Synecology and Traditional Autecology Flashcards
What is hypothesis
A proposed explanation for a phenomenon. Explanation based on previous observations when no scientific theory can explain the phenomenon. Must be testable
What is null hypothesis
A statement that there is no relationship between two conditions. Called H0 (null) until it is given statistical significance. Aim to disprove rather than prove within the scientific method
What is the goal of experimental design
The goal is to falsify, or disprove the hypothesis
What the controls of experimental design
Differentiate between experimental effects and other effects
What are the factors to consider from experimental design
Heterogeneity and replication Needs to collect environmental 'metadata' Sample size Sampling method Storage and transport
What is the analysis of experimental design
Experimental effects
Variables
Statistical approaches
What are microscopic cell counts
Cells are stained with a dye, usually, fluorescent. Filter on to membrane (known area). Count selection of representative fields (also known area)
What is flow cytometry
Objects can be detected by a flow cytometer because they scatter light. More often absorption and emission of light is used (fluorescence). Some organisms are fluorescent (auto-fluorescent), and others we stain with fluorescent dyes or proteins
What can flow cytometry detect
Flow cytometry can identify organisms using: Shape, Pigments (chlorophyll, phycoerythrin), DNA stains (heterotrophs, diving, and sexual cells), Respiration stains (live dead for bacteria, caspase for apoptosis in eukaryotes), and Cell structure stains (actin). Can be used to sort cells-cultivation, single-cell genomics
What is microbial biomass
Conversion factor from cell size and volume. Biochemical assays. Measure specific biomolecules. All microbial biomass has same amount of biochemical assayed. ATP + ADP + AMP (can tell growth state). Cell wall components: Muramic acid, Limulus assay (gram negative only), and Chitin (fungi only). DNA, protein, or lipid concentration
What is photosynthesis/primary production
Primary productivity: Uptake of radiolabeled 14CO2, Chlorophyll A content. Short time spans measure gross primary productivity. Long time spans measures net primary productivity (PP – Respiration
What are ‘potential’ activity measurements
Remove “bottom up” constraints by adding nutrients and/or substrate. Note – can be used to determine limiting nutrient. Measure and compare maximum possible rate of that activity for that system
What is Michaelis - Mentin Kinetics
- All members of the community respond in the same way to the substrate
- Uptake is functionally equivalent,
- Substrate utilized is representative of all substrates
What is thymidine uptake
DNA synthesized by growing cells at rate proportional to biomass. Use radiolabeled thymidine uptake rates to determine community growth rate. Assumes all bacteria take up thymidine and incorporate into DNA at the same rate
What is the functional gene amplicon sequencing
Do a bunch of sequence. Get a bunch of sequences. PCR amplification and amplify the sequence that is variable to differentiate between different organisms. Functional genes would be like the core genes
What are transcriptomes
All of the genes that are expressed under specific conditions. Must compare to control to understand regulation. Need genome sequence to understand context. Use reverse transcriptase to copy extracted RNA (usually enriched for mRNA) into cDNA. Sequence cDNA.
What are proteomics
All of the proteins synthesized under specific conditions. Same concerns as transcriptomes. Fragment isolated proteins, mass spec to determine fragment sequence, assemble using genome scaffold and/or overlapping fragments. Mostly developed over the last decade
What are metabolomics
All metabolites present (either inside or outside a cell) under specific conditions. End products of metabolic pathways, can be used to determine specific functions. Mass spec to characterize – very complex!
What are the supervised assembly-based metagenomics
Sequence analysis (taxonomic) relative to databased to known sequences
What are the unsupervised metagenome-assembled gneome
Sequence usage patterns and frequency of sequences in sample
What are the single-amplified genome
Cells sorted by flow cytometry or microfluids. DNA extracted from individual cells. Amplified by whole-genome amplification (introduces errors and contamination). Quality assessed as for MAGs
What is the comprehensiveness of assembly-based analysis
Can construct multiple whole genomes, but only for organisms with enough coverage to be assembled and binned
What is the community complexity of assembly-based analysis
In complex communities, only a fraction of the genomes can be resolved by assembly.
What is the novelty of assembly-based analysis
Can resolve genomes of entirely novel organisms with no sequenced relatively.