Standard methods for pollination research with Apis mellifera.

Authors: Keith S Delaplane, Arnon Dag, Robert G Danka, Breno M Freitas, Lucas A Garibaldi, R Mark Goodwin, Jose I Hormaza.

Table of contents

1. Introduction
2. Plant pollination requirements
   2.1. Determining plant mating system

      2.1.1. Testing for agamospermy (asexual reproduction through non-fertilized seeds)
      2.1.2. Testing for autogamy (auto- or self-pollination)
      2.1.3. Testing for geitonogamy (selfing within the same plant)
      2.1.4. Testing for xenogamy (reliance on out-crossing)
      2.1.5. Testing for mixed mating systems
   2.2. Testing for pollinating agents and pollination deficit
      2.2.1. Testing for wind pollination (anemophily)
      2.2.2. Testing for biotic (honey bee) pollination – single visits
      2.2.3. Testing for biotic (honey bee) pollination – multiple visits
      2.2.4. Fruit-setting experiments at the field level
3. Measuring pollen on bees and pollen deposition on stigmas
   3.1. Identifying and evaluating pollen quantity and quality transported by bees

      3.1.1. Microscopic pollen identification and making archival reference slides
      3.1.2. Pollen identification (palynology) with molecular methods
      3.1.3. Tracking pollen identity
      3.1.4. Pollen quantity Haemocytometers Alternative methods
      3.1.5. Pollen viability and quality Pollen viability Pollen germination and pollen tube growth in vitro
   3.2. Evaluating pollen identity, quantity and quality on stigmas
      3.2.1. Identifying pollen on stigmas
      3.2.2. Quantifying pollen deposited on a receptive stigma per visit or unit time
      3.2.3. Evaluating stigmatic receptivity
      3.2.4. Evaluating pollen germination and pollen tube growth in vivo
4. Measures of colony level pollination efficacy
   4.1. Proportion of foragers from a colony visiting a crop

      4.1.1. Pollen trapping Determining pollen trap efficiency Number of pollen traps Analysing pollen trap contents Effect of pollen traps on foraging
      4.1.2. Nectar collecting
      4.1.3. Proportion of colony bees collecting pollen
      4.1.4. Colony foraging rate
      4.1.5. Fraction of bees side-working flowers
   4.2. Relationship between foragers and their hives
      4.2.1. Marking bees in the crop
      4.2.2. Marking bees according to their hives
5. Determining crop-specific recommended pollinator densities
   5.1. Indirect extrapolations

      5.1.1. Bee densities in small field plots
      5.1.2. Field-scale transects
      5.1.3. Cage visitation rates
   5.2. Direct tests of whole fields to find the required number of colonies per hectare
   5.3. Appraising risk of competition between plants for pollination
   5.4. A cautionary note about recommendations
6. Measuring harvest and post-harvest effects of pollination
   6.1. Visual appearance
   6.2. Textural quality
   6.3. Flavour
   6.4. Nutritional value
7. Managing bee colonies for optimum pollination
   7.1. Bee attractants
   7.2. Feeding colonies

      7.2.1. Feeding syrup
      7.2.2. Feeding pollen
      7.2.3. Testing effects of feeding regimens on pollination performance
   7.3. Distribution of colonies within the crop
8. Conducting pollination research in greenhouses and tunnels
   8.1. Carbon dioxide (CO2) level
   8.2. Solar radiation
   8.3. Temperature and humidity
   8.4. Directed air flow
   8.5. Limited food resources
9. Pesticides and pollinators
10. Economic valuation of crop pollination by honey bees

   10.1. Determining yield in response to specific colony density
   10.2. Response variables and calculations
   10.3. Economic valuation at larger scales
11. Acknowledgements
12. References