Symbiont Diversity

Objectives

To figure out if ecologically succesful beetles have different fungal associated than less usccesful native ones.

Two approaches (=papers): Broad Sample of many species from many localities using simple culturing, and a Deep Sample targeting just a few species from a few localities, but environmental sequencing using SSU amplification and 454 sequencing.

We also need to do a test isolation from the gut, to see if other interesting fungal species appear!

Broad Sample

Localities: MA, NC, FL, hopefully MS or TX

Targeted species, Should be common and widespread.

Steps

1) collect targeted species, either in logs and rear out, or directly from the logs in the field, or from traps. Trapping likely more efficient in the spring. Targetting three collections of each species at each locality, at least two individuals of each species, ideally from diferent host species. The best would be to get conspecifics from both conifers as well as angiosperms. If there is any host-dependent variatioin in the symbiont, it would be there. Good species for that are X. ferrugineus, X. affinis, and X. saxeseni – frequently attacking any kind of hosts.

2) extract fungi from a) mycangium, b) the gut and c) surface-wash of the beetle.

3) save the beetle voucher for future genotyping (congruence between beetle relatedness and fungus community similarity)
(FL locality key: Palatka 1 – Palatka, FL; Palatka 2 – Savannah (pine); Welaka 1 – Myrtle B.; Welaka 2 – Lumber R.)

Deep Sample

Up to five beetles, representatives from different localities and hosts, or important species even if samples are limited, careful isolation from mycangia, and sequencing SSU library with 454.

Targeted species

Sampling