2. Mycorrhyzia
Word Mycorrhyzia was first used by german
researcher AB Frank in 1885
Myco-fungus Rhiza- Roots
Symbiotic relationship between special soil fungus
and fine plant roots
Mutualistic: both partners are benefited
Fungus: receive carbohydrate and growth factor
Plant: Increase nutrient absorption
Fungus: take role of plant root hair and extension
to root system
3. • Mycorrhzial association involve 3-
way interactions, between host
plant, Mutualistic fungi, and soil
factors
6. Role of Fungus in giving benefit to Tree
• Fungi increase supply of inorganic nutrients to trees
• P are insoluble in most soil
• Extrametrical hyphae extend over large volume of soil
than roots can
• Hormones produced by fungus
• Increase Tolerance of plant
• Increase resistant to infection by root pathogen-provide
physical barrier
7. Isolation and identification of
some Arbuscular Mycorrhiza (AM)
fungi for phytoremediation in soil
contaminated with paper mill
effluent
Dhitiman Chanda
9. Arbuscular Mycorrhiza
• Is a type of mycorrhizia in which fungus penetrates
the corticle cell of the roots of Vascular plants
• Characterized by formation of unique
structures,arbuscules and vesicles by fungi of
phylum glomerumycota
• Ubiquitous obligate mycobionts forming symbiosis
with terrestrial plant communities
• So far more than 170 species of AM fungi have
been recorded
10. Benefits of AM fungus
Increase rate of plant Survival
Reduce plant stress
Increase Plant nutrient acquisition
Increase Carbon and nitrogen deposition in soil
12. Research
• Arbuscular fungi enhance plant tolerance to varity of stresses including
nutirents, drought, metal toxicity, salinity and pathogens
• Study was taken to access the influence of paper mill effluent mycorrhizal
colonization and mycorrhyzial spore count and regression analysis revealed
that the mycorrhyzial colonization and mycorrhizal spore count are positively
correlated with physio chemical properties of polluted soil
• Glomus was dominant isolated mycorrhizal genus:
species Glomus fasciculatum Glomus macrocarpum, Glomus mosseae
• In this study we know the association between arbuscular mycorrhizal fungi in
plants growing in polluted soil and use of fungi as bioremediation agent for
polluted site
14. •Location of the study Area
•Collection of Soil Sample
•Collection of Root Samples
•Isolation of Micorrhizal spore
•Soil Physio-chemical analysis
15. Location of Study Area
The study was conducted at the polluted site inside the campus of Hindustan
Paper Corporation Limited, HPC, Assam, India where the solid sludge and
effluent have been dumped.
16. Collection of Soil Sample
• From the polluted soil,the rhizospheric soil samples were randomly selected
and then mixed together to obtain a composoil representative sample.
• The soil samplings were done from January 2012 to October 2013 in three
seasons, i.e., winter (November to February),summer (March to June) and
Monsoon or rainy (July to October)
• The soil samples were brought to the laboratory in sterile condition and stored
in a refrigerator at 4 0C until they were processed.
17. Collection of Root Samples
• Fine roots from plants of the same species were randomly collected and
mixed properly and a composoil root sample was obtained.
• Trypan blue method was followed for the determination of the intensity of root
colonization as described by Phillips and Hayman (1970).
18. Isolation of Micorrhizal spore
• Spore extraction from the soil was carried out using the Wet Sieving and
Decanting Technique by Gerdemann and Nicolson (1963).
• The isolated spores were mounted on glass slide using Polyvinyl Alcohol-
Lactic acid Glycerol (PVLG)
• observed under compound microscope (100- 1000X).Spores were identified
according to the manual of identification of VAM fungi by Schenek and Perez
(1990)
19. Wet Sieving and Decanting Technique by
Gerdemann and Nicolson (1963)
20. Soil Physio-chemical analysis
• The physical chracteristics of soil i.e., Moisture content,soil pH and soil
temperature were estimated for the collected polluted soil samples.
• The chemical chracteristic i.e., N,P,K,Organic C,Mg,Ca etc of the polluted soil
samples were estimated using the technique of Jackson (1985).Concentration
of trace metals.
• i.e.,Cu, Ni and Zn were determined by Atomic Absorption Spectrophotometer
21. Jackson Technique – Atomic Absorption
Spectroscopy
• Atomic-absorption (AA) Spectroscopy uses the absorption of light to measure
the concentration of gas-phase atoms
• Since samples are usually liquids or solids, the analyte atoms or ions must be
vaporized in a flame.
• The atoms absorbs ultraviolet or visible light and make transitions to higher
electronic energy levels
• The analyte concentration is determined from the amount of absorption
27. Linear Regression Analyses; to find out the
influence of various Edaphic factors on
Mycorrhizal colonization and Mycorrhizal spore
population.
28. Positive correlation coefficient (R) values
between Mycorrhizal spore population with Soil
moisture and Soil Temperature
Nitrogen 0.98
Potassium 0.84
Copper 0.97
Zinc 0.90
Nickel 0.97
29. Positive correlation coefficient (R) values between
Mycorrhizal spore population with Soil moisture
and Soil Temperature
Phosphorus 0.87
Magnesium 0.93
Calcium 0.69
31. Glomus fasiculatum
• 80-120 um in size
• Spore wall is 4.2 m thick,
• Yellow in colour
• the thicker walls often minutely
perforated with thick inward
projections
• hyphal walls occluded at maturity
32. Glomus macrocarpum
• red brown to dark brown (honey
colored)
• 120-400 m in width
• multilayered with cross channels in
walls with lignified in growth from outer
side of the wall(
33. Glomus mosseae
• Spores rarely filled with hyphae
• Yellow in colour
• Funnel shaped hyphae
• hyaline outer wall may not be obvious
34. Discussion
• The presence of trace metals in the polluted soil may be responsible for less
percentage of root colonization in the soil.
• The high alkalinity, pH and higher soil temperaure in the polluted soil is also
responsible for decrease in the number of mycorrhizal spores and root
infection
• The dominance of Glomus sp in the polluted soil is due to its higher metal
tolerance capacity as reported earlier by various workers
35. Conclusion
• AM vesicles are arbuscules can accumulate various trace metals and can
reduce a series of changes in plant physiology, nutrient availability and
microbial composition that may determine the outcome of a phytoremediation
attempt in the metal-stressed environment
• The various metal tolerant mycorrhizal fungi which are found to be evolved as
a trace metal-tolerance and thus they can play a very important role in the
phytoremediation of the polluted environment.
Notas do Editor
Mycorrhzial association involve 3 way interactions, between host plant, Mutualistic fungi and soil factors…
Ectomicorrhiza
Extracellular
Do not penetrate individual cell withen the roots
Endomicorrhyzae
Intracellular Penetrate cell was and cell membrane
Ectomicorrhiza
Extracellular
Do not penetrate individual cell withen the roots
Endomicorrhyzae
Intracellular Penetrate cell was and cell membrane
Ectomycorrhizal associations are mutualistic associations between higher fungi and Gymnosperms or Angiosperms in the plant families listed. ECM associations consist of a soil mycelium system, linking mycorrhizal roots and storage or reproductive structures. Ectomycorrhizal roots (formerly known as ectotrophic or sheathing mycorrhizas) are characterised by the presence of a mantle and Hartig net. An endomycorrhizal fungus forms hyphae that penetrate the cells of plant roots where they form balloon-like vesicles and branch out manifold to develop big surface areas dedicated to the exchange of minerals and carbohydrates. These structures gave rise to the name "arbuscular mycorrhizae" or AM (formerly known as "vesicular-arbuscular mycorrhizae"). Usually, an endomycorrhizal root cannot be easily distinguished from a nonmycorrhizal root without the help of a microscope and special dying techniques.
Among the various industries, paper and pulp industry is one of the notorious polluters of the environment. It has been categorized as one of the most polluting industries due to discharge of huge volumes of highly colored and toxic waste water (effluent) in the environment causing pollution of land (soil), air and water (Martin, 1998). Most of the paper and pulp industries discharge their insufficiently treated waste water into the river or stream which results in serious problems for aquatic life (Kesalkar et al., 2012)
The most important problem which the pulp and paper industry is facing today is the disposal of tremendous volumes of waste water. This waste water is rich in dissolved solids such as chlorides and sulphates of Na, Ca and varying amounts of suspended organic materials. In addition to these constituents, effluents also contain some trace metals like Hg, Pb, and Cr etc. The effluents are generally alkaline in reaction with high chemical and biological oxygen demands. Thus, the effluents discharge into the water systems make the water unfit for irrigation and potable use and create health hazards.
Treated industrial waste water could be used safely and effectively with proper precautions to increase the soil productivity (Chhonkar et al., 2000). However, despite being a useful source of plant nutrients (N, P, K, Ca etc.), the paper mill effluent often contains high amounts of various organic and inorganic materials, as well as toxic trace elements, which may accumulate in soils in excessive quantities under long term use. Subsequently, these toxic elements may cause severe problems to human beings and animals by entering into the food chains. Untreated industrial effluents contain higher amounts of Cd, Pb, Zn, Cu, Mn and Fe and enhance the concentration of the heavy metals in irrigated surface soils (Xiog et al., 2001). Significantly higher values of EC, organic carbon, available K, exchangeable cations ( Ca2+, Mg2+), exchangeable anion ( Cl-, HCO3-) along with micro-nutrient cation (Cu2+) have been reported in soils being irrigated by paper and pulp industry effluents (Singh et al., 2007). Similarly a decrease in germination percentage, seedling growth and their dry weight in different plants with an increase in paper mill effluent concentration had also been reported (Sundaramoorthy and Kunjithapatham, 2000). Zwieten et al. (2010) reported that the biochar formed due to slow pyrolysis of paper effluent affected the agronomic performance and soil fertility of some soils. Kumar et al. (2010) performed studies to determine the agronomical characteristics of Trigonella foenum-graecum irrigated with different concentrations of Paper mill effluent. Their study revealed that the effluent was rich in some plant nutrients and affected the agronomical characteristics of T. foenum-graecum (cv. Pusa early bunching) and physico-chemical characteristics of the soil as well.
The present investigation was undertaken with the objective to examine the characteristics of effluents from Paper and Pulp industry and to study the impact of lateral seepage on soil properties of agricultural fields adjoining main drains and changes in the elemental composition of plants therein.
Glomus was the most dominant isolated mycorrhizal genus of which three dominant species Glomus fasciculatum Glomus macrocarpum, Glomus mosseae have been identified. The study helps us to develop a protocol by studying the association of arbuscular mycorrhizal fungi in plants growing in poluted soil and the potential use of these AM fungi as a future bioremediation agent for rehabilitation of the polluted site contaminated with various trace metals.
The phsio-chemical properties of soil were estimated and the maximum moisture content, soil pH, and soil temperature were observed in the rainy seasons followed by summer and winter.The pH was found to be more alkaline in winter seasons in comparison to summer seasons.(Table 1).The all the chemical constituents soil N,K,Organic C (%), Copper (Cu) Nickel (Ni) and Zinc (Zn) were estimated maximum in rainy seasons followed by summer and winter seasons except soil Phosphorus(P),Magnesium(mg) and Calcium (Ca) recorded high values in winter followed by summer and rainy saesons (Table 2).The maximum number of mycorrhizal spore count and percentage of root infection were observed in the rainy seasons followed by summer.In winter seasons, lees number of mycorrhizal spore count and low percentage of root infection were reported