吐鲁番斗鸡MAO、COMT等候选基因的多态性与其攻击行为的关联分析

Turpan game fowl, which has been listed as national level protected varieties in livestock and poultry, is one of four game fowls in China, and it is famous for its extremely strong aggressiveness. Although the aggressive behavior of animal is supposed to relate to MAOA gene, COMT gene and 5-HT gene, the molecular genetic mechanism and formal genetics of this behavior have not been clarified completely. What's more, relevant researches also have not been reported. On the basis of the preliminary confirmation that MAOA have a specific contact with Turpan game fowl, this research construct the resource group which is used to research the phenotype genetics law of the aggressive behavior on game fowl by using the aggressive Turpan fowl and the no-aggressive rose comb fowl as parents. Through adopting PCR-SSCP to research polymorphism of candidate genes such as MAOA gene, and COMT gene,5-HT gene etc on the parents and their offspring, and combining with phenotypic analysis, we are able to screen the superior genotypes of the key genes that cause the aggressive behavior of Game fowl. Finally, testing the results by the technology of fluorescent quantitation and MAS. This research can verify the hereditary capacity of the aggressive behaviour on Turpan game fowl, select and control the major gene of the aggressive behaviour by MAS, and it offers foundation to people for the breeding of excellent game fowl and the further study of molecular mechanism of aggressive behavior in animals (including humans).

新疆的吐鲁番斗鸡是中国四大斗鸡之一，具有极强的攻击性，已被列入国家级畜禽保护品种。MAOA、COMT和5-HT等基因与动物攻击行为相关，但动物攻击行为的表型遗传和分子遗传机制尚未完全阐明，吐鲁番斗鸡的相关研究报道也很少。本研究在初步证实MAOA与吐鲁番斗鸡斗鸡攻击行为相关的基础上，以攻击性强的吐鲁番斗鸡和无攻击性的玫瑰冠鸡为亲本，构建研究斗鸡攻击行为表型遗传规律的资源群，采用PCR-SSCP、RFLP技术分析资源群亲本及各世代鸡MAOA、COMT等候选基因的多态性，结合吐鲁番斗鸡攻击行为表型的关联分析，筛选引起斗鸡攻击行为关键基因的优势基因型，并以荧光定量RT-PCR和MAS技术进行验证。探明吐鲁番斗鸡攻击行为的遗传特性，选定调控吐鲁番斗鸡攻击行为主效基因的MAS分子标记，为选育出攻击行为极强的吐鲁番斗鸡和进一步研究动物攻击行为（包括人类）分子机理提供科学的参考依据。

摘要:本项目已完成计划书中的各项计划和阶段目标（标题1.1-1.5），略有超额完成与计划书相关的研究内容（标题1.6-1.7）。按计划书研究了吐鲁番斗鸡攻击行为的表型特征与遗传规律，与斗鸡攻击行为相关的5-HT等7个基因的表达量和MAOA等4个基因的多态性、CORT等4种激素含量的分析。主要成果如下:.1.1“镜像自斗法”和“攻击行为量化谱”.此成果不仅打破了几百年来传统的斗鸡攻击行为“配对打斗”的选择方法，避免了“配对打斗”时造成斗鸡的损伤甚至报废，可有效保护斗鸡并准确选择优秀斗鸡。.1.2 攻击行为表型的遗传规律.对亲本斗鸡32只、F1代100只和F2代300只的检测发现吐鲁番斗鸡攻击行为表型的遗传特性遵循孟德尔的分离遗传规律。.1.3 体型外貌特性:吐鲁番斗鸡具有体躯紧凑，颈部、胸部与腿部肌肉发达，符合斗鸡喜打斗的体型特征。.1.4 分子标记.在MAOA、5-HT2CR、SLC6A4、COMT、NR2E1等4个基因的PCR-SSCP分析中各筛选出1个位点可作为吐鲁番斗鸡攻击行为选择的分子标记，表明斗鸡的攻击行为受多基因调控，为吐鲁番斗鸡分子标记辅助选择（MAS）提供了理论参考依据。.1.5 攻击行为相关基因的调控作用.5-HT、COMT、SLC6A4、NR2E1、TRH、5-HTT、TPH1、DTNBP1等7个基因表达量的结果进一步证实斗鸡攻击行为受多基因调控，且5-HT基因呈正相关，TPH1基因呈负相关。.1.6 攻击行为表型与基因型的关联分析.对吐鲁番斗鸡和F1代鸡各30只攻击行为表型与SLC6A4的基因型的关联分析验证了影响吐鲁番斗鸡攻击行为主效基因SLC6A4的分子标记。.1.7 甲状腺轴对斗鸡攻击行为的作用.攻击性强的吐鲁番斗鸡TRH阳性细胞数和TH含量始终显著高于攻击弱的新罗曼蛋鸡，表明下丘脑-垂体-甲状腺轴与斗鸡攻击行为相关。 .1.8 CORT等激素对吐鲁番斗鸡攻击行为的作用.分析皮质醇(CORT)等4种激素含量的结果表明吐鲁番斗鸡的攻击行为具有攻击性精神分裂症的表征，可将斗鸡作为精神分裂症研究的模式动物，对动物（包括人类）攻击行为的深入研究具有重要意义。.1.9 论文.发表中文核刊学术论文6篇与参会论文4篇(其中1篇获二等奖)。.1.10 人才培养.指导4名硕士生和8名本科生完成毕业论文(其中1篇获优秀毕业论文奖)。