You can perform a direct PCR using Plant Phire Direct PCR kit (Thermo Scientific) or you can extraxt DNA pollen using the classic Doyle & Doyle protocol (CTAB). I prefer the first option, it is very fast and I have the same result.

Best regards, Olga

I am not aware of a comprehensive manual, but I have Attached the urls of a number of pollen atlases that are accessible from the internet.

Extraction of DNA from pollen is not too difficult so long as you realise that the exine coating is very resilient and can last millions of years in extreme conditions. SO you will need an extraction technique that takes this into account and uses a combination of grinding, CTAB or xanthate or hot phenol.

Pollen Atlases

We have a paper under revision on this. We used the qiagen Animal kit for DNA extraction and we've got very good results. We also simultaneously compared the morphology using pollen atlases for the region of study.

I'm currently using this column based kit for GMO detection in honey. Despite the starting material is not user friendly at all, it works very well.

http://www.generon.it/dati/pdf/user_guide/exd001_en_user_guide.pdf

There is a nice book from Moore , Webb and Collinson, 1991, Pollen alalysis, Blackwell Science Std., 2nd edition. It is a field and laboratory manual of pollen analysis, covering morphological identification from Pollen of North Europe and North America.

Thanks a lot for your kind suggestions. We are studying some of them for further plans.

For general pollen morphology see the Springer-textbook M. Hesse et al., Pollen terminology (http://www.springer.com/life+sciences/plant+sciences/book/978-3-211-79893-5). In addition I can recommend the free database PALDAT: http://www.paldat.org/

Kind regards,

Matthias Svojtka

Hi,

There are a books that are the milestones of the palynology, such as Erdtman, G., 1969. Handbook of Palynology; Fægri, K., Kaland, P.E., Krzywinski, K., 1989. Textbook of Pollen Analysis, 4th ed.; Moore, P.D., Webb, J.A., Collinson, M.E., 1991. Pollen Analysis. Second Edition; Hesse et al., 2010. Pollen Terminology: An illustrated handbook.

These books can help you to understand the pollen morphology of several families and genera. There are also several Atlas but they, provide sometimes only photographs of the pollen grains without explanations or key.

For specific studies (on taxa or genera) I suggest you to look on several scientific journals such as Grana, Journal of Integrative Plant Biology, Plant systematics and evolution, and so on.

Regards,

Gabriella

The answer depends on your plant group. It could be that pollen morphology has beed described very well for the taxonomic unit of your study, but DNA sequences are not present in GeneBank except one-two species.

Depending from which region your pollen are coming (tropics versus Europe/USA) a problem for identification using DNA will also be that for most plant species only chloroplast sequences exist, while I assume you will be trying to sequence nuclear genes from your pollen. So finding hits for your regions might be hard, much worse than when you would have had cDNA.

I am used to identify fossil pollen grains with the photographic atlases by Reille, which comprises most of the European and North African plant species. The references are the following:

- Reille, M, (1992) Pollen et Spores d'Europe et d'Afrique du

Nord. Laboratoire de Botanique Historique et Palynologie,

Marseille, France.

- Reille M. 1995 Pollen et spores d'Europe et d'Afrique du Nord: supplement 1. Marseille: Laboratoire de Botanique Historique et Palynologie 274p. - illus.. ISBN 2950717519

- As far as I know, there is also a supplement 2 published in 1999.

As stated above, Moore et al. (1991) is also a classical book for pollen morphology from Europe, including pictures of the main pollen types and useful keys for identification.

I hope it helps!

Besides all the answers already given, RIKILT institute of food safety (Wageningen) has published an expert system for identifying 96 different airborne pollen types (http://www.wageningenur.nl/en/show/Determinator-Pollen-module-1.htm )

I found that even closely related species of Schoenae (Cyperaceae) showed clear differences in pollen morphology between species under electron microscopy. I didn't try any DNA analysis (Wheeler and Bruhl 2000).

One good book for study of pollen is "Pollen Biology: A Laboratory Manual" written by Shivan and Rangaswamy published by Springer.....

If you are looking for an overall work for the identification of pollen by morphology, then the online pollen atlases are good. A print reference that may be helpful is How to Know Pollen and Spores by Ronald O. Kapp, W. C. Brown & Co., Dubuque, Iowa. It is a little dated but is an easy-to-use and fairly comprehensive reference to get started.

Regards,

Jim Mickle