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机构地区:[1]中国海洋大学教育部海水养殖重点实验室,山东青岛266003
出 处:《水产学报》2004年第2期133-138,共6页Journal of Fisheries of China
基 金:国家自然科学基金资助(30170735);教育部留学回国人员科研启动基金
摘 要:研究了利用紫外线诱导栉孔扇贝雄核发育的条件。表明紫外线(254nm)对卵子染色质失活是有效的。在强度为2.8mW·cm-2·s 1的紫外线下照射20s的卵子与正常精子混合后能保持一定的受精率(60.2%),且D形幼虫发生率为0。染色体检查结果显示此时单倍体率最高(49.2%)。说明在强度为2.8mW·cm-2·s-1的紫外线下照射20s是获得雄核发育单倍体的适宜条件。研究发现受精率和D形幼虫发生率随照射时间的增加而下降,遗传失活的卵子与正常精子受精后其胚胎发育至D形幼虫前期停止。实验中各处理组均出现非整倍体。出现的原因可能由于紫外线照射剂量对卵子染色体遗传失活的作用程度不同以及DNA的光修复。Androgenesis is defined as all-paternal inheritance. Viable androgenetic diploids can be generated by inhibiting the first cleavage to double paternally derived chromosomes,after fertilization of genetically inactivated eggs with normal sperm. It is a technique that could facilitate the production of completely homozygous isogenic lines, examine sex determination, make genetic analysis, and protect endangered species. Gamma and X-rays inactivation are the usual methods used to inactivate egg nuclei. However, since special facilities are required to manage radioactivity safety, these methods are not practical for routine induction of androgenesis. In many fish species, efficient procedures of genetically inactivating eggs and restoring diploidy have been studied and achievements of successful diploid androgenesis were reported. In contrast, in mollusks, there were only a few reports of cytobiological studies on one species of spontaneous androgenetic Corbicula leana, artificial induction of androgenesis has rarely been studied.Chlamys farreri is a main cultured mollusk species in China, as well as an important object of study in mollusk breeding science. In this study, optimum conditions of ultraviolet (UV) ray irradiation for genetic inactivation of eggs were examined to develop simple and safe techniques to induce haploid androgenesis in C. farreri. Mature cultured C. farreri were collected locally in late March and early May from the coast of Weihai, Shandong. Eggs and sperm were obtained by artificially inducing spawning with the stimulations of dryness and raising water temperature from 16 to 20°C. Discharged eggs were collected by suction and rinsed in filtered seawater several times. Suspensions of sperm and egg were prepared at concentrations of 1.0×10~6 sperm per mL and 1.0×10~4 egg per mL by dilution with filtered seawater. Egg suspension (4 mL) was spread on a plastic Petri dish and treated with the UV rays (254 nm) at intensity of 2.8 mW·cm^(-2)·s^(-1) for 0, 5, 10, 15, 20, 25, 30, 35, 40, 45,
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