Cynipini Larval Sequencing Project

Hi (North American) gall hunters!

Every year for gall week, you all go out and find a ton of interesting galls, including many undescribed and understudied species. Most of the time, there's no easy path to making scientific use of those specimens. This year, we have an opportunity to make use of a large proportion of what you might find. Gallformers.org is partnering with the Forbes Lab at the University of Iowa and the Prior Lab at Binghamton University on a grant to study the oak gall wasp community in North America. This fall we're hoping to conduct a pilot study to test the procedure we hope to use over the next few years. The goal of the study is to barcode sequence all of the inducer, inquiline, and parasitoid larvae present in oak galls over the course of their development, so any gall inhabitant and any developmental stage is valuable.

If you're interested in participating, you can start collecting immediately--you don't need to wait for gall week. In fact, we're hoping to spread the arrivals out across the season. Feel free to get started now or wait until after gall week, whatever suits your schedule.

What we're asking you to do is this:

Find oak wasp (cynipini) galls on oaks and document them (and their host oak) on iNat

Collect galls and put them in a sealed container (ziploc, plastic cup with lid, etc)

Label each collection with its iNat record, collector name, collection date, location, and host tree species

Within 2-3 days of collection, mail to the Forbes lab:

Andrew Forbes
The University of Iowa
434A Biology Building
Iowa City, IA 52242

In the interest of economizing on postage, it would be ideal to send fewer packages with more galls instead of many packages with fewer galls. We don't have funds to pay for shipping this year (that would be part of the grant, if we get it), so it will be up to you to decide how you want to approach that. Just know that if you wait too long, the larvae we're looking for may not arrive intact.

If you're planning to host a gall week event, we'd love to have you consolidate a shipment of collections made by participants before and during that event.

If you want to make collections over a longer period and not mail them immediately, you are also welcome to dissect the galls yourself and preserve any larvae you find in 95% ethanol and store them in your freezer until shipment. We can potentially send you vials already filled with ethanol if you are interested in doing this. As always, just make sure they are clearly labelled.

DO NOT freeze galls or larvae dry; this will kill them and make it impossible to ship them intact.

As always, it’s your responsibility to make sure that you collect only where it is legally permissible to do so--make sure you avoid collecting in National Parks or similar locations. Most municipal parks should be fine but it’s always good to double check. Use your judgment in terms of collection effort, but generally speaking your ability to affect populations of these insects is negligible.

If you have questions, you can reply here, or tag Dr Forbes at @aforbes10 or contact him by email at andrew-forbes@uiowa.edu or tag Guerin Brown at @moneykittens. They'll be the ones receiving and processing your specimens. More info is available here: https://www.inaturalist.org/journal/megachile/83377-cynipini-and-associates-larval-sequencing-pilot-study

If you have any feedback about this procedure in advance or as you put it into action, please let us know. Working out the kinks of this process is our main goal this year.

Thank you all--looking forward to seeing what you all find!

Publicado el agosto 21, 2023 03:45 MAÑANA por megachile megachile

Comentarios

@hannawacker Plenty of galls on the Gambels in the Abajos and La Sals this time of year! There's some interesting ones on the oaks across the road from Newspaper Rock (on the path to the creek) as well.

Publicado por tepary hace 6 meses

Doing some collecting of Q austrina in October this year. Would it be better to rear out any galls we collect, or send them in for testing? We can do preservation in ethanol in-house.

Publicado por bunnymom20 hace 6 meses

I would say send them in first and then once we have the DNA we can rear after that?

Publicado por megachile hace 6 meses

Should each individual gall have its own iNat record?

Publicado por bayareawalker hace 6 meses

You can group multiple galls into a single collection observation if they share a host, location, date, etc.

Publicado por megachile hace 6 meses

We have a few galls collected Saturday that we'd like to mail in... But we're in Canada. Wondering if this is feasible, given the delay at the border. Also wondering if anyone is aware of regulations that would prohibit this kind of shipment.
@megachile @aforbes10

Thank you!

Publicado por woodhaunt hace 6 meses

Tagging @friesen5000 as well, as he's likely dealt with this!

Publicado por woodhaunt hace 6 meses

@woodhaunt I'm not sure what the regulations are regarding shipping from Canada, but I would say that if you aren't comfortable with it, don't send them. We're doing this as a pilot, as Adam says, such that next year we can take a bit more of an organized run at it and can be prepared for questions like this. If you don't send your galls, you could always rear them, or try to cut them open, photograph the larvae within, and then preserve those larvae in >95% EtOH until such time as we can figure out how to get them to us.

Publicado por aforbes10 hace 6 meses

I have mailed specimens in ethanol from the US into Canada without issue before but not the reverse so idk.

Publicado por megachile hace 6 meses

Thanks very much @aforbes10 and @megachile!

Publicado por woodhaunt hace 6 meses

@aforbes10 @moneykittens Would you be interested in adult specimens collected in the field for the project?

Publicado por obad-hai hace 6 meses

@obad-hai If they're hand catches then probably not. If you collect a gall and an adult emerges before you can mail it, then yes.

Publicado por moneykittens hace 6 meses

I'm happy to buy my own ethanol and I have lots of appropriate tubes, but I'm wondering about a good source of 95% ethanol for civilians that doesn't contain PCR inhibitors? Is Everclear 190 proof appropriate?

Publicado por hkibak hace 5 meses

@hkibak high proof everclear is fine if you're in a jurisdiction that allows sell of it. If you want to have a source for lab grade ethanol in the future, you can find it on Amazon (surprisingly; https://www.amazon.com/Laboratory-Grade-Denatured-Ethyl-Alcohol-500mL/dp/B0754NR5B8/ref=sr_1_3?crid=2MUKXJW7XY60J&keywords=lab+grade+ethanol&qid=1697207058&sprefix=lab+grade+ethanol%2Caps%2C91&sr=8-3). Its denatured so it isn't suitable to nucleotide precipitation but its perfect for just storing specimens until extraction.

Publicado por moneykittens hace 5 meses

Thanks for the link. Good to know that denatured is fine for storage followed by eventual DNA prep.

Publicado por hkibak hace 5 meses

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