Tracie M. Jenkins, Brian T. Forschler, Dan R. Suiter, Tyler D. Eaton What Multiple DNA Markers Revealed About Coptotermes formosanus Maternal Population Structure Tracie M. Jenkins, Brian T. Forschler, Dan R. Suiter, Tyler D. Eaton Coptotermes formosanus is a subterranean termite from Asia that has found its way to the US.

A Serious Invasive Pest First found on the US mainland in Charleston, SC in 1957. Likely reached the US mainland by military shipments from the Pacific in WWII. Interstate commerce is a major culprit in the spread of Coptotermes formosanus. Spotty distribution, but found in most southeastern states. Coptotermes formosanus was first found on the US mainland in Charleston, SC in 1957. It likely reached the mainland by military shipments form the Pacific in WWII. It has spread throughout the southeastern US most likely due to interstate commerce. In Georgia, it is suspected that Coptotermes formosanus first came into the state by the import of railroad crossties from New Orleans, LA. Its presence in crossties is especially problematic because many are used in landscaping around residential and commercial structures. The termites then migrate from the crossties into the structures. It has a spotty distribution but is found in most southeastern states, and it could continue to spread due to the transport of crossties and other wood products.

A Serious Invasive Pest Considered among the 100 worst invasive species. Highly destructive. Success could be related to the plasticity of the breeding structure (Vargo et al. 2006). Coptotermes formosanus is considered among the 100 worst invasive species. It is very destructive. I used to work in an eradication program for the FST in Georgia. We would visit homeowners and businesses with FST infestations. One homeowner had recently purchased his home for 250,000 and then discovered the FST infestation. He said that he had spent an additional 250,000 repairing the damage, so he basically paid for his home twice, and his finances are devastated. The success of this termite could be related to the plasticity of the breeding structure which would allow adaptation to a variety of conditions.

Question: How many Coptotermes formosanus maternal lines are present in the southeastern United States?

Insect Mitochondrial Genome In order to do that, we used mitachondrial DNA sequence markers because the mitochondrial genome is maternally inherited. This is an illustration of a typical mitachondrial genome although this does not hold for all species. The COI, 16srRna, and COII genes are often used in phylogenetic studies.

Methodology: Genes 16S - very conserved COI - conserved, but more variable COII- most variable Tue, 05/05/2009 - 12:33 — bioquicknews www.bioquicknews.com/node?page=16 www.cytochemistry.net/Cell-biology/mitoch2.htm, Fawcett, A Textbook of Histology, Chapman and Hall, 12th edition, 1994.

Methodology: Collections Termites collected from three southern states: LA, GA, SC Baton Rouge, LA – 1 site, 3 individuals New Orleans, LA - 18 sites, 59 individuals Sites could be structures or trees infested with the termites.

Methodology: Collections GA-4 areas: Atlanta (6 sites, 100 individuals), Columbus (1 site, 8 individuals), Cairo (1 site, 2 individuals), Savannah (1 site, 2 individuals) SC-Citadel Military College (1 site, 63 individuals) We collected a large number from Atlanta because we are planning to use these sample in other studies. Wood pile at Citadel.

Collection Data: Sites Charleston Baton Rouge Savannah Atlanta New Orleans Columbus Cairo

Methodology: Sequencing Extracted DNA using Qiagen Blood and Tissue kit. Amplified 16S, COI, and COII with primers from Simon et al, 1994. Sequence mtDNA genes to confirm haplotypes: COII-750bp, COI-840bp, 16S-390bp.

Results: Haplotypes 16S and COI sequences were identical for all samples. COII sequences separated into two haplotypes: GA1, C2.

Results: Haplotypes LA: Baton Rouge Haplotypes: GA1 Collection from structure only 1 site, 3 individuals

Results: Haplotypes LA: New Orleans Haplotypes: GA1, C2 Collections from in or around structures & from trees 18 sites, 59 individuals 7 sites were GA1 and 11 sites were C2

Results: Haplotypes GA: Atlanta, Columbus, Cairo, Savannah Haplotype: GA1 Collections from around or in structures & railroad ties 9 sites, 112 individuals

Results: Haplotypes SC: Haplotype: GA1 Collections from around or in a structure at the Citadel Military College 18 sites, 63 individuals

Results: BLAST Search Revealed that GA1 haplotype aligned 100% with sequences from samples collected in MS, FL, HI, and China. In addition, the C2 haplotype aligned 100% with sequences from samples collected in MS, HI, Japan, Taiwan and China.

GA1 GA1 GA1 GA1, C2 GA1 GA1 C2 GA1, C2 GA1

Future Research Use mtDNA sequence markers for determining maternal origins. Maternal origins traced for Coptotermes gestroi (Jenkins et al, 2007) and Reticulitermes flavipes (Austin et al, 2005).

Future Research Use SSR and AFLP markers. Studies using highly variable markers, such as microsatellites, should provide greater power in identifying likely source populations and the routes of introduction of invasive subterranean termites (Vargo and Husseneder, 2009). I mentioned the large number of samples collected in Atlanta. We are planning to study the diversity of these termites in greater detail using SSR and AFLP markers.

We could not have done this work without the support of colleagues and friends. It continues to be an adventure. Thank you