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Karyotype and Idiogram of the Axis Deer (Axia Axis, Cervidae) by Conventional Staining, GTG-, High-Resolution GTG-, and Ag-NOR-Banding Techniques

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Karyotype and Idiogram of the Axis Deer (Axia Axis, Cervidae) by Conventional Staining, GTG-, High-Resolution GTG-, and Ag-NOR-Banding Techniques © 2017 The Japan Mendel Society Cytologia 82(1) Special Issue: 91–98 Karyotype and Idiogram of the Axis Deer (Axia axis, Cervidae) by Conventional Staining, GTG-, High-Resolution GTG-, and Ag-NOR-Banding Techniques Hathaipat Khongcharoensuk1, Alongklod Tanomtong1*, Isara Patawang2, Praween Supanuam3, Somnuek Sornnok4 and Krit Pinthong5 1 Toxic Substances in Livestock and Aquatic Animals Research Group, Department of Biology, Faculty of Science, Khon Kaen University, Muang, Khon Kaen 40002, Thailand 2 Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand 3 Program in Biology, Faculty of Science, Ubon Ratchathani Rajabhat University, Ubon Ratchathani 34000, Thailand 4 Agricultural Technology Division, Department Technology and Industries, Faculty of Science and Technology, Prince of Songkla University (Pattani Campus), Muang, Pattani 94000, Thailand 5 Program in Biology, Department of Fundamental Science, Faculty of Science and Technology, Surindra Rajabhat University, Surin 32000, Thailand Received December 22, 2014; accepted April 8, 2015 Summary The standardized karyotype and idiogram of the axis deer (Axis axis, Cervidae) at Khon Kaen Zoo, Thailand were established. Blood samples were taken from two male and two female axis deer. After standard whole blood T-lymphocytes were cultured at 37°C for 72 h in the presence of colchicine, metaphase spreads were performed on microscopic slides and air-dried. Conventional staining, GTG-, Ag-NOR-banding and high-resolu- tion techniques were applied to stain the chromosome. The results showed that the diploid chromosome number of A. axis was 2n=66 and the fundamental number (NF) was 70 in both male and female. The types of autosomes observed were 2 large metacentric, 2 large submetacentric, 2 large telocentric, 6 medium telocentric and 52 small telocentric chromosomes. The X chromosome was a large telocentric chromosome, and the Y chromosome was a small telocentric chromosome. The GTG-banding and high-resolution techniques provided that the respective numbers of bands and locations of A. axis were 246 and 294, and each chromosome pair could be clearly dif- ferentiated. In addition, the subtelomeric q-arm of chromosome pair 2 and the telomeric q-arm of chromosome pair 4 showed clearly observable nucleolar organizer regions (NORs) and secondary constrictions. Our results are the first reports of GTG-, high-resolution GTG- and Ag-NOR-banding techniques on this species. The karyotype formula of A. axis is as follows: m sm t t t 2n (66) = L2 + L 2 + L 2 + M 6 + S 52 + Sex-chromosomes Key words Axis deer, Axis axis, Karyotype, GTG-banding, Ag-NOR-banding. The family Cervidae (infraorder Pecora, suborder Ru- which make them an ideal group for chromosomal evo- minantia, order Artiodactyla) has at least 90 species, 20 lution studies. It is proposed that the ancestral cervid genera, four tribes (Muntiacini, Cervini, Capreolini and karyotype is 2n=70 based on comparative karyotype Rangiferini) and two subfamilies (Cervinae and Capreo- analysis of many deer species (Neitzel 1987, Tanomtong linae) (Janis and Scott 1987, Nowak 1999, Groves 2006, et al. 2005, 2010, Huang et al. 2005). Wilson and Reeder 2006). The classification of deer in The axis deer (Axis axis), also known as chital deer or the family Cervidae use external morphology, biogeog- spotted deer, is a deer which commonly inhabits wooded raphy, physiology, cytogenetics and especially decidu- regions of South Asia including India, Sri Lanka, Ne- ous cranial appendages. According to the presence or pal, Bangladesh, Bhutan and Pakistan. The axis deer absence of antlers, all deer species were classified into skin is pinkish fawn, marked with white spots, and its antlered and antlerless deer. The extant deer species underparts are also white (Fig. 1). Its antlers, which it have diverse karyotypes; their diploid chromosome sheds annually, are usually three-pronged and curve in numbers range from 2n=6 in the female Indian muntjac a lyre shape and may extend to 75 cm. Compared to the (Muntiacus muntjak vaginalis) (Wurster and Benirschke hog deer (Hyelaphus porcinus), its close relative, the 1970) to 2n=80 in the Capreolus capreolus pygargus, axis deer has a more cursorial build. It also has a more advanced morphology with antler pedicles being pro- * Corresponding author, e-mail: [email protected] portionally short and its auditory bullae being smaller. DOI: 10.1508/cytologia.82.91 It also has large nares. Their lifespans are around eight 92 H. Khongcharoensuk et al. Cytologia 82(1) Special Issue Fig. 1. General characteristics of male (A) and female (B) axis deer (Axis axis). Fig. 2. Metaphase chromosome plate and karyotype of male (A) and female (B) axis deer (Axis axis) 2n=66 by conventional staining technique; scales indicate 10 µm. to 14 years. Within the axis deer were recognized comparison finding with previous reports. Moreover, the two subspecies: common axis deer (A. a. axis) and Sri finding obtained here is the first report on standardized Lankan axis deer (A. a. ceylonensis). These animals are karyotype and idiogram measurements by GTG-, high- widespread in many countries. This species is listed on resolution GTG- and Ag-NOR-banding techniques. This CITES appendix III and is an IUCN least concern spe- cytogenetic information will represent basic knowledge cies (Grzimek 2004, Groves 2006). and can be applicable to genetic diversity, taxonomy, To date, there are few cytogenetic reports of the Axis conservation and evolution. axis, such as those of Hsu and Benirschke (1974), Asher et al. (1999), Bonnet-Garnier et al. (2003) and Shanthi Materials and methods et al. (2008), which report karyotypes of the convention- al, RBG-, QFH-banding and chromosome painting of Blood samples of the Axis axis (two males and two this animal. Our present study shows a confirmation and females) were collected from Khon Kaen Zoo, Thailand 2017 Karyotype and Idiogram of the Axis Deer (Axia axis, Cervidae) by Conventional Staining 93 Table 1. Mean length of short arm chromosome (Ls), long arm chromosome (Ll), total arm chromosome (LT), relative length (RL), centromeric index (CI) and standard deviation (SD) of RL, CI from 10 metaphases of male axis deer (Axis axis) in Thailand, 2n=66. Chromosome pair Ls Ll LT RL±SD CI±SD Size Types 1 2.519 5.749 8.269 0.064±0.0059 0.438±0.017 Large Submetacentric 2* 0.000 7.556 8.156 0.058±0.0089 0.000±0.000 Large Telocentric 3 3.303 4.707 8.010 0.062±0.0076 0.702±0.007 Large Metacentric 4* 0.000 4.779 4.779 0.037±0.0047 0.000±0.000 Medium Telocentric 5 0.000 4.773 4.773 0.037±0.0033 0.000±0.000 Medium Telocentric 6 0.000 4.655 4.655 0.036±0.0027 0.000±0.000 Medium Telocentric 7 0.000 4.110 4.110 0.032±0.0028 0.000±0.000 Small Telocentric 8 0.000 3.904 3.904 0.030±0.0026 0.000±0.000 Small Telocentric 9 0.000 3.884 3.884 0.030±0.0019 0.000±0.000 Small Telocentric 10 0.000 3.804 3.804 0.029±0.0021 0.000±0.000 Small Telocentric 11 0.000 3.748 3.748 0.029±0.0019 0.000±0.000 Small Telocentric 12 0.000 3.664 3.664 0.028±0.0021 0.000±0.000 Small Telocentric 13 0.000 3.562 3.562 0.028±0.0018 0.000±0.000 Small Telocentric 14 0.000 3.524 3.524 0.027±0.0012 0.000±0.000 Small Telocentric 15 0.000 3.440 3.440 0.027±0.0012 0.000±0.000 Small Telocentric 16 0.000 3.355 3.355 0.026±0.0013 0.000±0.000 Small Telocentric 17 0.000 3.278 3.278 0.025±0.0013 0.000±0.000 Small Telocentric 18 0.000 3.204 3.204 0.025±0.0013 0.000±0.000 Small Telocentric 19 0.000 3.157 3.157 0.024±0.0012 0.000±0.000 Small Telocentric 20 0.000 3.087 3.087 0.024±0.0012 0.000±0.000 Small Telocentric 21 0.000 3.024 3.024 0.023±0.0012 0.000±0.000 Small Telocentric 22 0.000 2.988 2.988 0.023±0.0013 0.000±0.000 Small Telocentric 23 0.000 2.941 2.941 0.023±0.0012 0.000±0.000 Small Telocentric 24 0.000 2.891 2.891 0.022±0.0013 0.000±0.000 Small Telocentric 25 0.000 2.866 2.866 0.022±0.0012 0.000±0.000 Small Telocentric 26 0.000 2.810 2.810 0.022±0.0012 0.000±0.000 Small Telocentric 27 0.000 2.804 2.804 0.022±0.0014 0.000±0.000 Small Telocentric 28 0.000 2.751 2.751 0.021±0.0014 0.000±0.000 Small Telocentric 29 0.000 2.624 2.624 0.020±0.0011 0.000±0.000 Small Telocentric 30 0.000 2.545 2.545 0.020±0.0011 0.000±0.000 Small Telocentric 31 0.000 2.441 2.441 0.019±0.0013 0.000±0.000 Small Telocentric 32 0.000 2.285 2.285 0.018±0.0014 0.000±0.000 Small Telocentric X 0.000 6.393 6.393 0.049±0.0043 0.000±0.000 Small Telocentric Y 0.000 2.345 2.345 0.018±0.0017 0.000±0.000 Small Telocentric Remark: *=Nucleolar organizer region/NOR. Table 2. Mean length of short arm chromosome (Ls), long arm chromosome (Ll), total arm chromosome (LT), relative length (RL), centromeric index (CI) and standard deviation (SD) of RL, CI from 10 metaphases of female axis deer (Axis axis) in Thailand, 2n=66.
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