Pollination Biology - Science topic

Paint brush works fine. The following link talks about other methods:

Dear Ashutosh Saini,

This is very positive news.

Thank you, Regards,

Dariusz Prokopowicz

Thank you Amrul Jihadi and Michael Eickermann

Campaigns... … We report the results of a task force of the North American Pollinator Protection Campaign (NAPPC) that examined potential effects of vector management practices on pollinators, and how these programs could be adjusted to minimize negative effects on pollinating species … Ginsberg, H. S., Bargar, T. A., Hladik, M. L., & Lubelczyk, C. (2017). Management of arthropod pathogen vectors in North America: minimizing adverse effects on pollinators. Journal of medical entomology, 54(6), 1463-1475.

@Michal Filipiak Your question has one associated question as to whether pollinators really need conservation?? If so, then how many pollinator species have become extinct thus far, and whether new pollinator species have also evolved and added to our ecosystem??

Luis J. Castillo-Pérez Thank you for sharing it.

go through the listed papers-

1. Bird-pollinated flowers in an evolutionary and molecular context

Quentin Cronk, Isidro OjedaJournal of Experimental Botany, Volume 59, Issue 4, March 2008, Pages 715–727, https://doi.org/10.1093/jxb/ern009

2. Efficient avian pollination of Strelitzia reginae outside of South Africa

Author links open overlay panelF.HoffmannF.DanielA.Fortier1S.-S.Hoffmann-TsayShow morehttps://doi.org/10.1016/j.sajb.2010.09.018 .

3.Reproductive_biology_of_Strelitzia_nicolai_and_Strelitzia_reginae_in_the_conditions_of_a_greenhouse

I studied avocado pollinators, bees and non-bees, in their native land Central America and out of America. I investigated the avocado visitors' behavior, pollen carrying and pollination effectiveness. There are more works like mine, studying non-bee pollinators of various plant species.

Please see in : Pollinators of Avocado Chapter, Full-text available, January 2002

Thanks a lot for this great clarification, although I left Saudi Arabia 3 years ago and the questions was to figure out how larger carpenter bees is adapted to get nectar from Calotropis procera. The stems of the plans also were used by bees for nesting. I may try to find this work again when I will be back to Egypt after my research visit to Hungary.

Thanks again Christopher and I wish for you all the best.

Arvind Thanks for the interesting articles. Yet I don't have direct information regarding the number of pollen grains needed for fertilization in almond.

Yes, there are some intersting papers about sensing of pollen - with different results:

Klaus

It is also important to have controls so that you can compare the "health" of the ecosystem. You may also want to have baseline data for comparison. I guess this study may take time to see any significant changes, unless you create experiments that will alter the environment and reserves others for comparison.

Samuel:

This link might be useful:

Flowering plants face a pollen dilemma, that is they benefit if more pollen is for pollination of conspecific flowers than for attracting/rewarding flower visitors/pollinators. However, the competition for pollination is probably exclusively intraspecific and thus plays no role on the pollination market. But when some individual begin to use a mechanism which helps to allocate more pollen for pollination than for rewarding flower visitors, then the benefit will translate into reproductive success

Dear Koen,

Most probably it is a Rosaceae pollen and could be pollen of Crataegus or Prunus. I have already prepared modern pollen samples from Rosaceac- Creataegus collected from Zagros Mountains, Iran and its pollen looks like what you have posted here. Crataegus tree is a favorable source of nectar for Bumblebees.

Best

Dear Timo,

I would suggest to perform pairwise comparisons with Bonferroni or alternatively to run a chi-square test between pairwises.

Good luck and best wishes.

I really like the interpretation of Peter Bernhardt  and fan of his work. Currently, the trend is everything global analysis as above detail mentioned by Peter Bernhardt , without understandings the local ecology. We have a couple of papers from Nepal Himalaya 1500-4400m, under construction that shows that even the different guild of pollinators are active in the area, all of them are not  flower visitor and pollinator. So it's complex and understanding them at the local level is better for the first instant instead utilizing the work done in Europe or America in your local context.

Dear Gerardo,

many red-flowering Castilejas are pollinated by hummingbirds further north. Maybe your species has a mixed strategies with bees and hummingbirds as pollinators. It may also be that bees are no effective pollinators. You have to observe.

For collecting nectar I would use a standard protocol: Cover several flowers with fleeze during night and early morning and water the plants in the evening to make sure they have enough water. In the morning you will probably be able to collect nectar using micro capillaries (0.5 or 1 microliter). There are some data in the literature for some North American species having high sucrose contents >60% and giving also sugar concentration.

Best regards

Stefan

Dear Joao, check for the papers of Kenji Suetsugu, he is also present here on Research Gate; you might find some technical help, while here is an excerpt of references from my database (key word "video surveillance") I find useful:

I tried the following while recording pollinator assemblage and behavior on Satureja: I recorded insect visitors for 8 hours a day (several days) with Sonny HD camera with additional battery pack and two memory cards using my tripod. Afterwards, videos were digested with iSpy software with option "motion capture" when the software recognized visitor movements and segregates video clips... For me it worked, but it takes some time to evaluate the recordings.

Kindly also have a look at this link wherein (Chapter 9) the importance of Bees are stressed for self-pollinating crop of Mustard:

Best

Syed

Dear Carlos,

The article that pops into mind is an early one in the history of euglossine bee biology:  Williams, N. H., and C. H. Dodson.  1972.  Selective attraction of male euglossine bees to orchid floral fragrances and its importance in long distance pollen flow.  Evolution 26: 84-95.

There may be others, but I am sure that they would have cited this one.

Regards,

Jim

Dear colleagues,

thanks everybody for your advices. Thank you, Anja, for this useful contact.

Dear all,

I would like to mention toxicity of pollen which plays a major role for oligolectic bees; see for example: Praz C.J., Müller A., Dorn S. (2008) Specialized bees fail to develop on non-host pollen: Do plants chemically protect their pollen? Ecology 89, 795–

804.

Klaus

Well drained soil is that which allows water to percolate through it. Standing water or  water saturated soil deprives root oxygen. 

Loamy soil is that which has equal proportion of sand silt and clay. It is most ideal soil for plant growth.

Addition of organic matter like compost and composted manure makes the soil acidic.

Aluminium sulphate and sulphur coated urea can also be used to reduce the soil pH.

That´s so good!

My best wishes, Rachda!

Many possibilities.  Fruit may develop without seeds or with aborted seeds. Endosperm can develop without embryo. Fruit development may not occur.  In plants for fruit development fertilization is not always essential. Even a stimulus can promote fruit development.

Dear Prabhakaran

You can refer this book Practical Pollination Biology by Dafni. It may be helpful to you.

There are three very good books to start studying pollination:

1. Techniques for pollination biologists (Kearns, C. A .; Inouye, D. W., 1993).

2. Practical Pollination Biology (Chittka, L .; Kevan, P.G., 2005).

3. Pollination Biology Handbook of Experimental (Jone C.E .; Little, R. J., 1983).

These books are very good to learn techniques for the study of pollination systems.

I hope this information will be useful to you.

Regards

Cristian Martínez

Jayant:

Have a look at this link:

Best

Syed

Thanks for your feedback!

I worked with a group that had done some soil "traps" in order to increase the abundance of a single ground-nesting bee species. Based on that experience, I would say that it is much, much more difficult than for cavity nesters. Ground-nesting bees appear to be sensitive to the type, compaction, and capillarity of the soil. I would recommend other passive sampling techniques, such as pan traps (suggested above) or blue vane traps--see a comparison at the link provided.

Dear Hojat Abasi,

I agree with the previous answers. Just be sure that you freeze mature, fully dehydrated Lilium pollen grains directly in tubes filled with liquid N2. We are punching a small hole into the tube lid, so the vials don't explode in the -80°C freezer if there are rests of liquid N2 in the tube. Using this method, lily pollen showed germination and tube growth even after more than 10 years!

Cheers

Gerhard 

 Hi Cristian,

Thank you very much. It helps a lot.

Best,

Zhenghua

This is what I meant: the bees do not perceive polarization induced false colours by the main retina, because they have twisted rhabdoms. They certainly do not look into the flowers by the DRA. But the butterflies do suffer from this artifact. So heliotropism might be intended to facilitate flower recognition by those who suffer.

Definetly a Dpsacoideae strong morphological similarities to Cephalaria and Dipsacus.

Clarke, G.C.S., Jones, M.R. 1981. The Northwest European Pollen Flora, 21: Dipsacaceae. Review of Palaeobotany and Palynology 33: 1–25.

or paldat:

Best regards 

Johannes Martin Bouchal

Dear Reza and Karl,

Yes, all right, and it confirms the available data on the trophic relations of these bees (all from the genus Dasypoda, Melittidae).

Thank you for a quick and correct answer, as I myself would have spent a lot of time searching for such data (and time, as always, is not enough).

With best wishes,

Vladimir

Hi dears, i think this code is useful:

Flower Pollination Algorithm

This matlab demo code shows how to use the new flower algorithm to solve unconstrained global optimization problems.

Required Products

MATLAB 7.14 (R2012a)

In my opinion the above mentioned bee belongs to Amegilla sp. Dr Hema Somanathan,IISER,trivandrum is one of the bee expert in India,her email address is hsomanathan@iisertvm.ac.in

It very much depends on the way you plan to do the sampling, e.g. pan-trapping versus transect captures.  Some of the references in the attached might be helpful.

Thank you for the invitation.  I am interested in role of stingless bees in crop pollination. I am interested to your project .

K.Vijayakumar, Research Scholar

Dept. of Zoolog, KASC, Bharathiar University, Coimbatore, TN, India

Try using very young anthers or pollen cones (this depends on species used...I used maize and am now trying out on cycads (Zamia species)...with flowering plants, put directly in Farmer or Carnoy and with cycads try pretreatment at 1°C for 24 hours, then put in fixative...try collecting at 11:30 a. m. or  1:30 p.m...furthermore, just type the question on the internet and lots of papers dealing with the subject will be available to you...remember methods may vary depending on species, time of collection and age of simple...luck

I thought I'd provided an answer to this question but it's not showing.  Odd.  Anyway, I wanted to say that the kind of manipulation of floral parts that you are talking about has been quite commonly done and can be an effective test of floral function, as the examples here show.  He's one from my group (there's a PDF on ResearchGate):

 Lamborn, E. & Ollerton, J. (2000) Experimental assessment of the functional morphology of inflorescences of Daucus carota (Apiaceae): testing the “fly catcher effect”. Functional Ecology 14: 445-454

Also look for work by Scott Armbruster on Dalechampia, and Paul Wilson on Impatiens.

In my experience bees in these genera visit a wide range of flower colours.  Bees are quick learners and rapidly associate particular colours and scents with a reward, though there seem to be "innate" preferences in some groups that have been tested.  

Hi Anna Maria, 

perhaps these papers may help you

I photographed this plant in Ukraine (Kyiv Region, Borodianka District, Piskivka vill., in 2013)

Hello Marwa, 

I agree with Sunil's suggestion. If you want to collect mature pollen grains, you can simply remove open flowers (I use 10 for Arabidopsis) from the plant and place them in a eppi tube with PBS (or even water, depending on what you're doing next with the pollen). I vortex this tube for ~10 seconds to release pollen grains from anthers. Then you can centrifuge the tube, preferably not at full speed, so that flower parts stay in the supernatant and pollen grains form a small yellow pellet. You can vortex and spin down again to obtain more grains, if necessary. 

If you want to collect mature pollen in bulk, you can check out this paper describing the vacuum method, as Yuan-Yeu mentioned above 

Johnson-Brousseau, S.A. and McCormick, S. (2004) A compendium of methods useful for characterizing Arabidopsis pollen mutants and gametophytically-expressed genes. Plant J. 39, 761–775. 

I would only follow Reza's protocol if you want to collect the sporopollenin-based outer pollen wall from pollen grains. 

If you want to collect immature pollen grains, I could give you some suggestions too, but this is less easy to do.

Happy pollen collecting!

I don't mind having a look at your proposal. I'll be happy to give you some feedback.

Good ideas Peter! Why not also follow the causes for visiting various sources of pollen? Peters answer is somewhat restricted to pollen mixing during a single foaging trip, which might be caused by the rareness of some flowering plant species or signal standarsization (formerly Mullerian mimicry) in these plants. Recently Eckhardt M, Haider M, Dorn S & Müller A published an article termed Pollen mixing in pollen generalist solitary bees: a possible strategy to complement or mitigate unfavourable pollen properties? which arreared in J Anim Ecol. (2014) 83(3):588-97. doi: 10.1111/1365-2656.12168. showing that pollen mixing might be caused by toxic pollen. Other caused might be rareness of amino acids and other compounds in pollen of some plants. In these cases solitary bees are more affected than eusocial bees, the latter of which exhibit pollen mixing at the colony level.

Most available books on pollination techniques give you a range of techniques for determining when a stigma is receptive.  The peroxidase technique has been discussed above.  Check the literature and consider the use of the esterase test.  It works equally well on wet and dry-type stigmas, Dr. Ollerton because, technically, ALL stigmas are wet stigmas since pollen grains can't hydrate on a stigma surface until the pistil gives them just a little drinkie of water and sugar... right?  What we call dry stigmas do produce some fluids but they can not produce free-flowing fluid.  Even the thin, proteinaceous pellicle of a stigma produces a little water while the so-called "wet" stigma of a petunia is really composed primarily of lipids (grease) instead of water.  Virtually all stigmas release some sort of an esterase combination when they are ready to receive pollen (wet or dry) and that actually includes flowers that bloom underwater (they release esters into the water as a "stigmosphere."  If this old technique was good enough for the Heslop-Harrisons, Bruce Knox, Cameron Mconchie (spelling?), Jo' Kendrick and their colleagues/students it's good enough for YOU!

Hello Florian!

I need to buy fluorescent dye for pollination movement studies (~ 40 microns). You could direct me to a store ?

Thanks! Amanda

Thanks to all for the useful links and guidance

Neotropical species often forage in a different way to many old-world species, in addition to showing a much greater diversity that of old-world fruit bats. Feeding behaviour is likely to be influenced by numerous environmental factors (habitat density and moon-phase (relating to light levels and detectability by potential predators), distance from roost (and the availability of feeding roost sites) and even breeding behaviour in species which lek or have harems,

The body size of neotropical bats is also far more variable. Smaller species of phylostomid, such as Ectophylla will apparently forage in huge numbers around fruiting figs, but are unlikely to be able to eat on the wing (they are tiny), and so I imagine will settle in the tree itself or in temporary feeding roosts, and other phylostomid species certainly make use of feeding roosts to process larger seeds.

Few people have looked at this systematically, though Tuanjit has been looking at seed rain (and dispersers) in a deforested patch in Peninsula Thailand, and I believe some of the African bat groups have also done some research.

Cynopterus is said to be one of the main afforesters for Anak Krakatau; so looking at the floral record there and searching for any gaps between species which are eaten by bats, but not largely swallowed, and those swallowed and defecated might be one way of looking to see what fruits are being treated in different ways. But as is normally the case with bats-we need more bat researchers! 

I don't think this is a very good paper to base these questions from because it is not intended to be a review of native pollinators in coffee systems. The paper by Ngo et al. might be a better place to start.

Dear Klaus,

According to my unpublished data, the ability of pollen germination decreases sharply after its placement in skopa. After that pollen is much stronger sterilized by bee female under the formation a pollen ball in the cell (probably by adding secretory substances). This prevents further germination of pollen in a cell, for not spoiling the food for larvae.

Best regards,

Vladimir.

Hi Jeff Ollerton

We conducted a study in tundra (!) grasslands in Northern Norway and found the exclusion of herbivores to cause modification of species composition but not species richness or diversity. I have attached the publication. Cheers, Kari Anne

Well, I do not think that you will be able to sample a sufficient amount of faeces from living bees in cages at all (because of their special rectum). In our cage studies bees hardly ever defecated, although we kept them for more then four weeks. The only exception was, when bees were infected with gut parasites like Nosema apis or ceranae. I think you should go for your second suggestion and use cohort representatives for each measuring interval.

Dear Alejandro

Please note that this might be an effect of flower age.

Not all of the flowers of an inflorescence are of the same Age. Since nectar production is known to be dependent of Age this might explain your Observation.

But you might be interested in papers like

"Variation in rate of nectar production depends on floral display size: a pollinator manipulation hypothesis"

at http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2435.2004.00815.x/abstract

Regards

Nikola

1-3: Diptera, Tipuloidea ?

4-6: Coleoptera, Chrysomelidae, cf. Diabrotica

If you are looking to analyze the composition of what the glands contain in great detail, I suggest collecting some gland extract and making fractions, firstly by centrifuging the extract to precipitate proteins, then making further fractions of the smaller metabolites based on polarity. I would suggest analyzing the proteins using MALDI-ToF-MS, and using LC-MS to analyze the non-volatile metabolites (reverse phase LC for non-polar metabolite analysis and HILIC LC for polar metabolites), operating in both positive and negative mode to obtain maximal metabolite ionization yield. If you have the luxury of a UPLC-MS, then your metabolite profiles will be much richer. To analyze the most volatile components, I would suggest solid phase micro-extraction (SPME) and subsequent GC-MS. If you're interested in identifying correct chirality of molecules, then I suggest using cyclodextrin based columns. If you're looking for differences in gland extracts between certain bee treatments, then a principal component analysis will cluster the samples, and if you have a split in clusters between treatments, then you know the metabolite profiles vary, and you could look at the loading plot to identify the metabolites which contributed most to the variation, and hence are (usually) accountable for the differences. Hope this helps! Feel free to message me if I've been a bit too complicated.

Thanks for your answer Peter Bernhardt. I have considered doing the measurements myself, but unfortunately I'm a bit late for that, since the summer is almost gone and most of the species finished their flowering already. 

The King et al. paper significantly underestimates the amount of time and effort needed to assess pollinator effectiveness, it's not a trivial task for even moderately generalised plant species.  Interaction networks of flower visitation provide a first approximation and have similar properties to pollinator visitation networks (see:  Ollerton J., Johnson S. D., Cranmer, L. & Kellie, S. (2003) The pollination ecology of an assemblage of grassland asclepiads in South Africa. Annals of Botany 92: 807-834)

It's important to realise that visitation is an ecological interaction regardless of the outcome for the plant, though understanding the role of the flower visitor is important if we wish to study things like floral evolution.

Understanding the effect of flower visitation on animal fitness is much more complex and I don't know of any studies that have yet attempted it except for brood-site pollination systems such as figs and fig wasps, and yuccas and yucca moths.

Hi I have not done this myself but a colleague of mine used the following method http://ac.els-cdn.com/S1439179110000174/1-s2.0-S1439179110000174-main.pdf?_tid=6c67750a-e5f0-11e3-b566-00000aacb360&acdnat=1401230832_2b8bd93dec0a33e396e42ae91e99019b

to collect nectar from multiple plant species, some of which were also opened small flowers. Maybe worth a go?

Thank you!

The two 'things' at right may be pollen grains of Ulmus (verrucate exines, and possibly pores only along the profile); the other may be a pollen of Poaceae, grass pollen, that has a smooth surface (scabrate) and one pore (not visible).

You should take pictures at higher magnifications, or use less staining color to not cover the surface and apertures of pores, if possible.

The best way to take spectral reflectance readings of things like pan traps, flowers, fabrics, etc. is generally to use equipment and protocol similar to that employed in Chittka, L. & Kevan, P.G. (2005). Flower colour as advertisement. In Dafni, A., Kevan, P.G., Husband, B.C. (eds.) Practical Pollination Biology. Enviroquest Ltd., Cambridge , ON , Canada , pp. 157-196. (pdf here: http://chittkalab.sbcs.qmul.ac.uk/2005/ChittkaKevan05new.PDF ) - quite a lot of labs now have Ocean Optics or similar equipment and the capacity to measure reflectance between 300 and 700nm using a known light source and white standard. The overall setup costs about US$10-12K last time I checked, but it's not complicated to use if you are careful and precise and doesn't take a lot of time to do a few dozen samples. With flowers, it's important to lay the petals flat but with minimal damage and use the freshest material possible.

Polarisation measurements will require additional kit and I've not tried this myself. It may be worth looking at the floral iridescence work from the Whitney lab and some of the work on insect structural colour and iridescence from the Vukusic lab?

I should add, that the described methods are not completely the same. With acetolysis you will just see the sporopollenin, so just the outer walls of the pollen grains. With fuchsin jelly the pollen will stay "intact" with its cellular content. Under the light microscope the view will not be the same. If you already have some experience with pollen counting, I would choose the method you already know, if not it does not really matter.

Dear Francinaldo,

Before reading all the great papers suggested by my colleagues, I think you need a more basic introduction into the mechanisms of colour vision in bees (and other insects). I suggest to read the following two papers:

Chittka, L. & Kevan, P.G. (2005). Flower colour as advertisement. In Dafni, A., Kevan, P.G., Husband, B.C. (eds.) Practical Pollination Biology. Enviroquest Ltd., Cambridge , ON , Canada , pp. 157-196.

Chittka, L., & Wells, H. (2004). Color vision in bees: mechanisms, ecology and evolution. In: Prete, F.: Complex Worlds from simpler nervous systems; MIT Press, Boston pp. 165-191.

The first paper provides a step-by-step instruction how to measure spectral reflectance of (artificial) flower surfaces and how to calculate the respective colour loci (and distances) in a bee colour space. The second paper gives you a more general introduction into colour psychophysics and evolution of colour vision in bees. Both papers can be downloaded from Lars Chttka’s webpage:

Best wishes,

Johannes

There are many papers dealing with that problem. We recently published a couple of papers on the geographic variation in phenology and reproductive success in a columnar cacti. The answer is yes. There are clear geographic patterns in the composition of pollinator communities, both at the single species level (specific scale) and the community assemblage level. Check:

Bustamante, E., Casas, A. y Búrquez, A. 2010. Geographic variation in reproductive success of Stenocereus thurberi (Cactaceae): effects of pollination timing and pollinator guild. American Journal of Botany 97(12):2020–2030.

and

Bustamante, E. y Búrquez, A. 2008. Effects of Plant Size and Weather on the Flowering Phenology of the Organ Pipe Cactus (Stenocereus thurberi). Annals of Botany 102: 1019–1030

I can send you the pdfs if requested... Saludos

I used to use brightly colored dye from the Radiant corp. (which is mentioned in Kearns and Inouye techniques for pollination biologists, but the company seems to be out of business. I've obtained fluorescent dyes from a company called Risk Reactor, which sells them in short and long wave lengths (you need a UV light source to see the particles). Some studies on nectar-feeding birds suggest they can see brightly-colored dyes, so if you have those concerns, UV dyes in short-wave may be best. I haven't found any dyes with particle sizes of 100 microns, though; if anyone has, please let us know.