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Soil Microbiology ENVIRONMENTAL MICROBIOLOGY WHAT IS SOIL? Soils are excellent culture media for the growth of many kinds of microorganism Who is at home in the soil? Soils are excellent culture media for the growth of many kinds of microorganism Size of Soil Organisms Macro or large (>2 mm) Meso or mid-size (2–0.2 mm) Micro or small (<0.2mm) Mite Earthworm Alfalfa root Yeast Springtail Bacteria A single teaspoon of soil contains over 1,000,000,000 bacteria, about120,000 fungi and 25,000 algae. Organism Earthworms Mites Nematodes Protozoa Algae Fungi Actinomycetes Bacteria 1 Number per gram soil (~1 tsp) Biomass1 (lbs per acre 6”) – 1-10 10 – 100 up to 100 thousand up to 100 thousand up to 1 million up to 100 million up to 1 billion 100 – 1,500 5 – 150 10 – 150 20 – 200 10 – 500 1,000 – 15,000 400 – 5,000 400 – 5,000 Biomass is the weight of living organisms Microorganism in 1 acre of average fertility soil would weight as a medium sized-dairy cow Bacteria Fungi MOO! Actinomycetes Algae Protozoa The world of microorganisms in soil Bacteria Bacterial population of the soil exceeds other groups of microorganisms in number and variety. Plate count yields 1-10% of the total count (as compared to direct microscopic counts). Dominant genera : Arthrobacter, Pseudomonas, Agrobacterium, Flavobacterium and Bacillus. Animal pathogens : Clostridium, Bacillus, Coxiella and Streptococcus Plant pathogens : Agrobacterium, Erwinia, Corynebacterium, Pseudomonas and Xanthomonas. Different physiological groups are also present. Where are the microorganisms located in the soil? Usually the top 2-3 cms. Commonly found close to root surfaces, in dead roots, on soil particles or amongst aggregates of soil particles. Soils that are clayey have many bacteria because these soils have lot of small pores. Sandy soil is less suitable habitat. Fertile Soil Clay Soil Infertile Soil Sandy Soil Soil factors that affect microorganism growth Spherical Bacteria Organic matter Aeration (oxygen) Moisture and temperature Soil fertility and pH Rod-Shaped Bacteria Organic matter in the soil Organic matter = anything that contains carbon compounds that were formed by living organisms. Dead forms of organic material - mostly dead plant parts (85%) Living parts of plants - mostly roots (10%) Living microbes and soil animals Partly decayed organic matter is called humus Components of Soil Organic Matter Microbial degradation or organic matter Involves complex processes. Chemical alteration of organic matter, physical fragmentation and release of mineral nutrients. The dead organic matter is colonized by microbes and degraded with help of microbial enzymes. Macromolecules are broken down into simpler units and further degraded into constituent elements. Organic matter decomposition Everyone is involved Bacteria Population increases rapidly when organic matter is added to soil Quickly degrade simple compounds - sugars, proteins, amino acids Have a harder time degrading cellulose, lignin, starch Cannot get at easily degradable molecules that are protected Bacteria on fungal strands Spiral bacteria Rod bacteria Effect of soil temperature Microorganisms have been found growing in virtually all environments where there is liquid water, regardless of its temperature. Microorganisms are found growing at the sub zero temperatures of Antarctic soil to temperatures as high as 115oC in deep sea hydrothermal vents. General response of microorganisms to temperature and pH. SOIL MOISTURE CONTENT Availability of water (aw - known as water activity) is a critical factor that affects the growth of all cells. aw pure water = 1 range of m.o. = aw 1 to 0.7. aw agricultural soils : 0.9 to 1. Limiting aw for growth of certain procaryotes. Soil pH The pH or hydrogen ion concentration of natural environments varies from about 0.5 in the most acidic soils to about 10.5 in the most alkaline lakes. Most free living prokaryotes can grow over a range of pH. Depending on pH preferences microorganisms are classified into – acidophiles, neutrophiles and alkaliphiles. Sources of N Lightning Inorganic fertilizers Nitrogen Fixation Animal Residues Crop residues Organic fertilizers Forms of Nitrogen Urea CO(NH2)2 Ammonia NH3 (gaseous) Ammonium NH4 Nitrate NO3 Nitrite NO2 Atmospheric Dinitrogen N2 Organic N Roles of Nitrogen Plants and bacteria use nitrogen in the form of NH4+ or NO3 It serves as an electron acceptor in anaerobic environment Nitrogen is often the most limiting nutrient in soil and water. Nitrogen is a key element for amino acids nucleic acids (purine, pyrimidine) cell wall components of bacteria (NAM). NITROGEN CYCLE AMMONIFICATION NITRIFICATION DENITRIFICATION NITROGEN FIXATION Nitrogen in the air nitrogen fixing plant eg pea, clover animal protein plant made protein root nodules (containing nitrogen fixing bacteria) nitrates absorbed dead plants & animals urine & faeces denitrifying bacteria decomposition by bacteria & fungi ammonia nitrates bacteria nitrites bacteria (nitrifying bacteria) www.worldofteaching.com N2 N2O NH4 NO2 R-NH2 NO NO2 NO3 Ammonification or Mineralization N2 N2O NH4 NO2 R-NH2 NO NO2 NO3 Mineralization or Ammonification Decomposers: earthworms, termites, slugs, snails, bacteria, and fungi Uses extracellular enzymes initiate degradation of plant polymers Microorganisms uses: Proteases, lysozymes, nucleases to degrade nitrogen containing molecules Plants die or bacterial cells lyse release of organic nitrogen Organic nitrogen is converted to inorganic nitrogen (NH3) When pH<7.5, converted rapidly to NH4 Example: Urea NH3 + 2 CO2 Nitrification N2 N2O NH4 NO2 R-NH2 NO NO2 NO3 Nitrification Two step reactions that occur together : 1rst step catalyzed by Nitrosomonas 2 NH4+ + 3 O2 2 NO2- +2 H2O+ 4 H+ 2nd step catalyzed by Nitrobacter - 2 NO2 + O2 2 NO3 - Optimal pH is between 6.6-8.0 If pH < 6.0 rate is slowed If pH < 4.5 reaction is inhibited Denitrification N2 N2O NH4 NO2 R-NH2 NO NO2 NO3 Denitrification Removes a limiting nutrient from the environment 4NO3 + C6H12O6 2N2 + 6 H20 Inhibited by O2 Not inhibited by ammonia Microbial reaction Nitrate is the terminal electron acceptor Nitrogen Fixation N2 N2O NH4 NO2 R-NH2 NO NO2 NO3 Nitrogen Fixation Energy intensive process : N2 + 8H+ + 8e- + 16 ATP = 2NH3 + H2 + 16ADP + 16 Pi Performed only by selected bacteria and actinomycetes Performed in nitrogen fixing crops (ex: soybeans) Microorganisms fixing Azobacter Require the enzyme Beijerinckia nitrogenase Inhibited by oxygen Inhibited by ammonia (end product) Azospirillum Clostridium Cyanobacteria The nitrogen fixing bacteria are found inlumps on the roots called root nodules. The bacteria and the plant have a symbiotic relationship: the bacteria benefits by having food and shelter from the plant and the plant benefits by having nitrates produced by the bacteria. Roots of a legume plant (peas, beans and clover). www.worldofteaching.com