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Published September 18, 2014 | Supplemental Material + Published
Journal Article Open

The genomic substrate for adaptive radiation in African cichlid fish

Abstract

Cichlid fishes are famous for large, diverse and replicated adaptive radiations in the Great Lakes of East Africa. To understand the molecular mechanisms underlying cichlid phenotypic diversity, we sequenced the genomes and transcriptomes of five lineages of African cichlids: the Nile tilapia (Oreochromis niloticus), an ancestral lineage with low diversity; and four members of the East African lineage: Neolamprologus brichardi/pulcher (older radiation, Lake Tanganyika), Metriaclima zebra (recent radiation, Lake Malawi), Pundamilia nyererei (very recent radiation, Lake Victoria), and Astatotilapia burtoni (riverine species around Lake Tanganyika). We found an excess of gene duplications in the East African lineage compared to tilapia and other teleosts, an abundance of non-coding element divergence, accelerated coding sequence evolution, expression divergence associated with transposable element insertions, and regulation by novel microRNAs. In addition, we analysed sequence data from sixty individuals representing six closely related species from Lake Victoria, and show genome-wide diversifying selection on coding and regulatory variants, some of which were recruited from ancient polymorphisms. We conclude that a number of molecular mechanisms shaped East African cichlid genomes, and that amassing of standing variation during periods of relaxed purifying selection may have been important in facilitating subsequent evolutionary diversification.

Additional Information

© 2014 The Authors. This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported licence. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons licence, users will need to obtain permission from the licence holder to reproduce the material. Received 18 December 2013. Accepted 01 August 2014. Published online 03 September 2014. We would like to thank the Broad Institute Genomics Platform for sequencing of the 5 cichlid genomes and transcriptomes. Sequencing, assembly, annotation and analysis by Broad Institute were supported by grants from the National Human Genome Research Institute (NHGRI). Genome evolution, duplication and TE analysis, ILS and ancient variant analyses were also supported by Swiss National Science Foundation grant PBLAP3-142774 awarded to D.B. and by University of Oxford Nuffield Department of Medicine Prize Studentship to Y.I.L. TE and copy number variation analyses were supported by the German Science Foundation (DFG), and advanced grant 29700 ("GenAdap") by the European Research Council (ERC). CNE analysis and zebrafish functional assays were supported by the Biomedical Research Council of A*STAR, Singapore. MicroRNA sequencing and annotation was supported by ERC Starting Grant to E.A.M.; M.M. was supported by a fellowship from the Wellcome Trust. MicroRNA and target in situ hybridization was supported by grant 2R01DE019637-04 to J.T.S. Population genomics analyses were supported by Swiss National Science Foundation grants 31003A-118293 and 31003A-144046 to O.S. T.D.K., R.D.F., A.M., O.S., J.T.S., K.L.C., N.O., J.-F.B., D.J.P. and H.A.H. conceived the original tilapia white paper. F.D.P. , K.L.-T. and E.S.L. revised, planned and oversaw the genome project. D.J.P., W.S., H. S. G., M.E.S., O.S., K.L.C., T.D.K., G.H., O.E. and H.A.H. provided tissues and RNAs for sequencing. C.A. prepared the high molecular weight tilapia DNA. M.L. extracted genomic DNA for sequencing. L.W. prepared 40-kb libraries (Fossils) for Illumina sequencing. R.S. performed quality control of RNA. J.A., J.J. and F.D.P. oversaw the sequencing and assembly of genomes and transcriptomes as well as submissions of data. J.T.M. and P.R. performed quality control of assemblies and alignments of genomes. J.M.T. performed de novo assembly of transcriptomes. M.C. performed quality control of tilapia and M. zebra assemblies. A.B., Sa.Y., I.M., S.G., D.P., F.J.R., T.S., Sh.Y. and D.B.J. assembled the genome. F.G., R.G., M.R., J.-F.B., H.D'C., C.O.-C. contributed to the tilapia radiation hybrid map. F.B.-H. and N.A. analysed the OR and TAAR gene families. B.A., T.H. and S.S. annotated the tilapia genome. D.B. and Y.I.L. annotated the N. brichardi and the lake cichlids. D.B. performed gene expression, genome evolution, gene duplication and TE insertion analyses. Y.I.L. and L. S.-P. performed quality control of RNA-seq data and assemblies, gene evolution, incomplete lineage sorting and ancient variant analyses. S.F., Oleg S. and A.M., N.O., M.N. and H.N. analysed the TE landscape of cichlid genomes. S.F., Oleg S. and A.M. performed the TE burst history analysis and analysed copy number variants using read depth. E.B. and S.C.P.R. analysed duplications by comparative genomic hybridization (aCGH). H.A.H. and R.M.H. performed PCR to validate the transcriptome. A.Y.N., Z.W.L., A.P.L. and B.V. performed conserved CNE analysis and functional assays of cichlid CNEs. M.M. and E.M. performed microRNA sequencing and annotation from embryos of cichlid species as well as target identification. R.A., F.J.T. and R.D.F. annotated adult brain microRNAs in A. burtoni. R.B., N.S.H. and J.T.S. performed microRNA and target gene in situ hybridization. O.S. designed and oversaw the population genomics data analysis from Lake Victoria species; L.G., S.M. and I.K. generated the data; C.E.W., I.K., H.J.N. and O.S. analysed the data. F.D.P., K.L.-T. and O.S. wrote the manuscript with input from D.B., C.E.W. and Y.I.L., I.K., J.T.S., W.H., C.P.P. as well as additional authors. L.G. assisted with figure preparation and coordination. Competing financial interests The authors declare no competing financial interests. Genome assemblies and transcriptomes have been deposited in GenBank. The BioProject Identifiers are as follows. Genome sequencing: PRJNA59571 (SRP004171) for O. niloticus; PRJNA60365 (SRP004799) for N. brichardi; PRJNA60367 (SRP004869) for P. nyererei; PRJNA60369 (SRP004788) for M. zebra; and PRJNA60363 (SRP004787) for A. burtoni. Transcriptome sequencing (mRNAs): PRJNA78915 for O. niloticus; PRJNA77747 for N. brichardi; PRJNA83153 for P. nyererei; PRJNA77743 for M. zebra; and PRJNA78185 for A. burtoni. Additional SRA information for each tissue can be found in the Supplementary Informations. Transcriptome sequencing (microRNAs): PRJNA221867 (SRS489376) for O. niloticus; PRJNA222491 (SRS491903) for N. brichardi; PRJNA222489 (SRS491906) for P. nyererei; PRJNA221871 (SRS491904) for M. zebra; and PRJNA222490 (SRS491905) for A. burtoni. Cichlid microRNAs were deposited in miRBase.

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