Corresponding author: Uliana V. Gorobeyko (
Academic editor: A. Saifitdinova
The DNA-barcoding and chromosomal study of the eastern water bat,
Gorobeyko UV, Kartavtseva IV, Sheremetyeva IN, Kazakov DV, Guskov VYu (2020) DNA-barcoding and a new data about the karyotype of
The eastern water bat,
The morphological heterogeneity and the presence of two major groups of forms in
The DNA-barcoding based on 657 bp length sequences of cytochrome c oxidase I (
Karyotype features are essential diagnostic characteristics of many mammalian species (
For the genus
Only conventional staining karyotypes of
Thus, the aim of present paper is to study DNA barcodes and chromosomes of
There are 19 specimens of
In addition, the 26
Sampling localities and GenBank sequencing data of
Code | Locality | Coordinates | GenBank | Specimen | Sex | 2n/ |
X | Y | Chromosomal stainings |
---|---|---|---|---|---|---|---|---|---|
|
Primorsky Krai, Primorsky Velican Cave |
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3240 | m | - | |||
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3400 | f | 44/52 |
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- | ||||
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3865 | f | 44/52 |
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- | GTG, CBG | |||
– | 3867 | m | 44/52 |
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A | GTG, CBG | |||
|
3869 | f | - | ||||||
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Primorsky Krai, Spasskaya Cave |
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3258 | m | 44/52 |
|
A | |
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3259 | m | 44/52 |
|
A | ||||
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Khabarovsky Krai, Komsomolsk Nature Reserve |
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UG16-18 | m | - | |||
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UG21-18 | m | - | ||||||
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Khabarovsky Krai, Komsomolsk-on-Amur City |
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UG28-18 | m | - | |||
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UG36-18 | f | - | ||||||
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Amur Oblast, Zeya City |
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3332 | m | - | |||
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3333 | m | 44/52 |
|
A |
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|||
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3334 | f | - | ||||||
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3335 | f | - | ||||||
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3336 | f | 44/52 |
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- | ||||
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3337 | m | - | ||||||
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3338 | f | 44/52 |
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- | ||||
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3339 | f | - | ||||||
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Primorsky Krai, Priiskovaya Cave |
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m |
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Sakhalin Oblast |
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- |
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Transbaikal Krai |
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m |
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|||
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Transbaikal Krai |
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m |
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Mongolia |
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- |
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Tuva Republic |
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- |
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Altai Republic |
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m |
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Altai |
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f |
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Altai |
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2f |
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South Korea |
|
KW001-004 | - |
|
Total DNA was isolated from ethanol-fixed tissues by the method of saline extraction (
All sequences were aligned using the software program BioEdit, version 7.0.9.0 and deposited in the GenBank database. The accession numbers of our and sequences obtained from GenBank are reported in the Table
The interspecific nucleotide diversity (π) and haplotype diversity (
Chromosome preparations were obtained from
The results of differential staining were analyzed with an AXIOSKOP 2 Plus microscope (Zeiss). The microimage registration and adjustment was performed with a CCD camera with software (META Systems GmbH, Germany) of the Joint-Use Center «Biotechnology & Genetic Engineering» in the Federal Scientific Center of the East Asia Terrestrial Biodiversity Far East Branch Russian Academy of Sciences (Vladivostok, Russia).
The DNA barcodes are obtained for 18 from a total of 19
Maximal Likelihood tree of the cytochrome oxidase I gene.
A pairwise genetic distances between the specimens of
The nucleotide diversity for the whole species is amounted to 0.00227±0.00032 with the haplotype diversity
A total of 9
The relationship among a total of 9 haplotypes reflected in the median‐joining network (Fig.
Distributional range and
Haplotypes G7 and G8 form a separate branch on the network and are found only in 8 specimens from Tuva and the Altai. The G8 haplotype revealed in the one specimen from the Altai differed from G7 on one nucleotide substitution and from G1 on two nucleotide substitution. The spread of G7 and G8 is apparently coincides with the distribution of nominative subspecies.
The other differential branch on the network is a G9 haplotype differed from G1 on three nucleotide substitution. It is found only in 4 individuals from South Korea. The distinct subspecies for
Most of the haplotypes represented in the samples are separated by G1 just one mutation creating a starlike network characteristic for expanding populations that have been through a bottleneck or been founded recently. However,
The conventional staining karyotypes of eight
It was previously reported for the specimens from the Primorsky Krai the fundamental number of autosomal arms was 50 (
The X chromosome is biarmed and it was not possible to determine whether this is a submetacentric or metacentric. At the same time the previously examined individuals from the Primorsky Krai have shown clearly a metacentric X chromosome. It is possible that these karyotypic differences are due to the methodological difficulties, such as the various spiralization of metacentric chromosomes or the lack of metaphase plates on the preparation often occurred in the analysis of chromosome suspensions obtained
The patterns of
The sequential GTG- and AgNOR-banding of
Distribution of nucleolus organizer regions: mean value of active
ID | No cells | chromosomal arm no. | ||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 18 | 19 | 20 | 21 | 22 | 23 | 24 | 25 | ||
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0,41 | 0,5 | 0,27 | 0,14 | 0,09 | 0,27 | 0,86 | 0,55 | 0,27 | 0,27 | 0,27 | 0,09 | |||||
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0,78 | 0,93 | 0,48 | 0,2 | 0,33 | 0,45 | 1 | 0,8 | 0,8 | 0,13 | 0,2 | 0,2 | |||||
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0,16 | 0,38 | 0,13 | 0,19 | 0,17 | 0,27 | 0,36 | 0,33 | 0,38 | 0,38 | 0,14 | 0,2 | |||||
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0,34 | 0,63 | 0,41 | 0,45 | 0,32 | 0,68 | 0,9 | 0,88 | 0,56 | 0,18 | 0,1 | 0,06 |
On average only 4.7
The amounts and localizations of heterochromatin bands on chromosomes of three
Distribution of
Species | 2n |
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|
Chromosome arm no. |
|
Source | ||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 18 | 19 | 20 | 21 | 22 | 23 | 24 | 25 | |||||||
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44 | 52 | M | A | + | + | + | + | + | + | + | + | + | + | + | 11 |
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44 | 52 | M |
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+ | + | + | + | + | + | + | + | + | + | + | + | + | 13 |
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44 | 52 | M | A | + | + | + | + | + | 5 |
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44 | 52 | M | A | + | + | + | + | + | + | 6 |
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44 | 52 |
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A | + | + | + | + | + | + | + | + | + | + | + | + | 12 |
this study | |||||
44 | 52 |
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+ | + | + | 3 |
1) The male
2) The female
3) In karyotype of the female
The distinct telomeric heterochromatic segments found on several chromosomes of both individuals from the Primorsky Krai were previously described only for the Chinese
All individuals studied had the heteromorphic chromosome pairs. The similar intraspecific heteromorphism of several heterochromatic segments was previously observed in a few Eurasian
The individuals differing in the amounts and localizations of heterochromatin bands on chromosomes are also belonged in different
Comparison of C-banded karyotypes of far eastern
Intraspecific variations of heterochromatic material in karyotypes of
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1/2 | 3/4 | 5/6 | 16/17 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 18 | 19 | 20 | 21 | 22 | 23 | 24 | 25 | ||||||
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44 | 50 | o | o | o | + | x | o | o | + | o | + | o | x | o | + | + | o | o | x | + | ++ | o | + | - |
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44 | 50 | tel | tel | +, tel | +, tel | ++, tel | +, |
xx, tel | xx | +, |
xx, tel | +, tel | xx | +, tel | +, |
+, tel | o | xx | +, tel | + | x | ++ | + | •, A |
|
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44 | 50 | tel | tel | tel | +, |
o | xx | ++ | + | +, tel | + | + | o | + | + | +, tel | + | +, tel | + | + | + | x | + | - |
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44 | 52 | + | + | + | +, int | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + | +, |
+ | •, |
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44 | 52 | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + | int , |
•, |
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44 | 52 | ++ | ++ | ++ | ++ | ++ | + | ++ | ++ | ++ | ++ | ++ | + | + | + | + | ++ | + | + | x | x | arm, |
++ | - |
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44 | 52 | xx | + | xx | + | + | ++ | ++ | + | ++ | + | + | + | ++ | ++ | + | + | + | ++ | + | + | arm, |
++ | •, A |
|
The
The chromosomal characteristics of
We would like to thank Elena V. Ignatenko and Sergei Yu. Ignatenko (Zeisky Nature Reserve), Vadim V. Bobrovsky and Polina Van (Komsomolsky Nature Reserve) for their help in mounting the expeditions. This study was partly funded by RFBR according to the research project № 18-34-00285.
The other collecting data for
species data