Register      Login
Australian Journal of Botany Australian Journal of Botany Society
Southern hemisphere botanical ecosystems
RESEARCH ARTICLE (Open Access)

Improving floral nectar storage on filter paper for sugar recovery

Bianca Amato A , Sophie Petit https://orcid.org/0000-0002-7984-5123 A B * and Russell Schumann B
+ Author Affiliations
- Author Affiliations

A UniSA STEM, University of South Australia, Mawson Lakes, SA 5095, Australia.

B Kangaroo Island Research Station, Dudley West, SA 5222, Australia.

* Correspondence to: sophie.petit@unisa.edu.au

Handling Editor: Anthony O’Grady

Australian Journal of Botany 69(8) 585-595 https://doi.org/10.1071/BT21006
Submitted: 18 January 2021  Accepted: 21 July 2021   Published: 29 October 2021

© 2021 The Author(s) (or their employer(s)). Published by CSIRO Publishing. This is an open access article distributed under the Creative Commons Attribution-NonCommercial 4.0 International License (CC BY-NC)

Abstract

Nectar analysis has been used to understand pollination systems, but nectar storage methods have rarely been considered as potential sources of inaccuracy in the recovery of data. Prompt nectar sugar analysis is not always possible and storage methods can affect results. We aimed to develop an effective method to store nectar on filter paper. Nectars from two subspecies of Eremophila maculata (Scrophulariaceae) and Strelitzia reginae (Strelitziaceae) were spotted on filter papers. Nectars were redissolved and assayed by high-performance liquid chromatography to determine the masses of sugars recovered from the papers from Day 0 to Day 30. We evaluated the effects of the method of elution, paper type and size, and storage treatments on sugar recovery. Liquid nectars were also stored in the refrigerator. Sugars were best eluted from filter papers in 15 mL of water and agitated for 1 min. Nectar sugars stored on small papers tended to be recovered more successfully than those stored on larger papers (significantly for glucose). Paper performed better than nylon for glucose. Desiccant had a marginal positive effect on nectar sugar recovery, and filter paper performed better than did refrigeration of liquid nectar for storage. If highly accurate measurements are needed, nectars should be eluted with large volumes of water from small filter papers stored with desiccant within a few days of collection.

Keywords: nectar elution, nectar storage methods, nectar storage on filter paper, nectar sugar analysis, zeolite.


References

Amato B (2015) Evaluating field methods for the storage of floral nectar. BSc(Hons) thesis, University of South Australia, Mawson Lakes, SA, Australia.

Amato B, Petit S, (2017) A review of the methods for storing floral nectars in the field. Plant Biology 19, 497–503.
A review of the methods for storing floral nectars in the field.Crossref | GoogleScholarGoogle Scholar | 28303638PubMed |

Amorim FW, Galetto L, Sazima M (2013) Beyond the pollination syndrome: nectar ecology and the role of diurnal and nocturnal pollinators in the reproductive success of Inga sessilis (Fabaceae). Plant Biology 15, 317–327.
Beyond the pollination syndrome: nectar ecology and the role of diurnal and nocturnal pollinators in the reproductive success of Inga sessilis (Fabaceae).Crossref | GoogleScholarGoogle Scholar | 22823072PubMed |

Aronne G, Malara P (2019) Fiber‐optic refractometer for in vivo sugar concentration measurements of low‐nectar‐producing flowers. New Phytologist 224, 987–993.
Fiber‐optic refractometer for in vivo sugar concentration measurements of low‐nectar‐producing flowers.Crossref | GoogleScholarGoogle Scholar |

Bailey B, Ullucci P, Bauder R, Plante M, Crafts C, Acworth I (2014) ‘Carbohydrate analysis using HPLC with PAD, FLD, charged aerosol detection, and MS detectors.’ (ThermoFisher Scientific: Boston, MA, USA)

Baker HG (1977) Non-sugar chemical constituents of nectar. Apidologie 8, 349–356.
Non-sugar chemical constituents of nectar.Crossref | GoogleScholarGoogle Scholar |

Baker HG, Baker I (1973) Amino-acids in nectar and their evolutionary significance. Nature 241, 543–545.
Amino-acids in nectar and their evolutionary significance.Crossref | GoogleScholarGoogle Scholar |

Baker HG, Baker I (1983a) Floral nectar sugar constituents in relation to pollinator type. In ‘Handbook of experimental pollination biology’. (Eds C Jones, R Little) pp. 117–141. (van Nostrand Reinhold Co Inc.: New York, NY, USA)

Baker HG, Baker I (1983b) A brief historical review of the chemistry of floral nectar. In ‘The biology of nectaries’. (Eds B Bentley, T Elias) pp. 126–152. (Columbia University Press: New York, NY, USA)

Baskin SI, Bliss CA (1969) Sugar occurring in the extrafloral exudates of the Orchidaceae. Phytochemistry 8, 1139–1145.
Sugar occurring in the extrafloral exudates of the Orchidaceae.Crossref | GoogleScholarGoogle Scholar |

Brown M, Downs CT, Johnson SD (2010) Sugar preferences of a generalist nonpasserine flower visitor, the African speckled mousebird (Colius striatus. Auk 127, 781–786.
Sugar preferences of a generalist nonpasserine flower visitor, the African speckled mousebird (Colius striatus.Crossref | GoogleScholarGoogle Scholar |

Carter C, Thornburg RW (2000) Tobacco nectarin I purification and characterisation as a germin-like, manganese superoxide dismutase implicated in the defense of floral reproductive tissues. Journal of Biological Chemistry 275, 36726–36733.
Tobacco nectarin I purification and characterisation as a germin-like, manganese superoxide dismutase implicated in the defense of floral reproductive tissues.Crossref | GoogleScholarGoogle Scholar |

Carter C, Thornburg RW (2004) Is the nectar redox cycle a floral defense against microbial attack? Trends in Plant Science 9, 320–324.
Is the nectar redox cycle a floral defense against microbial attack?Crossref | GoogleScholarGoogle Scholar | 15231276PubMed |

Clement A, Yong D, Brechet C (1992) Simultaneous identification of sugars by HPLC using evaporative light scattering detection (ELSD) and refractive index detection (RI). Application to plant tissues. Journal of Liquid Chromatography 15, 805–817.
Simultaneous identification of sugars by HPLC using evaporative light scattering detection (ELSD) and refractive index detection (RI). Application to plant tissues.Crossref | GoogleScholarGoogle Scholar |

Corbet SA (2003) Nectar sugar content: estimating standing crop and secretion rate in the field. Apidologie 34, 1–10.
Nectar sugar content: estimating standing crop and secretion rate in the field.Crossref | GoogleScholarGoogle Scholar |

Corbet S, Unwin DM, Prys-Jones O (1979) Humidity, nectar and insect visits to flowers, with special reference to CrataegusTilia and Echium. Ecological Entomology 4, 9–22.
Humidity, nectar and insect visits to flowers, with special reference to CrataegusTilia and Echium.Crossref | GoogleScholarGoogle Scholar |

Coskun O (2016) Separation techniques: chromatography. Northern Clinics of Istanbul 3, 156–160.
Separation techniques: chromatography.Crossref | GoogleScholarGoogle Scholar | 28058406PubMed |

de Vega C, Herrera CM (2012) Relationships among nectar-dwelling yeasts, flowers and ants: patterns and incidence on nectar traits. Oikos 121, 1878–1888.
Relationships among nectar-dwelling yeasts, flowers and ants: patterns and incidence on nectar traits.Crossref | GoogleScholarGoogle Scholar |

Dhandhukia P, Thakker J (2011) Quantitative analysis and validation of method using HPTLC. In ‘High-performance thin-layer chromatography (HPTLC)’. (Ed. M Srivastava) pp. 203–221. (Springer: Berlin, Germany)

Dósa G (2008) Nectar collection and analysis with wick-sampling method. Acta Botanica Hungarica 50, 93–96.
Nectar collection and analysis with wick-sampling method.Crossref | GoogleScholarGoogle Scholar |

Dungan RJ, Beggs JR, Wardle DA (2004) A simple gravimetric technique for estimating honeydew or nectar production. New Zealand Journal of Ecology 28, 283–288.

Field A (2009) ‘Discovering statistics using SPSS’, 3rd edn. (Sage Publication Ltd: London, UK)

Galletto L, Bernardello G (2005) Nectar. In ‘Practical pollination biology’. (Eds A Dafni, PG Kevan, BC Husband) pp. 156–216. (Enviroquest: Cambridge, ON, Canada)

Gilbert MT (1987) ‘High performance liquid chromatography.’ (IOP Publishing: Bristol, UK)

Girard F, Antoni M, Sefiane K (2008) On the effect of Marangoni flow on evaporation rates of heated water drops. Langmuir 24, 9207–9210.
On the effect of Marangoni flow on evaporation rates of heated water drops.Crossref | GoogleScholarGoogle Scholar | 18671417PubMed |

Gottsberger G, Schrauwen J, Linskens HF (1984) Amino acids and sugars in nectar, and their putative evolutionary significance. Plant Systematics and Evolution 145, 55–77.
Amino acids and sugars in nectar, and their putative evolutionary significance.Crossref | GoogleScholarGoogle Scholar |

Guo X, Bruins AP, Covey TR (2006) Characterisation of typical chemical background interferences in atmospheric pressure ionisation liquid chromatography–mass spectrometry. Rapid Communications in Mass Spectrometry 20, 3145–3150.
Characterisation of typical chemical background interferences in atmospheric pressure ionisation liquid chromatography–mass spectrometry.Crossref | GoogleScholarGoogle Scholar | 16998786PubMed |

Heil M, Rattke J, Boland WJS (2005) Postsecretory hydrolysis of nectar sucrose and specialisation in ant/plant mutualism. Science 308, 560–563.
Postsecretory hydrolysis of nectar sucrose and specialisation in ant/plant mutualism.Crossref | GoogleScholarGoogle Scholar | 15845855PubMed |

Herrera CM, de Vega C, Canto A, Pozo MI (2009) Yeasts in floral nectar: a quantitative survey. Annals of Botany 103, 1415–1423.
Yeasts in floral nectar: a quantitative survey.Crossref | GoogleScholarGoogle Scholar | 19208669PubMed |

Jaroenkit T, Chen NJ, Paull RE (2008) Nectar secretion, mucilage production and mold growth on bird-of-paradise inflorescences. Postharvest Biology and Technology 49, 431–435.
Nectar secretion, mucilage production and mold growth on bird-of-paradise inflorescences.Crossref | GoogleScholarGoogle Scholar |

Karkacier M, Erbas M, Uslu MK, Aksu M (2003) Comparison of different extraction and detection methods for sugars using amino-bonded phase HPLC. Journal of Chromatographic Science 41, 331–333.
Comparison of different extraction and detection methods for sugars using amino-bonded phase HPLC.Crossref | GoogleScholarGoogle Scholar | 12935307PubMed |

Kearns CA, Inouye DW (1993) ‘Techniques for pollination biologists.’ (University Press of Colorado: Boulder, CO, USA)

Kingsolver J, Daniel T (1995) Mechanics of food handling by fluid-feeding insects. In ‘Regulatory mechanisms in insect feeding’. (Eds R Chapman, G de Boer) pp. 32–73. (Springer: Boston, MA, USA)

Lewis B, Smith F (1969) Sugars and derivatives. ‘Thin-layer chromatography’. (Ed. E Stahl) pp. 807–837. (Springer: Berlin, Germany)

Mallick SA (2000) Technique for washing nectar from the flowers of Tasmanian leatherwood (Eucryphia lucida Eucryphiaceae). Austral Ecology 25, 210–212.
Technique for washing nectar from the flowers of Tasmanian leatherwood (Eucryphia lucida Eucryphiaceae).Crossref | GoogleScholarGoogle Scholar |

McKenna MA, Thomson JD (1988) A technique for sampling and measuring small amounts of floral nectar. Ecology 69, 1306–1307.
A technique for sampling and measuring small amounts of floral nectar.Crossref | GoogleScholarGoogle Scholar |

Morrant DS, Schumann R, Petit S (2009) Field methods for sampling and storing nectar from flowers with low nectar volumes. Annals of Botany 103, 533–542.
Field methods for sampling and storing nectar from flowers with low nectar volumes.Crossref | GoogleScholarGoogle Scholar | 19074446PubMed |

Nichol P, Hall JL (1988) Characteristics of nectar secretion by the extrafloral nectaries of Ricinus communis. Journal of Experimental Botany 39, 573–586.
Characteristics of nectar secretion by the extrafloral nectaries of Ricinus communis.Crossref | GoogleScholarGoogle Scholar |

Nicolson SW, Thornburg RW (2005) Nectar chemistry. In ‘Nectaries and nectar’. (Eds SW Nicolson, M Nepi, E Pacini) pp. 215–263. (Springer: Dordrecht, Netherlands)

Nicolson SW, Van Wyk B-E (1998) Nectar sugars in Proteaceae: patterns and processes. Australian Journal of Botany 46, 489–504.
Nectar sugars in Proteaceae: patterns and processes.Crossref | GoogleScholarGoogle Scholar |

Pate JS, Peoples MB, Storer PJ, Atkins CA (1985) The extrafloral nectaries of cowpea (Vigna unguiculata (L.) Walp.). II. Nectar composition, origin of nectar solutes, and nectary functioning. Planta 166, 28–38.
The extrafloral nectaries of cowpea (Vigna unguiculata (L.) Walp.). II. Nectar composition, origin of nectar solutes, and nectary functioning.Crossref | GoogleScholarGoogle Scholar | 24241308PubMed |

Pate J, Shedley E, Arthur D, Adams M (1998) Spatial and temporal variations in phloem sap composition of plantation-grown Eucalyptus globulus. Oecologia 117, 312–322.
Spatial and temporal variations in phloem sap composition of plantation-grown Eucalyptus globulus.Crossref | GoogleScholarGoogle Scholar | 28307910PubMed |

Percival MS (1961) Types of nectar in angiosperms. New Phytologist 60, 235–281.
Types of nectar in angiosperms.Crossref | GoogleScholarGoogle Scholar |

Petit S, Rubbo N, Schumann R (2011) Nectar collected with microcapillary tubes is less concentrated than total nectar in flowers with small nectar volumes. Australian Journal of Botany 59, 593–599.
Nectar collected with microcapillary tubes is less concentrated than total nectar in flowers with small nectar volumes.Crossref | GoogleScholarGoogle Scholar |

Power EF, Stabler D, Borland AM, Barnes J, Wright GA, Durand P (2017) Analysis of nectar from low-volume flowers: a comparison of collection methods for free amino acids. Methods in Ecology and Evolution 9, 734–743.
Analysis of nectar from low-volume flowers: a comparison of collection methods for free amino acids.Crossref | GoogleScholarGoogle Scholar | 29938013PubMed |

Ruhlmann JM, Kram BW, Carter CJ (2010) CELL WALL INVERTASE 4 is required for nectar production in Arabidopsis. Journal of Experimental Botany 61, 395–404.
CELL WALL INVERTASE 4 is required for nectar production in Arabidopsis.Crossref | GoogleScholarGoogle Scholar | 19861655PubMed |

Shahidzadeh-Bonn N, Rafai S, Azouni A, Bonn D (2006) Evaporating droplets. Journal of Fluid Mechanics 549, 307–313.
Evaporating droplets.Crossref | GoogleScholarGoogle Scholar |

Shenoy M, Radhika V, Satish S, Borges RM (2012) Composition of extrafloral nectar influences interactions between the myrmecophyte Humboldtia brunonis and its ant associates. Journal of Chemical Ecology 38, 88–99.
Composition of extrafloral nectar influences interactions between the myrmecophyte Humboldtia brunonis and its ant associates.Crossref | GoogleScholarGoogle Scholar | 22234428PubMed |

Simpson BB, Neff JL (1981) Floral rewards: alternatives to pollen and nectar. Annals of the Missouri Botanical Garden 68, 301–322.
Floral rewards: alternatives to pollen and nectar.Crossref | GoogleScholarGoogle Scholar |

Staab M, Methorst J, Peters J, Blüthgen N, Klein AM (2017) Tree diversity and nectar composition affect arthropod visitors on extrafloral nectaries in a diversity experiment. Journal of Plant Ecology 10, 201–212.