45 3 Vol.45, No.3 2014 5 OCEANOLOGIA ET LIMNOLOGIA SINICA May, 2014 * 1 1 2 1, 3 1 4 1 1 (1. 266071; 2. 264200; 3. 100049; 4. 266109) (Scophthalmus maximus),, 14.8 15.5 C, 15min 67.5MPa 6min, 72%, 70%, 73%, ; ; ; Q953 doi: 10.11693/hyhz20130400028 (Scophthalmus maximus) (Scophthalmidae) (Scophthalmus) (Nelson, 2006),, 1992,,,,, (, 2010),,,, (, 2003), 1 (P<0.05), 8%(Cal et al, 2006),,,,,,,, (, 1999) 20 80,,, (Salmo gairdneri) (Chourrout, 1982) (Ictalurus punctatus) (Bidwell et al, 1985) (Oreochromis niloticus O. mossambicus)(myers, 1986) (Anstichthys nobilis) (, 1990) (Carassius auratus transparent colored variety)(, 1991) (Dicentrarbus labrax)(peruzzi et al, 2003) (Perca flavescens) (Malison et al, 1993) (Cynoglossus semilaevis)(, 2012) (Paralichthys olivaceus)(, 2012),,, * 863, 2012AA10A402 ; ( ), 200903046 ;, 41276171,,, E-mail: wzhwzh0123@163.com :,,, E-mail: youfeng@qdio.cas.cn : 2013-04-25, : 2013-06-28
658 45, 1 1.1, 40 60cm, 35 50cm,, 14.8 15.5 C, 4 1.2, 5, 20min 17.5min 15min 12.5min 10min, 67.5MPa, 6min,, 1.3, 15min, 5, 62.5MPa 65MPa 67.5MPa 70MPa 72.5MPa, 6min,, 1.4, 15min, 67.5MPa, 5, 4min 5min 6min 7min 8min,, 1.5, 3 3,, 20min 15min 10min; 62.5MPa 67.5MPa 72.5MPa; 4min 6min 8min, 1.6 DNA : 100 200, 0.075mol/L KCl ( : =3..1) 15%Giemsa, 30, DNA : 10, 260 DAPI (PARTEC, CCA- ) DNA DNA 100, DNA 200 1.7 1000, 600mL, : (%) = / 100% (%) = / 100% (%) = / 100% (%) = / 100% / 100%, SPSS16.0, Duncan s 2 2.1 ( 1), 15.0 15.5 C 110h, (100h ) 2n=44(Bouza et al, 1994), 88, ( 2), 100, 200,
3 : 659 1 ( ) ( ) Fig.1 Embryo development of turbot in control and induction groups, 4 8, 1mm Fig.2 2 ( ) ( ) (, 1000 ) ( ) Chromosome (above, 1000 ) and histograms of ploidy analysis (below) of turbot in control (left) and induction groups (right) 2.2 3 12.5min (P<0.05),, 15min, 15min ; 67.5MPa, 67.5MPa ;, 6min, 6min 2.3 1,, (P<0.05), 15min ( 2), 3,
660 45 Tab.1 表 1 大 菱 鲆 四 倍 体 人 工 诱 导 正 交 实 验 结 果 Results of orthogonal experiment in turbot tetraploid induction F Sig. F Sig. F Sig. F Sig. 2.064 0.153 4.374 0.027 3.749 0.041 3.327 0.057 2.605 0.099 1.564 0.234 1.492 0.249 1.050 0.368 0.250 0.781 0.498 0.615 0.508 0.610 0.229 0.797 F F, Sig 表 2 大 菱 鲆 四 倍 体 人 工 诱 导 正 交 实 验 结 果 分 析 Tab.2 Analysis of orthogonal experiment in turbot tetraploid induction K1 0.111 0.239 0.131 0.288 0.288 0.311 K2 0.299 0.163 0.213 0.400 0.352 0.330 K3 0.084 0.091 0.150 0.297 0.344 0.344 0.215 0.148 0.082 0.112 0.064 0.033 3 (a) (b) (c) Fig.3 Effects of initiation time, treatment pressure and treatment duration on fertility rate, survival rate of gastrula stage, hatching rate and tetraploid rate, He (2012) ( ) 鲌 ( ), (colchicine) B(cytochalasin B, CB),,, (Chourrout, 1984),,,,,,,,, (90% 100%), (Xu et al, 2008),, (P<0.05);,,,
3 : 661,,,,,, ;,,,, (Sakao et al, 2006),,,, (2012), 68.3%; (2012), 8 15cm 13.3%, 70%,,, (Wang et al, 2008),,,,,, 2010. (Scophthalmus maximus)., 41(3): 301 306, 1999.. :, 148 155,,, 2003.., 5: 3 6,,, 2012. (Paralichthys olivaceus)., 43(2): 382 388,,, 2012., 19(2): 196 201,,, 1997. ( ) ( )., 21 (3): 197 283, 1990.., 36 (1): 70 75,,, 1991..., 15(4): 333 341 Aldridge F J, Marston R Q, Shireman J V, 1990. Induced triploids and tetraploids in bighead carp, Hypophthalmichthys nobilis, verified by multi-embryo cytofluorometric analysis. Aquaculture, 87(2): 121 131 Bidwell C A, Chrisman C L, Libey G, 1985. Polyploidy induced by heat shock in channel catfish. Aquaculture, 51: 25 32 Bouza C, Sánchez L, Martínez P, 1994. Karotypic characterization of turbot (Scophthalmus maximus) with conventional, fluorochrome and restriction endonuclease-banding techniques. Marine Biology, 120(4): 609 613 Cal R M, Vidal S, Gómez C, Álvarez-Blázquez B et al, 2006. Growth and gonadal development in diploid and triploid turbot (Scophthalmus maximus). Aquaculture, 251: 99 108 Chourrout D, 1982. Tetraploidy induced by heat shocks in the rainbow trout (Salmo gairdneri R.). Reproduction Nutrition Development, 22: 569 574 Chourrout D, 1984. Pressure-induced retention of second polar body and suppression of first cleavage in rainbow trout: Production of all-triploids, all-tetraploids, and heterozygous and homozygous diploid gynogenetics. Aquaculture, 36(1 2): 111 126 He W, Qin Q, Liu S et al, 2012. Organization and variation analysis of 5S rdna in different ploidy level hybrids of red cruciancarp topmouthculter. PLoS ONE, 7(6): e38976. doi: 10.1371/journal.pone.0038976 Malison J A, Kayes T B, Held J A et al, 1993. Manipulation of ploidy in yellow perch (Perca flavescens) by heat shock, hydrostatic pressure shock and spermatozoa inactivation. Aquaculture, 110: 229 242 Myers J M, 1986.Tetraploid induction in Oreochromis spp. Aquaculture, 57: 281 287 Nelson J S, 2006. Fishes of the world, 4th ed. New York. John Wiley and Sons Peruzzi S, Chatain B, 2003. Induction of tetraploid gynogenesis in the European sea bass (Dicentrarbus labrax L.). Genetica, 119: 225 228 Refstie T, 1981. Tetraploid rainbow trout produced by cytochalasin B. Aquaculture, 25: 51 58 Sakao S, Fujimoto T, Kimura S, 2006. Drastic mortality in tetraploid induction results from the elevation of ploidy in masu salmon Oncorhynchus masou. Aquaculture, 252(2 4): 147 160 Wang W, You F, Xu J H, 2008. Genetic analysis of meio- and mitogynogenetic stocks of Paralichthys olivaceus with microsatellite markers. ACTA Oceanologica Sinica, 27(2): 149 156
662 45 Xu J, You F, Wu X, 2008. Induction of triploidy in large yellow croacker Pseudosciaena crocea (Richardson, 1846): effects of pressure shocks and growth performance in the first rearing year. Aquaculture Research, 39: 1369 1376 INDUCTION OF TETRAPLOID IN TURBOT SCOPHTHALMUS MAXIMUS WU Zhi-Hao 1, YOU Feng 1, SONG Zong-Cheng 2, HU Jin-Wei 1, 3, WANG Li-Juan 1, ZHU Xiang-Ping 4, TAN Xun-Gang 1, LI Jun 1 (1. Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao266071, China; 2. Shenghang Aquatic Science and Technology Co. Ltd., Weihai 264200, China; 3. University of Chinese Academy of Sciences, Beijing 100049, China; 4. College of Animal Science and Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China) Abstract Turbot Scophthalmus maximus is an important cultured marine fish in North China. The induction of tetraploid turbot is important in germplasm enhancement for large-scale applications of triploid. However, there is almost no report on artificial induction of tetraploid turbot. In this study, the tetraploid turbot was induced by hydrostatic pressure shock to prevent embryo cleavage. The results show that the optimal initiation time for pressure shock was 15 min before the first cleavage at 14.8 15.5 C. The optimal treatment pressure and treatment duration were 67.5MPa and 6 min. The hatching rate and tetraploid rate could reach 72% and 70%, respectively. This paper provides basic data for production of fertile tetraploid adults in turbot. Key words Turbot Scophthalmus maximus; tetraploid; artificial induction; hydrostatic pressure shock