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中国对虾及中华绒螯蟹多倍体诱导中6-DMAP在卵子中残留量的检测
吴长功, 张晓军, 崔朝霞, 周令华, 相建海
中国科学院海洋研究所
摘要:
海洋动物多倍体的诱导目前已成为海水养殖中品种改良的重要手段(楼允东,1984)。在美国,太平洋牡蛎和虹鳟的多倍体诱导已大规模应用于生产,并获得良好的经济和社会效益。我国也先后开展了鱼类(桂建芳等,1991;尤锋,1993)、中国对虾(戴继勋等,1993;相建海等,1992)、贝类(喻子牛等,1995)及中华绒螯蟹(陈立侨等,1997)等多种海洋动物多倍体诱导的研究,并取得了一些进展。6-二甲基氨基嘌呤(6-DMAP)由于其低毒、经济和高效,目前已成为海洋动物多倍体诱导中经常使用的诱导剂,但至今,国内外尚无关于6-DMAP在卵子内的残留的报道。在环境保护和生物安全倍受关注的今天,人们非常关心多倍体诱导食物的安全性问题,本文首次利用高压液相色谱技术检测了在中国对虾和中华绒螯蟹多倍体诱导处理后,卵子和幼体内6-DMAP的残留量,并对检测结果进行了分析讨论。
关键词:  多倍体诱导  残留  6-二甲基氨基嘌呤
DOI:
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基金项目:中国科学院海洋研究所调查研究报告第4258号;国家973项目资助1999012009号。
DETECTION OF 6-DMAP REMAINING IN TREATED EGGS OF PENAEUS CHINENSIS AND ERIOCHEIR SINENSIS IN POLYPLOID INDUCTION
Wu Changgong, Zhang Xiaojun, Cui Zhaoxia, Zhou Linghua, Xiang Jianhai
Institute of Oceanology, Chinese Academy of Sciences
Abstract:
The remained 6-DMAP in treated eggs during polyploid induction by 6-DMAP in Penaeus chinensis and Eriocheir sinensis was detected using high performance chromatography (HPLC). 100% triploid embryos were induced by inhibiting the second polar body using 300 μmol/L 6-DMAP in fertilized eggs in P.chinensis. Tetraploid of Eriocheir sinensis could be induced using 300μmol/L 6-DMAP inhibiting the first cleavage, and the highest ratio of tetraploid checked in blastosphere stage was 58%. In HPLC detection, Waters-NOVA PAK C18 (5mm× 15mm) was used, and methanol:water (1:1) was used as dilution. 6-DMAP can be detected at 254nm, the minimum of detected 6-DMAP was 0.003μg/ml. The treated eggs rinsed thrice in seawater, the remained 6-DMAP cannot be detected in HPLC at the given condition. During the hatching period, the remained 6-DMAP was diluted and rinsed again and again. 6-DMAP can be degraded in environment. The results indicated that using 6-DMAP to induce polyploid in the eggs of Penaeus chinensis and Eriocheir sinensis is effective and safe.
Key words:  polyploid induction  6-DMAP
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