Scientia Agricultura Sinica ›› 2023, Vol. 56 ›› Issue (3): 572-586.doi: 10.3864/j.issn.0578-1752.2023.03.014

• ANIMAL SCIENCE·VETERINARY SCIENCE • Previous Articles    

Genome-Wide Association Study of Egg Quality Traits in Longyan Shan-Ma Duck

SUN YanFa1,2(), WU Qiong1,2, LIN RuLong3, CHEN HongPing3, GAN QiuYun1, SHEN Yue1, WANG YaRu1, XUE PengFei1, CHEN FeiFan1, LIU JianTao1, ZHOU ChenXin1, LAN ShiShi1, PAN HaoZhe1, DENG Fan1,5, YUE Wen3, JIANG XiaoBing4(), LI Yan1,2()   

  1. 1College of Life Sciences, Longyan University, Longyan 364012, Fujian
    2Fujian Provincial Key Laboratory for the Prevention and Control of Animal Infectious Diseases and Biotechnology/Fujian Provincial Universities Key Laboratory of Preventive Veterinary Medicine and Biotechnology/Longyan University, Longyan 364012, Fujian
    3Longyan Shan-Ma Duck Original Breeding Farm, Agricultural Bureau of Xinluo District, Longyan 364031, Fujian
    4Fujian Provincial Animal Husbandry Headquarters, Fuzhou 350003
    5College of Animal Science (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002
  • Received:2021-10-20 Accepted:2022-11-16 Online:2023-02-01 Published:2023-02-14
  • Contact: JIANG XiaoBing, LI Yan E-mail:boysun2010@163.com;fzjxb@163.com;529783204@qq.com

Abstract:

【Objective】Single nucleotide polymorphisms (SNPs) and candidate genes for egg quality traits in duck were identified through genome-wide association study (GWAS) technology, so as to provide reference for molecular breeding to improve egg quality traits of Longyan Shan-Ma Duck.【Method】Egg quality traits of 235 female Longyan Shan-Ma Duck were measured, including egg weight (EW), egg shaped index (ESI), eggshell thickness (EST), eggshell strength (ESS), eggshell colour L* (ESCL), a* (ESCA), b* (ESCB), albumin height (AH), Haugh unit (HU), egg yolk colour (EYC), egg yolk weight (EYW), and egg yolk percentage relative to egg weight (EYP). Genetic parameters of these traits were estimated using multi-trait animal model by ASReml-R 4.1 software. Blood genomic DNAs of these ducks were genotyped using genotyping-by-sequencing (GBS) technology. The GWAS between egg quality traits of the late laying period and SNPs were performed. 【Result】 The heritability of EW, ESI, EST, ESL, ESA and AU was higher among the egg quality traits of Longyan Shan-Ma Duck, and ranged from 0.21 to 0.70. There was a strong positive genetic correlation (rg = 0.91±0.37) between EW and AU, a strong negative genetic correlation (rg = -0.98±1.03) between ESI and EYC. EST had a positive phenotypic correlation (rp = 0.41±0.06) with ESS, negative genetic and phenotypic correlations with ESA (rg = -0.86±0.25 and rp = -0.15±0.07), and positive genetic and phenotypic correlations with ESB (rg= 0.96±0.37 and 0.18±0.07). There were negative genetic and phenotypic correlations between ESA and ESB (rg = -0.64±0.28 and rp = -0.31±0.06). Results from the GWAS showed that seven SNPs were significantly associated with ESI, EST and yolk color (YC) at 5% Bonferroni-corrected genome-wide significance level (P<4.74×10-6), involving six candidate genes. One SNP, chr20:11135563: G:C, was associated with ESI, which was in leucine rich repeat containing 75A gene, located on chromosome 20. Two SNPs, chr13:5766560:A:G and chrZ:968819:C:T, were associated with EST, which were located on chromosome 13, downstream 6.86 Kb of LOC106014427 and in transcription factor 4 gene, respectively. Four SNPs were associated with EYC, one SNP chr2: 38155965:G:A in potassium voltage-gated channel subfamily H member 8 gene located on chromosome 2; three SNPs located on chromosome 9, two SNPs, chr9:22623156:G:A and chr9:22623155:T:C, in insulin receptor substrate 1 gene, and one SNP, chr9:22490158:A:T, in LOC106018641 gene. Eighty-one SNPs associated with egg quality traits reached at suggestive genome-wide significance level (P<9.48×10-5) were also found. Thirteen SNPs associated with YC were distributed in the 0.84 Mb (22.16-23.00 Mb) region of chromosome 9.【Conclusion】In this study, genetic parameters of egg quality traits of Longyan Shan-Ma Duck were estimated. Seven significant SNPs, six candidate genes, and one candidate region affecting ESI, EST and EYC traits were identified through GWAS. The findings from the present study provided a reference for the molecular breeding of egg quality traits in Longyan Shan-Ma Duck.

Key words: Longyan Shan-Ma Duck, egg quality traits, genetic parameter, SNPs, GWAS

Table 1

Descriptive statistics of egg quality traits"

性状
Trait
例数
N
平均值
Mean
最小值
Min
最大值
Max
标准差
SD
变异系数
CV(%)
蛋重EW (g) 235 69.70 51.30 84.30 5.28 7.57
蛋形指数ESI 235 1.33 1.15 1.50 0.06 4.52
蛋壳厚EST (mm) 235 0.33 0.20 0.44 0.04 10.68
蛋壳强度ESS (kg·cm-2) 233 3.40 0.61 6.00 1.04 30.68
蛋壳颜色L* 值ESCL 235 78.11 65.11 86.29 2.86 3.67
蛋壳颜色a*值ESCA 234 -3.04 -8.44 -0.53 1.52 -50.17
蛋壳颜色b*值ESCB 234 4.81 1.37 13.63 1.66 34.58
蛋白高度 AH (mm) 235 6.36 1.32 20.80 3.33 9.85
哈氏单位HU 235 74.84 38.22 95.47 10.70 14.29
蛋黄颜色EYC 235 7.37 3.00 13.00 1.22 16.52
蛋黄重 EYW (g) 232 24.14 11.70 37.30 3.70 15.33
蛋黄比例 EYP (%) 232 34.77 17.06 57.91 5.52 15.87

Table 2

Genetic parameter estimation of egg quality traits"

蛋重
EW
蛋形指数
ESI
蛋壳厚
EST
蛋壳强度
ESS
蛋壳颜色L*值
ESCL
蛋壳颜色a*值
ESCA
蛋壳颜色b*值
ESCB
蛋白高度
AH
哈氏单位
HU
蛋黄颜色
EYC
蛋黄重
EYW
蛋黄比例
EYP
蛋重
EW
0.21±
0.16
- - - - - - 0.91±
0.37*
- - - -
蛋形指数
ESI
- 0.29±
0.17
- - - - - - - -0.98±
1.03*
- -
蛋壳厚
EST
- - 0.41±
0.17
- - -0.86±
0.25**
0.96±
0.37**
- - - - -
蛋壳强度
ESS
- - 0.41±
0.06***
0.01±
0.10
- - - - - - - -
蛋黄颜色L*值
ESCL
- - - - 0.37±
0.18
- - - - - - -
蛋黄颜色a*值
ESCA
- -0.16±
0.07*
-0.15±
0.07**
- 0.25±
0.07***
0.70±
0.20
-0.64±
0.28*
- - - - -
蛋黄颜色b*值
ESCB
- - 0.18±
0.07**
0.18±
0.06*
-0.31±
0.06***
-0.31±
0.06***
0.16±
0.14
- - - - -
蛋白高度
AH
0.04±
0.07*
- - 0.20±
0.06**
- -0.09±
0.07*
- 0.24±
0.18
- - - -
哈氏单位
HU
-0.18±
0.07***
- - 0.22±
0.06**
- -0.12±
0.07*
- - 0.08±
0.14
- -
蛋黄颜色
EYC
-0.10±
0.07*
0.07±
0.06*
- - - - - - 0.18±
0.06**
0.07±
0.14
- -
蛋黄重
EYW
0.24±
0.06***
- - - - - - - - - 0.01±
0.12
-
蛋黄比例
EYP
-0.26±
0.06***
- - - - - - - - - 0.87±
0.02***
0.08±
0.12

Table 3

Distribution of after quality control and independent SNP markers by chromosome"

染色体
Chromosome
SNPs数量
No. of SNPs
独立SNPs数量
No. of independent SNPs
染色体
Chromosome
SNPs数量
No. of SNPs
独立SNPs数量
No. of independent SNPs
1 9911 1523 16 1132 172
2 7488 1250 17 30 9
3 6116 988 18 1420 230
4 4026 644 19 989 149
5 4252 706 20 1546 251
6 2311 435 21 1382 242
7 2120 335 22 1072 177
8 2479 402 23 278 63
9 2163 407 24 687 122
10 1242 202 25 974 186
11 2177 369 26 134 48
12 1028 152 27 819 189
13 2066 377 28 766 173
14 1607 307 29 506 80
15 1545 240 Z 440 119
总计Total 62706 10547

Fig. 1

Principal component analysis plot of population structure"

Table 4

Bonferroni-corrected 5% genome-wise significant SNPs"

性状
Trait
染色体
Chromosome
标记号
SNP ID
物理位置
Position (bp)
A1
A1
BETA值
BETA
P
P value
最近的基因
Nearest gene
蛋形指数ESI 20 chr20:11135563:G:C 11135563 C -0.05 1.48×10-6 within LRRC75A
蛋壳厚EST 13 chr13:5766560:A:G 5766560 A 0.02 1.36×10-6 6.86 kb D LOC106014427
Z chrZ:968819:C:T 968819 T -0.02 1.96×10-6 within TCF4
蛋黄颜EYC 2 chr2:38155965:G:A 38155965 A 1.05 1.97×10-6 within KCNH8
9 chr9:22490158:A:T 22490158 T -0.97 3.77×10-6 within LOC106018641
9 chr9:22623155:T:C 22623155 C -0.92 1.91×10-6 within IRS1
9 chr9:22623156:G:A 22623156 A -1.08 8.75×10-7 within IRS1

Table 5

SNPs significance at genome-wise level suggestive significant association"

性状
Trait
染色体
Chromosome
标记号
SNP ID
物理位置
Position (bp)
A1
A1
BETA值
BETA
P
P value
最近的基因
Nearest gene
蛋重EW 2 chr2:10782468:A:G 10782468 G 3.45 2.81×10-5 U 120.07 kb ADARB2
2 chr2:10790319:T:C 10790319 T 3.31 5.81×10-5 U 112.21 kb ADARB2
5 chr5:5547457:T:C 5547457 T 2.89 3.36×10-5 U 63.45 kb MIS18BP1
5 chr5:5547530:A:G 5547530 G 2.80 6.33×10-5 U 63.38 kb MIS18BP1
蛋形指数ESI 3 chr3:81386471:A:G 81386471 A -0.05 6.66×10-5 D 13.01 NT5E
5 chr5:7009743:G:A 7009743 G 0.06 2.77×10-5 within CDKL1
15 chr15:11227397:T:C 11227397 C 0.05 6.39×10-5 within AXIN1
15 chr15:11227409:G:A 11227409 A 0.05 7.59×10-5 within AXIN1
15 chr15:13294556:C:T 13294556 T 0.05 2.39×10-5 within UBN1
15 chr15:13395347:A:G 13395347 A 0.05 3.49×10-5 within LOC101799012
15 chr15:14536844:G:A 14536844 A 0.05 1.02×10-5 U 41.78 kb RBFOX1
16 chr16:2579629:A:T 2579629 T -0.04 5.07×10-5 within EP400
16 chr16:3098786:G:A 3098786 A -0.04 1.97×10-5 D 22.42 kb LOC110352440
25 chr25:6315960:T:C 6315960 C -0.06 5.29×10-5 D 26.17 kb LOC106020386
蛋壳厚度EST 2 chr2:22513511:C:A 22513511 A 0.03 2.51×10-5 within ZNF804B
2 chr2:22582514:A:G 22582514 G 0.03 4.43×10-5 within ZNF804B
2 chr2:22582670:G:A 22582670 A 0.03 6.51×10-5 within ZNF804B
6 chr6:7192663:G:A 7192663 A 0.03 6.46×10-5 U 18.98 kb KCNK18
6 chr6:12756206:T:A 12756206 A 0.02 7.06×10-5 within SH3PXD2A
12 chr12:140915:T:A 140915 A -0.02 9.44×10-5 within THSD4
12 chr12:198263:G:A 198263 A -0.02 5.31×10-5 within THSD4
13 chr13:5721234:C:T 5721234 T 0.02 7.27×10-5 within LOC106014427
19 chr19:4873329:A:G 4873329 G 0.02 2.69×10-5 within LOC113845541
蛋壳强度ESS 5 chr5:33699890:A:T 33699890 T 0.66 6.76×10-5 within LOC113843881
蛋壳颜色L*值
ESCL
7 chr7:3182113:T:A 3182113 A -3.45 6.81×10-5 within CTR9
7 chr7:3204268:G:A 3204268 A -3.31 6.08×10-5 U 0.65 kb EIF4G2
7 chr7:3204312:A:G 3204312 G -3.34 6.05×10-5 U 0.60 kb EIF4G2
7 chr7:3223217:A:G 3223217 G -3.55 1.05×10-5 U 14.01 kb LOC113844171
7 chr7:3223320:T:A 3223320 A -3.37 9.92×10-6 U 13.91 kb LOC113844171
14 chr14:9802965:A:G 9802965 G 2.04 5.95×10-5 within CLINT1
蛋壳颜色a*值
ESCA
1 chr1:160744676:G:C 160744676 G -1.09 9.13×10-5 D 3.53 kb PHF5A
1 chr1:174907611:C:T 174907611 T -0.89 5.86×10-5 U 19.87 kb GPR85
4 chr4:6332903:C:G 6332903 G -0.93 5.91×10-5 U 6.34 kb MANBA
8 chr8:10668874:C:A 10668874 A -0.97 8.83×10-5 D 3.43 kb LOC113844346
蛋壳颜色 b*值
ESCB
1 chr1:59043295:C:T 59043295 T -1.54 7.39×10-5 U 142.06 kb ARHGEF7
1 chr1:59706095:T:C 59706095 C 1.03 6.95×10-5 D 3.76 kb COL4A2
3 chr3:16497369:T:C 16497369 C -1.19 2.36×10-5 U 13.30 kb VSX1
4 chr4:7424076:C:G 7424076 G 1.83 6.35×10-5 within RAP1GDS1
8 chr8:33254427:C:T 33254427 T 1.20 3.93×10-5 within SLC30A7
蛋白高度AU 2 chr2:102420094:T:G 102420094 G -0.89 5.83×10-5 U 14.46 kb RBBP8
4 chr4:5482597:A:C 5482597 C 1.49 4.59×10-5 D 21.57 kb LOC106017184
4 chr4:71726051:A:G 71726051 G 0.90 2.81×10-5 within LOC106017926
7 chr7:7414460:C:T 7414460 T 0.80 3.39×10-5 D 1.45 kb LOC106019248
20 chr20:6093764:T:C 6093764 T 1.30 4.41×10-5 U 5.58 kb SGSM2
20 chr20:10720431:A:G 10720431 G 1.01 3.37×10-5 U 15.28 kb RTN4RL1
哈氏单位HU 1 chr1:111329343:T:C 111329343 C 12.09 8.38×10-5 within TAGLN3
4 chr4:75257160:A:C 75257160 A -8.15 3.42×10-5 U 91.88 kb LOC110353528
7 chr7:7414460:C:T 7414460 T 6.42 5.32×10-5 D 1.45 kb LOC106019248
20 chr20:6093764:T:C 6093764 T 10.22 8.41×10-5 U 5.58 kb SGSM2
20 chr20:10720431:A:G 10720431 G 8.18 4.19×10-5 U 15.28 kb RTN4RL1
蛋黄颜色EYC 1 chr1:196541696:G:T 196541696 T 0.75 5.41×10-5 within CAMK1D
2 chr2:2433073:A:G 2433073 G 0.94 2.65×10-5 within GJC2
2 chr2:37584936:A:G 37584936 G 1.01 1.99×10-5 D 60.71 kb SATB1
2 chr2:37585118:C:T 37585118 T 0.77 6.42×10-5 D 60.53 kb SATB1
2 chr2:57679783:T:C 57679783 C 0.71 6.82×10-5 within GLI3
2 chr2:57679906:A:G 57679906 G 0.70 9.04×10-5 within GLI3
2 chr2:115153719:T:C 115153719 C -0.97 5.50×10-5 within ZNF521
2 chr2:115178457:C:T 115178457 T -1.10 5.57×10-5 within ZNF521
3 chr3:51410494:C:T 51410494 T -0.80 5.15×10-5 within RMND1
3 chr3:51410551:A:G 51410551 G -0.80 5.15×10-5 within RMND1
6 chr6:6402767:G:A 6402767 A -0.85 8.86×10-5 within CACUL1
7 chr7:10854039:A:G 10854039 G -1.28 6.98×10-5 within MAP2
9 chr9:22162962:T:G 22162962 G -1.08 4.62×10-5 within NYAP2
9 chr9:22163003:A:G 22163003 G -1.07 5.51×10-5 within NYAP2
9 chr9:22163123:T:C 22163123 C -1.07 5.51×10-5 within NYAP2
9 chr9:22490118:C:T 22490118 C -0.96 6.18×10-6 within LOC106018641
9 chr9:22490249:A:G 22490249 A -0.83 2.13×10-5 within LOC106018641
9 chr9:22628656:T:C 22628656 C -0.97 5.97×10-5 within IRS1
9 chr9:22636905:C:T 22636905 T -1.04 4.55×10-5 within IRS1
9 chr9:22798030:T:C 22798030 C -0.92 1.94×10-5 within COL4A3
9 chr9:22841648:T:C 22841648 C -0.95 6.18×10-5 within COL4A3
9 chr9:23005131:A:G 23005131 G -1.04 7.05×10-5 within DAW1
14 chr14:1139032:G:A 1139032 A -0.90 2.55×10-5 within ZIC4
24 chr24:3527498:A:G 3527498 A 0.74 7.73×10-5 within EPB41
蛋黄重EYW 2 chr2:2865193:A:G 2865193 A -2.66 6.56×10-5 U 2.17 kb SEC22C
9 chr9:11219039:A:G 11219039 G 2.15 9.42×10-5 within TP63
9 chr9:11360616:G:A 11360616 A 2.27 4.28×10-5 within TP63
9 chr9:11360684:A:G 11360684 G 2.27 4.43×10-5 within TP63
蛋黄比例EYP 2 chr2:129285222:C:T 129285222 T -6.95 6.53×10-5 within RMDN1
5 chr5:8855150:G:A 8855150 A 5.41 6.83×10-5 D 19.17 kb LOC110352963
6 chr6:708965:T:C 708965 C 6.13 2.71×10-5 within DPYSL4

Fig. 2

Manhattan plot of genome-wide study association study for egg shape index The x axis represents the genomic position of SNP, and the y axis represents -log10-tansformed P value of genome-wide association study. Each dot represents one SNP. The red line represents the Bonferroni-corrected 5% genome-wise significant threshold line at -log10 (4.74×10-6). The black dotted line indicates the genome-wise suggestive association threshold line at -log10 (9.48×10-5). The same as below"

Fig. 3

Manhattan plot of genome-wide study association study for eggshell thickness SNPs"

Fig. 4

Manhattan plot of genome-wide study association study for yolk color"

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