arbscular Mycorhizal spore density/50gm soils

Species      richness

Site 1

30.73

6.67

2.07

119

5

Site 2

32.81

6.72

0.98

73

3

Site 3

38.76

6.55

2.90

31

3

Site 4

36.60

6.62

1.97

101

3

Site 5

35.76

6.78

1.39

33

5

Site 6

34.40

6.56

3.56

94

2

Site 7

37.70

6.71

1.34

50

4

Site 8

38.20

6.81

0.41

53

3

Site 9

33.31

6.69

3.31

84

3

Site 10

30.36

6.66

3.15

76

4

Site1 RDVV campus, Site 2ndSadar area, Site 3rdGorigath, Site 4thAdhartal area, Site 5thDumna park, Site 6thHathital, Site 7thGorabazar area, Site 8thRanital area, site 9th SFRI Jabalpur, Site 10th TFRI Jabalpur.

Table 2.  VAM fungi associated with Parthenium hysterophorus

Sites

VAM fungi identified from Parthenium hysterophorus

RDVV Campus

Glomus badium, Glomus geosporum, Sclerocystis corymeoides, Acaulospora laevis.

Sadar area

Glomus geosporum, glomus mosseae, Scetullospora jabalpurensis.

Gaurighat area

Glomus badium, Gigaspora margarita, Glomus fasciculatum, Acaulospora appendiculata, Glomus geosporum.

Adhartal area

Sclerocystis corymeoides, Glomus mosseae, Acaulospora rehmii, Gigaspora margarita.

Dumna park

Gigaspora margarita, Glomus fasciculatum, Glomus badium, Acaulospora laevis.

Figure 2. Pictures showing different species of VAM fungi isolated from Parthenium hystophorus

 

 

 

 

 

 

Glomus badium were isolated from RDVV campus area, Gauri ghat area and Dumna park Glomus geosporum were isolated from RDVV campus, Sadar area and Gauri ghat area. Sclerocystis corymeoides were isolated from RDVV campus  and Adhartal area. Acaulospora laevis from RDVV campus and Dumna park, while Glomus mosseae were isolated from Sadar area and Adhartal area. Scetullospora was recorded only from site Sadar area. Gigaspora margarita was found from site Gauri ghat, Adhartal and Dumna park. Glomus fasciculatum were found from Gauri ghat and Dumna park. Acaulospora apendiculata were to be seen only in Gauri ghat area. Acaulospora from Adhartal area only. Only genus Glomus seems to show high occurrences with this Parthenium species. It can be surveyed that the majority of species of genus Glomus may adapted a stronger mechanism of symbiosis with different plant hosts as some sort of a co-evolving mechanism. From the result Glomus species was the predominant genus, followed by Acaulospora Species. The predominant occurrence of Glomus is due to their potential to survive in any kind of area, which shows adaptation in any area of our Jabalpur region. Our findings were similar to the observations of Opik et al (2006). Our results also indicated that soil pH decreased with increasing soil depth, due to percolation of water in this region, leading to formation of acids. Spore population were having correlation with the PH of soil. Soil moisture also increased with increased depth. AMF fungi give negative relation with moisture content.

Conclusion

The highest VAM spore density was measured from RDVV Campus of approximately 119 density/50 gram soils and least was noticed from site Gauri ghat area. Species richness was maximum from RDVV Campus and from site Dumna park, while site Sadar, Gorabazar, Adhartal contains species richness of 3 each. Site Gorabazar area has species richness of 4 each. The result suggested that AM fungi are well distributed in Parthenium plant species in central Jabalpur area. Population of AM fungi, frequency of occurrences and their distribution varied with the soil depth, pH, and moisture content in that region.

References

Almås AR, Bakken LR, Mulder J. 2004. Changes in tolerance of soil microbial communities in Zn and Cd contaminated soils. Soil Biology and Biochemistry, 36(5): 805-813.

Chaturvedi S, Sharma AK. 2009. Arbuscular mycorrhizal fungal diversity in some medicinal plants. Myco News, 20(4):10-1.

Gerdemann JW, Nicolson TH. 1963. Spores of mycorrhizalEndogone extracted from soil by wet sieving and decanting. Transactions of the British Mycological Society, 46: 235-244.

Jamal A, Ayub N, Usman M, Khan AG. 2002. Arbuscular Mycorrhizal Fungi enhance Zinc and Nickle uptake from contaminated soil by soyabean and lentil. International Journal of Phytoremediation, 4(3): 203-221.

Joner EJ. Leyval C. 2003. Phytoremediation of organic pollutants using mycorrhizal plants: a new aspect of rhizosphere interactions. Agronomie, 23(5-6): 495-502.

Kubato M, Mc Gonigle TP, Hyakumachi M. 2005. Co-occurrence of Arum and Paris-type morphologies of arbuscular mycorrhiza in cucumber and tomato. Mycorrhiza, 15: 73-77.

Kumar R.  Prakash A,  Kumari S, Utkarshini, Kumari N, Kumar R, Sinha K, Kumar S. 2015. Cytotoxicity of ethanolic extracts of in vivo, in vitro and Biotized Grown Plants of Vernoniadivergens on EAC Cell Lines. International Journal of Pharmacognosy and Phytochemical Research, 6(4): 678-684.

Malcova R, Gryndler M. 2003. Amelioration of Pb and Mn toxicity to arbuscular mycorrhizal fungus Glomus intraradices by maize root exudates. Biologia Plantarum, 47: 297-299.

Miller RM. 1979. Some occurrence of vesicul ararbuscular mycorrhiza in natural and jhum fallow ecosystems of the Red Desert. Canadian Journal of Botany, 57: 619-623.

Mukhopadhyay S, Maiti SK. 2009. VAM fungi –A future prospect for biological reclamation of mine degraded lands. International Journal of Environmental Protection, 29(9): 801-809.

Neeraj SA, Mathew J, Varma AK. 1991. Occurrence of VA mycorrhizae within Indian semi-arid soild. Biology and Fertility of Soils, 11: 140-144.

Phiri S, Rao IM, Barrios  E, Singh BR. 2003. Plant growth, Mycorrhizal association, Nutrient uptake and phosphorus dynamics in a volcanic-ash.Soil in Colombia as affected by the establishment of Tithonia diverssifolia. Journal of Sustainable Agriculture, 21(3): 41- 59.

Requenta N, Jimenez I, Toro M, Barea JM. 1997. Interactions between plant growth promoting rhizobacteria (PGPR), Arbuscula rmycorrhizal fungi and Rhizobium spp. in the rhizosphere of Anthyllis cytisoides, a model legume for revegetation in Mediterranean semi-arid ecosystem. New Phytologist Journal, 136: 667-677.

Spain JL, Sieverding E, Oehl F. 2006. Appendicispora: a new genus in the arbuscular mycorrhiza-forming Glomeromycetes, with a discussion of the genus Archaeospora. Mycotaxon, 97: 163-182.

Manuscript Management System
Submit Article Subscribe Most Popular Articles Join as Reviewer Email Alerts