蒙古马主要性格性状及其候选基因的行为遗传学研究
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摘要
本研究以世界上最古老的马地方品种之一的蒙古马为研究对象,采用问卷调查和生理学测试方法,测试其几个主要性格行为性状。同时应用PCR-SSCP、PCR-RFLP等技术手段分析BDNF、DBH、SLC6A4、COMT、MAOB等性格候选基因8个片段的单核苷酸标记,测序后提交GenBank,登录号分别为:FJ913852～FJ913859。研究蒙古马主要性格行为性状及其候选基因之间的联系,以寻找蒙古马主要性格行为性状相关的分子标记,为建立蒙古马性格行为性状的分子标记辅助选育方法奠定理论基础。通过综合研究得出以下结论:
(1)问卷调查方法对于测试马匹的性格行为性状方法有效。评价者的回答之间一致性较强。
(2)母马的性格行为特征与其他马有明显区别,3岁以下的马学习能力较强,适宜开始进行训练。
(3)应用因子分析方法对马性格行为问卷调查的各行为指标进行归类,得到5个因子,第一主因子、第二主因子和第三主因子中所包含的行为变量高度相关。因子分析结果反映了蒙古马的行为模式和行为特点。
(4)本研究中实验房间(马厩)的中间位置悬挂黄色气球,墙壁上粘贴红、黄相间的条幅,在这样的新奇环境中马匹的绝大多数生理指标在实验刚开始时表现为突然增加,表明这样的评价系统已通过测试,用以评估焦虑状况是合理的。
(5)问卷调查统计分析结果中的因子1,即焦虑性状与马匹在生理学测试中的心率、△心率和排粪次数显著相关。
(6)心率变化与性别具有相关性,母马在新奇环境中心率上升明显比骟马和公马快。
(7)本研究中PCR-SSCP与PCR-RFLP对蒙古马性格候选基因的分析,结果表明DBH (1)、DBH (2)、MAOB基因片段为中度多态,具有合适信息。其他基因片段为低度多态,包含很少信息。DBH(1)、DBH(2)、MAOB基因片段成为较为理想的遗传标记,使连锁分析不但可行而且有效。
(8)计算各基因片段变异位点的χ2值结果表明仅SLC6A4(1)、SLC6A4(2)、SLC6A4(3)的多态位点处于Hardy-Weinberg平衡状态,提示SLC6A4基因上的三处多态位点已达到遗传平衡,在以后的进化过程中这些位点的基因型也会稳定遗传下去。
(9)相关性分析表明BDNF基因C222T和G261C显著影响蒙古马的咬人行为;DBH(1)基因G758A显著影响蒙古马的忍耐力和对其他马友好的行为,对理解力、沙浴、过河和游泳、识途性和群居性等行为影响极显著;COMT基因G217A显著影响蒙古马的竞争性行为;SLC6A4(1)基因T1362C显著影响蒙古马的神经质、恐慌性、惊慌型、胆怯性、对马具的反应和易捕捉度等行为,对独立性行为影响极显著;SLC6A4(3)基因AT1615-1616GC显著影响蒙古马的情绪性行为。
(10)蒙古马BDNF基因C222T和G261C和SLC6A4(1)基因T1362C主要影响问卷调查分析中的第一主因子,即焦虑性状;蒙古马DBH(1)基因G758A和COMT基因G217A主要影响问卷调查分析中的第二主因子,即训练能力性状;蒙古马SLC6A4(3)基因AT1615-1616GC和SLC6A4(1)基因T1362C影响问卷调查分析中的第三主因子,即亲和性性状。
This research chooses the Mongolian horse, one of the world's oldest breeds of horses, as the research object through the use of questionnaires and physiological testing methods to test the main characteristics of their behavior. At the same time, it applies technical means of PCR-SSCP and PCR-RFLP to analyze single nucleotide polymorphisms markers of candidate genes such as BDNF, COMT, SLC6A4 and MAOB. This research focuses on the linkage between the main temperament behavioral characteristics of Mongolian horses and the candidate genes in order to find molecular markers concerning Mongolian horses’behavior traits; thus theoretical foundation could be laid for Mongolian horses’behavior traits, mainly related to the molecular markers as well as to the assisted selection methods. The following conclusions have been made through the joint study:
(1) It is an effective questionnaires survey conducted to test a horse's characteristics of their behavior. The answers among the evaluators show obvious consistency.
(2) The characteristics of mare behavior are different from others, and a less-than 3-year-old horse has a strong learning ability, and is suitable for training.
(3) By applying the factor analysis method, the behavioral acts indicators in the horse characteristics shown in questionnaires survey are classified into five factors. The behavioral variables contained among the first, second and third main factors are highly correlated. Factor analysis results reflect the behavior patterns and behavior characteristics of the Mongolian horse.
(4) In this study, the experiment was taken in a room with yellow balloons hanging in the middle, and on the walls there waere pasted red and yellow banners. The vast majority of physiological indicators of horses showed a sudden increase in the beginning of the experiment in such a strange environment, which indicated that the evaluation system has been tested, and it is reasonable to evaluate the horse's state of anxiety.
(5) The statistical analysis in the questionnaires shows that the first factor, that is, the state of anxiety is significantly correlated with the horse's heart rate during the physiological test, the△heart rate and the number of defecography.
(6) The changes in heart rate are relevant to gender, for the mare’s heart rate is far faster than that of the castrated horses and the stallions.
(7) In this study, the analysis results of the Mongolian horse’s characteristic candidate genes by means of PCR-SSCP and PCR-RFLP show that genetic fragments such as DBH1, DBH2, MAOB are moderately polymorphic with appropriate information,while other genetic fragments are low-grade polymorphism with very little information. Genetic fragments of DBH1, DBH2, and MAOB are considered as better ideal genetic markers, making the linkage analysis not only feasible, but also effective.
(8)Calculation results of the value of X2 in the variation of gene fragments locus show that the polymorphic loci of SLC6A4(1), SLC6A4(2), SLC6A4(3) are in a state of Hardy-Weinberg equilibrium, and the genotype of these sites will be inherited stably in the future evolution.
(9) Correlation analysis showed that the BDNF genes as C222T and G261C have a significant impact on the Mongolian horse’s biting behavior; DBH1 gene as G758A has a significant effect on endurance in Mongolian horses and on other horse-friendly behavior; at the same time the DBH1 gene has remarkable influence on such acts as comprehension ability, sand bath, crossing the river and swimming, the ability to recognize roads and live in groups; COMT gene as G217A has a greatly significant impact on the competitiveness of the Mongolian horse; SLC6A4(1) gene as T1362C has a significant impact on nervousness, panic, timid, and the behavior such as the reaction to harness, the degree of the learning ability of the Mongolian horse. The gene significantly affects the independence acts of the horse; SLC6A4(3) gene as AT1615-1616GC has significant impact on the mood of the Mongolian horse.
(10) BDNF gene as C222T and G261C, and SLC6A4(1) gene as T1362C of the Mongolian horse mainly affects the first principal factor in the questionnaire survey analysis, that is, anxiety levels; DBH1 gene as G758A and COMT and G217A gene of the Mongolian horse mainly affect the second principal factor in the questionnaire survey analysis, that is, the training capacity traits; SLC6A4(3) gene as AT1615-1616GC and SLC6A4(1) gene as T1362C of the Mongolian horse impact the third main factor in the analysis of the questionnaire survey, that is, affinity traits.
(1)问卷调查方法对于测试马匹的性格行为性状方法有效。评价者的回答之间一致性较强。
(2)母马的性格行为特征与其他马有明显区别,3岁以下的马学习能力较强,适宜开始进行训练。
(3)应用因子分析方法对马性格行为问卷调查的各行为指标进行归类,得到5个因子,第一主因子、第二主因子和第三主因子中所包含的行为变量高度相关。因子分析结果反映了蒙古马的行为模式和行为特点。
(4)本研究中实验房间(马厩)的中间位置悬挂黄色气球,墙壁上粘贴红、黄相间的条幅,在这样的新奇环境中马匹的绝大多数生理指标在实验刚开始时表现为突然增加,表明这样的评价系统已通过测试,用以评估焦虑状况是合理的。
(5)问卷调查统计分析结果中的因子1,即焦虑性状与马匹在生理学测试中的心率、△心率和排粪次数显著相关。
(6)心率变化与性别具有相关性,母马在新奇环境中心率上升明显比骟马和公马快。
(7)本研究中PCR-SSCP与PCR-RFLP对蒙古马性格候选基因的分析,结果表明DBH (1)、DBH (2)、MAOB基因片段为中度多态,具有合适信息。其他基因片段为低度多态,包含很少信息。DBH(1)、DBH(2)、MAOB基因片段成为较为理想的遗传标记,使连锁分析不但可行而且有效。
(8)计算各基因片段变异位点的χ2值结果表明仅SLC6A4(1)、SLC6A4(2)、SLC6A4(3)的多态位点处于Hardy-Weinberg平衡状态,提示SLC6A4基因上的三处多态位点已达到遗传平衡,在以后的进化过程中这些位点的基因型也会稳定遗传下去。
(9)相关性分析表明BDNF基因C222T和G261C显著影响蒙古马的咬人行为;DBH(1)基因G758A显著影响蒙古马的忍耐力和对其他马友好的行为,对理解力、沙浴、过河和游泳、识途性和群居性等行为影响极显著;COMT基因G217A显著影响蒙古马的竞争性行为;SLC6A4(1)基因T1362C显著影响蒙古马的神经质、恐慌性、惊慌型、胆怯性、对马具的反应和易捕捉度等行为,对独立性行为影响极显著;SLC6A4(3)基因AT1615-1616GC显著影响蒙古马的情绪性行为。
(10)蒙古马BDNF基因C222T和G261C和SLC6A4(1)基因T1362C主要影响问卷调查分析中的第一主因子,即焦虑性状;蒙古马DBH(1)基因G758A和COMT基因G217A主要影响问卷调查分析中的第二主因子,即训练能力性状;蒙古马SLC6A4(3)基因AT1615-1616GC和SLC6A4(1)基因T1362C影响问卷调查分析中的第三主因子,即亲和性性状。
This research chooses the Mongolian horse, one of the world's oldest breeds of horses, as the research object through the use of questionnaires and physiological testing methods to test the main characteristics of their behavior. At the same time, it applies technical means of PCR-SSCP and PCR-RFLP to analyze single nucleotide polymorphisms markers of candidate genes such as BDNF, COMT, SLC6A4 and MAOB. This research focuses on the linkage between the main temperament behavioral characteristics of Mongolian horses and the candidate genes in order to find molecular markers concerning Mongolian horses’behavior traits; thus theoretical foundation could be laid for Mongolian horses’behavior traits, mainly related to the molecular markers as well as to the assisted selection methods. The following conclusions have been made through the joint study:
(1) It is an effective questionnaires survey conducted to test a horse's characteristics of their behavior. The answers among the evaluators show obvious consistency.
(2) The characteristics of mare behavior are different from others, and a less-than 3-year-old horse has a strong learning ability, and is suitable for training.
(3) By applying the factor analysis method, the behavioral acts indicators in the horse characteristics shown in questionnaires survey are classified into five factors. The behavioral variables contained among the first, second and third main factors are highly correlated. Factor analysis results reflect the behavior patterns and behavior characteristics of the Mongolian horse.
(4) In this study, the experiment was taken in a room with yellow balloons hanging in the middle, and on the walls there waere pasted red and yellow banners. The vast majority of physiological indicators of horses showed a sudden increase in the beginning of the experiment in such a strange environment, which indicated that the evaluation system has been tested, and it is reasonable to evaluate the horse's state of anxiety.
(5) The statistical analysis in the questionnaires shows that the first factor, that is, the state of anxiety is significantly correlated with the horse's heart rate during the physiological test, the△heart rate and the number of defecography.
(6) The changes in heart rate are relevant to gender, for the mare’s heart rate is far faster than that of the castrated horses and the stallions.
(7) In this study, the analysis results of the Mongolian horse’s characteristic candidate genes by means of PCR-SSCP and PCR-RFLP show that genetic fragments such as DBH1, DBH2, MAOB are moderately polymorphic with appropriate information,while other genetic fragments are low-grade polymorphism with very little information. Genetic fragments of DBH1, DBH2, and MAOB are considered as better ideal genetic markers, making the linkage analysis not only feasible, but also effective.
(8)Calculation results of the value of X2 in the variation of gene fragments locus show that the polymorphic loci of SLC6A4(1), SLC6A4(2), SLC6A4(3) are in a state of Hardy-Weinberg equilibrium, and the genotype of these sites will be inherited stably in the future evolution.
(9) Correlation analysis showed that the BDNF genes as C222T and G261C have a significant impact on the Mongolian horse’s biting behavior; DBH1 gene as G758A has a significant effect on endurance in Mongolian horses and on other horse-friendly behavior; at the same time the DBH1 gene has remarkable influence on such acts as comprehension ability, sand bath, crossing the river and swimming, the ability to recognize roads and live in groups; COMT gene as G217A has a greatly significant impact on the competitiveness of the Mongolian horse; SLC6A4(1) gene as T1362C has a significant impact on nervousness, panic, timid, and the behavior such as the reaction to harness, the degree of the learning ability of the Mongolian horse. The gene significantly affects the independence acts of the horse; SLC6A4(3) gene as AT1615-1616GC has significant impact on the mood of the Mongolian horse.
(10) BDNF gene as C222T and G261C, and SLC6A4(1) gene as T1362C of the Mongolian horse mainly affects the first principal factor in the questionnaire survey analysis, that is, anxiety levels; DBH1 gene as G758A and COMT and G217A gene of the Mongolian horse mainly affect the second principal factor in the questionnaire survey analysis, that is, the training capacity traits; SLC6A4(3) gene as AT1615-1616GC and SLC6A4(1) gene as T1362C of the Mongolian horse impact the third main factor in the analysis of the questionnaire survey, that is, affinity traits.
引文
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