Also please extract DNA from muscles of Xyleborus horridus from Honduras (Caroline’s stuff).
Articles posted by jirihulcr
We received about 30 beetle samples from a collaborator in Italy, they are in a box in the lab freezer. Some of them are very important, please stop by and I will explain. I would like you to do two things with them:
1) Incorporate them into our database and our -80 freezer: record the samples into the database (see spreadsheet in an email), put new labels in, write our vial numbers on the tubes (side and top), and put them in the freezer.
2) Choose ONE well-preserved specimen of Xyleborus eurygraphus from one of three vials that contain it and extract DNA from it. Use either the Qiagen protocol for tissues or the new OmniPrep (should be slightly better). The extract will then be amplified with three primer pairs that you get from Caroline, for a phylogenetic study that she is doing. Before you do that, I will show you how to record a DNA extraction event and a PCR event into the database.
Here is a summary of what we talked about, plus a few more things that i need your help with:
– list of most common tree/forest pests in FL and insecticides approved for them
– get a black twig borer ASAP and isolate fungi from it for Polly’s experiment
– please pull together a small collection of beetles for an extension event: Xyleborus glabratus, Xylosandrus crassiusculus, Dendroctonus frontalis, Dendroctonus terebrans. Make LAAARGE labels for them (in plain English), put them in a specimen mailing box, and ship them to:
UF-IFAS Extension at Seminole County
250 W. County Home Rd.
Sanford, FL 32773-6197
– please mail the framed beetle collection in my student’s office to:
Florida Forest Service
3125 Agriculture Center Drive
St. Augustine, Florida 32092
…please pack it carefully, it’s glass…
– get to Archbold
– plan a trip to Austin
– make and updated list of Cryphalini classification
– measure the amount of DNA in a single individual of a small cryphaline (use OmniPrep, MoBio, Qiagen, and Ex-n-Amp to compare).
– assemble all sequences currently available for Cryphalini (GenBank, BOLD, Jordal, Cognato)
A couple more things:
The #3 Send ’em in will also have a movie box.
Right side of the map:
– title “More interesting stuff here:”
–What is this project all about, anyway?
–Why do beetles
–What are these “bark beetles”?: These beetles are much more that just what the name says: yes, many live in bark, but many also live inside trees and grow gardens of fungi for food (those are called ambrosia beetles). Many live in a strange family system with one brother an many sisters that have babies together. Several species are the world’s most destructive forest pests which, withe help of climate change, are turning Canada into grasslands. Want o know more? Click on the species that you trapped on the map, or visit our website: www.ambrosiasymbiosis.org.
–Several cool beetle species:
—the redbay ambrosia beetle, a foreigner from Asia that’s destroying Florida’s forests and the avocado industry: https://edis.ifas.ufl.edu/in886
—the Southern Pine Beetle: a native pest that once used to kill thousands of acres of pines yearly (http://edis.ifas.ufl.edu/in333) but now it is a rare insect!
—the granulated ambrosia beetle: the most common beetle in your trap is probably not a native insect, but this invader from Asia: http://edis.ifas.ufl.edu/in288
There is an empty box with vials by the printer. could you please fill it up with cryovials 3/4 full of ethanol? Thank you!
Polly, the address of the Clerid guys is here:
Qld Primary Industries Insect Collection
DAFF Biosecurity Queensland
Level 2C East – Ecosciences Precinct
GPO Box 267
Brisbane Qld 4001
phone: 07 3255 4357
Please send it via regular mail – I don’t think you will need a project number for that.
The beetles that will go to the synoptic collections will be pinned, but don’t do it yet. As we discussed today, we may need to re-organize our collection first. For now, put them all in ethanol. You don’t even have to identify them right now. Sorting the backlog is a higher priority.
Tings to look into on the otherwise awesome site (i’m looking at it in Chrome):
– mouse arrow disappears when touching links
– can the tabs “Join project” etc be numbered 1 to 3?
– the MAP YOUR BEETLES” should perhaps be “4: AND SEE WHAT LIVES IN YOUR BACKYARD” I just really want it to be a clear sequence of steps towards the map.
– replace ethanol with “Purell alcohol-based hand sanitizer”
– the tab “catch beetles” disappears after a while
– Supplies: soda bottle (2 liter), hand sanitizer (alcohol-based, Purell is best), string, paint brush, small ziploc bag, padded envelope for shipping
– the link to the printable PDF is a good idea, but a heads up: we will have to use a different one that reflects this simpler method, based on hand sanitizer. Good for now though…
The lower field, that contains the map:
– Can the map be big? I mean as big as is convenient/possible? Ideally across 2/3 or 3/4 of that field.
– The remaining portion of that field should have the heading “What is this project about” that’s good. And I think underneath that we will have to include a couple of educational or explanatory items, or links to them. For example, on of them should be: Why do beetles fall into hand sanitizer? The explanation is: “When a tree dies, the process of wood degradation produces alcohol. Because bark and ambrosia beetles evolved attraction to alcohol, because need to find dead trees to reproduce. Hand sanitizers contain lots of alcohol, and that’s we can use them to attract bark beetles!” Should we make a pop-up? Or some other type or short paragraph-type records? There will be more of those…
Polly, when you have a minute, could you send ONE specimen of Xylosandrus amputatus collected in Georgia to E. R. Hoebeke at the University of Georgia? We already sent him specimens before.
E. R. Hoebeke
Collection of Arthropods
Georgia Museum of Natural History
University of Georgia
Athens, GA 30602
I think the vial is somewhere in the backlog, recently added. Caroline should know more.
Thank you in advance for your help with Latasha. Here is a summary of what we will do with Latasha on her first visit:
If she has beetles, we will identify them, and grind them up to extract symbionts. Here is how. We will plate them and wait a week.
If she doesn’t have beetles, or only species that are not easy to tell a story about, we will use our fungi.
– Raffaelea subfusca from Craig
– Ambrosiella hartigii from Martin Sigut
– Fusarium sp. from Martin Sigut
– Raffaelea lauricola from Marc
– Diplodia from oaks as a free-living control (or some better species?)
If these fungi are in decently clean cultures, we can subculture them directly into a competitive assay. If they need some cleanup, we will first subculture them alone.
Competitive assay: We will record single-direction reactions: each field has a record of the reactions of the fungus in the upper row. We also need reactions of each to itself, and a growth rate without a competitor. Can we do two replicates of each assay?
|R sub||R lau||A har||F sp||Dipl|
|R sub||same-growth decrease?||R lau reaction||A har reaction||F sp reaction||Dipl reaction|
|R lau||R sub reaction||same-growth decrease?||A har reaction||etc||etc|
|no competitor||R sub growth||etc|
Better ideas welcome!
How should we score it? How about Strong inhibition, medium, weak, no effect? Or estimated percentage of growth decrease compared to the no-competitor control??