Genome and transcriptome mechanisms driving cephalopod evolution.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
04 05 2022
04 05 2022
Historique:
received:
15
04
2021
accepted:
28
03
2022
entrez:
4
5
2022
pubmed:
5
5
2022
medline:
7
5
2022
Statut:
epublish
Résumé
Cephalopods are known for their large nervous systems, complex behaviors and morphological innovations. To investigate the genomic underpinnings of these features, we assembled the chromosomes of the Boston market squid, Doryteuthis (Loligo) pealeii, and the California two-spot octopus, Octopus bimaculoides, and compared them with those of the Hawaiian bobtail squid, Euprymna scolopes. The genomes of the soft-bodied (coleoid) cephalopods are highly rearranged relative to other extant molluscs, indicating an intense, early burst of genome restructuring. The coleoid genomes feature multi-megabase, tandem arrays of genes associated with brain development and cephalopod-specific innovations. We find that a known coleoid hallmark, extensive A-to-I mRNA editing, displays two fundamentally distinct patterns: one exclusive to the nervous system and concentrated in genic sequences, the other widespread and directed toward repetitive elements. We conclude that coleoid novelty is mediated in part by substantial genome reorganization, gene family expansion, and tissue-dependent mRNA editing.
Identifiants
pubmed: 35508532
doi: 10.1038/s41467-022-29748-w
pii: 10.1038/s41467-022-29748-w
pmc: PMC9068888
doi:
Substances chimiques
RNA, Messenger
0
Types de publication
Journal Article
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2427Subventions
Organisme : Austrian Science Fund FWF
ID : P 30686
Pays : Austria
Organisme : NCATS NIH HHS
ID : UL1 TR000430
Pays : United States
Organisme : NCATS NIH HHS
ID : UL1 TR002389
Pays : United States
Informations de copyright
© 2022. The Author(s).
Références
Shigeno, S., Andrews, P. L. R., Ponte, G. & Fiorito, G. Cephalopod brains: an overview of current knowledge to facilitate comparison with vertebrates. Front. Physiol. 9, 952 (2018).
pubmed: 30079030
pmcid: 6062618
doi: 10.3389/fphys.2018.00952
Wang, Z. Y. & Ragsdale, C. W. Cephalopod nervous system organization. in Oxford Research Encyclopedia of Neuroscience (Oxford University Press, 2019).
Putnam, N. H. et al. The amphioxus genome and the evolution of the chordate karyotype. Nature 453, 1064–1071 (2008).
pubmed: 18563158
doi: 10.1038/nature06967
Holland, P. W. H., Garcia-Fernàndez, J., Williams, N. A. & Sidow, A. Gene duplications and the origins of vertebrate development. Development 1994, 125–133 (1994).
doi: 10.1242/dev.1994.Supplement.125
Albertin, C. B. et al. The octopus genome and the evolution of cephalopod neural and morphological novelties. Nature 524, 220–224 (2015).
pubmed: 26268193
pmcid: 4795812
doi: 10.1038/nature14668
Hallinan, N. M. & Lindberg, D. R. Comparative analysis of chromosome counts infers three paleopolyploidies in the mollusca. Genome Biol. Evol. 3, 1150–1163 (2011).
pubmed: 21859805
pmcid: 3194838
doi: 10.1093/gbe/evr087
Garrett, S. & Rosenthal, J. J. C. RNA editing underlies temperature adaptation in K+ channels from polar octopuses. Science 335, 848–851 (2012).
pubmed: 22223739
pmcid: 4219319
doi: 10.1126/science.1212795
Rosenthal, J. J. C. The emerging role of RNA editing in plasticity. J. Exp. Biol. 218, 1812–1821 (2015).
pubmed: 26085659
pmcid: 4487009
doi: 10.1242/jeb.119065
Liscovitch-Brauer, N. et al. Trade-off between transcriptome plasticity and genome evolution in cephalopods. Cell 169, 191–202.e11 (2017).
pubmed: 28388405
pmcid: 5499236
doi: 10.1016/j.cell.2017.03.025
Alon, S. et al. The majority of transcripts in the squid nervous system are extensively recoded by A-to-I RNA editing. eLife. https://elifesciences.org/articles/05198 (2015).
Sakurai, M., Okada, S., Ueda, H. & Yang, Y. Discovering A-to-I RNA editing through chemical methodology “ICE-seq” ICE-seq. in RNA Editing: Methods and Protocols (eds. Picardi, E. & Pesole, G.) 113–148 (Springer US, 2021).
Tan, M. H. et al. Dynamic landscape and regulation of RNA editing in mammals. Nature 550, 249–254 (2017).
pubmed: 29022589
pmcid: 5723435
doi: 10.1038/nature24041
Schwiening, C. J. A brief historical perspective: Hodgkin and Huxley. J. Physiol. 590, 2571–2575 (2012).
pubmed: 22787170
pmcid: 3424716
doi: 10.1113/jphysiol.2012.230458
Hodgkin, A. L. & Huxley, A. F. A quantitative description of membrane current and its application to conduction and excitation in nerve. J. Physiol. 117, 500–544 (1952).
pubmed: 12991237
pmcid: 1392413
doi: 10.1113/jphysiol.1952.sp004764
Williams, L. W. The anatomy of the common squid: Loligo pealii, Lesueur (Library and printing-office late E.J. Brill, Leiden, Holland, 1909).
Rosen, H. E. & Gilly, W. F. Myogenic activity and serotonergic inhibition in the chromatophore network of the squid Dosidicus gigas (family Ommastrephidae) and Doryteuthis opalescens (family Loliginidae). J. Exp. Biol. 220, 4669–4680 (2017).
pubmed: 29061686
Marian, J. E. A. R. et al. Male alternative reproductive tactics and associated evolution of anatomical characteristics in loliginid squid. Front. Physiol. 10, 1281 (2019).
pubmed: 31680998
pmcid: 6803530
doi: 10.3389/fphys.2019.01281
Oshima, M. et al. Peripheral injury alters schooling behavior in squid, Doryteuthis pealeii. Behav. Process. 128, 89–95 (2016).
doi: 10.1016/j.beproc.2016.04.008
Koenig, K. M., Sun, P., Meyer, E. & Gross, J. M. Eye development and photoreceptor differentiation in the cephalopod Doryteuthis pealeii. Development 143, 3168–3181 (2016).
pubmed: 27510978
Holt, A. L., Sweeney, A. M., Johnsen, S. & Morse, D. E. A highly distributed Bragg stack with unique geometry provides effective camouflage for Loliginid squid eyes. J. R. Soc. Interface 8, 1386–1399 (2011).
pubmed: 21325315
pmcid: 3163417
doi: 10.1098/rsif.2010.0702
Messerli, M. A. et al. Construction and composition of the squid pen from Doryteuthis pealeii. Biol. Bull. 237, 1–15 (2019).
pubmed: 31441702
pmcid: 7340512
doi: 10.1086/704209
Crawford, K. et al. Highly efficient knockout of a squid pigmentation gene. Curr. Biol. 30, 3484–3490.e4 (2020).
pubmed: 32735817
pmcid: 7484294
doi: 10.1016/j.cub.2020.06.099
Dawe, E. G., Hendrickson, L. C., Colbourne, E. B., Drinkwater, K. F. & Showell, M. A. Ocean climate effects on the relative abundance of short-finned (Illex illecebrosus) and long-finned (Loligo pealeii) squid in the northwest Atlantic Ocean. Fish. Oceanogr. 16, 303–316 (2007).
doi: 10.1111/j.1365-2419.2007.00431.x
Hinegardner, R. Cellular DNA content of the Mollusca. Comp. Biochem. Physiol. A Physiol. 47, 447–460 (1974).
doi: 10.1016/0300-9629(74)90008-5
Gao, Y. M. & Natsukari, Y. Karyological studies on seven cephalopods. Venus Jpn. J. Malacol. 49, 126–145 (1990).
Wang, J. & Zheng, X. Comparison of the genetic relationship between nine Cephalopod species based on cluster analysis of karyotype evolutionary distance. Comp. Cytogenet. 11, 477–494 (2017).
pubmed: 29093799
pmcid: 5646656
doi: 10.3897/compcytogen.v11i3.12752
Belcaid, M. et al. Symbiotic organs shaped by distinct modes of genome evolution in cephalopods. Proc. Natl Acad. Sci. USA 116, 3030–3035 (2019).
pubmed: 30635418
pmcid: 6386654
doi: 10.1073/pnas.1817322116
Tanner, A. R. et al. Molecular clocks indicate turnover and diversification of modern coleoid cephalopods during the Mesozoic Marine Revolution. Proc. R. Soc. B Biol. Sci. 284, 20162818 (2017).
doi: 10.1098/rspb.2016.2818
Anderson, F. E. & Lindgren, A. R. Phylogenomic analyses recover a clade of large-bodied decapodiform cephalopods. Mol. Phylogenet. Evol. 156, 107038 (2021).
pubmed: 33285289
doi: 10.1016/j.ympev.2020.107038
Kröger, B., Vinther, J. & Fuchs, D. Cephalopod origin and evolution: a congruent picture emerging from fossils, development and molecules. BioEssays 33, 602–613 (2011).
pubmed: 21681989
doi: 10.1002/bies.201100001
Feschotte, C. Transposable elements and the evolution of regulatory networks. Nat. Rev. Genet. 9, 397–405 (2008).
pubmed: 18368054
pmcid: 2596197
doi: 10.1038/nrg2337
Kretschmer, R., Ferguson-Smith, M. A. & de Oliveira, E. H. C. Karyotype evolution in birds: from conventional staining to chromosome painting. Genes 9, 181 (2018).
pmcid: 5924523
doi: 10.3390/genes9040181
Zhang, Y. et al. The genome of Nautilus pompilius illuminates eye evolution and biomineralization. Nat. Ecol. Evol. 5, 927–938 (2021).
pubmed: 33972735
pmcid: 8257504
doi: 10.1038/s41559-021-01448-6
Simakov, O. et al. Deeply conserved synteny resolves early events in vertebrate evolution. Nat. Ecol. Evol. 4, 820–830 (2020).
pubmed: 32313176
pmcid: 7269912
doi: 10.1038/s41559-020-1156-z
Wang, S. et al. Scallop genome provides insights into evolution of bilaterian karyotype and development. Nat. Ecol. Evol. 1, s41559-017-0120–017 (2017).
Adachi, K., Ohnishi, K., Kuramochi, T., Yoshinaga, T. & Okumura, S.-I. Molecular cytogenetic study in Octopus (Amphioctopus) areolatus from Japan. Fish. Sci. 80, 445–450 (2014).
doi: 10.1007/s12562-014-0703-4
Wang, Z. Y. & Ragsdale, C. W. Cadherin genes and evolutionary novelties in the octopus. Semin. Cell Dev. Biol. 69, 151–157 (2017).
pubmed: 28627384
doi: 10.1016/j.semcdb.2017.06.007
Styfhals, R., Seuntjens, E., Simakov, O., Sanges, R. & Fiorito, G. In silico Identification and expression of protocadherin gene family in Octopus vulgaris. Front. Physiol. 9, 1905 (2019).
pubmed: 30692932
pmcid: 6339937
doi: 10.3389/fphys.2018.01905
Rubinstein, R. et al. Molecular logic of neuronal self-recognition through protocadherin domain interactions. Cell 163, 629–642 (2015).
pubmed: 26478182
pmcid: 4624033
doi: 10.1016/j.cell.2015.09.026
Chen, W. V. & Maniatis, T. Clustered protocadherins. Development 140, 3297–3302 (2013).
pubmed: 23900538
pmcid: 3737714
doi: 10.1242/dev.090621
Tomarev, S. I., Chung, S. & Piatigorsky, J. Glutathione S-transferase and S-crystallins of cephalopods: Evolution from active enzyme to lens-refractive proteins. J. Mol. Evol. 41, 1048–1056 (1995).
pubmed: 8587103
doi: 10.1007/BF00173186
Tomarev, S. I. & Piatigorsky, J. Lens crystallins of invertebrates. Eur. J. Biochem. 235, 449–465 (1996).
pubmed: 8654388
doi: 10.1111/j.1432-1033.1996.00449.x
Sweeney, A. M., Des Marais, D. L., Andrew Ban, Y.-E. & Johnsen, S. Evolution of graded refractive index in squid lenses. J. R. Soc. Interface 4, 685–698 (2007).
pubmed: 17293312
pmcid: 2373386
doi: 10.1098/rsif.2006.0210
van Giesen, L., Kilian, P. B., Allard, C. A. H. & Bellono, N. W. Molecular basis of chemotactile sensation in octopus. Cell 183, 594–604.e14 (2020).
pubmed: 33125889
pmcid: 7605239
doi: 10.1016/j.cell.2020.09.008
Hirayama, T. & Yagi, T. Clustered protocadherins and neuronal diversity. Prog. Mol. Biol. Transl. Sci. 116, 145–167 (2013).
pubmed: 23481194
doi: 10.1016/B978-0-12-394311-8.00007-8
Zipursky, S. L. & Sanes, J. R. Chemoaffinity revisited: dscams, protocadherins, and neural circuit assembly. Cell 143, 343–353 (2010).
pubmed: 21029858
doi: 10.1016/j.cell.2010.10.009
Sanes, J. R. & Zipursky, S. L. Synaptic specificity, recognition molecules, and assembly of neural circuits. Cell 181, 536–556 (2020).
pubmed: 32359437
doi: 10.1016/j.cell.2020.04.008
Goodman, K. M. et al. γ-Protocadherin structural diversity and functional implications. eLife. https://elifesciences.org/articles/20930 (2016).
Cai, J., Townsend, J. P., Dodson, T. C., Heiney, P. A. & Sweeney, A. M. Eye patches: protein assembly of index-gradient squid lenses. Science 357, 564–569 (2017).
pubmed: 28798124
pmcid: 5682922
doi: 10.1126/science.aal2674
Cai, J. & Sweeney, A. M. The proof is in the pidan: generalizing proteins as patchy particles. ACS Cent. Sci. 4, 840–853 (2018).
pubmed: 30062112
pmcid: 6062823
doi: 10.1021/acscentsci.8b00187
Nishikura, K. Functions and regulation of RNA editing by ADAR deaminases. Annu. Rev. Biochem. 79, 321–349 (2010).
pubmed: 20192758
pmcid: 2953425
doi: 10.1146/annurev-biochem-060208-105251
Basilio, C., Wahba, A. J., Lengyel, P., Speyer, J. F. & Ochoa, S. Synthetic polynucleotides and the amino acid code, v. PNAS 48, 613–616 (1962).
pubmed: 13865603
pmcid: 220825
doi: 10.1073/pnas.48.4.613
Shoshan, Y., Liscovitch-Brauer, N., Rosenthal, J. J. C., Eisenberg, E. & O’Connell, M. Adaptive Proteome Diversification by Nonsynonymous A-to-I RNA Editing in Coleoid Cephalopods. Mol. Biol. Evol. 38, 3775–3788 (2021).
pubmed: 34022057
pmcid: 8382921
doi: 10.1093/molbev/msab154
Moldovan, M., Chervontseva, Z., Bazykin, G. & Gelfand, M. S. Adaptive evolution at mRNA editing sites in soft-bodied cephalopods. PeerJ 8, e10456 (2020).
pubmed: 33312772
pmcid: 7703385
doi: 10.7717/peerj.10456
Jiang, D. & Zhang, J. The preponderance of nonsynonymous A-to-I RNA editing in coleoids is nonadaptive. Nat. Commun. 10, 1–10 (2019).
doi: 10.1038/s41467-019-13275-2
Athanasiadis, A., Rich, A. & Maas, S. Widespread A-to-I RNA editing of Alu-containing mRNAs in the human transcriptome. PLoS Biol. 2, e391 (2004).
pubmed: 15534692
pmcid: 526178
doi: 10.1371/journal.pbio.0020391
Orecchini, E., Frassinelli, L. & Michienzi, A. Restricting retrotransposons: ADAR1 is another guardian of the human genome. RNA Biol. 14, 1485–1491 (2017).
pubmed: 28640667
pmcid: 5785221
doi: 10.1080/15476286.2017.1341033
Rosenthal, J. J. C. & Bezanilla, F. Extensive editing of mRNAs for the squid delayed rectifier K+ channel regulates subunit tetramerization. Neuron 34, 743–757 (2002).
pubmed: 12062021
doi: 10.1016/S0896-6273(02)00701-8
Blow, M., Futreal, P. A., Wooster, R. & Stratton, M. R. A survey of RNA editing in human brain. Genome Res. 14, 2379–2387 (2004).
pubmed: 15545495
pmcid: 534661
doi: 10.1101/gr.2951204
Köhler, M., Burnashev, N., Sakmann, B. & Seeburg, P. H. Determinants of ca2+ permeability in both TM1 and TM2 of high affinity kainate receptor channels: diversity by RNA editing. Neuron 10, 491–500 (1993).
pubmed: 7681676
doi: 10.1016/0896-6273(93)90336-P
Palavicini, J. P., O’connell, M. A. & Rosenthal, J. J. C. An extra double-stranded RNA binding domain confers high activity to a squid RNA editing enzyme. RNA 15, 1208–1218 (2009).
pubmed: 19390115
pmcid: 2685509
doi: 10.1261/rna.1471209
Cosson, B. et al. Oligomerization of EDEN-BP is required for specific mRNA deadenylation and binding. Biol. Cell 98, 653–665 (2006).
pubmed: 16836486
doi: 10.1042/BC20060054
Levanon, E. Y. et al. Systematic identification of abundant A-to-I editing sites in the human transcriptome. Nat. Biotechnol. 22, 1001–1005 (2004).
pubmed: 15258596
doi: 10.1038/nbt996
Kim, D. D. Y. et al. Widespread RNA editing of embedded alu elements in the human transcriptome. Genome Res. 14, 1719–1725 (2004).
pubmed: 15342557
pmcid: 515317
doi: 10.1101/gr.2855504
Crookes, W. J. et al. Reflectins: the unusual proteins of squid reflective tissues. Science 303, 235–238 (2004).
pubmed: 14716016
doi: 10.1126/science.1091288
Guerette, P. A. et al. Nanoconfined β-sheets mechanically reinforce the supra-biomolecular network of robust squid sucker ring teeth. ACS Nano 8, 7170–7179 (2014).
pubmed: 24911543
doi: 10.1021/nn502149u
Tan, Y. et al. Infiltration of chitin by protein coacervates defines the squid beak mechanical gradient. Nat. Chem. Biol. 11, 488–495 (2015).
pubmed: 26053298
doi: 10.1038/nchembio.1833
da Fonseca, R. R. et al. A draft genome sequence of the elusive giant squid, Architeuthis dux. GigaScience 9, 152 (2020).
doi: 10.1093/gigascience/giz152
Fiorito, G. et al. Guidelines for the care and welfare of cephalopods in research—a consensus based on an initiative by CephRes, FELASA and the Boyd Group. Lab. Anim. 49, 1–90 (2015).
pubmed: 26354955
doi: 10.1177/0023677215580006
Fiorito, G. et al. Cephalopods in neuroscience: regulations, research and the 3Rs. Invert. Neurosci. 14, 13–36 (2014).
pubmed: 24385049
pmcid: 3938841
doi: 10.1007/s10158-013-0165-x
Lopes, V. M. et al. Cephalopod biology and care, a COST FA1301 (CephsInAction) training school: anaesthesia and scientific procedures. Invertebr. Neurosci. 17, 8 (2017).
doi: 10.1007/s10158-017-0200-4
Burton, J. N. et al. Chromosome-scale scaffolding of de novo genome assemblies based on chromatin interactions. Nat. Biotechnol. 31, 1119–1125 (2013).
pubmed: 24185095
pmcid: 4117202
doi: 10.1038/nbt.2727
Schmidbaur, H. et al. Emergence of novel cephalopod gene regulation and expression through large-scale genome reorganization. Nat. Commun. 13, 2172 (2022).
pubmed: 35449136
doi: 10.1038/s41467-022-29694-7
Edgar, R. C. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res. 32, 1792–1797 (2004).
pubmed: 15034147
pmcid: 390337
doi: 10.1093/nar/gkh340
Larsson, A. AliView: a fast and lightweight alignment viewer and editor for large datasets. Bioinform. Oxf. Engl. 30, 3276–3278 (2014).
doi: 10.1093/bioinformatics/btu531
Minh, B. Q. et al. IQ-TREE 2: new models and efficient methods for phylogenetic inference in the genomic era. Mol. Biol. Evol. 37, 1530–1534 (2020).
pubmed: 32011700
pmcid: 7182206
doi: 10.1093/molbev/msaa015
Kalyaanamoorthy, S., Minh, B. Q., Wong, T. K. F., Haeseler, Avon & Jermiin, L. S. ModelFinder: fast model selection for accurate phylogenetic estimates. Nat. Methods 14, 587–589 (2017).
pubmed: 28481363
pmcid: 5453245
doi: 10.1038/nmeth.4285
Hoang, D. T., Chernomor, O., von Haeseler, A., Minh, B. Q. & Vinh, L. S. UFBoot2: improving the ultrafast bootstrap approximation. Mol. Biol. Evol. 35, 518–522 (2018).
pubmed: 29077904
doi: 10.1093/molbev/msx281
Sanderson, M. J. r8s: inferring absolute rates of molecular evolution and divergence times in the absence of a molecular clock. Bioinformatics 19, 301–302 (2003).
pubmed: 12538260
doi: 10.1093/bioinformatics/19.2.301
Shu, S., Goodstein, D. & Rokhsar, D. PERTRAN: genome-guided RNA-seq read assembler. https://www.osti.gov/biblio/1241180 (2013).
Haas, B. J. et al. Improving the Arabidopsis genome annotation using maximal transcript alignment assemblies. Nucleic Acids Res. 31, 5654–5666 (2003).
pubmed: 14500829
pmcid: 206470
doi: 10.1093/nar/gkg770
Yeh, R. F., Lim, L. P. & Burge, C. B. Computational inference of homologous gene structures in the human genome. Genome Res. 11, 803–816 (2001).
pubmed: 11337476
pmcid: 311055
doi: 10.1101/gr.175701
Salamov, A. A. & Solovyev, V. V. Ab initio gene finding in Drosophila genomic DNA. Genome Res. 10, 516–522 (2000).
pubmed: 10779491
pmcid: 310882
doi: 10.1101/gr.10.4.516
Flynn, J. M. et al. RepeatModeler2 for automated genomic discovery of transposable element families. Proc. Natl Acad. Sci. USA 117, 9451–9457 (2020).
pubmed: 32300014
pmcid: 7196820
doi: 10.1073/pnas.1921046117
Smit, A. et al RepeatMasker Open-4.0. http://www.repeatmasker.org (2013).
Sievers, F. et al. Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol. Syst. Biol. 7, 539 (2011).
pubmed: 21988835
pmcid: 3261699
doi: 10.1038/msb.2011.75
Price, M. N., Dehal, P. S. & Arkin, A. P. FastTree 2-approximately maximum-likelihood trees for large alignments. PLoS ONE 5, e9490 (2010).
pubmed: 20224823
pmcid: 2835736
doi: 10.1371/journal.pone.0009490
Camacho, C. et al. BLAST+: architecture and applications. BMC Bioinform. 10, 421 (2009).
doi: 10.1186/1471-2105-10-421
Dobin, A. et al. STAR: ultrafast universal RNA-seq aligner. Bioinformatics 29, 15–21 (2013).
pubmed: 23104886
doi: 10.1093/bioinformatics/bts635
Li, H. et al. The sequence alignment/Map format and SAMtools. Bioinformatics 25, 2078–2079 (2009).
pubmed: 19505943
pmcid: 2723002
doi: 10.1093/bioinformatics/btp352
Cingolani, P. et al. A program for annotating and predicting the effects of single nucleotide polymorphisms, SnpEff: SNPs in the genome of Drosophila melanogaster strain w1118; iso-2; iso-3. Fly 6, 80–92 (2012).
pubmed: 22728672
pmcid: 3679285
doi: 10.4161/fly.19695
Katoh, K. & Toh, H. Recent developments in the MAFFT multiple sequence alignment program. Brief. Bioinform. 9, 286–298 (2008).
pubmed: 18372315
doi: 10.1093/bib/bbn013
Krogh, A., Larsson, B., von Heijne, G. & Sonnhammer, E. L. L. Predicting transmembrane protein topology with a hidden markov model: application to complete genomes. J. Mol. Biol. 305, 567–580 (2001).
pubmed: 11152613
doi: 10.1006/jmbi.2000.4315
Guo, Y. et al. A chromosomal-level genome assembly for the giant African snail Achatina fulica. GigaScience 8, giz124 (2019).
pubmed: 31634388
pmcid: 6802634
doi: 10.1093/gigascience/giz124