培育品系中鸡青胫性状的遗传规律分析及分子基础初探

doi:10.11843/j.issn.0366-6964.2022.06.007

摘要/Abstract

摘要： 深受中国消费者青睐的青胫性状受隐性伴性基因或常染色体显性基因控制。为探明培育品系W系青胫性状的遗传规律及分子基础，本研究首先选取W系黑羽青胫与洛岛红黄羽黄胫的成熟公、母鸡(1♂∶5♀)进行正反交试验，然后对后代中青、黄胫母雏各4只的胫部皮肤进行转录组测序分析。结果表明，正交后代有青胫293只(♂172只，♀121只)，黄胫51只(♂20只，♀31只); 反交后代有青胫256只(♂156只，♀100只)，黄胫73只(♂29只，♀44只)，正或反交后代均有两种胫色，且青胫显著多于黄胫，表明W系青胫性状属于常染色体显性遗传。将有胫色分离的杂交组合后代按胫色、羽色统计，青、黄胫个体分别为黑、黄羽，青、黄胫分离比符合1∶1(121∶117)，因此青、黄胫性状或黑、黄羽性状可能受1对等位基因控制。雏鸡的胫部皮肤转录组测序分析表明，差异表达基因显著富集于黑色素生成通路(P < 0.01)，Mc1r基因和已知参与黑色素合成的基因如Tyr、Tyrp1在青、黄胫皮肤中的表达量存在极显著差异。另外，一些尚无报道参与胫色形成的基因如Wnt16、Wnt3a、Fzd10等也存在极显著差异。青胫性状的形成除涉及显著富集的黑色素生成通路外，还涉及细胞外基质与受体信号、信号传导、细胞骨架与迁移、细胞黏附、鞘脂与糖脂代谢。综合遗传分析与转录组分析，本研究推定培育品系W系中的青胫性状受Mc1r基因控制，呈常染色体显性遗传，青胫性状的形成涉及多个信号通路，这为青胫性状形成机制的阐明奠定了基础。

关键词: 鸡, 胫色, 显性遗传, 常染色体遗传, Mc1r

Abstract: The black shank trait favored by Chinese consumers is controlled by recessive sex-related gene or autosomal dominant gene. To explore the inheritance and molecular basis of black shank trait in a cultivated W strain, this study first conducted reciprocal crossing test between mature male and female chickens (1♂∶5♀) from the W line (black feather black shank) and the Rhode Island Red line (yellow feather yellow shank), followed by transcriptome analysis on the shank skin samples collected from the female offspring (4 chicks with black shank and 4 chicks with yellow shank) of the reciprocal crosses. The results showed that there were 293 chicks with black shank (♂172, ♀121) and 51 chicks with yellow shank (♂20, ♀31) in the cross of the W line with the Rhode Island Red line, there were 256 chicks with black shank (♂156, ♀100) and 73 chicks with yellow shank (♂29, ♀44) in the reciprocal cross, the offspring of both crosses had two shank colors, and the number of the chicks with black shank was significantly higher than that with yellow shank in each cross, indicating that the black shank trait of the W line belongs to autosomal dominant inheritance. Based on the statistics of shank color and feather color in the offspring of the breeding groups with shank color separation, it was found that the chicks with black shank had black feathers and the chicks with yellow shank had yellow feathers, the number of the chicks with black shank to that with yellow shank (121∶117) conformed to the separation ratio of 1∶1, suggesting that the black vs. yellow shank traits in the cultivated strain were controlled by a pair of alleles. The transcriptome analysis of shank skin samples showed that the differentially expressed genes (DEGs) were significantly enriched in Melanogenesis pathway (P < 0.01), the expression of Mc1r and the genes, such as Tyr and Tyrp1, which are known to be involved in the regulation of melanin synthesis was significantly different between black and yellow shank skin samples, and some other genes, such as Wnt16, Wnt3a, Fzd10, that are not previously reported to be involved in shank color formation, also showed significant difference. In addition to the Melanogenesis pathway, black shank formation was also involved in extracellular matrix and receptor signaling, signal transduction, cell cytoskeleton and migration, cell adhesion, sphingolipid and glycolipid metabolisms. In conclusion, based on genetic analysis and transcriptome analysis, this study inferred that the black shank trait in the cultivated W strain was regulated by Mc1r gene, was inherited in autosomal dominant mode, the formation of black shank trait involved multiple signaling pathways, which laid a foundation for clarifying the mechanism underlying formation of black shank trait.

Key words: chicken, shank color, dominant inheritance, autosomal inheritance, Mc1r

中图分类号:

S831.2

赵超, 徐尚立, 李硕, 穆继安, 李方博, 张瑾麒, 赵敏孟, 刘龙, 龚道清, 耿拓宇. 培育品系中鸡青胫性状的遗传规律分析及分子基础初探[J]. 畜牧兽医学报, 2022, 53(6): 1723-1734.

ZHAO Chao, XU Shangli, LI Shuo, MU Ji'an, LI Fangbo, ZHANG Jinqi, ZHAO Minmeng, LIU Long, GONG Daoqing, GENG Tuoyu. Genetic Analysis and Molecular Basis of Black Shank Trait in Cultivated Strain of Chicken[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(6): 1723-1734.

参考文献 34

[1]	ZHOU X. Study of model of gene regulation of shank color conversion[D]. Wuhan: Huazhong Agricultural University, 2020. (in Chinese)周鑫. 胫色转化的基因调控模式研究[D]. 武汉: 华中农业大学, 2020.
[2]	SUN G R, LI G X, KANG X T, et al. Genetic analysis of shank color in Gushi and Anka chickens[J]. Chinese Journal of Animal Science, 2010, 46(19): 26-29. (in Chinese)孙桂荣, 李国喜, 康相涛, 等. 固始鸡与安卡鸡资源群胫色遗传分析[J]. 中国畜牧杂志, 2010, 46(19): 26-29.
[3]	WAKAMATSU K, ZIPPIN J H, ITO S. Chemical and biochemical control of skin pigmentation with special emphasis on mixed melanogenesis[J]. Pigm Cell Melanoma Res, 2021, 34(4): 730-747.
[4]	TSUKAMOTO K, JACKSON I J, URABE K, et al. A second tyrosinase-related protein, TRP-2, is a melanogenic enzyme termed DOPAchrome tautomerase[J]. EMBO J, 1992, 11(2): 519-526.
[5]	LIU Q, JIANG Y, HE Z Y, et al. Advances in research on the effect of tyrosinase on melanin synthesis in silky fowl[J]. Heilongjiang Animal Science and Veterinary Medicine, 2018(11): 93-95. (in Chinese)刘青, 江彦, 何资颖, 等. 酪氨酸酶对乌骨鸡黑色素合成影响的研究进展[J]. 黑龙江畜牧兽医, 2018(11): 93-95.
[6]	BATESON W, PUNNETT R C. On gametic series involving reduplication of certain terms[J]. J Genet, 1911, 1(4): 293-302.
[7]	KNOX C W. The inheritance of shank color in chickens[J]. Genetics, 1935, 20(6): 529-544.
[8]	MCGIBBON W H. Green shanks and adult mortality in chickens[J]. J Hered, 1979, 70(1): 44-46.
[9]	DORSHORST B, MOLIN A M, RUBIN C J, et al. A complex genomic rearrangement involving the Endothelin 3 locus causes dermal hyperpigmentation in the chicken[J]. PLoS Genet, 2011, 7(12): e1002412.
[10]	ARORA G, MISHRA S K, NAUTIYAL B, et al. Genetics of hyperpigmentation associated with the Fibromelanosis gene (Fm) and analysis of growth and meat quality traits in crosses of native Indian Kadaknath chickens and non-indigenous breeds[J]. Br Poult Sci, 2011, 52(6): 675-685.
[11]	SHINOMIYA A, KAYASHIMA Y, KINOSHITA K, et al. Gene duplication of endothelin 3 is closely correlated with the hyperpigmentation of the internal organs (Fibromelanosis) in silky chickens[J]. Genetics, 2012, 190(2): 627-638.
[12]	LI H, QIU X P, LONG J R. Genetic study about the color of feather and skin on the Black-bone Fowls[J]. Chinese Journal of Animal Science, 2002, 38(6): 45-46. (in Chinese)李华, 邱祥聘, 龙继蓉. 乌骨鸡羽色及肤色的遗传研究现状及展望[J]. 中国畜牧杂志, 2002, 38(6): 45-46.
[13]	KERJE S, LIND J, SCHVTZ K, et al. Melanocortin 1-receptor (MC1R) mutations are associated with plumage colour in chicken[J]. Anim Genet, 2003, 34(4): 241-248.
[14]	TAKEUCHI S, SUZUKI H, HIROSE S, et al. Molecular cloning and sequence analysis of the chick melanocortin 1-receptor gene[J]. Biochim Biophys Acta Gen Subjects, 1996, 1306(2-3): 122-126.
[15]	DROZDZ M M, WANG J, PISKOUNOVA E, et al. A nuclear cAMP microdomain functions as a tumor suppressor in melanoma[J]. J Invest Dermatol, 2019, 139(5 Suppl 1): S146.
[16]	KIJAS J M H, WALES R, TÖRNSTEN A, et al. Melanocortin receptor 1 (MC1R) mutations and coat color in pigs[J]. Genetics, 1998, 150(3): 1177-1185.
[17]	LIU X N, JING J J, LI B J, et al. Advance in the research of ASIP and MC1R Genes[J]. China Herbivore Science, 2015, 35(1): 42-46. (in Chinese)刘晓妮, 景炅婕, 李宝钧, 等. ASIP和MC1R基因的研究进展[J]. 中国草食动物科学, 2015, 35(1): 42-46.
[18]	BOROVANSKÝ J, RILEY P A. Melanins and melanosomes: biosynthesis, biogenesis, physiological, and pathological functions[M]. Weinheim, Germany: Wiley-VCH Verlag GmbH & Co. KGaA, 2011.
[19]	ZU P Y, LI W, LIN J D, et al. Polymorphism and bioinformatics analysis of MC1R gene in Chishui Black-Bone chickens[J]. Journal of Hunan Agricultural University: Natural Sciences, 2020, 46(1): 84-92. (in Chinese)祖盘玉, 李维, 林家栋, 等. 赤水乌骨鸡MC1R基因多态性及其生物信息学分析[J]. 湖南农业大学学报: 自然科学版, 2020, 46(1): 84-92.
[20]	GUO X L, LI X L, LI Y, et al. Genetic variation of chicken MC1R gene in different plumage colour populations[J]. Br Poult Sci, 2010, 51(6): 734-739.
[21]	HU Z C, MENG Q D, ZHANG M W, et al. Sequence analysis of MC1R gene in Hetian chicken and White-Velvet Silky chicken[J]. Heilongjiang Animal Science and Veterinary Medicine, 2019, (17): 29-33, 183-184. (in Chinese)胡卓成, 孟庆铎, 张梦文, 等. 河田鸡与白绒乌骨鸡MC1R基因序列分析[J]. 黑龙江畜牧兽医, 2019, (17): 29-33, 183-184.
[22]	XI S W, ZHOU M F, CHENG D, et al. The variation and haplotype of MC1R gene and its effect on the plumage coloration of indigenous chicken breeds in Jiangxi province, China[J]. Acta Agriculturae Universitatis Jiangxiensis, 2021, 43(2): 378-388. (in Chinese)席苏望, 周明芳, 成笛, 等. 探究MC1R基因在江西地方鸡中的变异、单倍型及其对羽色的影响[J]. 江西农业大学学报, 2021, 43(2): 378-388.
[23]	GUO X L. Study on selection breeding and variation of MCIR gene in Hebei Domestic Chicken[D]. Baoding: Hebei Agricultural University, 2009. (in Chinese)郭秀丽. 河北柴鸡MC1R基因变异研究[D]. 保定: 河北农业大学, 2009.
[24]	TIAN M. Gene mapping and candidate gene expression analysis for dermal hyperpigmentation loci in the chicken[D]. Beijing: China Agricultural University, 2013. (in Chinese)田明. 鸡黑色素过度沉积性状基因的定位与候选基因的表达分析研究[D]. 北京: 中国农业大学, 2013.
[25]	NAM I S, OH M G, NAM M S, et al. Specific mutations in the genes of MC1R and TYR have an important influence on the determination of pheomelanin pigmentation in Korean native chickens[J]. J Adv Vet Anim Res, 2021, 8(2): 266-273.
[26]	SCHWOCHOW D, BORNELÖV S, JIANG T, et al. The feather pattern autosomal barring in chicken is strongly associated with segregation at the MC1R locus[J]. Pigm Cell Melanoma Res, 2021, 34(6): 1015-1028.
[27]	KABIR M H, TAKENOUCHI A, HAQANI M I, et al. Discovery of a new nucleotide substitution in the MC1R gene and haplotype distribution in native and non-Japanese chicken breeds[J]. Anim Genet, 2020, 51(2): 235-248.
[28]	EDELMAN A M, BLUMENTHAL D K, KREBS E G. Protein serine/threonine kinases[J]. Annu Rev Biochem, 1987, 56: 567-613.
[29]	D'MELLO S A N, FINLAY G J, BAGULEY B C, et al. Signaling pathways in melanogenesis[J]. Int J Mol Sci, 2016, 17(7): 1144.
[30]	GAJOS-MICHNIEWICZ A, CZYZ M. WNT signaling in melanoma[J]. Int J Mol Sci, 2020, 21(14): 4852.
[31]	XU Y Y, WANG C, HE Z, et al. MAGEA1 promotes the metastasis of malignant melanoma by activating Wnt/β-catenin signaling pathway[J]. Academic Journal of Chinese PLA Medical School, 2021, 42(2): 182-188, 196. (in Chinese)许媛媛, 王晨, 贺赞, 等. MAGEA1通过激活Wnt/β-catenin信号通路促进恶性黑色素瘤的转移[J]. 解放军医学院学报, 2021, 42(2): 182-188, 196.
[32]	DORSKY R I, RAIBLE D W, MOON R T. Direct regulation of nacre, a zebrafish MITF homolog required for pigment cell formation, by the Wnt pathway[J]. Gene Dev, 2000, 14(2): 158-162.
[33]	DORSKY R I, MOON R T, RAIBLE D W. Control of neural crest cell fate by the Wnt signalling pathway[J]. Nature, 1998, 396(6709): 370-373.
[34]	DUNN K J, BRADY M, OCHSENBAUER-JAMBOR C, et al. WNT1 and WNT3a promote expansion of melanocytes through distinct modes of action[J]. Pigm Cell Res, 2005, 18(3): 167-180.