07 Mycologic Research

This article will take you into the deep exploration of the modern mycology research toolkit, from basic field surveys to cutting-edge laboratory techniques.I will share 30 years of practical experience accumulated in field research and laboratory work, providing practical tips you can apply immediately while explaining the scientific principles behind it.

Random sample formula is suitable for uniform habitats, while the specimen band rule is more suitable for areas with obvious environmental gradients.In my study of Cascade Mountains in Washington, I set up a 500-meter-long specimen along the altitude gradient, and set up a 2×2-meter specimen every 50 meters, successfully recording the pattern of fungal communities changing with altitude.

Practical Case: In the primeval forests of the Oregon Coast Mountains, we compared the efficiency of random sample and systematic sample banding method.The results show that in complex terrain, combining two methods—first randomly selecting the region and then setting up a system demographic within the region—can obtain the most comprehensive list of species.

GPS precise positioning skills:

Environmental Data Record Standardization:

1. Base integrity: Use professional excavation tools to carefully dig out the entire bacteria to ensure that the bacterial troughs and cords are not destroyed.The base of mushrooms contains key identification characteristics, especially for toxic species such as Amanita.

2. Developmental stage representative: Collect specimens of different maturity levels - young, mature and elderly individuals.This helps study developmental changes and is crucial for accurate identification.

3. Eco Photography Agreement:

Expert recommendation: Always wear gloves to deal with unknown fungi.Even non-toxins can cause allergic reactions.

Gold standard for outdoor notes:

Comparison of drying technology:

| Methods | Applicable Scenarios | Advantages and Disadvantages |

|------|-----------|--------|

| Air dry | Small, thin fleshy fungi | Low cost but low humidity environment |

| Silicone drying | Most meaty fungi | Fast speed, good conformal shape, reusable |

| Food dehydrator | Large-scale collection | Controllable temperature and high efficiency |

| Professional drying box | Research institute | Precise temperature and humidity control |

Liquid Immersion Preservation Formula:

Tag information integrity checklist:

Environmental DNA technology is revolutionizing fungal diversity research.By analyzing DNA fragments in soil, water or air filters, we can detect fungi that are difficult to cultivate or observe.

Practical Case: In the geothermal area of ​​Yellowstone National Park, we used the eDNA method to discover new fungal lineages that were not detected in traditional investigations.These microorganisms survive in extreme environments at 80°C, expanding our understanding of fungal heat resistance.

eDNA Sampling Professional Process:

1. Use sterile tools to collect soil or litter

2. Put it in a sterile container immediately

3. Cold chain transport to the laboratory (preferably within 24 hours)

4. -80°C for a long time storage until DNA extraction

Choosing fresh, healthy tissue blocks is key.I prefer to take samples from the junction of the cap and the stalk, which is a low contamination rate and strong growth vitality.

Medium Selection Guide:

Practical experience in temperature control: Different fungi have specific temperature requirements.Wood-decaying fungi usually grow best at 25-28°C, while soil fungi may prefer 20-25°C.Establishing a temperature gradient experiment can determine the optimal growth conditions.

Short-term storage: Refrigerate at 4°C, transfer every 3-6 months

Medium-term storage: Covered with mineral oil, stored for 1-2 years

Long-term storage: -80°C ultra-low temperature freezing or liquid nitrogen storage

Sperm library management: Establish a detailed strain database, including source, identification information, growth characteristics and metabolic characteristics.

Professional procedure for microscope use:

1. Anatomical microscopy: Observe the macroscopic structure and prepare spore printing

2. Composite microscope: Detailed study of microscopic characteristics

Professional method for slice technology:

Dyeing technology selection table:

| Dyeing agent | Usage | Effect |

|--------|------|-----|

| KOH (Potassium Hydroxide) | General Observation | Transparent Tissue, Enhanced Contrast |

| Melzer Reagents | Starchy detection | Starchy spores turn blue and black |

| Congo Red | Cell Wall Staining | Enhanced Cell Wall Visibility |

| Cotton Blue | Mycelium Observation | Blue contrast, easy to observe |

Sample quality assessment:

Extraction method comparison:

Quality Control Inspection:

Primary selection strategy:

PCR optimization experience:

Pollution Prevention and Control:

Technical Selection Guide:

| Technology Type | Applicable Scenarios | Data Features |

|------------------------------------------------------------------------------------------------------------------------------

| Sanger Sequencing | Single Gene, Pure Culture | High Accuracy, Long Reading |

| Illumina NGS | Metagenome, Diversity | High Throughput, Short Reading |

| PacBio | Full length ITS, genome | Long read length, high error rate |

| Nanopore | Real-time, field sequencing | Portable, long-lasting reading |

Practical Case: We used Illumina sequencing to analyze fungal communities in forest soil in Montana, and detected more than 10,000 operating taxonomies (OTUs) in a single run, far beyond the capabilities of traditional methods.

Basic Analysis Process:

1. Sequence quality control and trimming

2. OTU clustering or ASV analysis

3. Taxonomy allocation (using UNITE or GenBank database)

4. Diversity analysis (α and β diversity)

Phylogenetic Analysis:

Data Submission Specification:

Solvent Selection Science:

Extraction efficiency improvement technology:

TLC Quick Filter Solution:

HPLC method development:

GC special application:

Mass Spectrometry Technology Combination:

NMR complete process:

Standardized test for antibacterial activity:

Antioxidation capacity assessment:

Cytotoxicity professional assessment:

Study on Enzyme Inhibitory Activities:

Diverity Index Selection Guide:

Spatial Distribution Analysis:

Time Dynamic Research Design:

Decomposition rate determination:

Mycorrhizal Function Research:

Quantitative Circulation:

Complete Soil Analysis Solution:

Meteorological Data Integration:

Advanced Statistical Analysis:

Multi-omics data integration strategy:

System Biology Methods:

Single-cell sequencing applications:

Single-cell imaging progress:

CRISPR in mycology:

Prospects of synthetic biology:

1. Basic Skills Training (6-12 months):

2. Intermediate Capacity Building (1-2 years):

3. Advanced Professional Development (3-5 years):

Basic Essentials ($500-1000):

Intermediate Extensions ($2000-5000):

Advanced Professional ($10,000+):

Field Investigation Error:

Laboratory Error:

Data analysis error:

Mycologic research is in an unprecedented stage of rapid development.From citizen scientists holding smartphones to professional labs equipped with the latest sequencing equipment, everyone can contribute to this field.The key is to master the correct methodology, maintain scientific rigor, and maintain connection with the global mycology community.

Action steps now:

1. Identify your research interests and available resources

2. Start with basic methods and gradually build skills

3. Join professional organizations (such as the American Fungal Society)

4. Participate in citizen science projects (such as iNaturalist)

5. Find mentors and collaborators

Remember that every carefully collected specimen and every accurately recorded data point are important bricks and stones for building the mycologic knowledge building.Start your fungus research journey – countless scientific discoveries await keen eyes and well-trained hands.