Indian Journal of Animal Research

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Research Article

Indian Journal of Animal Research, volume 54 issue 2 (february 2020) : 138-142

Analysis on correlation between polymorphism of MyoG gene exon I and body size traits of sheep

Jun. Yan. Bai1,*, Heng.Cao1, You. Bing .Yang1, Yi .Zhang2, Xin.Yue.Li1, Zi.Heng.Li1, Wei.Guang.Hao1, Fei.Yang.Zheng1
1College of Animal Science and Technology, Henan University of Science and Technology, Luoyang-471 003, China.
2College of Animal Science and Technology, China Agricultural University, Beijing-100 193, China.
Cite article:- Bai Yan. Jun., Heng.Cao, .Yang Bing You., .Zhang Yi, Xin.Yue.Li, Zi.Heng.Li, Wei.Guang.Hao, Fei.Yang.Zheng (2019). Analysis on correlation between polymorphism of MyoG gene exon I and body size traits of sheep . Indian Journal of Animal Research. 54(2): 138-142. doi: 10.18805/ijar.B-1081.

ABSTRACT

Myogenin (MyoG) is responsible for centering control over myocyte differentiation process and can influence the meat production of animals directly. Large tailed Han sheep, small tailed Han sheep, Yuxi fatty tailed sheep, Lanzhou large tailed sheep, Mongolia sheep and Tong Sheep were used as the experimental materials in this study. Polymorphism of MyoG gene exon I was tested by native polyacrylamide gel electrophoresis. The correlation between polymorphism of MyoG gene exon I and body size traits was analyzed. Influences of MyoG gene exon I on growth traits of sheep were discussed. Results demonstrated that 2 alleles (A, B) and 3 genotypes (AA, BB and AB) were detected in MyoG gene exon I of all 6 sheep varieties. The A allele frequencies in MyoG gene exon I of small tailed Han sheep, large tailed Han sheep, Yuxi fatty tailed sheep, Lanzhou large tailed sheep, Mongolia sheep and Tong Sheep were 0.5167, 0.2500, 0.4376, 0.6500, 0.5750 and 0.7125, while the B allele frequencies were 0.4833, 0.7500, 0.5625, 0.3500, 0.4250 and 0.2875, respectively. Chest width and neck length of the AA genotype of MyoG gene exon I were significantly higher than those of the AB genotype (P<0.01) and BB genotype (P<0.05). Body length and rump length of the AA genotype were significantly higher than those of the AB genotype (P<0.05). The chest depth and hip width of the AA genotype was far lower than that of the AB genotype (P<0.01). The hip height of the AA genotype was far lower than that of the BB genotype (P<0.05).

INTRODUCTION

Myogenin (MyoG) gene is responsible for centering control over myocyte differentiation process and can influence the meat production of animals directly. Hasty et al., (1993) and Nabeshima et al., (1993) discovered that MyoG is one of important members of MRFs family. Further studies demonstrated that heritable variation of this gene is related with growth rate of MyoG and quantity of muscle fibers in beasts and birds (Tepas et al., 2000; Gao et al., 2005; Anton et al., 2006). It is an excellent candidate gene that many affect characteristics of muscle fiber and carcass quality and growth traits, thus influencing meat quality and meat production of beasts and birds (Kim et al., 2009; Bhutan et al., 2009; Wang et al., 2007; Xue et al., 2007; Zhao et al., 2005; Wei et al., 2016; Chai et al., 2018; Wei et al., 2017). At present, the research on sheep mainly focuses on satellite marker polymorphism (Bai et al., 2015), gene polymorphism (Bai et al., 2016; Bai et al., 2017a; Abdelmoneim et al., 2017), genome-wide selection and so on (Wang, 2018). Many scholars have analyzed the polymorphism of MyoG gene in goats and sheep (Gao et al., 2009; Liu et al., 2009; Liu et al., 2010; Bai et al., 2017b). This study analyzed the polymorphism of MyoG gene in five sheep breeds,the correlation between polymorphism of  MyoG gene exon I and body size traits of sheep was analyzed.

MATERIALS AND METHODS

Genome DNA extraction
 
Large tailed Han sheep (40), small tailed Han sheep (40), Yuxi fatty tailed sheep (40), Lanzhou large tailed sheep (40), Mongolia sheep (40) and Tong Sheep (40) were collected as animal samples.
 
PCR amplification
 
Pre-denaturation at 94 for 4 min, then denaturation at 94 for 40 s, annealing at 60 for 1 min, annealing at 72 for 20 seconds, denaturation, annealing and elongation were carried out for 35 cycles, then elongation at 72 and finally the reaction was completed and cooled and preserved at 4°C.
 
SSCP
 
15% non denaturing poly acrylamide gels were used to detect the products. The 6 uL PCR amplification products were mixed with 6 uL denaturation solution, added with paraffin wax oil seal and heated in 10 min at 98 degree water bath. The amplified products were sampled at 10 Ul point and Marker at the appropriate swimming lane point. When the blue indicator strip runs to the bottom of the electrophoresis cell, turn off the power supply. Silver nitrate dyeing method is used for dyeing, mainly through fixation, oxidation, dyeing, color rendering, photography and other links.
 
Data analysis
 
Allele frequency and genotype frequency of MyoG gene exon I were calculated by PopGen32. Correlation between MyoG gene exon I and body size traits was analyzed by the general linear model in SPSS17.0. Gender and month age were non-genetic factors which were considered in the reference model. Genotypes of MyoG gene exon I were used as the genetic factor.

RESULTS AND DISCUSSION

Detection of PCR products and SSCP
 
The PCR amplification products of MyoG gene exon I were detected by agarose gel electrophoresis. All of the six sheep breeds gave clear target bands at 325bp (Fig 1). SSCP test showed that three genotypes (AA, AB and BB) were detected in MyoG gene exon I of all six sheep breeds (Fig 2).
 

Fig 1: PCR product detection of exon I of MyoG gene.


 

Fig 2: Sscp detection of exon I of MyoG gene in sheep.


 
Polymorphism analysis of MyoG Gene Exon I
 
Genotype frequency, gene frequency and polymorphism information contents of MyoG gene exon I in six sheep breeds were shown in Table 1. The frequency of AA genotype was the highest in Tong Sheep (0.5750) and that of AA genotype was the lowest in large tailed Han sheep (0.1250). The frequency of AB genotype was the highest in Mongolia sheep (0.4500) while it was the lowest in Lanzhou large tailed sheep (0.2000). The frequency of BB genotype was the highest in Large tailed Han sheep (0.6250) while it was the lowest in Tong sheep (0.1500). Chu et al., (2005) concluded that the genotype AA of MyoG gene exon I had the highest frequency in four sheep breeds. Liu et al., (2007) pointed out that there were three genotypes (AA, AB and BB) in amplification fragment of exon I. The allele frequency of A was far higher than that of B. Polymorphism fragments sequencing of exon I demonstrated that a single base substitution mutation (T®C) at 305 bp of MyoG gene resulted in changes of coding amino acid from cysteine to arginine. Han et al., (2016) discovered base mutation of A®C at 109 bp of MyoG gene exon I, which led to missense mutation of glutamic acid (GAG)® alanine (GCG) at 37 aa of MyoG protein. In this experiment, two alleles (A, B) and three genotypes (AA, AB, BB) were detected in MyoG gene exon I, which were consistent with previous research results.
 

Table 1: Polymorphic information of exon I of MyoG gene.


 
MyoG gene exon I showed the highest heterozygosity (0.4994), quantity of effective allele (1.9978) and polymorphism information content (0.3747) in small tailed Han sheep. On the contrary, large tailed Han sheep showed the lowest heterozygosity (0.3750), quantity of effective allele (1.6000) and polymorphism information content (0.3047). According to polymorphism information content, MyoG gene exon I could be regarded as moderate polymorphism site. Large tailed Han sheep is a local sheep breed of Henan Province, it mainly distributes in areas surrounding Pingdingshan. Due to smaller body size and poorer growth and meat output than small tailed Han sheep, the culture quantity of large tailed Han sheep is becoming fewer and fewer. They have the characteristics of pure blood lineage, low genetic mutation, small heterozygosity and weak selection potentials. Oppositely, small tailed Han sheep has large body size, large culture quantity and hybridization with other varieties, whcih has relatively high genetic mutation, large heterozygosity and selection potentials.
 
Significance test indicated that Large tailed Han sheep showed similar genotype distribution with Yuxi fatty tailed sheep (P>0.05) (Table 2), but had extremely significant differences with small tailed Han sheep, Lanzhou large tailed sheep, Mongolia sheep and Tong sheep (P<0.01). There were significant differences of genotype distribution between Yuxi fatty tailed sheep and Lanzhou large tailed sheep and Mongolia sheep (P<0.05). In addition, no significant difference of genotype distribution between the rest two sheep breeds was observed (P>0.05).
 

Table 2: The difference of genotypes of exon I of MyoG gene in different species distribution.


 
Association between MyoG gene exon I polymorphism and body size traits
 It could be concluded from Table 3 that chest width and neck length of the AA genotype of MyoG gene exon I were significantly higher than those of the AB genotype (P<0.01) and BB genotype (P<0.05). Body length and rump length of the AA genotype were significantly higher than those of the AB genotype (P<0.05). The chest depth and hip width of the AA genotype was far lower than that of the AB genotype (P<0.01). The hip height of the AA genotype was far lower than that of the BB genotype (P<0.05). MyoG gene exon I had significant effect on the seven traits investigated in this study. Niu et al., (2015) showed that the mRNA level of MyoG gene in horse was the highest at 60 and 90 days of age,  significantly higher than that of other age groups (P<0.01). Yang et al., (2014) showed that the hip height of AA genotype of MyoG gene was significantly higher than that of AB genotype in Wandong yellow cattle (P<0.05). Han et al., (2016) showed that the weight, height and length of CC genotype of MyoG exon I were significantly higher than those of AA genotype in Eurasian sheep population of Henan province (P<0.05). The results of this study were similar to those of Niu et al., (2015) and Yang et al., (2014).
 

Table 3: Analysis of the relationship between the exon I of MyoG gene and the ulnar characters of sheep.

CONCLUSION

The study showed that the extron I of MyoG gene had effects on Chest width, neck length, Body length, rump length, chest depth, hip width, hip height.

ACKNOWLEDGEMENT

Sincere gratitude goes to the sponsor of National Natural Science Foundation (31201777).

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