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Nucleolar Organizer Regions Polymorphism and Karyological Analysis of Black Lancer, Bagrichthys Majusculus (Siluriformes, Bagridae) in Thailand

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Nucleolar Organizer Regions Polymorphism and Karyological Analysis of Black Lancer, Bagrichthys Majusculus (Siluriformes, Bagridae) in Thailand O2018 Thc Japon Mcndcl Socicly C)tolosia A}Q)t 193 199 Nucleolar Organizer Regions Polymorphism and Karyological Analysis of Black Lancer, Bagrichthys majusculus (Siluriformes, Bagridae) in Thailand Weerayuth Supiwong 1*, Nuntaporn Getlakha2, Somkid Chaiphech3, Krit Pinthong4, Sumalee Phimphan5 and Alongklod Tanomtong 5 rF$ulty ofApplicd scicuc. and Enginecring, Khon Kicrr tJnive6ity, Nons Khai(nmpus, Muans, Nong Khai.13000, Thailand rD€partment ofBiology, faculty ofS(ience and Technology, Mubar Chombueng Rajabhat Uriversity. ChombuenB. Ratchaburi 70150.'l'hniland rDepartmonr ofAnimal Sui(nce. Faculw ofAgricultue. RajamanEala Univelsiry ol Tccho.r,osy Srivijaya, Nrkom Sri Tharnmdral, Saiyai Cumpus. Thtlnlsong, Naklon Si Thm naral 80110, Thliland rDepa(me,rt ofl-und.mental Scienoc, Faculty ofSuience and'ltchnok,8y, Surntdra Raj$hat tJnivErsify. Muang, Surin 32000, Thiiland 'Toxic S ub\lances in Liveslock and Aq ualic An imnls Rescarch Group. D(panment o l Biology. Foculty ofSciencd, Khon Kacn tlnivcrsity, Muang, Knon Kacn 40002, Thailand Receiwd De.entber 21, 2017; arcetued Febtuar.t 20, 2Ul8 Summary Nuclcolar organizcr regions (NORS) and karyotypc in thc blaok lanu(J, Bagrichlhts tnaj $culus fiom Nakhon Phranom and Sing Buri provinccs. Thailand, wcrc invcstigatcd. Thc nlitotic chromosomc prcpara- tions wcre directly obtained from kidney tissues of eight male and eight fcmale spccinrens. Conventional and Ag-NOR staining tcchniques wcrc applied to the chromosomcs. The chromosomes numbcr of B. ntojlsc lus \Nas 2n:50 and the fundnnrcntal numbel (NF) ofall specimens was 96. The klryotypc consists ofsix large metacen- tric, six large submctaccntric, two largc acrocentric, eight medium mctaccntric. l2 mcdium submetacentic. two Nedium acrocentric. six sm0ll nretacentric, four sruall subllretaceDtric und four small teloceotric chtromosoDes. Scx chromosome was not obscrvcd. Morcovcr. thc intcrstitial NORs wclc clearly obscrved at the long arn1 ofthc metaccntric chromosome pair l. Th('rcsult levcaled that a hcleromorphic NOR typc was appearcd in onc malc and onc ttmale fish and other fish have homozygous NOR in the pair I chromosomcs. Key rlords Ragrichth.v-s majusdrh.r, Karyotype, NOR variation. Polymorphisrn, C-hromosomc nrrnber Freshwater fishes are especially important as thcy of species and interrelationships within fish families providc a sourcc of high quality protcin as wcll as food (Campiranont 2003, Taoomlong 20ll). Thc studics of source lbr peoplc who livc near a river basin. Fishes of the karyotypes help to invesligate thc aquatic structure the Bagridae tbmily belong to the order Siluriformes and of the species population in each habitat, so it can de- are of great importance as fbod lish. While. sorte spe- termine what species are related to each other in an ac- cies are kept as aquarium 6slres, nrany bagrids have re- curate manncr. This may hetp to facilitate the hybridiza- cently bccomc a boorning iu aquaculturc (Nclson 2006). tion bctv,,ccn them in thc f'uturc tbr strain improvcrnclt Therc arc six gcncra and approximatcly 25 spccics in thc (Sofy e/ a/. 2008), brccding practiccs of organisms by Bagridae family in Thailand (Rainboth 1996, Vidthaya- using chromosome set managcm€nt such as polyploidy non 2005, Ferraris 2007). B. maju.tculus is onc spccics inducing in catfishcs, Clarias bdtrdchu,t (Na-Nakorn ol thc lumily considcring as thc Tltai ecunolniu spccics et ol. 1980\ and rainbow tto\l loncorh.vnchu-t ,[,kis.t), rused for foods and in an aquarium (Vidthayanon 2005). brood stock selection of giant catfish (MengamPan el 41. Thc karyotypc is thc chromosourc con'tplcnrcnt of an 2004). Cytogcnctic knowledgc can also providc increas- individual or related group of intlividuals, as defined by ing useful data for future studies on chromosome evolu- chromosome sizc, morphology and number. Though for tion. The rcports of chromosome evolution in thc family all somatic cclls ofall individuats ofspccics, thc numbcr Channidac (snackhcnd 6sh) indicatcd that ccntric f'usion, of chromosonrcs is used as an indicalor of classification pcriccntric inversion and polyploidy ars the processes which have important roles on chromosome rearrange- rnent in the snackhead fish's phylogeny (Supiwong el a/. * Corrcsponding uulhor, e-mai! : supiwong(rghot,nail.eom DOI: 10.15o8/cylologia.83. 193 2009, Tanomtong e/ d/. 2014). I9.l Cytologia 83(2) arc oftcn ttscd Chromosqmc number and karyotypc A N as cssential data for the taxonomy of those animals with troubles on morphological classitication aspect (Giessmann 2002). tn fishes, chromosome banding data was used to support for classificatio[ in lhc CyPrinidac (Amemiya and Gold 1988, Gold and Li 1994, Boron 2001, Bcllafrontc e/ d/. 2005). Up to the present. 46 species of the Bagridae have bccn cytogcnctically studicd. The chromosome numbcr ranging from 2n=44 in Coreobogrus brevicorpus (Kint et ol. 1982) to 2rr=80 in Butusio havnolleri (Magtoon and Donsakul 2009) have been reported. However. the predominant diploid numbers represent 2r:52 and 2r=56 (Arai 20ll ). In the present study, chromosolne of thc B. najus.ulus ftorn Thailand was firstly investigatcd using conventional staining and Ag-NOR banding tcch- niques. Matcrials an(l mcthods The specimcns of B. najusutlus wcrc collected from 0 the Mekong rivcr, Nakhon Phranom province, northeasl = Sam pling Sites of Thailand (fivc males and five fcmalcs) and thc Chao I Phraya Rivcr, Sing Buri province, ccntral part o[ Thai- land (three malcs and $ree f'emales). Thc sampling sitr.'s o B and the characteristics of B. mojrccuhr.r are shown in l'... Fig. l. The fishes were transt'erred to laboratory aquaria and were kept under standard conditions for seven days prior to the cxpcriments. Proccdurc of chromosornc prcparation was following modified protocol of Supi- wong et al. (2009). The 0.05% colchicine (l mL to l00g 0 body weight) was injected to abdominal cavity and lcft for 45 min to I h. Chromosomes were prcpared from kid- ney cells by squash technique. Kidney tissues were cut Fig. l. Thailar mnp showirg lhe s.rnpling siles (A) rnd the gen- into small picccs, then mixcd with hypotonic solution cral charactoristic of8- ,r(rrlrrcllrr.r (B). (0.075 M KCI) and incubatect for 30rnin. Cells were fixed in fresh cool fixative (3 absolute methanol: I glacial ace- et aL1989, Lakra and Rishi 1991, Khuda-Bukhsh el o/. tic acid). The propared chromosomcs werq staincd with 1995, Donsakul 2001, Das and Khuda-BLrkhsh 2007a, I0% Giemsa tbr 30min, or 50% AgNOi and.2Yo gcla- Supiwong et a1.2013a,2014a, b). The NF of B. majrrs- tin lbr Ag-NOR staining technique (Howell and Black culus was 96 in both males and t'emales. Although our 1980). The metaphase figures were analyzed according result presented the same chromosome number as in to the chromosome classification following by Turpin previous studies, the NF is different (Donsakul 2002, and Lcjcune (l9651and Chaiyasut (1989). Magtoon and Arai 1988). Donsakul (2002) rcported that B. majusculus showcs NF=92 whercas Magtoon and Results and rliscnssion Arai ( 1988) reporled NF=94. These differences may be duc to different critcria uscd tbr the chromosome clas- Chromosome number, fundantetttal nunber and kar.yo- sificalion and/or inter-populalional variation in this spc- ,,pe o/8. majusculus cies. The NF in the family Bagridae varies tiom 72 to Cytogenctic studics havc bccn pclfornrcd on sixtccn 128 (Arai 201 l). Comparisons among the gcncra in thc specimens of B. naiusculus from Thailand. The results Bagridae revealed that the Hemibagtus showes higher rcvcaled thal thc diploid chromosontc numbcr was NF variation than others. lt varies from 72 to I14. Ghi- 2n=50, in both males and fcmalcs (Figs. 2, 3). This is gliotti et a/. (2007) assumcd that spccics with a largcr accordancc with the prcvious studics of Magtoon and NF are more advanced in evolutionary terms. Such Arai (1988) and Donsakul (2002). However. the 2a is changes in NF appear lo be related to the occurrence of diflerent from other species of the Bagridae (Barat and pericentric inversions, which are among the most com- Khurla-Bukhsh 1986, Sharma and Tripathi 1986. Yu mon nrutlificalions cuntributing to karyotypic rearrangc- 2018 NOR Polymorphism.nd Krryologicrl Anitlysis uf Bugtk hthr! ttfur{ulux in Thailand 195 I{.Ir' rr rr rr rr r. ll lI rt rr rr rt .r .r ml r 2 ! a 5 6 't7 t lr3as51a tt0 910 la II TT rl ll aa la tI II r,l t, lll4 r7 lt "t 1l l2 t3 td l5 t6 l? t8 lt t9 l0 2t t9 70 2l r lrr al 23 77 2-1 24 25 24 15 B FiB. 2. Karyotypes of male (A) and fcmale (B) of B. ,nai$culus (2n=50) by convcntional straining tcchnique. Arrows indicalc seoondary conslriction. Scale bars=5tlm- Tsble l. Mean length (,!m) ofshort chromosome arm (Ls), mean length oflong chrumosornc ann (Ll). total lcngth olchromosome anns (LT). rclarivc lcnglh (RL). ccnlromcric ilxlcx (Cl) and slandard dcvi.tion (S.D.) of RL and Cl from mctaphlsc chromosomcs of 20 cclls ;n B. nuj6crhts (2n=so). Chromosomc pail Ls LI L'I RLIS D CllS.l) Type Sizc 47.59 55.45 103.04 0.029410.0022 0.519+0.027 L 2 19.39 5r.05 90.44 0.0157r0.002.1 0.566i0.0-'t2 L 3 34.80 44.59 79.18 0.022510.0019 0.560i0.030 L 4 31.0-'i 19.65 7t.68 0.010510.0009 0.553r0.010 M 5 30.14 67.41 0.0112i0.0011 0.554t0.02t) M 6 29.4t 34.85 64.26 0.0183 i0.000(, 0.544r0.028 M 1 27 _96 33.21 6!.t7 0.0174i0.000it 0.542L0.1)26 M I 24.90 3t.37 56.28 0.0161 10.0012 0.559r0.029 m s 9 23.15 29.10 52.25 0.0150r0.0012 0.557+0.024 m s t0 20.64 27.X3 4ti.46 0.0138r0.0011 0.57 5 !,0.022 s ll 36.98 76.48 I13.46 0.012110.0017 0.673r0.058 L t2 13.98 62.14 96.12
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