Bagged flower yeast sampling (Ranunculaceae)

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Method synopsis

Fine-mesh bags were used to control pollinator access to flower stalks, allowing comparison of yeast densities between virgin and pollinator-exposed flowers. Nectar was extracted using calibrated microcapillary tubes and processed for yeast enumeration.

Synthesized from method descriptions across 1 paper using this protocol.

Procedure from a recent peer-reviewed implementation

Steps below were extracted from the most recent peer-reviewed implementation of this protocol in the corpus — Nectar Yeasts in the Tall Larkspur <i>Delphinium barbeyi</i> (Ranunculaceae) and Effects on Components of Pollinator Foraging Behavior (2014), PloS one. Implementations in other papers (listed below) may differ.

1. Bag flower stalks to control pollinator access
   Whole stalks on individual plants were bagged using fine-mesh bags constructed of bridal veil to prevent flower visitation by pollinators or other insects and allow for nectar accumulation. Care was taken to mark whether flowers were open or virgin (enlarged bud) prior to bagging, in addition to their sexual phase. Flowers were scored for their sex as either male (pollen-dehiscing anthers) or female (dried anthers and stigmas were exserted).
   Quantities: N = 2-25 flowers per stalk, flowers sampled individually in Zip-loc bags for processingDuration: Sampling occurred between 12 July and 4 August 2013Conditions: Summer field conditions at three different sites
   Equipment: fine-mesh bags, bridal veil material, Zip-loc bags
2. Extract nectar samples from flowers
   Nectar was extracted from flowers with calibrated microcapillary tubes and recorded the volume of each sample. Each sample was diluted with 9 μl of 30% lactophenol cotton blue solution for microscopic visualization of yeasts.
   Quantities: Each sample diluted with 9 μl of 30% lactophenol cotton blue solutionDuration: All processing occurred within 12 hours of collectionConditions: Laboratory processing within 12 hours of field collection
   Equipment: calibrated microcapillary tubes, 30% lactophenol cotton blue solution

Typical Equipment

• fine-mesh bags
• bridal veil material
• Zip-loc bags
• calibrated microcapillary tubes

Output Measurements

• yeast cell density
• nectar volume
• flower sex phase

Papers Using This Protocol (1)

Species Studied With This Protocol

Places Where Used