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转“全鱼”生长激素基因黄河鲤生物学特征的初步研究
Alternative TitlePrimary studies on the biology of ‘all-fish’ growth hormone transgenic common carp (Cyprinus carpio L.)
李德亮
Subtype博士
2008-06-16
Degree Grantor中国科学院水生生物研究所
Place of Conferral水生生物研究所
Keyword鲤鱼 转基因鱼 生态风险 权衡 生长速率 存活率 性腺发育 游泳能力
Abstract世界首例转基因鱼的诞生开辟了鱼类遗传育种的新纪元。转基因鱼由于具有生长速度快、饵料转化效率高、抗病和抗逆等优良养殖性状而显示出强大的应用潜力。转基因鱼被认为将在转基因动物中率先实现商业化。然而,由于政府和民众对转基因鱼潜在的生态风险的关注,致使迄今仍然无任何一例具有优良养殖性状的转基因鱼成功的实现商业化。因此,全面衡量和评估转基因鱼的生态风险是其商业化进程中的重要一环。 本研究围绕转基因鱼的生态风险这一重要科学问题,有针对性地对影响转“全鱼”生长激素基因黄河鲤种群适合度的重要生物学特征进行了初步的研究。与此同时,本研究也以快速生长转基因黄河鲤为模型,探讨鱼类生长速率与其存活率、繁殖特征以及游泳能力之间的权衡关系。主要研究结果如下: 1. 建立了快速生长转“全鱼”生长激素基因黄河鲤家系。通过测交的方式证实,转植基因在该家系 F2、 F3、 F4 和 F5 代中的分离比均为 50%。F2 代转基因鱼的平均体重是对照鱼的 1.8-2.5 倍,体重的特定生长率比对照鱼高出 10-20%。结果表明:“全鱼”生长激素转植基因有效整合在转基因黄河鲤基因组中,并能够在所建立的转基因鱼家系的各个世代间以孟德尔方式稳定遗传,且表现出明显的促生长效应。 2. 在 22.4 ± 0.09℃ 的水温环境下,转基因黄河鲤仔胚的孵化速率比对照仔胚快大约 2 个小时。与同一尾对照雌鱼杂交,转基因雄鱼与对照雄鱼后代的孵化率之间没有显著差异。内源性营养阶段的转基因仔鱼标准长、标准长特定生长率与对照鱼相似。外源摄食阶段,转基因仔鱼的体重、体重特定生长率、标准长、标准长特定生长率均显著的大于对照鱼。在人工养殖模式下,转基因黄河鲤在其仔、幼鱼阶段的死亡率显著的高于对照鱼。该结果进一步证实硬骨鱼类的快速生长与死亡率之间存在权衡关系。 3. 转基因鱼的体重和标准长分别是同龄对照鱼的 1.3-2.2 倍和 1.1-1.4 倍。实验期间,转基因雌鱼的性腺指数均显著的低于对照鱼,转基因雄鱼在不进行繁殖活动时的性腺指数也均显著的低于对照鱼,但在自然繁殖后的前两个月内,则与对照鱼没有显著差异。转基因鱼性腺的绝对重量显著的低于或等于对照鱼。转基因鱼性腺发育的速率慢于对照鱼。结果表明,具有快速生长效应的转基因黄河鲤首次性成熟年龄推迟,其快速生长效应以降低繁殖投资 (性腺指数) 为代价。 4. 转基因黄河鲤的绝对临界游泳速度和相对临界游泳速度的平均值分别比对照鱼低 22% 和 24%。这暗示着具有快速生长效应的转基因黄河鲤表现出低劣的游泳能力,同时也证实了鱼类生长速率和游泳能力之间存在权衡,即同一物种的个体生长率越高,临界游泳速度则越低。 5. 转基因黄河鲤与对照鲤的体型之间存在显著的差异。较对照鱼而言,转基因鱼具有较大的头部深度、较大的尾柄长度,但其体高和尾柄深度却较小。形态特征对实验鱼的临界游泳速度具有显著的影响。临界游泳速度比较高的鱼类通常具有较大的体高和尾柄深度,较小的尾柄长度。这些结果说明,形态特征的变化或许是造成转基因鱼临界游泳速度较低的因素之一。 6. 转基因黄河鲤的红细胞长度、宽度、表面积和体积均显著的小于对照鱼,其红细胞表面积与体积比显著的高于对照鱼,但其红细胞长度与宽度比与对照鱼之间没有显著差异。转基因鱼的日常代谢率显著的高于对照鱼,最大代谢率与对照鱼相似。正常状态下,转基因鱼的血细胞比容、血红蛋白浓度、血浆葡萄糖浓度、血浆乳酸浓度、血浆胆固醇浓度、血浆蛋白浓度均与对照鱼没有显著差异,但其平均血细胞血红蛋白浓度和血浆甘油三酯浓度则分别显著的低于和高于对照鱼。疲劳运动对两种鱼的血细胞比容、血浆蛋白浓度以及平均红细胞血红蛋白浓度没有造成显著的影响。疲劳运动后转基因鱼和对照鱼的血浆葡萄糖浓度和乳酸浓度均显著的增加,并分别经过 4h 恢复到了正常状态,表现出典型的次级应激反应。转基因鱼与对照鱼的血浆葡萄糖浓度应对疲劳运动的方式相似,但转基因鱼的血浆乳酸浓度应对疲劳运动刺激的反应强度较弱。疲劳运动后,转基因鱼的血浆胆固醇浓度和甘油三酯浓度均显著的升高,2h 后恢复到正常状态,而对照鱼的上述两种指标没有表现出明显的应答。转基因鱼血浆蛋白浓度对疲劳运动没有明显应答,但是对照鱼的血浆蛋白浓度从运动后 2h 后开始显著的低于对照鱼。结果表明:转基因鱼较对照鱼具有较小的红细胞形态、较高的日常代谢率、相似的最大活动代谢率和血液载氧能力,但是其血液生理指标 (血浆乳酸、胆固醇、甘油三酯和蛋白) 在应对疲劳运动的方式上较对照鱼发生了明显的变化。
Other AbstractThe production of first batch transgenic fish marks the advent of new era for fish genetics and breeding. Transgenic fish show great advantages for application to aquaculture due to their improvement on growth rate, feed conversion efficiency, resistance to disease and extreme condition. Fish may be considered the best candidate for the first marketable transgenic animal for human consumption. However, till date, no transgenic fish have been released into natural water body for commercialized cultivation as food, some of the reasons for this are the possible ecological risk posed by transgenic fish. Hence, carefully evaluation of the ecological risks posed by transgenic fish is one of the important aspects for achieving their commercialization. In order to evaluate their possible ecological risks, the present work primarily studied the biological traits of ‘all-fish’ growth hormone transgenic common carp (Cyprinus carpio L.) which could had great effects on their population fitness. At the same time, we used the ‘all-fish’ growth hormone transgenic common carp as a model system to explore the trade-offs between growth rate and survival, reproductive characteristics and swimming performance. The main results are presented as follows: 1. A line of fast-growing ‘all-fish’ growth hormone transgenic common carp was produced. Separating rates of the transgene in F2, F3, F4 and F5 were all 50%, examined by test cross. F2 transgenic fish had a mean body weight 1.8-2.5-fold heavier, and a mean specific growth rate (SGR) value 10%-20% higher than the controls. The results showed that ‘all-fish’ growth hormone transgene has effectively integrated, transmitted stably in Mendelian ratios in the homozygous line of transgenic common carp and could prompt the growth rate of common carp. 2. At 22.4 ± 0.09 °C, the average hatch time were significantly advanced about 2 h for transgenic carp compared to their nontransgenic siblings. There was not any significantly differences in hatch rate between the offsprings of transgenic males and control males hybrided with an control female. There were not any significantly differences in standard length and specific growth rate of standard length between transgenic fry and nontransgenic siblings at endogenous nutrition stage. At exogenous nutrition stage, transgenic fry had significantly higher body weight, specific growth rate of body weight, standard length and specific growth rate of standard length than nontransgenic siblings. In condition of artificial cultivation,transgenic fry and juvenile carp had significantly higher mortality than nontransgenic siblings. The result supported that there was a trade-off between growth rates and survival, i.e. faster-growing individuals had lower survival. 3. The mean body weight and standard length of transgenic carp were 1.3-2.2 times and 1.1-1.4 times than that of their nontransgenic siblings respectively. During the sampling period, transgenic females had significantly lower gonad index than nontransgenic sibling females. The gonad index of transgenic males was significantly lower than that of nontransgenic sibling males when spawn did not occur, but the differences were not significantly any more at the first two monthes after spawn. Transgenic fish had significantly lower or similar absolute gonad weight than their siblings. Compared to nontransgenic siblings, the gonad development of transgenic fish was significantly delayed. The present results showed the ‘all-fish’ growth hormone transgenic carp mature later, and their rapid growth was at the cost of reduced reproductive investment. 4. Transgenic carp had a mean absolute Ucrit (cm/s) value 22% or mean relative Ucrit (BL/s) value 24% lower than the controls. It suggested that fast-growing ‘all-fish’ GH-transgenic carp were inferior swimmers. It is also supported that there was a trade-off between growth rates and swimming performance, i.e. faster-growing individuals had lower critical swimming speed. 5. Principal component analysis identified differences in body shape between transgenic carp and nontransgenic controls, with transgenic fishes having significantly larger head depth, longer caudal length and smaller depth of the body and caudal region. Morphology had significant effects on swimming performance. Higher critical swimming speeds are related to the combination of deeper body, deeper caudal region and shorter caudal length. These findings suggested that morphological variations in GH-transgenic carp were responsible for their lower swimming abilities in comparison with nontransgenic controls. 6. Transgenic carp had significantly shorter cell length and width, smaller erythrocyte surface and volume, unchanged cell length – width ratio and larger cell surface – volume ratio. Transgenic fish had significantly higher routine metabolism, but their maximum metabolism was similar to controls. There were not significant differences in hematocrit, blood hemoglobin concentrations, plasma glucose, lactate, and cholesterol and protein concentrations between transgenic carp and nontransgenic controls at rest, but transgenic fish had significantly lower mean resting corpuscular hemoglobin concentrations and higher plasma triglyceride concentrations. Exhaustive exercise did not induced significant changes of hematocrit, hemoglobin concentrations and mean corpuscular hemoglobin concentrations in both strains. Plasma glucose and lactate increased significantly following exhaustive exercise and lasted for 4h. The manners of plasma glucose concentrations responding to exhaustive exercise were similar between two strains, but transgenic fish exhibited reduced response in plasma lactate concentrations compared to controls. Exhaustive exercise evoked significantly higher plasma cholesterol and triglyceride concentrations for 1h in transgenics which were not found in nontransgenic controls. Plasma protein concentrations were not altered by exhaustive exercise in transgenics but lower from 2h post-exercise in nontransgenic controls. These findings indicated that ‘all-fish’ growth hormone transgenic carp had significantly smaller erythrocyte appearance, higher routine metabolism, similar maximum metabolism, oxygen transport characteristics and changed physiological responses (plasma lactate, cholesterol, triglyceride and protein) to exhaustive exercise compared with controls.
Pages148
Language中文
Document Type学位论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/12282
Collection学位论文
Recommended Citation
GB/T 7714
李德亮. 转“全鱼”生长激素基因黄河鲤生物学特征的初步研究[D]. 水生生物研究所. 中国科学院水生生物研究所,2008.
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