Polymorphism of Growth-Correlated Genes Associated
with Fatness and Muscle Fiber Traits in Chickens
M. Lei, C. Luo, X. Peng, M. Fang, Q. Nie, D. Zhang, G. Yang, and X. Zhang1
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science,
South China Agricultural University, Guangzhou 510642, Guangdong, China
ABSTRACT Thirty single nucleotide polymorphisms
(SNP) and one 6-bp insertion-deletion (indel) from 8
genes of somatotropic axis were used to study the associa-
tion with chicken fatness and muscle fibers. The allele
frequency difference between Xinghua and White Plym-
outh Rock chickens was observed, and their effects on
fatness and muscle fiber traits were also evaluated by
linkage analyses. The G143831A (G+1705A) SNP of the
growth hormone (GH) gene was related to fat width,
and the G144762A (G+119A) SNP of the GH gene was
significantly associated with abdominal fat pad weight,
abdominal fat pad ratio, and crude fatty content of the
breast muscle. The 6-bp indel of the growth hormone
secretagogue receptor (GHSR) gene was significantly
linked with the fat traits. The C51978309T SNP of the
Key words: fatness, muscle fiber trait, linkage disequilibrium, single nucleotide polymorphism, linkage analysis
INTRODUCTION
Meat quality is a complex structural and functional
process that depends on species, genetic background,
metabolic status of the antemortem animal, the protein
complement of the muscle, and environmental factors.
Meat quality relies on several important characteristics,
including appearance, color, taste, fat content, texture,
and tenderness. Fatness and muscle fiber traits are the
major components of meat quality. The QTL mapping for
meat quality such as fatness and muscle fiber traits was
widely studied in the past decade (Ovilo et al., 2002; Nii
et al., 2005; Stearns et al., 2005). In chickens, several stud-
ies on meat quality QTL mapping were completed with
linkage analyses using microsatellite DNA (Jennen et al.,
2004, 2005; Abasht et al., 2006; Lagarrigue et al., 2006).
Not only linkage analyses with markers were used in
studies on meat quality, but also candidate approaches
were applied (Amills et al., 2005; Guyonnet-Dupérat et
©2007 Poultry Science Association Inc.
Received December 4, 2006.
Accepted January 16, 2007.
1
Corresponding author: xqzhang@scau.edu.cn
835
insulin-like factor-I (IGF-I) gene was significantly linked
with the transversal area of the leg muscle fiber and trans-
versal area of the breast muscle fiber. There was signifi-
cant linkage between the insulin (INS) gene and 2 traits
of the transversal area of transversal area of the leg muscle
fiber and transversal area of the breast muscle fiber. Asso-
ciation of 30 SNP and one 6-bp indel from 8 genes of
somatotropic axis with chicken fatness and muscle fiber
traits was analyzed in the present study. The GH, GHSR,
and leptin receptor genes were significantly related to
chicken fatness. The INS and IGF-I genes were linked with
muscle fiber density. Therefore, the genes of somatotropic
axis not only affected chicken growth and body composi-
tion but also were associated with fatness and muscle
fiber traits.
2007 Poultry Science 86:835–842
al., 2006). Compared with linkage analyses, candidate
approaches were less widely used.
The genes of somatotropic axis play a central role in
the regulation of growth and development (Mao et al.,
1997; Buyse and Decuypere, 1999; Vasilatos-Younken et
al., 2000). Previous studies showed that variation of these
genes affected gene expression at the transcription and
translation levels (Lo et al., 2003; Wyszynska-Koko et al.,
2006). Variation in the genes of somatotropic axis could
function as candidates for the evaluation of their effects on
chicken growth and development traits. Previous studies
have shown that some single nucleotide polymorphisms
(SNP) of the somatotropic axis genes indeed affected
growth traits significantly (Feng et al., 1997; Kuhnlein et
al., 1997; Amills et al., 2003; Lei et al., 2005; Nie et al.,
2005b; Fang et al., 2006; Qiu et al., 2006). On the other
hand, recent studies have shown that there was significant
association of growth and body composition with meat
quality characteristics (Le Bihan-Duval et al., 2001; Zer-
ehdaran et al., 2004), especially fat deposition and muscle
fiber density and sizes (Bruns et al., 2004; Scheuermann
et al., 2004). In human, mutations in prepoghrelin/ghrelin
gene were associated with obesity (Ukkola et al., 2001).
However, few studies on association of growth-correlated
genes with meat quality have been reported in chickens.
 836

Table 1. Details of single nucleotide polymorphism markers, genes, and primers1

Location2 Annealing Genotyping

Gene Chromosome (nt) Primer temperature (°C) method Enzyme
GH 27 G143831A 5′-tcccaggctgcgttttgttactc-3′; 5′-acgggggtgagccaggactg-3′ 60 PCR-RFLP Eco RV
G143978A 5′-gccctggcagccctgttaacc-3′; 5′-caccccaccatcgtatcccatc-3′ 60 PCR-RFLP Pag I
G142167T 5′-cccaacagtgccacgattccatg-3′; 5′-tgcgcaggtggatgtcgaacttg-3′ 59 PCR-RFLP Bsh136 I
G145086A 5′-atccccaggcaaacatcctc-3; 5′-cctcgacatccagctcacat-3′ 58 PCR-RFLP Msp I
A144762G 5′-atccccaggcaaacatcctc-3; 5′-cctcgacatccagctcacat-3′ 57 PCR-RFLP Msp I
GHR Z G6622190A 5′-cccttccattatgcattttatc-3; 5′-gggggtacactctagtcacttg-3′ 58 PCR-RFLP Eco72 I
C6622516T 5′-gcaacatcagaatcgctttt-3; 5′-tcccatcgtacttgaatatcc-3′ 58 PCR-RFLP Bsu R
A6626579G 5′-gaacccaggctctcaacagtg-3; 5′-tggaggttgaggtttatctgtc-3′ 56 PCR-RFLP Eco105 I
GHSR 9 G18790036A 5′-gtcgcctgcgtcctcctctt-3; 5′-acgggcaggaaaaagaagatg-3′ 61 PCR-RFLP Msp I
T18791236C 5′-cccacaaagttagctgcagac-3; 5′-cacctctccatctggctcat-3′ 60 PCR-RFLP Hin6 I
18792785 to 1879075913 5′-aggtggaaaaactgcaaaaag-3; 5′-aggcaccccataacttttcag-3′ 58 PCR-RFLP Csp6 I
C18793056T 5′-tggttgaaaagagagaatgct-3; 5′-ccacacgtctccttttatattc-3′ 58 PCR-RFLP Bsp119 I
C18793131T 5′-tggttgaaaagagagaatgct-3; 5′-ccacacgtctccttttatattc-3′ 57 PCR-RFLP Hin6 I
IGF-I 1 C51978309T 5′-ctgggctacttgagttactacat-3; 5′-cacggaaaataagggaatg-3′ 58 PCR-RFLP Tas I
C51983354T 5′-ctgggctacttgagttactacat-3; 5′-cacggaaaataagggaatg-3′ 59 PCR-RFLP Bsp119 I
C51978771T 5′-gccacccgaaagttaaccagaat-3; 5′-tccattgcggctctatct-3′ 60 PCR-RFLP Hinf I
C52028084T 5′-tgaaagggtctggccaaaaca-3; 5′-gggaagagtgaaaatggcagagg-3′ 59 PCR-RFLP Mph1103
IGFBP-2 7 G23967395T 5′-tttggttgagtcctaggcttg-3; 5′-aggcgtactacactgcagag-3′ 64 PCR-SSCP
T23966786A 5′-accggtctgagagcatccttg-3; 5′-gggaaaaagggtgtgcaaaag-3′ 64 PCR-SSCP
C23966654T 5′-gggcatttatatctgaggaacac-3; 5′-ggcaaagagcaacccaacac-3′ 60 PCR-RFLP Eco72 I
LEI ET AL.

T23966559G 5′-tttgttgtcgtggcttttttcag-3; 5′-gcttgtcacagttggggaag-3′ 55 PCR-RFLP Bsh136 I

G23966484A 5′-tttgttgtcgtggcttttttcag-3; 5′-gcttgtcacagttggggaag-3′ 56 PCR-RFLP Pvu I
INS 5 G11303145A 5′-cgtgtctcctttgcttcctac-3; 5′-tggagctttctgtgacaattc-3′ 58 PCR-RFLP Nde I
C11304264T 5′-tgttctgcatttggcccatac-3; 5′-gcagaatgtcagctttttgtcc-3′ 59 PCR-RFLP Msp I
T11306685C 5′-ctccatgtggcttccctgta-3; 5′-ggcttcttggctagttgcagt-3′ 60 PCR-RFLP Msp I
C11306451T 5′-ggtatctgaaaagcgggtctc-3; 5′-aatgctttgaaggtgcgatag-3′ 61 PCR-RFLP Msp I
LEPR 8 T28573025C 5′-atgctgcttgattcttcctcct-3; 5′-ccctaggcaaatggtaatgaac-3′ 56 PCR-RFLP Bsh136 I
A28573100G 5′-atgctgcttgattcttcctcct-3; 5′-ccctaggcaaatggtaatgaac-3′ 58 PCR-RFLP Bsh136 I
TSH-β 26 C2541870G 5′-cccttcttcatgatgtctctcc-3; 5′-ggtccttagttccatctgtgc-3′ 60 PCR-RFLP Csp6 I
C2543216T 5′-gagcacggtgagcattactgg-3; 5′-ggaggtacatttctgccacgt-3′ 61 PCR-RFLP Hin6 I
A2543276T 5′-gagcacggtgagcattactgg-3; 5′-ggaggtacatttctgccacgt-3′ 59 PCR-RFLP Msp I
1
GH = growth hormone gene; GHR = growth hormone receptor gene; GHSR = growth hormone secretagogue receptor gene, IGF-I = insulin-like growth factor-I gene; IGFBP-2 = insulin-like
growth factor binding protein-2 gene; INS = insulin gene; LEPR = leptin receptor gene; TSH-β = thyroid-stimulating hormone beta subunit gene; and SSCP = single strand conformational polymorphism.
2
Location on chromosome where gene is found (http://mgc.ucsc.edu/cgi-bin/hgBlat) (2004).
3
There was one insertion-deletion GGTACA for 18792785 to 187907591 in the chicken GHSR gene.
 GROWTH-CORRELATED GENES AND CHICKEN FATNESS 837
Table 2. Single nucleotide polymorphism allele frequencies and the chi-square test

Gene1 Loci Allele XH2 WRR3 F-value

GH G142167T G 0.53 0.78 15.512**
G143831A G 0.67 1.00 36.000**
G143978A G 0.79 0.85 17.379**
A144762G A 1.00 0.64 7.433*
G145086A G 0.74 0.57 0.130
GHR G6622190A G 1.00 1.00 0.000
C6622516T C 1 1 0
A6626579G A 0.38 0.94 31.482**
GHSR G18790036A G 0.93 1.00 5.373*
T18791236C T 0.75 0.53 10.697**
18792785 to 1879075914 I 0.74 0.65 1.070
C18793056T C 0.92 0.79 4.810
C18793131T C 0.76 0.60 5.560
IGF-I C51978309T C 0.50 0.50 0.000
C51983354T C 0.88 0.46 32.112**
C51978771T C 0.96 0.78 12.083**
C52028084T C 0.50 0.50 0.000
IGFBP-2 G23967395T G 0.74 0.89 5.230
T23966786A T 1.00 1.00 0.000
C23966654T C 0.31 0.67 20.355**
T23966559G T 1.00 1.00 0.000
G23966484A G 0.65 0.79 5.130
INS G11303145 A G 1.00 1.00 0.00
C11304264T C 0.65 0.82 8.5895*
T11306685C T 0.82 0.86 0.65
C11306451T C 0.57 0.70 3.21
LEPR T28573025C T 0.58 1.00 51.428**
A28573100G A 0.46 1.00 38.297**
TSH-β C2541870G C 0.78 0.83 2.940
C2543216T C 0.79 0.93 6.923**
A2543276T A 0.77 0.78 16.639**
1
GH = growth hormone; GHR = growth hormone receptor; GHSR = growth hormone secretagogue receptor;
IGF-I = insulin-like growth factor-I; IGFBP-2 = insulin-like growth factor binding protein-2; INS = insulin; LEPR =
leptin receptor; and TSH-β = thyroid-stimulating hormone beta subunit.
2
XH = Xinghua chickens.
3
WRR = White Plymouth Rock chickens.
4
There was one insertion-deletion GGTACA for 18792785 to 187907591 in the chicken GHSR gene.
*P < 0.05; **P < 0.01.

The purpose of the present study was to observe the
effect of the growth-correlated genes on fatness and mus-
cle fiber traits in chickens. Thirty SNP and one 6-bp indel
were selected from 8 genes of the somatotropic axis, the
growth hormone (GH), growth hormone receptor (GHR),
growth hormone secretagogue receptor (GHSR), insulin-
like growth factor-I (IGF-I), insulin-like growth factor
binding protein-2 (IGFBP-2), insulin (INS), leptin recep-
tor (LEPR), and thyroid-stimulating hormone beta sub-
unit (TSH-β). The linkage of the SNP with fatness and
muscle fiber traits was evaluated with linkage analyses
and linkage disequilibria in 2 unrelated populations.
MATERIALS AND METHODS
Chicken Populations and the Observation
of Chicken Fatness and Muscle Fiber Traits
A F2 resource population was constructed by crossing
the White Plymouth Rock chickens (WRR) with Xinghua
chickens (XH; Lei et al., 2005). Nine WRR males were
crossed to 9 XH females, and 6 WRR females were crossed
to 6 XH males, producing 17 F1 families and 454 F2 full-
sib individuals (221 males and 233 females). The resource
population was from 6 hatches. Ten fatness and muscle
fiber traits [abdominal fat pad weight (AFW), abdominal
fat pad ratio (AFPR), fat thickness under skin (FTS), fat
width (FW), transversal area of the leg muscle fiber
(TALMF), transversal area of the breast muscle fiber
(TABMF), CP content of the breast muscle (CPCBM), CP
content of the leg muscle (CPCLM), crude fatty content
of the breast muscle (CFCBM), and crude fatty content of
the leg muscle (CFCLM)] were measured.
Two unrelated populations, consisting of 36 XH indi-
viduals and 36 WRR individuals, respectively, were sam-
pled for a genetic diversity investigation in the present
study. The XH and WRR were parents of the F2 resource
population, both from Guangdong Wens Foodstuff Cor-
poration Ltd. (Guangdong, China). The XH was a Chinese
native breed with slow growth rate, and WRR was a
breed with fast growth rate. There were significant differ-
ences in fatness and muscle traits between the XH and
WRR chickens.
SNP Markers from the 8 Growth-Correlated
Genes and Genotyping
Thirty SNP and one 6-bp indel from the 8 growth-
correlated genes (Table 1) were selected to genotype the
838 LEI ET AL.
Table 3. Linkage analyses of 30 single nucleotide polymorphisms and one 6-bp insertion-deletion with chicken fatness and muscle fiber traits

Gene1 Loci AFW2 AFPR FTS FW TALMF TABMF CPCBM CPCLM CFCBM CFCLM
GH G142167T 0.209 0.227 0.023* 0.807 0.669 0.574 0.443 0.55 0.447 0.343
G143831A 0.277 0.371 0.093 0.000** 0.898 0.778 0.82 0.33 0.226 0.205
G143978A 0.789 0.71 0.783 0.148 0.000** 0.297 0.594 0.547 0.757 0.010**
G145086A 0.002** 0.005** 0.622 0.378 0.068 0.788 0.043* 0.585 0.531 0.32
A144762G 0.524 0.318 0.948 0.67 0.000** 0.045* 0.085 0.569 0.632 0.446
GHR G6622190A 0.105 0.121 0.861 0.003 0.322 0.458 0.725 0.994 0.504 0.375
A6626579G 0.496 0.712 0.020* 0.652 0.007** 0.000** 0.024* 0.468 0.833 0.000**
GHSR G18790036A 0.275 0.189 0.801 0.000** 0.853 0.058 0.114 0.566 0.279 0.609
T18791236C 0.932 0.886 0.48 0.662 0.725 0.866 0.759 0.951 0.603 0.308
18792785 to 1879075913 0.034* 0.017* 0.032* 0.482 0.027* 0.285 0.773 0.848 0.376 0.000**
C18793056T 0.835 0.729 0.609 0.826 0.796 0.44 0.74 0.725 0.126 0.763
C18793131T 0.42 0.491 0.427 0.253 0.013* 0.729 0.133 0.717 0.125 0.219
IGF-I C51978309T 0.354 0.521 0.521 0.7 0.000** 0.011* 0.698 0.591 0.459 0.197
C51983354T 0.561 0.618 0.812 0.52 0.249 0.796 0.69 0.303 0.013* 0.94
C51978771T 0.501 0.397 0.865 0.326 0.859 0.85 0.935 0.707 0.689 0.481
C52028084T 0.91 0.814 0.781 0.751 0.308 0.758 0.396 0.000** 0.67 0.001**
IGFBP-2 G23967395T 0.812 0.775 0.267 0.077 0.74 0.755 0.489 0.623 0.457 0.917
T23966786A 0.873 0.853 0.697 0.752 0.902 0.443 0.827 0.644 0.734 0.377
C23966654T 0.33 0.323 0.448 0.437 0.693 0.068 0.71 0.825 0.791 0.935
T23966559G 0.832 0.731 0.21 0.875 0.808 0.755 0.133 0.033* 0.757 0.194
G23966484A 0.787 0.545 0.038* 0.864 0.673 0.446 0.023* 0.007** 0.744 0.237
INS G11303145A 0.672 0.536 0.438 0.524 0.002** 0.016* 0.507 0.268 0.634 0.581
C11304264T 0.729 0.695 0.642 0.729 0.804 0.587 0.428 0.909 0.786 0.609
T11306685C 0.748 0.829 0.315 0.002** 0.782 0.881 0.851 0.834 0.835 0.000**
C11306451T 0.707 0.532 0.531 0.668 0.835 0.943 0.951 0.836 0.614 0.043*
LEPR T28573025C 0.027* 0.197 0.402 0.007** 0.306 0.651 0.784 0.853 0.461 0.582
A28573100G 0.137 0.164 0.618 0.11 0.614 0.669 0.778 0.531 0.434 0.796
TSH-β C2541870G 0.26 0.235 0.277 0.776 0.832 0.745 0.467 0.609 0.759 0.437
C2543216T 0.71 0.661 0.494 0.326 0.501 0.614 0.089 0.929 0.236 0.515
A2543276T 0.513 0.59 0.589 0.612 0.934 0.895 0.549 0.725 0.076 0.379
1
GH = growth hormone; GHR = growth hormone receptor; GHSR = growth hormone secretagogue receptor; IGF-I = insulin-like growth factor-
I; IGFBP-2 = insulin-like growth factor binding protein-2; INS = insulin; LEPR = leptin receptor; and TSH-β = thyroid-stimulating hormone beta
subunit.
2
AFW = abdominal fat pad weight (g); AFPR = abdominal fat pad ratio; FTS = fat thickness under skin (mm); FW fat width = (mm); TALMF =
transversal area of the leg muscle fiber; TABMF = transversal area of the breast muscle fiber; CPCBM = CP content of the breast muscle; CPCLM =
CP content of the leg muscle; CFCBM = crude fatty content of the breast muscle; and CFCLM = crude fatty content of the leg muscle.
3
There was one insertion-deletion GGTACA for 18792785 to 187907591 in the chicken GHSR gene.
*P < 0.05; **P < 0.01.

454 F2, 31 F1, and 30 parental chickens by RFLP and single
strand conformational polymorphism (SSCP).
The PCR was performed in a final volume of 25 ␮L
containing 1 ␮L of genomic DNA (2.5 ng/␮L), 0.25 ␮L
of each primer (25 ␮M), 0.5 ␮L of deoxynucleotide tri-
phosphates (10 ␮M) mixture, 1.5 ␮L of MgCl2 (25 mM),
0.2 ␮L of DNA polymerase (5 U/␮L; Takara, Tokyo, Ja-
pan), and 2.5 ␮L of 10 × reaction buffer on an ABI 2700
(Applied Biosystems, Foster City, CA) Thermal Cycle
with the following profile: initial denaturation at 94°C for
4 min, 35 cycles of 94°C for 30 s, Y°C for 30 s, 72°C for
30 s, and a final elongation at 72°C for 5 min where Y
refers to a different annealing temperature for each
primer (Table 1). Eight-microliter PCR products were di-
gested with 3.0 U of enzyme at 37°C overnight. Restriction
patterns were visualized in a 2 to 4% agarose gel electro-
phoresis stained with ethidium bromide. The A23966559T
and G23966484A SNP were genotyped with SSCP by a
12% polyacrylamide gel electrophoresis.
Statistical Analyses
The difference of allele frequencies between the 2 unre-
lated chicken populations was tested using SAS 8.1 FREQ
(SAS Institute Inc., Cary, NC). The linkage disequilibria
D′ value between each pair of SNP and the haplotype
structure of SNP within each gene were estimated by
Haploview (Daly et al., 2001). Linkage analyses of single
SNP with chicken fatness and muscle fiber traits were
performed with SAGE/SIBPAL package (http://darwin-
.cwru.edu/sage/index.php) (SAGE, 2006).
RESULTS
Allele Frequency in the
Unrelated Populations
Allele frequencies of 30 SNP and one 6-bp indel in
the 2 populations are shown in Table 2. Allele A of the
G18790036A SNP of the GHSR gene, allele A of the
G143831A SNP of the GH gene, allele C of the T28573025C
SNP, and allele G of the A28573100G SNP of the LEPR
gene were all absent in WRR chickens. In the XH and
WRR chickens, allele A of the G11303145A SNP of the
INS gene, allele A of the G6622190A SNP and allele T of
the C6622516T SNP of the GHR gene, and allele A of the
T23966786A SNP and allele G of the T23966559G SNP of
the IGFBP-2 gene were not found.
There was a significant difference for allele frequencies
of C11304264T of the INS gene between the XH and WRR
 GROWTH-CORRELATED GENES AND CHICKEN FATNESS 839
chickens, but there was no difference for the 18792785 to
187907591 indel, and the C18793056T and C18793131T
SNP of the GHSR gene. Highly significant differences
were found for the C2543216T and A2543276T SNP of
the TSH-β gene and the A6626579G SNP of the GHR gene.
No significant difference was observed in the G143978A
SNP of the GH gene, and C51978309T SNP and the
C52028084T SNP of the IGF-I gene (P > 0.05). A highly
significant difference was found in the C23966654T SNP
of the IGFBP-2 gene (P < 0.01).

Linkage Disequilibria of the SNP

in the 8 Growth-Correlated Genes
To further define the haplotype structures of the 8
growth-correlated genes and multiloci association of each
gene, haplotype blocks were analyzed between the XH
and WRR chickens using the Haploview program. Only
2 SNP were genotyped so that haplotype blocks and
multiloci association were not analyzed for the GHR and
LEPR genes. According to the 4-gamete testing, different
haplotype blocks appeared between the XH and WRR
chickens excluding SNP deviation from Hardy-Weinberg
equilibrium separately within each population, but only
the GH and IGFBP-2 genes were interesting. For the
IGFBP-2 gene, there was 1 main block in the XH chickens,
which showed that no recombination was observed in
the C23966654T and G23967395T SNP (D′ = 1), and a
different block appeared in WRR chickens, which showed
that the G23966484A, C23966654T, and G23967395T SNP
were linked (Figure 1). For the IGF-I gene, the G143831A
deviated from Hardy-Weinberg equilibrium in the WRR
chickens, but not in the XH chickens.

Linkage Analyses
Results from the 2-point linkage analyses are shown in
Table 3. There were significant associations of the
G11303145A SNP of the INS gene with TALMF and
TABMF. Positive additive genetic effects were observed
at a highly significant level for the TALMF, and highly
significant negative additive genetic effects were ob-
served for TABMF (Table 4). The A6626579G SNP of the
GHR gene was highly significantly associated with the
fatness and muscle fiber traits (P < 0.01). The G145086A
SNP of the GH gene was related to fatness traits such as
AFW and AFPR. The C51978309T of the IGF-I gene was
related to chicken TALMF and TABMF. The 6-bp indel of
the GHSR gene was significantly associated with fatness
traits such as AFW, AFPR, FTS, and CFCLM. There were
highly significant positive additive effects for the fatness
traits and CFCLM.
DISCUSSION
In the present study, 30 SNP and one 6-bp indel selected
from the 283 SNP in 12 genes of somatotropic axis (Nie
et al., 2005a) were associated with some fatness and mus-
cle fiber traits. Analyses of variance revealed highly sig-
Figure 1. Haplotype structures of the chicken growth hormone (GH)
and insulin-like growth factor binding protein-2 (IGFBP-2) gene, as
estimated by using the Haploview soft package. |D| values × 100 are
shown in the boxes, with empty boxes being 100 (or |D| = 1). A. Haplo-
type structure of the GH gene. The G143831A SNP deviated from Hardy-
Weinberg equilibrium in White Plymouth Rock (WRR) chickens, but
not in the Xinghua (XH) chickens. B. Haplotype structure of the IGFBP-
2 gene. The G23966484A SNP was located in different block in the XH
and WRR chickens.
nificant additive genetic effects. In 14 of the 30 SNP and
one 6-bp indel, there were significant differences for allele
frequencies between the XH and WRR chickens. Such
differences suggested that the 14 SNP could be associated
with chicken fatness and muscle fiber traits. Among the
14 SNP, 8 were linked significantly with one or more
chicken fatness and muscle fiber traits. The G142167T and
G143831A SNP of the GH gene and the G18790036A SNP
of the GHSR gene were associated with 1 trait, such as
FTS, FW, and FW, and the C51983354T SNP of the IGF-
I gene were related with CFCBM. The remaining 4 SNP
840 LEI ET AL.
Table 4. Gene effects on chicken fatness and muscle fiber traits

A6626579G of GHR2 18792785 to 187907591 indel3 of GHSR G11303145A of the INS

Additive Dominance Additive Dominance Additive Additive
Trait1 effect ± SE4 effect ± SE5 effect ± SE effect ± SE effect ± SE effect ± SE

AFW −2.20 ± 0.177 8.20 ± 0.026 29.70 ± 0.187 −4.15 ± 0.018 −5.30 ± 1.017 10.75 ± 0.057
AFPR 0.13 ± 0.013 1.56 ± 0.003 2.11 ± 0.013 −0.29 ± 0.001 −0.18 ± 0.071 0.18 ± 0.004
FTS 0.19 ± 0.015 1.32 ± 0.002 4.76 ± 0.025 −1.58 ± 0.002 −0.98 ± 0.117 −0.05 ± 0.005
FW −0.97 ± 0.033 −1.69 ± 0.004 7.66 ± 0.041 4.82 ± 0.005 −5.79 ± 0.444 −5.61 ± 0.019
TALMF −234.10 ± 75.664 −481.45 ± 20.45 632.70 ± 71.354 −495.75 ± 11.683 3,498.90 ± 577.448 1,992.20 ± 41.03
TABMF 4,521.60 ± 96.305 259.60 ± 22.086 −497.50 ± 71.236 3,368.45 ± 15.401 −2,869.40 ± 764.012 3,465.80 ± 40.698
CPCBM −0.59 ± 0.016 −0.78 ± 0.004 −0.19 ± 0.011 0.71 ± 0.002 −0.27 ± 0.065 0.05 ± 0.007
CPCLM 0.16 ± 0.009 −0.51 ± 0.01 0.18 ± 0.01 0.16 ± 0.003 1.36 ± 0.175 0.09 ± 0.007
CFCBM −0.02 ± 0.006 0.70 ± 0.003 0.81 ± 0.01 0.03 ± 0.001 0.81 ± 0.098 −0.56 ± 0.006
CFCLM −1.03 ± 0.019 0.58 ± 0.007 3.28 ± 0.035 0.50 ± 0.002 2.36 ± 0.191 −2.01 ± 0.019
1
AFW= abdominal fat pad weight (g); AFPR = abdominal fat pad ratio; FTS = fat thickness under skin (mm); FW fat width = (mm); TALMF =
transversal area of the leg muscle fiber; TABMF = transversal area of the breast muscle fiber; CPCBM = CP content of the breast muscle; CPCLM =
CP content of the leg muscle; CFCBM = crude fatty content of the breast muscle; and CFCLM = crude fatty content of the leg muscle.
2
GHR = growth hormone receptor gene; GHSR = growth hormone secretagogue receptor gene; and INS = insulin gene.
3
There was one insertion-deletion GGTACA for 18792785 to 187907591in the chicken GHSR gene.
4
Estimated by subtracting the solution for the BB genotype from that for the AA genotype, A = A and B = G for the A11303145G SNP, and the
A11303145G SNP of the INS gene, and A = A and B = G for the G6626579A SNP of the GHR gene, and A = I and B = D for the 18792785 to
187907591 indel of the GHSR gene.
5
Estimated by subtracting the average of solutions for homozygous genotypes from that for the heterozygous genotype.

were associated with 2 or more traits. The G143978A SNP
of the GH gene was related with TALMF and CFCLM,
and the T28573025C SNP of the LEPR gene was associated
with AFW and FW. The G145086A SNP of the GH gene
was related with AFW, AFPR, and CPCBM. The
A6626579G SNP of the GH gene was associated with FTS,
TALMF, TABMF, CPCBM, and CFCLM. Considering
some associations might be false positives, allele fre-
quency differences partially supported the accuracy of
the association analysis.
It is known that the genes of the growth axis played
crucial roles in the regulation of the growth, development,
and differentiation. There was an important association
of the meat quality and growth and body composition
(Le Bihan-Duval et al., 1998). Therefore, the genes of the
growth axis probably affect the meat quality traits of
the animals. The G143831A (G+1705A) SNP of the GH
gene was significantly associated with growth traits (Nie
et al., 2005b) but was only related with fat width. The
G144762A (G+119A) SNP of the GH gene was signifi-
cantly related to AFW, AFPR, and CPCBM in the present
study, which showed that there was a positive genetic
correlation with BW and meat quality traits (Le Bihan-
Duval et al., 2001; Nie et al., 2005b). The C51978309T SNP
of the IGF-I gene was linked with TALMF and TABMF,
and the correlation (r) of TALMF and TABMF was 0.65.
As an important candidate gene that affected the chicken
muscle cell development and reproduction, the IGF-I gene
was associated with BW, breast weight, and breast yield
(Amills et al., 2003). Myofiber numbers and myofiber den-
sities were related to BW, breast weight, and breast yield
(Scheuermann et al., 2003, 2004), which suggested that
the C51978309T SNP of the IGF-I gene could affect the
chicken muscle fiber growth. There were different haplo-
type structures for the IGFBP-2 gene in the XH and WRR
chickens, which showed the G23966484A (G+738A) SNP
of the IGFBP-2 gene was important. In the present study,
the G23966484A SNP of the IGFBP-2 gene was associated
with CPCBM and CPCLM. This SNP, located in the exon
2, possible affected the expression of the IGFBP-2 gene
at the transcription and translation levels (Lo et al., 2003;
Wyszynska-Koko et al., 2006). These results were consis-
tent with previous results that suggested a potential asso-
ciation of the G23966484A SNP of the IGFBP-2 gene with
growth and carcass traits (Besnard et al., 2001; Lei et al.,
2005). In the present study, the T28573025C SNP of the
LEPR gene was significantly associated with AFW and
FW. Schenkel et al. (2005) found that there was important
association of SNP within the leptin gene with fatness
(fat yield and subcutaneous fat). Ovilo et al. (2005) also
showed that the possible QTL was identified on chromo-
some 8 where the LEPR gene was located. As known,
there was a significant correlation of AFW and FW (r =
0.63). Therefore, association of the T28573025C SNP of
the LEPR gene with fatness possibly was reliable. The
A6626579G SNP of the GHR gene was associated with 5
traits (FTS, TALMF, TABMF, CPCBM, and CFCLM). To
test the accuracy of association of the A6626579G SNP of
the GHR gene with traits, a larger population should
be used.
Recently, some QTL that affected meat quality traits
have been detected in chickens by use of many kinds of
molecular markers. The QTL for AFW were found on
chromosomes 1, 3, 5, 7, 15, and 28 (Ikeobi et al., 2002;
Jennen et al., 2004; Lagarrigue et al., 2006). Some of these
QTL with effect on AFW were located on chromosome
7, which contains the IGFBP-2 gene (Jennen et al., 2004,
2005). Meanwhile, QTL for AFW and percentage abdomi-
nal fat on chromosome 1 where the IGF-I gene is located
were found (Suzuki et al., 2004; Jennen et al., 2005). The
QTL affecting fatness in male chickens were mapped to
less than 8 Mbp at the distal part of the chromosome 5,
which was close to the chicken INS gene (Abasht et al.,
2006). All these showed the locations of underlying QTL
 GROWTH-CORRELATED GENES AND CHICKEN FATNESS
affecting fatness and muscle fiber traits were often
mapped to, or close to, regions harboring candidate func-
tional genes of somatotropic axis. The candidate genes of
the somatotropic axis may affect chicken fatness deposi-
tion and muscle fiber traits.
Linkage disequilibria in the 2 unrelated populations
were analyzed for the 8 genes. Only the GH and IGFBP-
2 genes appeared as haplotype blocks in the XH and WRR
chickens by using Haploview software package (Daly et
al., 2001). For the other genes, no haplotype blocks were
found in both the XH and WRR chickens and, therefore,
fail to exhibit disequilibrium with fatness and muscle
fiber traits. However, haplotype blocks in the XH and
WRR chickens were consistent with the association.
Meanwhile difference for haplotype blocks in the XH
and WRR chickens suggested that difference of the gene
structures could be present between the XH and WRR
chickens and these could affect gene expression level.
In summary, association of 30 SNP and one 6-bp indel
from 8 genes of somatotropic axis with chicken fatness
and muscle fiber traits was analyzed in the present study.
Three genes, GH, GHSR, and LEPR, were significantly
related to the chicken fatness. Two genes, INS and IGF-
I, were linked with the muscle fiber density. In conclusion,
the genes of the somatotropic axis not only affected
chicken growth and body compositions but also were
associated with fatness and muscle fiber traits.
ACKNOWLEDGMENTS
The current work was funded by projects under the
Major State Basic Research Development Program China,
project No. 2006CB102100. Danlin He and Zhijun Peng
gave excellent technical assistance in the observations of
chicken fatness and muscle fiber traits.
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