公猪7号染色体繁殖性状QTL的精细定位和位置候选基因MEA1的鉴别研究

doi:10.11843/j.issn.0366-6964.2014.03.004

畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (3): 363-374.doi: 10.11843/j.issn.0366-6964.2014.03.004

公猪7号染色体繁殖性状QTL的精细定位和位置候选基因MEA1的鉴别研究

赵科伟，赵雪艳，张志燕，洪渊，范寅，任军，幸宇云^*

（江西农业大学动物生物技术国家重点实验室培育基地，南昌 330045）

收稿日期:2013-08-22 出版日期:2014-03-23 发布日期:2014-03-23
通讯作者: 幸宇云，副研究员，Tel/Fax：0791-83813080，E-mail：xingyuyun9@hotmail.com
作者简介:赵科伟（1987-），女，河北唐山人，硕士，主要从事动物遗传育种研究，E-mail：zhaokewei3856@126.com；赵雪艳（1987-），女，山东烟台人，博士生，主要从事动物遗传育种研究，E-mail：zhaoxueyan0102@163.com。二者同为第一作者
基金资助:
国家自然科学基金项目（31100891）

Fine Mapping of QTL and Identification of MEA1 as a Positional Candidate Gene Affecting Boar Reproduction Traits on SSC7

ZHAO Ke-wei，ZHAO Xue-yan，ZHANG Zhi-yan，HONG Yuan，FAN Yin，REN Jun，XING Yu-yun^*

(Candidate of National Key Laboratory for Animal Biotechnology，Jiangxi Agricultural University，Nanchang 330045， China)

Received:2013-08-22 Online:2014-03-23 Published:2014-03-23

摘要/Abstract

摘要：

本研究旨在精细定位公猪7号染色体（SSC7）影响附睾重和血浆睾酮浓度的数量性状位点（QTL），鉴别影响公猪繁殖性状的位置候选基因。以205头白色杜洛克×二花脸资源家系F₂公猪群体为研究材料，在初步定位的QTL区间内增加14个SNPs对该群体进行扫描以精细定位QTL。在精细定位QTL区间内筛选雄性增强抗原1（MEA1）为位置候选基因，采用标准关联分析和F-drop检验分析该基因的多态位点与公猪繁殖性状的关联性，通过实时定量PCR（RT-PCR）和差异表达分析其表达情况。结果表明：①影响附睾重和血浆睾酮浓度的QTL置信区间显著缩小。②MEA1基因的5个SNPs与公猪附睾重、血浆睾酮浓度和射精量极显著或显著相关（P<0.01或P<0.05）；单倍型与表型关联性分析显示，G-C-C-G-C单倍型与左、右侧附睾重极显著相关（P<0.01），A-C-C-G-G单倍型与左、右侧附睾重显著相关（P<0.05）；F-drop分析显示，g.-1168 A>G和g.1115 A>G位点可能为影响公猪血浆睾酮浓度的主效突变位点（QTN）或与QTN紧密连锁；差异表达分析表明，附睾重与MEA1在附睾中的表达量成负相关（相关系数为-0.300，P=0.1）；MEA1在成年公猪的18种组织中均有表达。本研究通过精细定位缩小了7号染色体影响公猪附睾重和血浆睾酮浓度的QTL置信区间；同时发现MEA1基因与公猪附睾重、血浆睾酮浓度和射精量显著相关，其中g.-1168 A>G和g.1115 A>G位点可能是影响公猪血浆睾酮浓度的QTN或与QTN紧密连锁，这2个位点可作为选育性欲较强公猪的分子标记。

Abstract:

This aim of this study was to fine map the QTL affecting boar epididymal weight and serum testosterone concentration on SSC7 and to identify the positional candidate genes affecting boar reproduction traits.The materials used in this study were 205 F₂ boars from a White Duroc× Erhualian resource population.For fine mapping the QTL affecting epididymal weight，14 SNPs in the region preliminarily mapped were increased as markers to scan the boar population.Male-enhanced antigen 1 (MEA1) was chosen as a positional candidate gene in the fine mapping region.Association and F-drop analyses were implemented to reveal the relationship between MEA1 polymorphisms and boar reproduction traits.RT-PCR and differential expression assays were used to detect the MEA1 expression.The results showed that：①Fine mapping analyses revealed that the QTL region affecting epididymal weight and serum testosterone concentration was significantly narrowed.② Association analyses revealed that five SNPs of MEA1 were extremely significantly or significantly associated with boar epididymal weight，serum testosterone concentration and semen volume (P<0.01 or P<0.05).G-C-C-G-C haplotype was extremely significantly associated with boar left and right epididymal weight (P<0.01)，and A-C-C-G-G haplotype was significantly associated with boar left and right epididymal weight (P<0.05).F-drop test showed that g.-1168A>G and g.1115A>G could be quantitative trait nucleotides (QTN) or be complete linkage disequilibrium with QTN affecting testosterone concentration.Differential expression analysis showed that epididymal weight had negative relationship with MEA1 expression level in this tissue (r²=-0.300,P=0.1).qRT-PCR assay detected the expression of MEA1 gene in all 18 selected tissues of a mature boar.QTL regions affecting epididymal weight and serum testosterone concentration were narrowed through fine mapping.Significant association was found between MEA1 gene and boar epididymal weight，serum testosterone concentration and semen volume.g.-1168 A>G and g.1115 A>G of MEA1 could be QTN or be complete linkage disequilibrium with QTN for boar serum testosterone concentration，and these two SNPs can be used as molecular markers for selecting boars with high libido.

中图分类号:

S828
S813.3

赵科伟，赵雪艳，张志燕，洪渊，范寅，任军，幸宇云. 公猪7号染色体繁殖性状QTL的精细定位和位置候选基因MEA1的鉴别研究[J]. 畜牧兽医学报, 2014, 45(3): 363-374.

ZHAO Ke-wei，ZHAO Xue-yan，ZHANG Zhi-yan，HONG Yuan，FAN Yin，REN Jun，XING Yu-yun. Fine Mapping of QTL and Identification of MEA1 as a Positional Candidate Gene Affecting Boar Reproduction Traits on SSC7[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(3): 363-374.

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公猪7号染色体繁殖性状QTL的精细定位和位置候选基因MEA1的鉴别研究

Fine Mapping of QTL and Identification of MEA1 as a Positional Candidate Gene Affecting Boar Reproduction Traits on SSC7

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