Seagrass Microbiome Sampling

Recently the Seagrass Microbiome group has been wrapped up in sending (and receiving!) microbiome sampling kits. These kits are part of a larger collaborative project focused on re-sequencing of Zostera marina samples in conjunction with sequencing of additional marine and freshwater Alismatid species and their microbiomes. JGI recently sequenced and released the Zostera marina genome, and we are hoping to build on their efforts and explore population level variation within Zostera marina, as well as differences in genome content and structure between Zostera and other Alismatids, in conjunction with microbiome sequencing.

The sampling kits sent by the seagrass microbiome group have focused on the microbial aspect of this project. We have asked members of the Zostera Experimental Network (ZEN) as well as additional collaborators to sample both plant tissue for sequencing (coordinated through Jay Stachowicz and Jeanine Olsen) and microbiome samples. We are extremely excited about this sample set, as it covers populations of Zostera marina across many different environments, for which we already have extensive metadata through the ZEN group! We are requesting root, sediment (within the rhizosphere), and leaf tissue, as detailed in the diagram below (courtesy of Jeanine Olsen).
microbiome

Collaborators are also sampling at two depths per site (deep and shallow), so that we can examine microbiome differences that may correlate with population depth. We are sampling 24 individuals per site, 12 per depth.

The kits are relatively straightforward and simple to both make and use, even if you’re not an experienced field microbiologist. We followed the kit and samplingl details we previously used (https://seagrassmicrobiome.org/protocols/microbial-sampling-kit/), with a few updates.

The kits now contain:
– 1 5cc syringe (for sediment collections)
– Tubes filled with Zymo buffer (DNA/RNA Shield)
– Plastic forceps
– Plastic spatula
– Parafilm
– Ethanol wipes

Here are a few photos of kit production:

Vann and Firl putting together kits in lab, photo from Katie Dahlhausen (@PhDKD)
Alana Firl and Laura Vann putting together kits in lab, photo from Katie Dahlhausen (@PhDKD)

 

Tubes all ready to go!
Tubes all ready to go!

 

Close up of kit
Close up of kit

 

Completed kits, ready to go!
Completed kits, ready to go!

We have sent out all of the kits, and have already started receiving some completed samples in the mail. Here is a close up of some of the samples from Kotzebue, AK.

Samples for one individual from Kotzebue, AK. From left to right: root, sediment, and leaf.
Samples for one individual from Kotzebue, AK. From left to right: root, sediment, and leaf.

A huge thanks to our collaborators for sampling, and to everyone from the Eisen lab who has helped make and send kits. Stay tuned for updates on sample processing and data !

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