“Pollens Online” research team aims at developing baseline protocols to predict climate change impacts on pollens and associated allergies. The very first step of protocols development is allergenic pollen isolation and collection in a state-of-the-art method from flowers with natural microflora only. Collected pollens are subject to morphological identification. Morphological identification of pollens includes light microscopy, Fourier-Transform Infrared Spectroscopy (FTIR). Morphological identification is followed by molecular analysis of allergenic pollen by extracting its DNA and proteins. Allergenic genes of pollens are screened through PCR, and proteins are studied through enzyme linked immuno-sorbent assay (ELISA), sodium dodecyl sulphate poly acrylamide gel electrophoresis (SDS-PAGE) and western blotting. Additionally, lipid contents of pollens are being studied through light chromatography mass spectrometry (LCMS).

Pollen-associated microbial identification is through making pollen suspensions and culturing suspensions on media plates, carrying out BIOLOG microbial identification and characterization, and genome sequencing. All allergenic plants found in different parts of the country are mapped through Arc-GIS software to draw maps of allergenic plants and assess allergies types and their timings.

Summary of protocols developed are as follow:

1. Pure pollen collection protocol.
2. Pollen identification and differentiation from spores.
3. Pollen associated bacteria culturing protocol and Biolog base identification
4. Pollen DNA extraction protocol
5. Pollen-protein extraction protocol
6. Pollen-allergen identification protocol
7. Climate change effect on morphology and metabolome protocol
8. Protocol for Climate change impact study on future plant growth distribution through MaxEnt modeling