Scientia Agricultura Sinica ›› 2011, Vol. 44 ›› Issue (3): 570-578 .doi: 10.3864/j.issn.0578-1752.sas-2010-06640

• ANIMAL SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Identification of MicroRNA SNPs of HSF1 Gene and Their Association with Heat Tolerance in Chinese Holstein

LI Qiu-ling,JU Zhi-hua, JIA Xiang-jie, HUANG Jin-ming, LI Jian-bin, LI Rong-ling, LI Fang,WANG Chang-fa, ZHONG Ji-feng
   

  1. （山东省农业科学院奶牛研究中心）
  • Received:2010-06-10 Revised:2010-08-09 Online:2011-02-01 Published:2011-02-01
  • Contact: WANG Chang-fa, ZHONG Ji-feng

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Abstract:

【Objective】 Heat shock transcription factor1 (HSF1) is the key protein in regulating heat stress response. 【Method】The polymorphisms of HSF1 gene and their association with heat tolerance in 638 Chinese Holstein cattle were investigated for the purpose of providing molecular marker information to facilitate the breeding efficiency of thermo tolerant cows. Primers were designed to amplify HSF1 gene fragment. DNA sequencing, PCR-SSCP and PCR-RFLP methods were used to analyze the polymorphisms. MicroRNA SNPs of HSF1 gene were localized by blasting to miRBASE database. Linkage disequilibrium and haplotype were analyzed using SHEsis software. The association of these polymorphisms with heat tolerance index including potassium content in erythrocytes (PCE), decrease rate of milk production (R), rectal temperature (RT) and heat-tolerance coefficient (HTC) was analyzed by SAS software. The transcriptional level of HSF1 mRNA under heat stress conditions was studied by fluorescent quantitative RT-PCR technology. 【Result】 Two novel microRNA SNPs, T909C and G4693T, were found. The cows with CC and CT genotype showed higher heat tolerance than those with TT genotype at T909C locus (P＜0.05). The cows with TT genotype showed higher heat tolerance than those with GG genotype at G4693T locus (P<0.05). Four haplotype and 10 haplotype combinations were found. The cows with H2H4 haplotype combination had lower PCE, RT and higher HCT than those with H1H3 haplotype combination (P<0.05). The cows with H2H4 haplotype combination had lower R than those with H1H1 haplotype combination (P<0.05). H2H4 was the heat tolerant haplotype combination. The transcriptional level of HSF1 mRNA under heat stress conditions was different in different tissues. The highest is that in heart, which is 11.24-fold enrichment than that in muscle (P<0.05). 【Conclusion】 microRNA SNPs of HSF1 gene could be chosen as the molecular markers for choosing thermo tolerant dairy cattle in breeding program.

Key words: Chinese Holstein, HSF1, thermal tolerance, microRNA, polymorphism

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