摘要
转“全鱼”GH基因鲤(转基因鲤)是将鲤鱼β-肌动蛋白基因启动子和草鱼GH基因融合而成的“全鱼”基因重组元件显微注射到黄河鲤受精卵内而得到的能够稳定遗传的转基因鱼,目前已经形成一个稳定的品系。转基因鱼的生态风险一直是制约转基因鱼产业化的瓶颈,种群适合度问题是评价转基因鱼生态安全的关键。转植GH基因除了促进快速生长和提高饵料转换效率外,还可能对转基因鱼形态、生理、代谢和行为等方面的许多表型产生直接或间接的影响,这些表型特征的改变又可能导致转基因鱼与对照鱼在生存方式、摄食特征和繁殖策略等方面的差异,进而导致转基因鱼种群存活率和繁殖率发生改变,最终引起转基因鱼适合度的变化。
摄食和呼吸功能与鱼类的适应性密切相关,摄食器官和呼吸器官的形态发育及其与功能的适应性是鱼类适合度评价的重要方面,本论文于2009年4月—2010年10月对转基因鲤的的呼吸器官的LM、SEM结构和摄食器官形态的进行测定分析,并对正常及低氧状态下鳃形态和鳃组织Na+、K+-ATP酶活性的变化进行比较,主要研究结果如下：
1.对转基因鲤和普通鲤呼吸器官(鳃)形态进行测量,包括对鳃丝数、鳃丝长、鳃小片数、鳃小片密度、鳃小片面积的测定,对各形态参数以体重为协变量进行协方差分析,结果表明：转基因鲤的单个鳃丝长、鳃丝总长、鳃小片总数、呼吸面积显著大于对照组,鳃丝数、鳃小片密度、单个鳃小片面积与对照鲤没有显著性差异。
2.对转基因鲤和普通鲤摄食器官形态进行测量,包括对头长、吻长、眼距、口裂宽、口裂高、第一鳃弓长、最长鳃耙长、鳃耙间距的测定,对各形态参数以体重为协变量进行协方差分析,结果表明：转基因鲤的头长、吻长、第一鳃弓长、鳃耙间距显著小于对照鲤,口裂高、口裂宽和鳃耙数等与对照鲤无显著性差异。
3.对转基因鲤和普通鲤在低氧条件下(0.8±0.1mg/L)处理7d,而后恢复充氧(7.5±0.5mg/L)3d,取样观测结果表明：低氧引起转基因鲤和对照鲤的鳃形态发生巨大变化,鳃小片间细胞团萎缩,鳃小片面积增大,7d后转基因鲤呼吸面积增大了96.3%,对照鲤呼吸面积增大了83.9%,转基因鲤的呼吸面积显著大于对照鲤;当溶氧恢复正常时,鳃小片间细胞团重新包埋鳃小片,鳃小片面积亦恢复,转基因鲤的恢复了78.4%,对照鲤恢复了65.3%,转基因鲤和对照鲤的呼吸面积已经没有显著性差异。
4.低氧引起转基因鲤和普通鲤的鳃组织Na-、K--ATP酶活性降低。转基因组Na-、K--ATP酶水平较对照组降低的快,恢复的也迅速,但两组之间并没有显著性差异。
"All-fish" GH transgenic common carp (transgenic fish) has been a stable line, produced by microinjection of "all-fish" gene included the grass carp GH drivend by theβ-actin gene promoter of common carp into the fertilized eggs of the common carp. The ecological safety has always been the indispensable step on the commercialization of transgenic fish, the key point is to evaluate the fitness of the transgenic fish. Besides accelarating growth performance and increasing food conversion efficiency of the fish, transgenic GH genes could also have direct or indirect effects on the morphology, physiology and behaviour and so on, which could change the survival, feeding and breeding of the fish, then led to the variation of the species survival and breeding rate, finally change the finess of the transgenic fish.
Focusing on the fitness of respiratory organs and feeding organs of transgenic fish, in this study we undertook the histological examination and SEM examination of the gill and measured the morphology of the feeding organs. Then we measured the morphology of the gill and the Na+、K+-ATPase activity of the gill tissue in hypoxia. The main results were listed as follows:
1. The gill morphology of the transgenic and nontransgenic (control) common carp was meansured, including the number of filaments, total filament length, total number of lamella, spacing of lamella, area of individual lamella and total lamella area. They were compared using one-way analysis of covariance with body weight as the covariate. It was shown that single filament length, total filament length, total number of lamella and total lamella area of the transgenic fish were significantly larger than those of control fish. But, there were no significant differences in all the other variables.
2. The feeding organs morphology of the transgenic and control fish was meansured, including the head length, snout length, spacing of eyes, mouth gape width, mouth gape length, length of fist gill arch, gill raker length and distance between gill raker. They were compared using a one-way analysis of covariance with body weight as the covariate. The results were that the head length, snout length, distance between gill raker were significantly smaller than those of control carp. There were no significant differences in all the other variables.
3. The exposure to hypoxia triggered the reduction of the ILCM and the gills displayed protruding lamella in transgenic and control fish. After 7 days of hypoxia, the total lamella area of transgenic carp incresed by 96.3%, was significantly larger than that of control fish. However, this morphological change was reversile. After 3 days recovery, the total lamella area of transgenic carp decreased by 78.4%, and 65.3% in control fish.
4. Hypoxia trigged the reduction of Na+、K+-ATPase activity in transgenic and control carp. That of transgenic carp changed faster than that of control fish, but there was no significant differerce between the two groups all over the expoure.
引文
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