Here, bioassays designed to monitor the development of a fungal pathogen, Colletotrichum fioriniae, in the presence of blueberry or cranberry floral extracts on glass coverslips are described. Water-, chloroform-, and field rainwater- based floral extraction techniques are detailed as well as insight into how this information can be applied.
Total time
~3-5 days (extract preparation 1-2 days, bioassay incubation 2-3 days, analysis 1 day)
Steps
1
Prepare fungal isolates and spore suspensions
Culture Colletotrichum fioriniae isolates and generate spore suspensions for use in subsequent bioassays. Adjust spore concentration to appropriate density for consistent inoculation.
▶ 00:27
2
Extract floral compounds using chloroform solvent
Perform chloroform-based extraction of blueberry and cranberry floral tissues to isolate lipophilic compounds. Prepare extracts for bioassay application.
▶ 01:51
3
Collect rainwater from blueberry flowers
Gather natural rainwater that drains through blueberry floral structures to capture water-soluble floral metabolites. Filter and prepare for bioassay use.
▶ 03:14
4
Collect rainwater from cranberry flowers
Gather natural rainwater from cranberry flowers to obtain field-derived floral extract. Prepare samples for comparison with laboratory-extracted materials.
▶ 04:40
5
Conduct bioassay with water-based floral extracts
Inoculate glass coverslips with Colletotrichum fioriniae spores and expose to water-based blueberry or cranberry floral extracts. Incubate and monitor fungal development over time.
▶ 05:49
6
Conduct bioassay with chloroform-based floral extracts
Inoculate glass coverslips with fungal spores and treat with chloroform-extracted floral compounds. Incubate and assess pathogen development under these conditions.
▶ 08:10
7
Analyze fungal development across extract treatments
Compare Colletotrichum fioriniae growth and morphology across water-based, chloroform-based, and rainwater extract treatments. Quantify development metrics and evaluate extract effects on pathogen progression.
▶ 09:40