Short Communication |
Corresponding author: Susette Castañeda-Rico ( susetteazul@gmail.com ) Academic editor: Nilton Cáceres
© 2023 Susette Castañeda-Rico, Lillian D. Parker, Evelyn Sánchez, Sheccid Rivas-Trasvina, Melissa T. R. Hawkins, Cody W. Edwards, Jesús E. Maldonado.
This is an open access article distributed under the terms of the CC0 Public Domain Dedication.
Citation:
Castañeda-Rico S, Parker LD, Sánchez E, Rivas-Trasvina S, Hawkins MTR, Edwards CW, Maldonado JE (2023) Novel genomic resources contribute to the systematics of threatened arboreal deer mice of the genus Habromys Hooper & Musser, 1964 (Cricetidae, Neotominae) within a neotomine–peromyscine phylogeny. ZooKeys 1179: 157-168. https://doi.org/10.3897/zookeys.1179.108759
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The Crested-tailed deer mouse, Habromys lophurus, is one of seven arboreal species within the genus Habromys. Species of this genus are monotypic, relatively rare, and occur in low densities. Their geographical distribution is highly fragmented due to being restricted to montane cloud forest in Mesoamerica and they are of conservation concern. All Habromys species are endemic to Mexico, except H. lophurus, which is also distributed in Guatemala and El Salvador. In this study, we obtained and characterized the first mitogenome and several thousand nuclear ultraconserved elements (UCEs) of H. lophurus to determine its phylogenetic position within neotomine–peromyscine mice. Its mitogenome sequence (16,509 bp) is only the second complete mitogenome obtained for this poorly known genus. We also obtained the first nuclear genomic data for H. lophurus, including 3,654 UCE loci, as well as a partial mitogenome of H. simulatus (6,349 bp), and 2,186 UCE for the outgroup Holochilus sciureus. Phylogenetic analyses that included our newly generated genomic data coupled with previously published data from other neotomine–peromyscine mice confirm the placement of H. lophurus, H. simulatus, and H. ixtlani within a highly supported clade. The Habromys clade was nested within a clade that also contains members of the genus Peromyscus and provides further support for the hypothesis of the paraphyly of Peromyscus. These genomic resources will contribute to future phylogenomic studies that aim to further elucidate the evolutionary history of this rare and critically endangered genus of rodents.
Conservation, Habromys lophurus, Mesoamerica, mitogenomes, museomics, phylogenomics, ultraconserved elements
The genus Habromys Hooper & Musser, 1964, considered a relict, comprises seven monotypic species (H. simulatus (Osgood, 1904), H. chinanteco (Robertson & Musser, 1976), H. delicatulus Carleton, Sánchez & Urbano-Vidales, 2002, H. schmidlyi Romo-Vázquez, León-Paniagua & Sánchez, 2005, H. lophurus (Osgood, 1904), H. lepturus (Merriam, 1898), and H. ixtlani (Goodwin, 1964)) which inhabit montane regions of Mesoamerica, ranging from central-southern Mexico to north-eastern El Salvador (
The crested-tailed deer mouse, Habromys lophurus, is distributed in Nuclear Central America, from Chiapas, Mexico to Guatemala and El Salvador (
We used a tissue sample of Habromys lophurus associated with a male museum voucher specimen (Fig.
Voucher specimen of Habromys lophurus (USNM 569392) from which the mitogenome and ultraconserved elements were obtained in this study. This species may be identified by its crested tail and usually by the absence of a white underside of the tail. Currently, there are no pictures available of live individuals of this species. Photograph by Katie Sayers.
DNA was extracted from the tissue sample (ca 1.5 mm2 of frozen internal organ) using the DNeasy Blood and Tissue Kit (QIAGEN, Inc., Valencia, CA, USA), following the manufacturer’s protocol, and quantified using a Qubit (Thermo Fisher, Waltham, MA, USA) fluorometer with a 1× dsDNA HS assay kit. DNA was sheared to an average length of 250 base pairs (bp) using a Bioruptor Pico sonicator (Diagenode Inc., Denville, NJ, USA) with a pulse of 30 s on, 30 s off for 90 cycles. A dual-indexed library with TruSeq-style indices (
For mitogenomes, read quality analysis of the FASTQ format files was conducted using FastQC v0.11.5 (
For the UCEs, raw data were processed following the PHYLUCE v1.6.7 pipeline (
To determine the phylogenetic position of H. lophurus, a Bayesian Inference (BI) analysis was performed using MrBayes v3.2.6 (
Specimens examined in this study using mitogenomes and UCEs with species name, accession number collection/ID study, GenBank accession number, and GenBank BioProject. Smithsonian Institution’s National Museum of Natural History (USNM), Museum of Texas Tech University (TTU/TK), Museo de Zoología “Alfonso L. Herrera” Facultad de Ciencias UNAM (MZFC), University of Michigan Museum of Zoology (UMMZ), and Museum of Vertebrate Zoology, University of California, Berkeley (MVZ).
Species | Number Scientific Collection/ID | Mitogenome (GenBank Number) | UCE (GenBank BioProject) |
---|---|---|---|
Habromys lophurus | USNM569392/USNM569392 | OP936043 | PRJNA907399 |
Habromys simulatus | MZFC10104/HBR031 | Figshare | PRJNA606805 |
Habromys ixtlani | – /TK93158 | KY707304 | – |
Peromyscus hooperi | USNM340233/USNM340233 | OP432689 | PRJNA880321 |
Peromyscus megalops | USNM340233/USNM340233 | ON528115 | PRJNA838631 |
Peromyscus attwateri | TTU143738/TK185663 | ON528112 | PRJNA838631 |
Peromyscus aztecus | USNM569848/USNM569848 | ON528113 | PRJNA838631 |
Peromyscus polionotus | USNM585473/USNM585473 | ON528117 | PRJNA838631 |
Peromyscus crinitus | TTU146966/TK193714 | ON528114 | PRJNA838631 |
Peromyscus mekisturus | UMMZ88967/UMMZ88967 | MT078818 | PRJNA606805 |
Peromyscus melanophrys | MZFC3907/MQ1229 | MT078816 | PRJNA606805 |
Peromyscus perfulvus | – /MCP119 | MT078817 | PRJNA606805 |
Peromyscus pectoralis | MZFC10465/FCR176 | MT078819 | PRJNA606805 |
Peromyscus mexicanus | MZFC11150/MRM030 | – | PRJNA606805 |
TTU82723/TK93144 | KY707303 | – | |
Podomys floridanus | TTU97866/TK92501 | ON528118 | PRJNA838631 |
Neotomodon alstoni | TTU82668/TK93098 | ON528110 | PRJNA838631 |
Onychomys leucogaster | TTU146304/TK171574 | ON528111 | PRJNA838631 |
Reithrodontomys mexicanus | TTU138428/TK178510 | ON528119 | PRJNA838631 |
Isthmomys pirrensis | USNM565924/USNM565924 | ON528108 | PRJNA838631 |
Neotoma mexicana | TTU104969/TK150189 | ON528109 | PRJNA838631 |
Holochilus sciureus | MVZ190356/MVZ190356 | OL685394 | PRJNA818347 |
The assembled complete mitogenome sequence of Habromys lophurus is 16,509 bp long (GenBank accession number OP936043), which covers 100% of the reference genome sequence. The average coverage per base was 2,701X with 347,009 unique sequence reads mapped to the reference genome. The sequence contained the standard features present in a vertebrate mitochondrial genome including one non-coding control region, 2 rRNA genes (12S rRNA and 16S rRNA), 22 tRNA genes, and 13 PCGs including NADH dehydrogenase (ND1, ND2, ND3, ND4, ND4L, ND5 and ND6), cytochrome c oxidase (COX1, COX2, and COX3), ATP synthase (ATP6 and ATP8) and cytochrome b. The base composition was 34.4% A, 23.8% C, 12.7% G, 29.1% T and the GC content was 36.5%. The mitogenome map (Fig.
Mitogenome map of the crested-tailed deer mouse Habromys lophurus (USNM 569392, GenBank accession number OP936043) depicting the gene organization. The inner circle shows the CG content along the mitogenome.
The UCE processing recovered 209,371 and 556,910 contigs for H. lophurus and Holochilus sciureus after Trinity assembly, respectively. A total of 3,654 UCE loci were obtained for H. lophurus and 2,186 for H. sciureus. The incomplete matrix contained 4,415 UCE loci (N = 20; average = 3,145; min = 1,087; max = 3,859). The 90% matrix included 438 UCE loci with an average of 15 informative sites and 649 bp length per locus.
Our phylogenetic analyses support the placement of H. lophurus, H. simulatus, and H. ixtlani within a clade with high nodal support (Fig.
Bayesian phylogenetic tree of neotomine–peromyscine mice using (A) complete mitogenomes and (B) ultraconserved elements (UCEs). Nodal support is provided with posterior probabilities values. GenBank accession numbers follow species names for the mitogenome-based phylogeny and the accession number collection for the UCE-based phylogeny.
Within Habromys, the mitogenome-based phylogeny showed that H. lophurus is sister to H. ixtlani, and both are sisters to H. simulatus. This relationship was not supported by the UCE-based phylogeny due to a lack of UCE data for H. ixtlani. A close relationship between H. lophurus and H. ixtlani is supported by previous morphological studies that suggest Habromys includes two main lineages: one containing the smallest and more arboreal species (H. simulatus, H. schmidlyi, H. delicatulus, and H. chinanteco) and the other including the largest and less arboreal species (H. ixtlani, H. lepturus, and H. lophurus) (
All Habromys species are of conservation concern given the increasing annual deforestation rates for montane cloud forest (
Efforts to determine the correct status of these species have been stifled because of the very limited number of studies that have focused on this genus. Furthermore, a recent study looking at distributional patterns and conservation of the genus Habromys (
We would like to thank the specimen collectors, collection managers, curators, and all museum staff of the Smithsonian Institution’s National Museum of Natural History (NMNH) for the loaned tissue sample used in this study. We thank Katie Sayers for the photograph of the museum specimen.
The authors have declared that no competing interests exist.
We used a tissue sample (USNM 569392) that was requested from the Smithsonian Institution’s National Museum of Natural History (NMNH) Biorepository after deposition in the National Collection, as such no Animal Care and Use Committee (ACUC) was required as this study did not collect any new specimen. This request was approved by the Division of Mammals under transaction #2089501. This species is not listed in CITES and is classified as Near-Threatened by the IUCN.
This project was funded by the Center for Conservation Genomics, Smithsonian National Zoo and Conservation Biology Institute and Smithsonian-Mason School of Conservation, George Mason University. This project also received federal support from the Latino Initiatives Pool, administered by the Smithsonian Latino Center. Publication of this article was funded by the University of California, Santa Barbara (UCSB)-Smithsonian Scholars Program.
SC-R, LDP, CWE, and JEM designed and conceptualized the study. SC-R, CWE, and JEM secure funding and resources. MTRH performed the specimen sampling. SC-R, LDP, MTRH, ES, and SR-T performed formal analyses. SC-R conducted laboratory work, archived the data, produced figures, and wrote the original draft. All authors contributed to writing, editing, and approving the manuscript.
Susette Castañeda-Rico https://orcid.org/0000-0002-4301-3579
Lillian D. Parker https://orcid.org/0000-0003-3370-9473
Melissa T. R. Hawkins https://orcid.org/0000-0001-8929-1593
Cody W. Edwards https://orcid.org/0000-0001-5912-6693
Jesús E. Maldonado https://orcid.org/0000-0002-4282-1072
The mitochondrial genome sequences are available in GenBank at https://www.ncbi.nlm.nih.gov/genbank/ under the accession number OP936043 and Figshare (https://doi.org/10.25573/data.23563929). The associated BioProject, BioSample and SRA numbers are PRJNA907399, SAMN31979917, and SRR22493489, respectively.