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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

Here, we present a protocol for collecting the floral fragrance volatiles from blooming flowers, using a non-destructive sampling procedure.

Abstract

Fragrances of many flower families have been sampled and the volatiles analyzed. Knowing the compounds that make up the fragrances can be an important step to conservation of flowers that are threatened or endangered. Because floral fragrance is critical for attracting pollinators, this method could be used to better understand or even enhance pollination. We present a protocol using a portable charcoal air filter and vacuum to collect floral fragrance volatiles, which are then analyzed by a GC-MS. By using this method, fragrance volatiles can be sampled using a non-destructive method with a machine that is easily transported. This methodology uses a rapid sampling procedure, cutting sampling time down from 2-3 hours to approximately 10 minutes. Using GC-MS, the fragrance compounds can be identified individually, based on authentic standards. The steps used for collecting fragrance and control data are presented, from material setup to collecting the data output.

Introduction

Flowers typically produce a fragrance used to attract pollinators. These fragrances are made up of many chemical compounds all acting together as a floral blend1,2,3. Without these fragrances, flowers would be less likely to pass on their genetic information using pollinators. Floral fragrance has been documented in many flowering plant families, with Orchidaceae being one of the more common families studied4. To understand the role of floral fragrance in pollination, it is important to nondestructively collect and analyze the chemical compoun....

Protocol

NOTE: Perfumes or scented lotions and products must not be worn during any of these procedures.

1. Flower selection

NOTE: Flowers used can be either naturally growing in the environment or kept under artificial environmental conditions. Temperature, humidity, and light level during collection can vary based on the specific flower species used, and what type of data is being collected. For example, data has been collected during the day and at night for the same flower.......

Representative Results

Representative data from the GC-MS are shown as a chromatogram in Figure 1. In addition to the chromatogram, a data file of results is also provided (Supplementary File 1). This data file provides the retention time for each peak (RT), and an identification of what compound that peak is (Library/ID). Peaks between 10:00 and 15:00 minutes are floral volatiles, due to the molecular weight of the compounds10. The numbers above the peaks signify the reten.......

Discussion

Though this technique is incredibly valuable for its sampling speed and portability, one limitation is using it for epiphytic species, or those growing on trees and not from the ground. In the original study10, one of the flowers sampled was epiphytic. Because the machine is too heavy to hang freely, a stable, elevated base must be made for sampling. Additionally, the machine can either be plugged in to an electrical outlet or battery powered, so if there is prolonged field sampling, there must be.......

Acknowledgements

USDA-ARS Research Project number 6036-22000-028-00D. The use of trade, firm, or corporation names in this publication is for the information and convenience of the reader. Such use does not constitute an official endorsement or approval by the United States Department of Agriculture or the Agriculture Research Service of any product or service to the exclusion of others that may be suitable. Additionally, the University of Florida Biology Department-Lewis and Varina Vaughn Fellowship in Orchid Biology (2017), and a University of Florida Graduate Research Fellowship (2014-2018) provided funding as well. We also thank Cindy Bennington from Stetson University for the orc....

Materials

NameCompanyCatalog NumberComments
Bulkhead UnionCole-PalmerUX-06390-10
FEP tubingCole-PalmerUX-06407-60
Gas ChromatographyHewlett Packard6890
Glass Wool, SilanizedSigma-Aldrich20411
Inlet linerAgilent5062-3587
Mass SpectrometerHewlett Packard5973
Reynolds oven bagReynolds Consumer ProductsTurkey size
Tenax Porous Polymer AdsorbentSigma-Aldrich11982

References

  1. Knudsen, J. T., Tollsten, L., Bergstrom, L. G. Floral scents- A checklist of volatile compounds isolated by head-space techniques. Phytochemistry. 33, 253-280 (1993).
  2. Dudareva, N. A., Pichersky, E. . Biology of floral scent. , (2....

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Floral FragranceVolatile CollectionGC MSThermal DesorptionPollinationFlower VolatilesPheromonesHeadspace Sampling

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