卵形鲳鲹生肌调节因子基因家族的鉴定及在胚胎中的表达

doi:10.3969/j.issn.1004-1524.2022.04.05

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (4): 695-705.DOI: 10.3969/j.issn.1004-1524.2022.04.05

卵形鲳鲹生肌调节因子基因家族的鉴定及在胚胎中的表达

余艳玲¹(), 罗洪林¹, 罗辉², 冯鹏霏¹, 潘传燕¹, 宋漫玲¹, 肖蕊¹, 张永德¹^,^*()

1.广西水产科学研究院广西水产遗传育种与健康养殖重点实验室,广西南宁 530021
2.西南大学动物科学学院,重庆 400715

收稿日期:2020-10-13 出版日期:2022-04-25 发布日期:2022-04-28
作者简介:*张永德,E-mail: yondar@126.com
余艳玲(1976—),女,陕西咸阳人,硕士,工程师,主要从事水产养殖研究。E-mail: 553850756@qq.com
通讯作者: 张永德
基金资助:
广西创新驱动发展专项资金(桂科AA17204080-3);广西创新驱动发展专项资金(桂科AA18242031-2);广西水产遗传育种与康养殖重点实验室自主课题(17-A-01-02);广西水产遗传育种与康养殖重点实验室自主课题(19-A-01-05)

Genome-wide identification and expression of MRF gene family in embryonic development of Trachinotus ovatus

YU Yanling¹(), LUO Honglin¹, LUO Hui², FENG Pengfei¹, PAN Chuanyan¹, SONG Manling¹, XIAO Rui¹, ZHANG Yongde¹^,^*()

1. Guangxi Key Laboratory of Aquatic Genetic Breeding and Healthy Aquaculture, Guangxi Academy of Fishery Sciences, Nanning 530021, China
2. College of Animal Science, Southwest University, Chongqing 400715, China

Received:2020-10-13 Online:2022-04-25 Published:2022-04-28
Contact: ZHANG Yongde

摘要/Abstract

摘要：

为研究卵形鲳鲹生肌调节因子(MRF)在胚胎发育中所发挥的重要作用,对卵形鲳鲹MRF基因家族进行了全基因组鉴定和生物信息学分析,并对其在13个胚胎发育阶段的基因表达进行了定量分析。结果表明:在卵形鲳鲹基因组中共鉴定出5个MRF基因家族成员:MyoD1、MyoD2、Myf5、Myf6和MyoG,分别编码297、263、240、231、250个氨基酸。MRF家族基因存在典型的BASIC与HLH结构域,其中MyoD1、MyoD2与MyoG分别定位于9号、1号与2号染色体,而Myf5与Myf6基因均定位于22号染色体且位于同一基因座。MRF家族基因胚胎定量表达研究表明,MyoG在受精卵到胚体形成期表达量极低,在眼囊、耳囊与心脏跳动期表达量迅速升高,而到晶体出现期表达量又有所降低,推测其可能在眼囊、耳囊与心脏跳动期发挥主要的生肌调节作用,而后期在维持肌肉形态或肌肉发育中仍发挥重要作用。MyoD1、MyoD2、Myf5与Myf6在受精卵到原肠中期表达量较低,而从原肠末期到晶体出现期表达量迅速升高(P<0.05),表明其主要从原肠末期开始发挥生肌调控作用。结果表明,MRF家族成员在胚胎发育过程中发挥重要的作用,但不同的MRF家族成员发挥作用的时间及功能可能不同。

关键词: 卵形鲳鲹, 生肌调节因子(MRF), MRF基因家族, 基因表达, 胚胎发育

Abstract:

In order to study the important role of myogenic regulator (MRF) in the embryonic development of Trachinotus ovatus, the whole genome identification and bioinformatics analysis of MRF gene family in T. ovatus were carried out, and the gene expression in 13 embryonic stages was quantitatively analyzed. Five MRF gene family members, MyoD1, MyoD2, Myf5, Myf6 and MyoG, encoding 297, 263, 240, 231 and 250 amino acids, respectively, were identified in T. ovatus genome data. The MRF family genes had a typical BASIC and HLH domains, where MyoD1, MyoD2 and MyoG were located on chromosome 9, 1 and 2 respectively, while Myf5 and Myf6 were located on chromosome 22 and at the same locus. Quantitative expression studies in embryos showed that the expression level of MyoG was very low during the period from zygote to embryo formed stage, while increased rapidly during the optic vesicle stage, otocyst vesicle stage and heart pulsation stage, but decreased at the formation of eye lens stage. It was speculated that MyoG may play a major role in the regulation of myogenesis during optic vesicle stage, otocyst vesicle stage and heart pulsation stage, but still play an important role in maintaining muscle morphology or muscle development in the later period. The expression levels of MyoD1, MyoD2, Myf5 and Myf6 genes were lower from the zygote to the mid-gastrulation stage, but increased rapidly from the late gastrulation to the formation of eye lens stage (P<0.05), indicating that they began to play an important role in the regulation of myogenesis from late gastrulation stage. The results showed that MRF family members played an important role in the process of embryonic development of T. ovatus, but different MRF family members may play different roles in different time and functions.

Key words: Trachinotus ovatus, myogenic regulatory factors (MRF), MRF gene family, gene expression, embryo

中图分类号:

S917.4

余艳玲, 罗洪林, 罗辉, 冯鹏霏, 潘传燕, 宋漫玲, 肖蕊, 张永德. 卵形鲳鲹生肌调节因子基因家族的鉴定及在胚胎中的表达[J]. 浙江农业学报, 2022, 34(4): 695-705.

YU Yanling, LUO Honglin, LUO Hui, FENG Pengfei, PAN Chuanyan, SONG Manling, XIAO Rui, ZHANG Yongde. Genome-wide identification and expression of MRF gene family in embryonic development of Trachinotus ovatus[J]. Acta Agriculturae Zhejiangensis, 2022, 34(4): 695-705.

图/表 10

参考文献 37

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引物名称Primer	上游引物Forward primer (5'-3')	下游引物Reverse primer (5'-3')
MyoD1	AACGCCATCAGCTACATCGAG	ACAGGTGGGGCCGTTAAAATC
MyoD2	CGGCCTACTGAAGCCCGAA	CTCATGGTTGCTGCTTTTCTCC
Myf5	AAGAACGAGAGTTTGGGCGA	AGGACGTGGTATATGGGCCT
Myf6	TCAGCTACATCGAGCGGTT	TCCAGTGGTACTCATGACCG
MyoG	CTTCTACGAGGGAGGGGACA	GCTGGATGGAGAGGCTTTGT
18S rRNA	GACTCGGGGAGGTAGTGACG	AGATACGCTATTGGAGCTGGAA

引物名称Primer	上游引物Forward primer (5'-3')	下游引物Reverse primer (5'-3')
MyoD1	AACGCCATCAGCTACATCGAG	ACAGGTGGGGCCGTTAAAATC
MyoD2	CGGCCTACTGAAGCCCGAA	CTCATGGTTGCTGCTTTTCTCC
Myf5	AAGAACGAGAGTTTGGGCGA	AGGACGTGGTATATGGGCCT
Myf6	TCAGCTACATCGAGCGGTT	TCCAGTGGTACTCATGACCG
MyoG	CTTCTACGAGGGAGGGGACA	GCTGGATGGAGAGGCTTTGT
18S rRNA	GACTCGGGGAGGTAGTGACG	AGATACGCTATTGGAGCTGGAA

蛋白质名称 Protein name	蛋白质编号 Locus ID	蛋白质长度 Size/aa	相对分子质量 Molecular weight/u	等电点 Pi	不稳定指数 Instability index	疏水性均值 GRAVY
ToMyoD1	EVM0022498	297	32 206.38	5.33	71.51	-0.591
ToMyoD2	EVM0018159	263	28 731.83	5.87	66.57	-0.634
ToMyf5	EVM0020882	240	26 187.14	6.23	89.12	-0.594
ToMyf6	EVM0005068	231	25 542.27	5.93	65.35	-0.849
ToMyoG	EVM0008148	250	27 421.64	6.33	65.97	-0.650

蛋白质名称 Protein name	蛋白质编号 Locus ID	蛋白质长度 Size/aa	相对分子质量 Molecular weight/u	等电点 Pi	不稳定指数 Instability index	疏水性均值 GRAVY
ToMyoD1	EVM0022498	297	32 206.38	5.33	71.51	-0.591
ToMyoD2	EVM0018159	263	28 731.83	5.87	66.57	-0.634
ToMyf5	EVM0020882	240	26 187.14	6.23	89.12	-0.594
ToMyf6	EVM0005068	231	25 542.27	5.93	65.35	-0.849
ToMyoG	EVM0008148	250	27 421.64	6.33	65.97	-0.650

物种 Species	分类 Taxonomy	基因名称 Gene name	蛋白质编号 Locus ID	蛋白质长度 Size/aa
高体鰤	鲹形目Carangiformes;	SdMyoD1	ENSSDUP00000023227	298
S. dumerili	鲹科Carangidae;	SdMyoD2	ENSSDUP00000025164	262
	鰤属Seriola	SdMyf5	ENSSDUP00000025182	240
		SdMyf6	ENSSDUP00000025208	238
		SdMyoG	ENSSDUP00000002433	250
黄尾鰤	鲹形目Carangiformes;	SlMyoD1	ENSSLDP00000028917	298
S. lalandi dorsalis	鲹科Carangidae;	SlMyoD2	ENSSLDP00000027557	263
	鰤属Seriola	SlMyf5	ENSSLDP00000024335	240
		SlMyf6	ENSSLDP00000024364	239
		SlMyoG	ENSSLDP00000009264	250
半滑舌鳎	鲽形目Pleuronectiformes;	CsMyoD1	ENSCSEP00000003405	296
C. semilaevis	舌鳎科Cynoglossidae;	CsMyoD2	ENSCSEP00000016973	214
	舌鳎属Cynoglossus	CsMyf5	ENSCSEP00000006862	246
		CsMyf6	ENSCSEP00000006878	241
		CsMyoG	ENSCSEP00000007229	243
大菱鲆	鲽形目Pleuronectiformes;	SmMyoD1	ENSSMAP00000005978	297
S. maximus	菱鲆科Scophthalmidae;	SmMyoD2	ENSSMAP00000010597	265
	菱鲆属 Scophthalmus	SmMyf5	ENSSMAP00000007369	297
		SmMyf6	ENSSMAP00000007390	269
		SmMyoG	ENSSMAP00000010312	246

卵形鲳鲹生肌调节因子基因家族的鉴定及在胚胎中的表达

Genome-wide identification and expression of MRF gene family in embryonic development of Trachinotus ovatus

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