ppt课件-nitrogenfixationlab-westernoregonuniversity.pptVIP

Culture-Dependent UnitLab Project TwoNitrogen Cycle Nitrogen Fixation (17.28) N2 fixed to NH3 and NH2 groups - COSTS ATP ONLY done by Prokaryotes - Bacteria and Archaea Conserved O2-intolerant Nitrogenase/Fe-Mo cofactor Assayed via acetylene reduction assay - understand Acetylene and N2 both share triple bond substrates. Microbial Diversity O2 Strategies - Table 17.10 Azotobacter: fastest aerobic ETC uses O2, capsule Cyanobacteria: O2-resistant heterocysts Methanogens anoxygenic phototrophs: live anoxic Symbionts: co-produce O2-binding leghemoglobin” You will view both phototrophs during that lab. Symbiotic Nitrogen Fixation (19.22) Legumes: root hairs secrete inducer molecules Symbionts: aerobic soil Rhizobium (Proteobacteria) Separate - no fixation; together - leghemoglobin Bacteroids: misshapen Rhizobium in nodules Non-leguminous Azolla/Anabaena, Alnus/Frankia; fine line between bacteroids and organelles… Rhizobium Enrichment Dissect, clean, surface-sterilize nodule, comparing Antiseptic: tissue-safe alcohol, H2O2 (Tables 20.3-4) Disinfectant: not tissue-safe, bleach (Tables 20.3-4) Media HAS nitrogen and simple sugar - capsule Hospital antiseptics/disinfectants first used by Lister, 1976. Azotobacter Enrichment (12.9, 17.28, and 18.1) Understand Beijerink’s enrichment, late 1800’s Media lacks nitrogen, has simple sugar - capsule Nitrification and Denitrification Prominent in soil, water, sewage/feces, and GI tract Nitrification, Aerobic Chemolithotrophy (17.12) Two-step process - ammonia then nitrite oxidation NH3 + O2 yields NO2 + H2O + ATP, Nitrosomonas NO2 + O2 yields NO3 + ATP, Nitrobacter Both are Proteobacteria (12.3); packed with lamellae Soil enrichment using NH3 media with aeration; measure either NH3 depletion or NO3 appearance. Anaerobic Nitrate Reduction (17.14) EITHER denitrifying (to N2) or nitrifying (to fixed N) NO3 + CH2O yields N2 + CO2 + ATP, Pseudomonas NO3 + CH2O yields NH3 + CO2 + ATP, Enterics Review