卵形鲳鲹选育群体微卫星标记、生长比较、形态性状与体重相关性分析和生态养殖研究
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摘要
为开发卵形鲳鲹优良品种,促进其养殖业健康、稳定、可持续性发展,本文对卵形鲳鲹基础选育群体进行了分子标记、生长比较以及主要形态性状与体重的相关性研究,并对其健康养殖模式进行了探讨。主要结果如下:
1卵形鲳鲹养殖群体的微卫星多态性分析
运用6对微卫星引物分别对海南、深圳、福建3个地区卵形鲳鲹(Trachinotus ovatus)养殖群体的遗传差异进行了分析。数据采用Popgen32软件分析,并用MEGA4.0作图,结果得出3个群体平均等位基因数(N_a)为3.67~3.83,平均有效等位基因数(N_e)2.43~3.03,平均观测杂合度(H_o)为0.48~0.66,平均期望杂合度(H_e)为0.56~0.64,平均多态信息含量(PIC)为0.49~0.55,可见3个养殖群体的遗传多样性较高。Hardy-weinberg平衡遗传偏离指数(d)显示海南和深圳群体杂合子过剩(0.10和0.12),而福建群体则出现杂合子缺失(-0.08),统计检验发现三个群体中各位点的遗传偏离并不显著(p>0.05)。根据3个群体的遗传相似度及遗传距离信息进行聚类分析表明,福建与深圳群体的亲缘关系较近,两者与海南群体的亲缘关系较远,三群体的确切来源还有待进一步研究,为卵形鲳鲹群体选育提供了指导。
2卵形鲳鲹选育群体形态性状与体重的相关性分析
跟踪测量了卵形鲳鲹(T. ovatus)选育群体1月龄,4月龄,7月龄,10月龄和13月龄个体的全长(x_1),体长(x_2),体高(x_3)和体重(y)。运用相关分析,多元回归分析和通径分析的方法,计算了以体重为依变量,其他形态性状为自变量的相关系数,通径系数和决定系数,并剖分各性状对体重的影响,并建立最优多元回归方程。结果表明:在不同的生长阶段,所测4个性状间的相关系数在0.017—0.960之间,除4月龄个体体高与其他性状间相关性不显著外((P>0.05),其他月龄群体各性状两两之间的相关性均达到了极显著水平(P<0.01);1月龄和4月龄个体全长对体重的直接作用最大,7月龄,10月龄和13月龄个体则是体高对体重的影响最大;各性状对体重决定程度分析与通径分析变化趋势一致,各月龄群体全长、体长、体高估计体重的多元线性回归方程的拟合度良好(86.9%-90.6%),说明影响体重的主要形态因素已找出,选育时期不同,所选择的形态性状也应有所差异,为卵形鲳鲹个体选育提供了理论依据。
3卵形鲳鲹3个养殖群体的生长比较
采集了海南、深圳和福建3个地区卵形鲳鲹养殖群体构建了亲本选育的基础群体,并跟踪测量了各群体的全长、体长、体高和体重,对各群体不同时期的生长速度,生长变异情况,6个月的绝对增重率、特定生长率,以及主要形态性状与体重的相关关系进行了分析。结果表明:经过6个月的人工选择后,各群体生长速度均较对照组有显著提高。福建群体的绝对增重率和特定生长率比海南群体分别高出0.23g/天和2.79%,比深圳群体分别高出0.49g/天和9.41%,且差异显著(p<0.05)。在生长早期(2月龄),福建群体体重变异程度最大,深圳群体次之,海南群体变异相对较平稳。变异程度相近时(6月龄),海南群体体重分布范围较广(200g-600g),而福建群体分布较为集中,体重在450-500克之间的个体占43.33%。综合说明选择的效果初步得到体现,福建群体比海南和深圳群体具备更强的生长优势,可做进一步选育。
4卵形鲳鲹滩涂多级综合生态养殖研究
滩涂综合养殖以其较强的可操作性,稳定性和灵活性受到越来越多学者的关注。本研究在广东省汕尾长沙湾地区构建了滩涂多级综合养殖体系,通过对体系中卵形鲳鲹(T. ovatus)、南美白对虾(Penaeus vannamei Boone)、锯缘青蟹(Scylla serrata)和近江牡蛎(Crassostrea)的生长与收获情况的分析以及养殖过程中水质监测分析来评估该模式的经济和环境效益。卵形鲳鲹、南美白对虾和锯缘青蟹混养在池塘中,比例为1.2:90:1.3;近江牡蛎单养在入水沟和围基池中,放养密度为2.3个/㎡。水流随着潮汐变化将残余的营养物质从高营养级物种输送给低营养级物种利用。体系外的3个鱼虾蟹混养池和3个牡蛎单养池设为对照组。同一检测时间内实验组和对照组温度、盐度和pH差异不显著(p>0.05),而其他水质指标则存在差别;进、排水沟水质也无明显差异;氨氮和活性磷的去除率分别为53.38%和58.02%;体系中各养殖品种的产量显著高于对照组(p<0.05),其中卵形鲳鲹实验组产量为4236.2 kg/ha,而对照组仅为3943.4kg/ha。体系中的总成本与对照组相差不大,但总收益和总利润却分别高出48.97%和123.48%。可见该养殖模式不仅能降低污染,而且节能高效。
In order to empolder the excellent varieties of Trachinotus ovatus and promote the healthy, stable, sustainable development of aquaculture, molecular and morphological markers were applied on the selective breeding program for T. ovatus, and healthy cultivation of T. ovatus was also carried out in this research. The results were showed as follows:
1 Genetic polymorphism of cultured populations of Trachinotus ovatus as revealed by microsatellites
Amplifications with 6 pairs of microsatellite primers we determined the genetic polymorphism of genomic DNA in three cultured populations of goldern pompano(T. ovatus), which were collected from Hainan,Shenzheng and Fujian province respectively. POPGEN ver.32 software package was used to analyze the data and phylogenetic trees were also constructed in MEGA ver.4.0. The result showed that in all three populations the value of average number of allele(sN_a)was 3.67~3.83, the value of average effective number of alleles(N_e) was 2.43~3.03, the oberved value of mean heterozygosity(H_o) varied from 48% to 66% and the expected value of mean heterozygosity(H_e) varied from 56% to 64%, the value of average polymorphism information content(PIC) was 0.49~0.55, a high genetic diversity among three populations could be concluded from the datas mentioned above. Additionally, we computed the Hardy-Weinberg index to access the distribution of genotypes, and the values in Hainan,Shenzhen and Fujian population were 0.10, 0.12 and -0.08, respectively. The Bonferroni Correction test showed that all the loci reflected no significant genetic disequilibrium in three populations(p>0.05). Cluster analysis was also applied based on the Nei's genetic identity and genetic distance. The result revealed that the phylogenetic relationship between the population FJ and SZ was closer, and the resource of these populations may request further studies.
2 Correlation analysis of morphometric traits and body weight on base population of T. ovatus
Morphometric traits of T. ovatus at 1-month-old stage, 4-month-old stage, 7-month-old stage, 10-month-old stage and 13-month-old stage were measured, including total length(x_1), body length(x_2), body height(x_3) and weight(y). Correlation coefficients, path coefficients and determination coefficients of each trait to body weight were calculated through path analysis and multiple linear regression. Effects of each morphometric trait on body weight were also dissected and then the optimal regression equation was established. The results showed that the Pearson correlations among 4 traits ranged from 0.017 to 0.960, and all reached significant level(p< 0.01) at different stages except that at 4-month-old stage. At 1-month-old stage and 4-month old stage, the trait with strongest direct effect on body weight was the total length, but that at other stages was body height. The result of determinant coefficients analysis was consistent with that of path analysis, and the fitness of equations at different stages were good(86.9%-90.6%). It indicated that the main morphometric traits affecting body weight were found and traits for selection should be thoroughly considered at different stages, which provided theoretical evidence for selective breeding for T. ovatus.
3 Growth comparation of three cultured populations of Trachinotus ovatus
Base population for selective breeding of T. ovatus had been constructed by three cultured populations, which were collected form Hainan, Shenzhen and Fujian respectively. And the total length, body length, body height and body weight of T. ovatus in each group were measured too. Variance analysis of the growth rate in different stages, growth variation, absolute growth rate, specific growth rate, and the correlation relationship between main morphological traits and body weight were also conducted. The results showed that growth rate in each experiment group was significantly higher than that in control group after beening selected for 6 months. The absolute growth rate and specific growth rate in Fujian group was 0.23g/d and 2.79% higher than that in Hainan group respectively, 0.49g/d and 9.41% higher than that in Shenzhen group(p<0.05). In the earlier stage(2-month-old), the largest weight variation was found in Fujian group, followed by Shenzhen group, and that in Hainan group was placid. With the same weight variation(6-month-old), the distribution of weight in Hainan group was wider(200g-600g), and that in Fujian group was concentrate, 43.33% of T. ovatus were weighted between 450-500 gram. It could be concluded that the selection effect was initially reflected, and Fujian group had a greater growth advantage than Hainan and Shenzhen group, it was a good candidate for further studies.
4 Foundational studies on integrated multi-trophic aquaculture(IMTA) in intertidal zone
Intertidal mudflat integrated aquaculture has been drawing reasearchers’interests in China for its controllability, sustainable management and flexible operation. The study was conducted to mearsure the potential environmental and economic efficiency of integrated multi-trophic aquaculture (IMTA) in intertidal zone by testing the effects of what on growth, survival and yield of T. ovatus, Penaeus vannamei Boone, Scylla serrata and Crassostrea in the IMTA system in Changsha Bay, Shanwei, Guangdong Province, China. Two culture modes were applied in the experiment. T.ovatus was polycultured with P. vannamei Boone and S. serrata in all the ponds at a ratio of 1.2 fish:90 shrimp:1.3 crab / m2, where oyster was monocultured in the channel and enclosure at the same density of 2.3 individuals / m2. Water in the system flowed and transferred with the change of tide to recycle waste nutrients from higher trophic-level species into production of lower trophic-level species of commercial value. Three ponds for polyculture of T.ovatus, P. vannamei Boone and S. serrata, and enclosures for oyster monoculture were set as control groups. No significant differences in temperature, salinity and pH were obeserved among all the treatments throughout the trial (p>0.05), where other water parameters were not the same case. There were also no significant differences in all the water parameters between infall and outfall of the IMTA system. The mean reduction efficiency of total ammonia nitrogen and reactive orthophosphate in the IMTA system were 53.38% and 58.02%, respectively. The yields of T.ovatus, shrimp, Scylla serrata, oyster in control groups were significantly lower than those in IMTA system(T.ovatus: 3943.4kg/ha vs 4236.2kg/ha, p<0.05). Total costs in IMTA system were slightly lower than that in controls, but benefits and net incomes was very higher by 48.97% and 123.48% respectively. It may be concluded that adopting an IMTA configuration in intertidal zone can not only mitigate the pollution to coastal waters from ponds, but also save energy and improve the economic efficiency.
1卵形鲳鲹养殖群体的微卫星多态性分析
运用6对微卫星引物分别对海南、深圳、福建3个地区卵形鲳鲹(Trachinotus ovatus)养殖群体的遗传差异进行了分析。数据采用Popgen32软件分析,并用MEGA4.0作图,结果得出3个群体平均等位基因数(N_a)为3.67~3.83,平均有效等位基因数(N_e)2.43~3.03,平均观测杂合度(H_o)为0.48~0.66,平均期望杂合度(H_e)为0.56~0.64,平均多态信息含量(PIC)为0.49~0.55,可见3个养殖群体的遗传多样性较高。Hardy-weinberg平衡遗传偏离指数(d)显示海南和深圳群体杂合子过剩(0.10和0.12),而福建群体则出现杂合子缺失(-0.08),统计检验发现三个群体中各位点的遗传偏离并不显著(p>0.05)。根据3个群体的遗传相似度及遗传距离信息进行聚类分析表明,福建与深圳群体的亲缘关系较近,两者与海南群体的亲缘关系较远,三群体的确切来源还有待进一步研究,为卵形鲳鲹群体选育提供了指导。
2卵形鲳鲹选育群体形态性状与体重的相关性分析
跟踪测量了卵形鲳鲹(T. ovatus)选育群体1月龄,4月龄,7月龄,10月龄和13月龄个体的全长(x_1),体长(x_2),体高(x_3)和体重(y)。运用相关分析,多元回归分析和通径分析的方法,计算了以体重为依变量,其他形态性状为自变量的相关系数,通径系数和决定系数,并剖分各性状对体重的影响,并建立最优多元回归方程。结果表明:在不同的生长阶段,所测4个性状间的相关系数在0.017—0.960之间,除4月龄个体体高与其他性状间相关性不显著外((P>0.05),其他月龄群体各性状两两之间的相关性均达到了极显著水平(P<0.01);1月龄和4月龄个体全长对体重的直接作用最大,7月龄,10月龄和13月龄个体则是体高对体重的影响最大;各性状对体重决定程度分析与通径分析变化趋势一致,各月龄群体全长、体长、体高估计体重的多元线性回归方程的拟合度良好(86.9%-90.6%),说明影响体重的主要形态因素已找出,选育时期不同,所选择的形态性状也应有所差异,为卵形鲳鲹个体选育提供了理论依据。
3卵形鲳鲹3个养殖群体的生长比较
采集了海南、深圳和福建3个地区卵形鲳鲹养殖群体构建了亲本选育的基础群体,并跟踪测量了各群体的全长、体长、体高和体重,对各群体不同时期的生长速度,生长变异情况,6个月的绝对增重率、特定生长率,以及主要形态性状与体重的相关关系进行了分析。结果表明:经过6个月的人工选择后,各群体生长速度均较对照组有显著提高。福建群体的绝对增重率和特定生长率比海南群体分别高出0.23g/天和2.79%,比深圳群体分别高出0.49g/天和9.41%,且差异显著(p<0.05)。在生长早期(2月龄),福建群体体重变异程度最大,深圳群体次之,海南群体变异相对较平稳。变异程度相近时(6月龄),海南群体体重分布范围较广(200g-600g),而福建群体分布较为集中,体重在450-500克之间的个体占43.33%。综合说明选择的效果初步得到体现,福建群体比海南和深圳群体具备更强的生长优势,可做进一步选育。
4卵形鲳鲹滩涂多级综合生态养殖研究
滩涂综合养殖以其较强的可操作性,稳定性和灵活性受到越来越多学者的关注。本研究在广东省汕尾长沙湾地区构建了滩涂多级综合养殖体系,通过对体系中卵形鲳鲹(T. ovatus)、南美白对虾(Penaeus vannamei Boone)、锯缘青蟹(Scylla serrata)和近江牡蛎(Crassostrea)的生长与收获情况的分析以及养殖过程中水质监测分析来评估该模式的经济和环境效益。卵形鲳鲹、南美白对虾和锯缘青蟹混养在池塘中,比例为1.2:90:1.3;近江牡蛎单养在入水沟和围基池中,放养密度为2.3个/㎡。水流随着潮汐变化将残余的营养物质从高营养级物种输送给低营养级物种利用。体系外的3个鱼虾蟹混养池和3个牡蛎单养池设为对照组。同一检测时间内实验组和对照组温度、盐度和pH差异不显著(p>0.05),而其他水质指标则存在差别;进、排水沟水质也无明显差异;氨氮和活性磷的去除率分别为53.38%和58.02%;体系中各养殖品种的产量显著高于对照组(p<0.05),其中卵形鲳鲹实验组产量为4236.2 kg/ha,而对照组仅为3943.4kg/ha。体系中的总成本与对照组相差不大,但总收益和总利润却分别高出48.97%和123.48%。可见该养殖模式不仅能降低污染,而且节能高效。
In order to empolder the excellent varieties of Trachinotus ovatus and promote the healthy, stable, sustainable development of aquaculture, molecular and morphological markers were applied on the selective breeding program for T. ovatus, and healthy cultivation of T. ovatus was also carried out in this research. The results were showed as follows:
1 Genetic polymorphism of cultured populations of Trachinotus ovatus as revealed by microsatellites
Amplifications with 6 pairs of microsatellite primers we determined the genetic polymorphism of genomic DNA in three cultured populations of goldern pompano(T. ovatus), which were collected from Hainan,Shenzheng and Fujian province respectively. POPGEN ver.32 software package was used to analyze the data and phylogenetic trees were also constructed in MEGA ver.4.0. The result showed that in all three populations the value of average number of allele(sN_a)was 3.67~3.83, the value of average effective number of alleles(N_e) was 2.43~3.03, the oberved value of mean heterozygosity(H_o) varied from 48% to 66% and the expected value of mean heterozygosity(H_e) varied from 56% to 64%, the value of average polymorphism information content(PIC) was 0.49~0.55, a high genetic diversity among three populations could be concluded from the datas mentioned above. Additionally, we computed the Hardy-Weinberg index to access the distribution of genotypes, and the values in Hainan,Shenzhen and Fujian population were 0.10, 0.12 and -0.08, respectively. The Bonferroni Correction test showed that all the loci reflected no significant genetic disequilibrium in three populations(p>0.05). Cluster analysis was also applied based on the Nei's genetic identity and genetic distance. The result revealed that the phylogenetic relationship between the population FJ and SZ was closer, and the resource of these populations may request further studies.
2 Correlation analysis of morphometric traits and body weight on base population of T. ovatus
Morphometric traits of T. ovatus at 1-month-old stage, 4-month-old stage, 7-month-old stage, 10-month-old stage and 13-month-old stage were measured, including total length(x_1), body length(x_2), body height(x_3) and weight(y). Correlation coefficients, path coefficients and determination coefficients of each trait to body weight were calculated through path analysis and multiple linear regression. Effects of each morphometric trait on body weight were also dissected and then the optimal regression equation was established. The results showed that the Pearson correlations among 4 traits ranged from 0.017 to 0.960, and all reached significant level(p< 0.01) at different stages except that at 4-month-old stage. At 1-month-old stage and 4-month old stage, the trait with strongest direct effect on body weight was the total length, but that at other stages was body height. The result of determinant coefficients analysis was consistent with that of path analysis, and the fitness of equations at different stages were good(86.9%-90.6%). It indicated that the main morphometric traits affecting body weight were found and traits for selection should be thoroughly considered at different stages, which provided theoretical evidence for selective breeding for T. ovatus.
3 Growth comparation of three cultured populations of Trachinotus ovatus
Base population for selective breeding of T. ovatus had been constructed by three cultured populations, which were collected form Hainan, Shenzhen and Fujian respectively. And the total length, body length, body height and body weight of T. ovatus in each group were measured too. Variance analysis of the growth rate in different stages, growth variation, absolute growth rate, specific growth rate, and the correlation relationship between main morphological traits and body weight were also conducted. The results showed that growth rate in each experiment group was significantly higher than that in control group after beening selected for 6 months. The absolute growth rate and specific growth rate in Fujian group was 0.23g/d and 2.79% higher than that in Hainan group respectively, 0.49g/d and 9.41% higher than that in Shenzhen group(p<0.05). In the earlier stage(2-month-old), the largest weight variation was found in Fujian group, followed by Shenzhen group, and that in Hainan group was placid. With the same weight variation(6-month-old), the distribution of weight in Hainan group was wider(200g-600g), and that in Fujian group was concentrate, 43.33% of T. ovatus were weighted between 450-500 gram. It could be concluded that the selection effect was initially reflected, and Fujian group had a greater growth advantage than Hainan and Shenzhen group, it was a good candidate for further studies.
4 Foundational studies on integrated multi-trophic aquaculture(IMTA) in intertidal zone
Intertidal mudflat integrated aquaculture has been drawing reasearchers’interests in China for its controllability, sustainable management and flexible operation. The study was conducted to mearsure the potential environmental and economic efficiency of integrated multi-trophic aquaculture (IMTA) in intertidal zone by testing the effects of what on growth, survival and yield of T. ovatus, Penaeus vannamei Boone, Scylla serrata and Crassostrea in the IMTA system in Changsha Bay, Shanwei, Guangdong Province, China. Two culture modes were applied in the experiment. T.ovatus was polycultured with P. vannamei Boone and S. serrata in all the ponds at a ratio of 1.2 fish:90 shrimp:1.3 crab / m2, where oyster was monocultured in the channel and enclosure at the same density of 2.3 individuals / m2. Water in the system flowed and transferred with the change of tide to recycle waste nutrients from higher trophic-level species into production of lower trophic-level species of commercial value. Three ponds for polyculture of T.ovatus, P. vannamei Boone and S. serrata, and enclosures for oyster monoculture were set as control groups. No significant differences in temperature, salinity and pH were obeserved among all the treatments throughout the trial (p>0.05), where other water parameters were not the same case. There were also no significant differences in all the water parameters between infall and outfall of the IMTA system. The mean reduction efficiency of total ammonia nitrogen and reactive orthophosphate in the IMTA system were 53.38% and 58.02%, respectively. The yields of T.ovatus, shrimp, Scylla serrata, oyster in control groups were significantly lower than those in IMTA system(T.ovatus: 3943.4kg/ha vs 4236.2kg/ha, p<0.05). Total costs in IMTA system were slightly lower than that in controls, but benefits and net incomes was very higher by 48.97% and 123.48% respectively. It may be concluded that adopting an IMTA configuration in intertidal zone can not only mitigate the pollution to coastal waters from ponds, but also save energy and improve the economic efficiency.
引文
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