Quantifying pollen deposition with macro photography and 'stigmagraphs'

Authors

  • Gail MacInnis McGill University
  • Jessica Forrest University of Ottawa

DOI:

https://doi.org/10.26786/1920-7603(2017)six

Abstract

The pollen deposited during a single visit by a flower visitor (“single-visit deposition”; SVD) is often measured by removing the stigma from the flower and counting the pollen grains deposited under a microscope. This process precludes study of any subsequent interactions between the flower and later visitors (such as pollen removal from the stigma). Furthermore, if the stigma is excised too soon after the pollinator visit, the flower may be rendered infertile, such that any analyses of fruit or seed yield in relation to pollen deposition must be done indirectly. Here, a method of pollen deposition measurement was developed using macro photography and the open-source image-analysis software program ImageJ/Fiji. Using colour segmentation options within the program, the pollen grains can be distinguished from the background stigmatic surface, and the percentage of stigma coverage can be calculated. This pollen deposition measurement method leaves the sampled flower in the field to develop into fruit, allowing any subsequent yield or quality analyses to be conducted directly.

Author Biographies

Gail MacInnis, McGill University

PhD Student

Department of Natural Resource Sciences

McGill University

Montreal, QC

Jessica Forrest, University of Ottawa

Assistant Professor

Biology Department

University of Ottawa

References

Adler LS, Irwin RE (2006) Comparison of pollen transfer dynamics by multiple floral visitors: experiments with pollen and fluorescent dye. Annals of Botany 97(1):141-150. DOI: https://doi.org/10.1093/aob/mcj012

Ariza MT, Soria C, Medina-Mínguez JJ, Martínez-Ferri E (2012) Incidence of misshapen fruits in strawberry plants grown under tunnels is affected by cultivar, planting date, pollination, and low temperatures. HortScience 47(11):1569-1573. DOI: https://doi.org/10.21273/HORTSCI.47.11.1569

Conner JK, Davis R, Rush S (1995) The effect of wild radish floral morphology on pollination efficiency by four taxa of pollinators. Oecologia 104(2):234-245. DOI: https://doi.org/10.1007/BF00328588

Costa CM, Yang S (2009) Counting pollen grains using readily available, free image processing and analysis software. Annals of Botany 104:1005-1010. DOI: https://doi.org/10.1093/aob/mcp186

Cresswell JE (2000) Manipulation of female architecture in flowers reveals a narrow optimum for pollen deposition. Ecology, 81(11):3244-3249. DOI: https://doi.org/10.1890/0012-9658(2000)081[3244:MOFAIF]2.0.CO;2

Dafni A., Kevan, PG, & Husband, BC (2005) Practical pollination biology. Cambridge, Ontario, Canada. Enviroquest, Ltd.

Falque M, Vincent A, Vaissiere B, Eskes A (1995) Effect of pollination intensity on fruit and seed set in cacao (Theobroma cacao L.). Sexual Plant Reproduction 8(6). DOI: https://doi.org/10.1007/BF00243203

Ferreira T, Rasband W (2012) ImageJ User Guide - ImageJ/Fiji 1.46r. [online] URL: https://imagej.nih.gov/ij/docs/ guide/user-guide.pdf (accessed April 2015).

Flanagan RJ, Mitchell RJ, Knutowski D, Karron JD (2009) Interspecific pollinator movements reduce pollen deposition and seed production in Mimulus ringens (Phrymaceae). American Journal of Botany 96(4):809-815. DOI: https://doi.org/10.3732/ajb.0800317

Garibaldi LA, Steffan-Dewenter I, Winfree R, Aizen MA, Bommarco R, Cunningham SA, … Klein AM (2013) Wild pollinators enhance fruit set of crops regardless of honey bee abundance. Science 339(6127):1608–11. DOI: https://doi.org/10.1126/science.1230200

Hall M, Frank E, Holmes G, Pfahringer B, Reutemann P, Witten IH (2009) The WEKA Data Mining Software: An Update. ACM SIGKDD Explorations Newsletter 11(1). [online] URL: http://dx.doi.org/10.1145/1656274.1656278 (accessed May 2015). DOI: https://doi.org/10.1145/1656274.1656278

Harder LD, Thomson JD, Cruzan MB, Unnasch RS (1985) Sexual reproduction and variation in floral morphology in an ephemeral vernal lily, Eyythronium Americanum. Oecologia 67(2):286-291. DOI: https://doi.org/10.1007/BF00384301

Howitt AJ, Pshea A, Carpenter WS (1965) Causes of deformity in strawberries evaluated in a plant bug control study. Michigan Agricultural Experiment Station Quarterly Bulletin 48:161-166.

Javorek SK, Mackenzie KE, Vander Kloet SP (2002) Comparative pollination effectiveness among bees (Hymenoptera: Apoidea) on lowbush blueberry (Ericaceae: Vaccinium angustifolium). Annals of the Entomological Society of America 95(3):345-351. DOI: https://doi.org/10.1603/0013-8746(2002)095[0345:CPEABH]2.0.CO;2

Johnson D (2008) How to Do Everything: Digital Camera, 5th edition. McGraw-Hill Osborne Media, New York.

Kearns CA, Inouye DW (1993) Techniques for Pollination Biologists. University Press, Colorado.

King C, Ballantyne G, Willmer PG (2013) Why flower visitation is a poor proxy for pollination: measuring single‐visit pollen deposition, with implications for pollination networks and conservation. Methods in Ecology and Evolution 4(9):811-818. DOI: https://doi.org/10.1111/2041-210X.12074

Ledesma N, Sugiyama N (2005) Pollen quality and performance in strawberry plants exposed to high-temperature stress. Journal of the American Society for Horticultural Science 130(3):341-347. DOI: https://doi.org/10.21273/JASHS.130.3.341

MacIvor J, Albert J, Mirwan H, Wilkes M, MacInnis G, … Button L (2013) Tips and tricks for pollination biologists. NSERC-CANPOLIN publications. [online] URL: https://www.uoguelph.ca/canpolin/New/ Tips%20and%20Tricks%20Guide%20for%20Pollination%20Biologists.pdf (accessed May 2015).

McGregor SE (1976) Insect pollination of cultivated crop plants (Vol. 496). Agricultural Research Service, US Department of Agriculture.

Motten AF, Campbell DR, Alexander DE, Miller HL (1981) Pollination effectiveness of specialist and generalist visitors to a North Carolina population of Claytonia virginica. Ecology 62(5):1278-1287. DOI: https://doi.org/10.2307/1937292

Ne'eman G, Jürgens A, Newstrom‐Lloyd L, Potts SG, Dafni A (2010) A framework for comparing pollinator performance: effectiveness and efficiency. Biological Reviews 85(3):435-451. DOI: https://doi.org/10.1111/j.1469-185X.2009.00108.x

Nitsch JP (1950) Growth and morphogenesis of the strawberry as related to auxin. American Journal of Botany 211-215. DOI: https://doi.org/10.1002/j.1537-2197.1950.tb12183.x

Schemske DW, Horvitz CC (1984) Variation among floral visitors in pollination ability: a precondition for mutualism specialization. Science 225:519–521. DOI: https://doi.org/10.1126/science.225.4661.519

Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nature Methods 9(7):671–675. DOI: https://doi.org/10.1038/nmeth.2089

Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, … Cardona A (2012) Fiji: an open-source platform for biological-image analysis. Nature Methods 9(7):676-682. DOI: https://doi.org/10.1038/nmeth.2019

Schlindwein C, Wittmann D, Martins CF, Hamm A, Siqueira JA, Schiffler D, Machado IC (2005) Pollination of Campanula rapunculus L. (Campanulaceae): How much pollen flows into pollination and into reproduction of oligolectic pollinators? Plant Systematics and Evolution 250(3-4):147–156. DOI: https://doi.org/10.1007/s00606-004-0246-8

Snow AA (1982) Pollination intensity and potential seed set in Passiflora vitifolia. Oecologia 55(2):231–237. Waites AR, Ågren J (2004) Pollinator visitation, stigmatic pollen loads and among-population variation in seed set in Lythrum salicaria. Journal of Ecology 92(3):512–526. DOI: https://doi.org/10.1007/BF00384492

Waser NM, Price MV (1991) Reproductive costs of self-pollination in Ipomopsis aggregata (Polemoniaeae): are ovules usurped. American Journal of Botany 1036-1043. DOI: https://doi.org/10.1002/j.1537-2197.1991.tb14511.x

Young HJ, Stanton ML (1990) Influences of Floral Variation on Pollen Removal and Seed Production in Wild Radish. Ecology 71(2):536–547 DOI: https://doi.org/10.2307/1940307

Žebrowska J (1995) The viability and storage of strawberry pollen. Plant Breeding 114(5):469-470. DOI: https://doi.org/10.1111/j.1439-0523.1995.tb00837.x

Cucurbita pepo L. stigmagraph

Downloads

Published

2017-02-01

How to Cite

MacInnis, G., & Forrest, J. (2017). Quantifying pollen deposition with macro photography and ’stigmagraphs’. Journal of Pollination Ecology, 20, 13–21. https://doi.org/10.26786/1920-7603(2017)six

Issue

Section

Notes on Methodology