Corbiculate Bees

The Controversy of the Phylogyeny of Corbiculate Beesand what could cause it.: 

The Controversy of the Phylogyeny of Corbiculate Bees and what could cause it. Lisa Booth

Outline : 

Outline Background About Corbiculate Bees Behavioral Phylogeny Morphological Phylogeny Molecular Phylogeny Methods and Results Alignment Outgroup Gene Conclusions

Corbiculate Bees: Why are they studied?: 

Corbiculate Bees: Why are they studied? Include economically important species Form complex eusocial societies

Corbiculate Bees: What are they?: 

Corbiculate Bees: What are they? Family: Apidae Subfamily: Apinae Monophyletic group of Long-tongued bees Comprised of 4 monophyletic extant tribes Euglossini: Orchid bee Bombini: Bumble bee Apini: Honey bee Meliponini: Stingless bee

Euglossini: Orchid bee: 

Euglossini: Orchid bee Pollinate orchids by collecting chemicals on their legs Do not possess eusocial behavior Most are solitary www.ufv.br/dbg/bee/euglossa.htm

Bombini: Bumble bee: 

Bombini: Bumble bee Important for crop and wildflower pollination Are social Form small colonies www.sankey.ws/bombini.jpg www.malawicichlidhomepage.com/macro_nature/

Apini: Honey bee: 

Apini: Honey bee Only 6-11 species Developed social behavior and structure 30 million years ago Maintain the same society today Highly eusocial www.durhamsbeefarm.com/photobee.htm www.sphoto.com/medium/wkbeeandamp;cells76.jpg

Meliponini: Stingless bee: 

Meliponini: Stingless bee Bite, but do not sting Some produce honey, but not enough to harvest in most cases Are highly eusocial www.ecuador-images.net/insect-bee18.jpg encarta.msn.com/.../Stingless_Bee_in_Amber.html

The Behavioral Phylogeny: 

The Behavioral Phylogeny Noll’s Research 42 Behavioral characteristics Characters are based on Nest construction Food storage Worker reproduction Female-female interaction Food provisioning

The Behavioral Phylogeny: 

The Behavioral Phylogeny Orchid Bumble Honey Stingless

The Behavioral Phylogeny: 

The Behavioral Phylogeny Characters that separate the bumble bees from the stingless bees. 6: Hygienic Habits 13: Brood Cell Construction 16: Brood Cell Construction 18: Cell Construction and Oviposition 20: Cell Provisioning 24: Brood Food Origin 28: Foraging Behavior 29: Division of Non-reproductive behavior 30: Food exchange among adults 31: Nectar exchange among the individuals before being stored 36: Colony foundation 37: Queen foraging 40: Dominant female or queen disappearance 41: Morphological differences between castes

The Morphological Phylogeny: 

The Morphological Phylogeny Serrao’s Research Proventricular structure Widened Apex Triangular Apex Spine-like hairs Long columnar folds

The Morphological Phylogeny: 

The Morphological Phylogeny Stingless Honey Bumble Orchid

The Molecular Phylogeny: 

The Molecular Phylogeny Cameron’s research Used Molecular data from 4 genes Nuclear DNA 28S Opsin Mitochondrial DNA 16S Cytochrome b

The Molecular Phylogeny: 

The Molecular Phylogeny Honey Orchid Bumble Stingless

The Molecular Phylogeny: 

The Molecular Phylogeny Orchid Bumble Stingless Honey

The Molecular Phylogeny: 

The Molecular Phylogeny Honey Orchid Bumble Stingless

The Molecular Phylogeny: 

The Molecular Phylogeny Honey Bumble Stingless Orchid

The Molecular Phylogeny: 

The Molecular Phylogeny Honey Orchid Bumble Stingless

Controversy: 

Controversy Both Noll’s data and Serrao’s data support the single origin of eusocial bees Cameron’s data supports the dual origin of eusocial bees www.entomology.cornell.edu

Methods: Repeat Noll’s Analysis: 

Methods: Repeat Noll’s Analysis Copy data matrix from Noll’s paper into a NEXUS format Use PAUP 4.0 beta for a Parsimony analysis Use PAUP 4.0 beta to find Bootstrap values Looked at each character individually

Results: Behavioral Analysis: 

Results: Behavioral Analysis

Results: Behavioral Analysis: 

Results: Behavioral Analysis Single Origin Characteristics 4: Nest Coating 6: Hygienic Habits 9: Cell Walls 10: Brood Cell Architecture 13: Brood Cell Construction 16: Brood Cell Construction 18: Cell Construction and ovipositition 20: Cell Provisioning 24: Brood Food Origin 28: Foraging Behavior 29: Division of Nonreproductive behavior 30: Food exchange among adults 31: Nectar exchange among the individuals before being stored 40: Dominant female or queen disappearance 41: Morphological differences between castes Dual Origin Characteristics 7: Wax envelope covering the brood area 8: Materials employed in cell construction 12: Reuse of cells or space where larvae are reared 25: Brood food composition 27: Food storage 35: Removal of cell walls after cocoon spinning 38: Social regulation

Methods: Repeat Cameron’s Analysis: 

Methods: Repeat Cameron’s Analysis Found Cameron’s aligned data on-line Realigned data using ClustalX Gap penalty of 10/0.05 Aligned Ingroup species first Aligned Outgroup sequences to the Ingroup Used PAUP 4.0 beta for Parsimony and Likelihood analyses Used Mr. Bayes for a Bayesian analysis

Results: Repeat Cameron’s Analysis: 

Results: Repeat Cameron’s Analysis 28S Bumble Stingless Orchid Honey

Results: Repeat Cameron’s Analysis: 

Results: Repeat Cameron’s Analysis Opsin Honey Bumble Orchid Stingless

Results: Repeat Cameron’s Analysis: 

Results: Repeat Cameron’s Analysis 16S Bumble Orchid Honey Stingless

Results: Repeat Cameron’s Analysis: 

Results: Repeat Cameron’s Analysis Cytochrome b Honey Orchid Bumble Stingless

Results: Repeat Cameron’s Analysis: 

Results: Repeat Cameron’s Analysis Concatenated Honey Orchid Bumble Stingless

Methods: Look at another gene: 

Methods: Look at another gene Found a new gene on GenBank, Elongation Factor 1-alpha (nuclear) Aligned DNA sequences with ClustalX Gap penalty of 10/0.05 Aligned Ingroup first Aligned Outgroup sequences to Ingroup Used PAUP for Parsimony and Likelihood analyses Used Mr. Bayes for a Bayesian analysis

Results: Look at new gene: 

Results: Look at new gene Elongation Factor Bumble Stingless Honey Orchid

Methods: Investigate Outgroups: 

Methods: Investigate Outgroups Redefined the Outgroup using Cameron’s outgroup species Melissodes Centris Anthophora Habropoda Xylocopa Found new outgroup species for the Cytochrome b and Elongation Factor genes Allodape Braunsapis Ceratina

Results: Investigate Outgroups (Parsimony Trees): 

Results: Investigate Outgroups (Parsimony Trees)

Results: Investigate Outgroups (Likelihood Trees): 

Results: Investigate Outgroups (Likelihood Trees)

Results: Investigate Outgroups (Bayesian Trees): 

Results: Investigate Outgroups (Bayesian Trees)

Conclusions: Alignment: 

Conclusions: Alignment Not able to repeat Cameron’s alignment for all genes. Calls into question Cameron’s alignments Alignment does affect the tree Concatenated Tree 16S Tree

Conclusions: Gene Choice: 

Conclusions: Gene Choice The different genes do not agree. 28S and 16S have similar tree topologies of (AE)(BM). Both are ribosomal non-coding Opsin and Cytochrome b have similar tree topologies of A(E(BM)). Both are genes for enzymes EF has a topology of (AE)(BM). Enzyme gene but does not match other enzyme genes.

Conclusions: Outgroups: 

Conclusions: Outgroups Outgroup choice is very important Xylocopa and Centris seem to affect the tree the most Braunsapis, Allodape and Ceratina seem to be good outgroups, but I wasn’t able to find much data to test them fully Cameron’s outgroups didn’t group together, they mixed with the ingroup which makes them a poor outgroup

Conclusions: Model of Analysis: 

Conclusions: Model of Analysis Slight differences between the different tree options (parsimony, likelihood, and Bayesian) 16S and Opsin were the most sensitive to the Model of Analysis

Conclusions: Controversy: 

Conclusions: Controversy Not an easily solved problem Based on majority it seems that the most likely topology is A(E(BM)). The Red blocks in the tables Least sensitive to all the variants Alignment Gene Outgroup

Acknowledgements: 

Acknowledgements Thanks to Dennis Pearl and John Wenzel for all their suggestions and help. Thanks also to Jeff Pan for his help with computer programs.

References: 

References Danforth, B.N., J. Fang, S. Sipes, S.G. Brady andamp; E. Almeida (2004). Phylogeny and molecular systematics of bees (Hymenoptera: Apidea). Cornell University, Ithaca, NY http://www.entomology.cornell.edu/BeePhylogeny/ 'Euglossini.' Wikipedia, The Free Encyclopedia. 20 Jun 2006, 23:38 UTC. Wikimedia Foundation, Inc. 16 Aug 2006 andlt;http://en.wikipedia.org/w/index.php?title=Euglossiniandamp;oldid=59714591andgt;. 'Bumblebee.' Wikipedia, The Free Encyclopedia. 5 Aug 2006, 07:19 UTC. Wikimedia Foundation, Inc. 16 Aug 2006 andlt;http://en.wikipedia.org/w/index.php?title=Bumblebeeandamp;oldid=67786525andgt;. 'Honeybee.' Wikipedia, The Free Encyclopedia. 16 Aug 2006, 03:37 UTC. Wikimedia Foundation, Inc. 16 Aug 2006 andlt;http://en.wikipedia.org/w/index.php?title=Honeybeeandamp;oldid=69944973andgt;. 'Stingless bee.' Wikipedia, The Free Encyclopedia. 24 Jul 2006, 14:15 UTC. Wikimedia Foundation, Inc. 16 Aug 2006 andlt;http://en.wikipedia.org/w/index.php?title=Stingless_beeandamp;oldid=65555172andgt;. Noll, Fernando B. 'Behavioral Phylogeny of Corbiculate Apidae (Hymenoptera; Apinae), with Special Reference to Social Behavior.' Cladistics 18 (2002): 137-153. Winston, Mark L. and Charles D. Michener. 'Dual origin of highly social behavior among bees.' Proc. National Academy of Science 74.3 (Mar 1977): 1135-1137. Cameron, Sydney A. and Patrick Mardulyn. 'Multiple Molecular Data Sets Suggest Independent Origins of Highly Eusocial Behavior in Bees (Hymenoptera:Apinae).' Systematic Biology 50.2 (Apr 2001): 194-214. Serrao, J.E. 'A comparative study of the proventricular structure in corbiculate apinae (Hymenoptera, Apidae).' Micron 32 (2001): 379-385.