I)M)S) Add caterpillarsMsc -- Hajibabaei lab at UofGPrivileged to share my master’s thesis About exploring barcode splits of morphologically cryptic species of Lepidoptera
I)What are cryptic species?M)really a bi-product of how we detect species which has historically been by morphological identification S) as we integrate different datasets like natural history data (specifically for leps, caterpillar morphology, host plant specificity, genitalic morphology, molecular data what we thought was a single evolutionary lineage can change.
I)Specific examples of cryptic species that were initially underscored by DNA barcodes are:M) Ten species in one, Perichares, S) . Key discoveries in the Hesperiidae family include the once polyphagousAstraptesfulgerator, now divided into 10 cryptic species based on DNA barcode divergences, distinct food-plants, subtle morphological traits and ecological preferences(Hebert et al., 2004).Pericharesphiletesis now divided into four species described by DNA barcode divergences as well as caterpillars, pupae and food-plant differences (Burns et al., 2008).Most recently, Burns et al. discovered crypsis within Porphyrogenespeterwegei, using DNA barcode divergences, caterpillar developmental changes, food-plant conservatism and host-specific parasitism (2010).
To further investigate this case we expanded our sampling to include ind from geographically distributed populationsTo try and assertain if DHJ01/DHJ02 are sister taxa, which would suggest this incongruence to be incomplete lineage sorting of nuclear alleles. However if they turn out not to be sister taxa then the ACG is a secondary contact zone for these populations and would suggest interbreeding.
The COI data suggests a
-ITS2,sanger reads not 454, poor quality due to non-homogenized repeats.-character analysis shows that DHJ01 and DHJ02 share nucleotide characters that are most similar to N American grouping and different from S american grouping.
Basically ACG represents the place where the two, long-separated lineages meet... there they do not seem to have full reproductive isolation as revealed by introgression of nuclear markers, but they do remain parapatric because of ecological preferences as you suggested - and females being very sedentary, then most if not all of the introgression is male-driven - which explains that nuclear alleles filter through the barrier whereas there's no mitochondrial exchanges...
Claudia Bertrand - Invertebrates Plenary
Exploring barcode splits among morphologically cryptic species of Lepidoptera through examiningalternative loci and next-generation sequencing Claudia Bertrand, Rodolphe Rougerie, Daniel H. Janzen, Winnie Hallwachs and Mehrdad Hajibabaei 1
outline1. Cryptic species & DNA barcodes2. Methods3. Case study 1: M. clusoculis, B. perses, E.satellitia4. Case study 2: U.belli5. Case study 3: E. imperialis 2
What are cryptic species? “species that are hidden under a single taxonomic name because they are morphologically indistinguishable” (Bickford et al., 2007) Delineation •Morphology •Ecology •Geography •Behaviour •Genetic 3janzen.sas.upenn.edu
Astraptes fulgerator complex 4Hebert, P. D. N., et al, 2004, Ten species in one: DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator: PNAS
52% Morphological or Ecological Correlates 48% Lack Morphological or Ecological Correlates 5Janzen, D. et al, 2009, Integration of DNA barcoding into an ongoing inventory of complex tropical biodiversity: Molecular Ecology Resources
Case study 1: Bardaxima perses 98 N = 32 1 N = 30 99 1 99 0.05% 1% 1% 0.5%B. persesDHJ01B. persesDHJ02 Wolbachia absentCOI cytb EF1a ITS2 13
Case study 1: Eumorpha satellitia 98 N = 10 0.2% 96 74 84 N = 10 5% 5%E. satellitiaDHJ01 0.1%E. satellitiaDHJ02 Wolbachia absentE. satellitiaDHJ03 COI cytb EF1a ITS2 14
What then, are these mt-lineages?1. Recent speciation event • Retention of ancestral polymorphisms in nuclear markers • Find a character that represents species boundaries2. Under-detected heteroplasmy or pseudogenes • 454-sequence base approaches for deep sequencing of intra- individual variability3. Reticulation instead of speciation • Representative of a past isolation event • Sequence individuals that are allopatrically distributed 15
outline1. Cryptic species & DNA barcodes2. Methods3. Case study 1: M. clusoculis, B. perses, E.satellitia4. Case study 2: U.belli5. Case study 3: E. imperialis 16
Case study 2: Urbanus belli U.belliDHJ03 U.belliDHJ02 U. belliDHJ01 100 Rainforest 100 100 1% Dryforest 17
Case study 2: Urbanus belli 100 W 100 N=10 W 100 N=11 100 W 100 W N=12 W 93 100 100 W 100 W 100 100 70 W 1% 0.5% 0.5% 71U. belliDHJ03U. belliDHJ02 Wolbachia presentU. belliDHJ01 COI cytb EF1a ITS2 18
Wolbachia infection: Unidirectional % Identity Match in WSP &Marker Allele MLST Database wsp 115 100% gatB 71 98% coxA 67 100% ftsZ 65 99% hcpA 74 99% fbpA 6 98% Supergroup B Strain: Werren et al ., 2008, Nature Reviews Lep species from Ecuador 19
ITS2 secondary structure: Compensatory base changes (CBC) I IV II (marked by U-U mismatch) All compensatory base changes III were found in helices II and III Interspecific CBC >> Intraspecific CBC51% consensus of aligned structures with gaps 20
outline1. Cryptic species & DNA barcodes2. Methods3. Case study 1: M. clusoculis, B. perses, E.satellitia4. Case study 2: U.belli5. Case study 3: E. imperialis 21
Case study 3: Eacles imperialis100 DHJ02 Rainforest DHJ01 100 Semi-deciduous lowland forest Dryforest 1% 22
Case study 3: Eacles imperialis 2 1. Retention of ancestral polymorphisms 1% 0 0 2% 2. Interbreeding 0 2% 0.2% Wolbachia absentE. imperialisDHJ01E. imperialisDHJ02 COI cytb EF1a ITS2 23
Case study 3 COI DHJ02 1. non-sister status South American 2. ACG secondary contact zone North American DHJ011% 24
Case study 3 EF1a Eacles imperialisDHJ01/DHJ021% 25
Conclusions to date:• ACG secondary contact zone• Interbreeding• Parapatric – female ecological preferences• Male-driven – COI lineages 27
AcknowledgementHajibabaei Lab Collaborators FundingShadi Shokralla Committee Genome CanadaJoel Gibson Dr. Dan Janzen Ontario Genomics InstituteIan King Dr. Winnie Hallwachs NSERC CanadaSaina Taidi Dr. Rodolphe RougerieClaudia Bertrand Dr. Alex SmithJennifer Spall Dr. Teresa CreaseStephanie BoilardSteven Van KonynenburgMelissa BraschelJessica KlawunnVanessa Patterson-DohertyMehrdad Hajibabaei 28