不同物候期藏绵羊肉品质、营养成分及肉质相关基因表达特征分析

doi:10.13304/j.nykjdb.2024.0235

摘要/Abstract

摘要：

为分析不同物候期（返青期和青草期）牧草营养成分对放牧藏绵羊肉品质、营养成分及肉质相关基因表达特征的影响，通过测定返青期（5月）和青草期（8月）牧草的营养成分及藏绵羊背最长肌、前腿肌和后腿肌的肉品质、营养成分及肉质相关基因表达量。结果表明，青草期牧草的粗蛋白和粗脂肪含量显著高于返青期，返青期牧草的酸性洗涤纤维含量显著高于青草期。青草期藏绵羊背最长肌的剪切力、pH_{1 h}（屠宰后1 h 肉样的pH）显著高于返青期，熟肉率显著高于返青期，失水率极显著低于返青期；肌肉的粗脂肪含量显著高于返青期，水分含量显著低于返青期。藏绵羊LPL和FABP3基因在青草期的表达量显著高于返青期，SCD、ADSL、CAST和MSTN基因在返青期的表达量显著高于青草期。相关性分析表明， ADSL基因表达量与蛋白含量呈极显著负相关（r=-0.995，P<0.01），与粗脂肪含量呈极显著正相关（r=0.865，P<0.01）。以上研究结果为不同物候期牧草的合理利用和藏绵羊肉品质改善提供重要参考。

关键词: 物候期, 肉品质, 营养成分, 基因表达, 藏绵羊

Abstract:

To examine the impact of forage nutrients during different phenological stages （regreen stage and grassy stage） on the quality， nutrient composition and gene expression characteristics related to meat quality in pastured Tibetan sheep meat， the nutrient composition of herbage during the regreen stage （May） and grassy stage （August）， as well as the meat quality， nutrient composition and expression of genes related to meat quality in the longissimus dorsi muscle， foreleg muscle， and hind leg muscle of Tibetan sheep were analyzed. The results indicated that there were significant increases in crude protein and crude fat contents during the grassy stage compared to the regreen stage， while acid detergent fiber content was significantly higher in the regreen stage than that in the grassy stage. Compared to the regreen stage， the shear force and pH_{1 h} （pH after slaughter 1 h） of the longissimus dorsi muscle of Tibetan sheep were significantly higherin the grassy stage. Additionally， the cooked meat rate was significantly higher in the grassy stage than that in regreen stage， whereas the water loss rate was significantly lower than that in the regreen stage. The crude fat content of muscle in grassy stage was significantly higher than that in regreen stage， while the water content was significantly lower in grassy stage. Additionally， the expression levels of LPL and FABP3 genes of Tibetan sheep were significantly higher in grassy stage than those in regreen stage， whereas the expression levels of SCD， ADSL， CAST， and MSTN genes were also significantly higher in regreen stage than those in grassy stage. Correlation analysis revealed that ADSL was significantly negative correlation with protein content （r=-0.995， P<0.01）， as well as significantly positive correlation with crude fat content （r=0.865， P<0.01）. Above results provided valuable insights for optimizing forage utilization and enhancing meat quality of Tibetan sheep during different phenological stages.

Key words: phenological stage, meat quality, nutrient composition, gene expression, Tibetan sheep

中图分类号:

S826.8+3

陈小伟, 沙玉柱, 刘秀, 邵鹏阳, 王翻兄, 谢转回, 杨文鑫, 陈倩玲, 高敏, 黄薇. 不同物候期藏绵羊肉品质、营养成分及肉质相关基因表达特征分析[J]. 中国农业科技导报, 2025, 27(7): 161-171.

Xiaowei CHEN, Yuzhu SHA, Xiu LIU, Pengyang SHAO, Fanxiong WANG, Zhuanhui XIE, Wenxin YANG, Qianling CHEN, Min GAO, Wei HUANG. Analysis of Gene Expression Characteristics Associated with Quality， Nutrient Composition， and Meat Quality of Tibetan Sheep Meat During Different Phenological Stages[J]. Journal of Agricultural Science and Technology, 2025, 27(7): 161-171.

图/表 8

参考文献 48

[1]	JING X P, WANG W J, DEGEN A A, et al.. Small intestinal morphology and sugar transporters expression when consuming diets of different energy levels:comparison between Tibetan and small-tailed Han sheep [J/OL]. Animal, 2022,16(3):100463 [2024-02-20]. .
[2]	LIU X, SHA Y Z, LYU W B, et al.. Multi-omics reveals that the rumen transcriptome,microbiome,and its metabolome co-regulate cold season adaptability of Tibetan sheep [J/OL]. Front. Microbiol., 2022,13: 859601 [2024-02-20]. .
[3]	JIAO J X, WANG T, ZHOU J W, et al.. Carcass parameters and meat quality of Tibetan sheep and Small-tailed Han sheep consuming diets of low-protein content and different energy yields [J]. J. Anim. Physiol. Anim. Nutr., 2020, 104(4):1010-1023.
[4]	董轩.羊肉品质评价指标、影响因素及不同烹调方式对品质的影响[J].现代食品,2020,12(23):16-19.
	DONG X. Evaluation indexes, influencing factors and effects of different cooking methods on mutton quality [J]. Mod. Food, 2020, 12(23):16-19.
[5]	卫智军,杨静,苏吉安,等.荒漠草原不同放牧制度群落现存量与营养物质动态研究[J].干旱地区农业研究,2003,21(4):53-57.
	WEI Z J, YANG J, SU J A, et al.. A study on the standing forages and nutrient dynamics of community on Stipa breviflora grassland under different systems [J]. Agric. Res. Arid Areas, 2003, 21(4):53-57.
[6]	YANG C T, GAO P, HOU F J, et al.. Relationship between chemical composition of native forage and nutrient digestibility by Tibetan sheep on the Qinghai-Tibetan Plateau [J]. J. Anim.Sci., 2018, 96(3):1140-1149.
[7]	贺福全,陈懂懂,李奇,等.三江源高寒草地牧草营养时空分布[J].生态学报,2020, 40(18): 6304-6313.
	HE F Q, CHEN C J, LI Q, et al.. Temporal and spatial distribution of herbage nutrition in alpine grassland of Sanjiangyuan [J]. Acta Ecol. Sin., 2020, 40(18): 6304-6313.
[8]	ZHANG X, HAN L, HOU S, et al.. Metabolomics approach reveals high energy diet improves the quality and enhances the flavor of black Tibetan sheep meat by altering the composition of rumen microbiota [J]. Front. Nutr., 2022, 9: 915558-915572.
[9]	PRACHE S, SCHREURS N, GUILLIER L.Review:Factors affecting sheep carcass and meat quality attributes [J/OL].Animal, 2022,16(S1):100330 [2024-02-20]..
[10]	刘秀,魏红,史浩, 等. 草地放牧藏绵羊不同月份肉品质及脂肪酸特征分析[J].家畜生态学报,2024,45(5):35-43.
	LIU X, WEI H, SHI H . et al .. Analysis of meat quality and fatty acid characteristics of Tibetan sheep grazed on grassland in different months [J]. J. Livest. Ecol., 2024, 45(5): 35-43.
[11]	王莉梅,梁俊芳,王德宝,等.自然放牧条件下不同月龄乌珠穆沁羊的肉品质分析[J].食品科技,2018,43(12):118-124, 130.
	WANG L M, LIANG J F, WANG D B, et al.. Effect of different age on meat quality in Ujumqin sheep under natural grazing condition [J]. Food Sci. Technol., 2018, 43(12):118-124, 130.
[12]	曹阳,赵雪,于永生,等.肉羊心型脂肪酸结合蛋白基因第2外显子多态性与肉质性状相关分析[J].东北农业大学学报,2013,44(9):30-34.
	CAO Y, ZHAO X, YU Y S, et al.. Correlation analysis on polymorphisms and meat quality trait at the exon 2 of H-FABP gene in sheep [J]. J. Northeast. Agric. Univ., 2013, 44(9):30-34.
[13]	曲桂娟,杨连玉,秦贵信,等.不同杂交品种肉牛背最长肌LPL基因表达的发育性变化及其对IMF的影响[J].中国兽医学报,2011,31(2):272-274.
	QU G J, YANG L Y, QIN G X, et al.. Developmental changes of LPL gene expression of longissimusdorsi and their effects on IMF in different crossbred cattle breeds [J]. Chin. J. Vet. Sci., 2011, 31(2):272-274.
[14]	张静静,宋玉芹,李桢,等.中国西门塔尔牛SCD1基因多态性与肉质性状的相关分析[J].华北农学报,2013,28(1):54-57.
	ZHANG J J, SONG Y Q, LI Z, et al.. Polymorphisms of SCD1 gene and its association with meat quality traits in Chinese Simmental [J]. Acta Agric. Boreali Sin., 2013, 28(1):54-57.
[15]	张丽英.饲料分析及饲料质量检测技术[M].3版.北京:中国农业大学出版社,2007:1-435.
[16]	刘素英,尤华,刘勇军,等. 畜禽肉质的测定: [S].北京:中国标准出版社,2007.
[17]	中华人民共和国农业部. 肉嫩度的测定剪切力测定法: [S].北京:中国农业出版社,2006.
[18]	陈泽勇,杨万颖,罗美中,等. 肉与肉制品水分含量测定: [S].北京:中国标准出版社,2008.
[19]	国家卫生和计划生育委员会,国家食品药品监督管理总局. 食品安全国家标准食品中蛋白质的测定: [S].北京:中国标准出版社,2017.
[20]	国家质量监督检验检疫总局,中国国家标准化管理委员会. 肉与肉制品总脂肪含量测定: [S].北京:中国标准出版社,2008.
[21]	国家质量监督检验检疫总局,中国国家标准化管理委员会. 肉与肉制品总灰分测定: [S].北京:中国标准出版社,2009.
[22]	LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-△△Ct method [J]. Methods, 2001, 25(4):402-408.
[23]	扎西卓玛,王宏博,李桂香,等.高寒草甸草场牧草营养成分测定与分析[J].湖北农业科学,2018,57(1):109-111.
	Zhaxizhuoma, WANG H B, LI G X, et al.. Determination and analysis of forage nutrients in alpine meadow [J]. Hubei Agric. Sci., 2018, 57(1):109-111.
[24]	程财. 乌拉盖草原牧草营养价值评定及不同饲喂水平对肉牛生产性能的影响[D].呼和浩特:内蒙古农业大学, 2020.
	CHENG C. Nutritional value assessment and different feeding levels of forage grass in Wulagai Grassland effect on the performance of beef cattle [D]. Hohhot: Inner Mongolia Agricultural University, 2020.
[25]	DESTEFANIS G, BRUGIAPAGLIA A, BARGE M T, et al.. Relationship between beef consumer tenderness perception and Warner-Bratzler shear force [J].Meat Sci., 2008,78(3):153-156.
[26]	曾志宏.肉的品质与pH值的相关性[J].肉类工业,2001,1(1):31-32.
[27]	MOON S S, YANG H S, PARK G B, et al.. The relationship of physiological maturity and marbling judged according to Korean grading system to meat quality traits of Hanwoo beef females [J]. Meat Sci., 2006, 74(3):516-521.
[28]	赵丽, 沈继源, 包高良, 等. 藏绵羊不同部位骨骼肌营养成分、肌纤维特性及与相关基因表达的关联分析[J]. 西北农业学报, 2023, 32(6): 825-834.
	ZHAO L, SHEN J Y, BAO G L, et al.. Correlation analysis of nutrient components and muscle fibers in skeletal muscle of Tibetan sheep and their gene expression [J]. Acta Agric. Bor-Occid. Sin., 2023, 32(6): 825-834.
[29]	卜振鲲,都帅,降晓伟,等.饲草比例对羔羊生产性能、肌肉营养成分和脂肪酸组成与含量的影响[J].黑龙江畜牧兽医,2021(11):92-96, 102.
	BU Z K, DU S, JIANG X W, et al.. Effect of forage ratio on production performance, muscle nutritional component and fatty acid composition and content of lambs [J]. Heilongjiang Anim. Sci. Vet. Med., 2021(11): 92-96, 102
[30]	BU Z K, GE G T, JIA Y S, et al.. Effect of hay with or without concentrate or pellets on growth performance and meat quality of Ujimqin lambs on the Inner Mongolian Plateau [J/OL]. Anim.Sci. J., 2021,92(1):e13553 [2024-02-20]. .
[31]	何林芳,李宁.不同饲养方式对南江黄羊肉营养成分的影响[J].畜禽业,2022,33(7):18-20.
[32]	吕亚宁, 叶文文, 兰旅涛. 牛肌内脂肪沉积影响因素及相关基因研究进展[J]. 中国草食动物科学, 2019, 39(4): 55-57.
	LYU Y N, YE W W, LAN L T, et al.. Research progress on influencing factors and related genes of fat deposition in bovine muscle [J]. Chin. J. Herbiv. Sci., 2019, 39(4): 55-57.
[33]	乔永,黄治国,李齐发,等.绵羊肌肉LPL基因表达的发育性变化及其对肌内脂肪含量的影响[J].中国农业科学,2007,40(10):2323-2330.
	QIAO Y, HUANG Z G, LI Q F, et al.. Developmental changes of the LPL mRNA expression and the effect on IMF content in sheep muscle [J]. Sci. Agric. Sin., 2007, 40(10):2323-2330.
[34]	马晓燕,祖玲玲,吐来力江·哈木太,等.新疆也木勒白羊MSTN基因在不同组织中的表达定量研究[J].中国草食动物科学,2015,35(5):4-7.
	MA X Y, ZU L L, Hamutai Tulailijiang, et al.. Study on the expression of MSTN gene in different tissues of yemule lambs [J]. China Herbiv. Sci., 2015, 35(5):4-7.
[35]	李度, 章晓炜, 袁琼雨, 等. 7种地方鸡屠宰性能、肉品质及肌苷酸相关基因表达的比较研究[J]. 中国畜牧杂志, 2023, 59(9): 215-219.
[36]	方钱海, 郭帅, 任红艳, 等. 猪SCD1和SCD5双基因敲除细胞系的构建及其对脂肪沉积的影响[J]. 农业生物技术学报, 2024,32(4):795-806.
	FANG Q H, GUO S, REN H Y, et al.. Construction of porcine (SUS scrofa) SCD1 and SCD5 double knockout cell lines and their effects on fat deposition [J]. J. Agric. Biotechnol., 2024,32(4):795-806.
[37]	陈春华,赵生国,雷赵民,等.甘肃肉牛CAST基因的多态性及其与肉质性状的相关性研究[J].中国农学通报,2013,29(35):17-22.
	CHEN C H, ZHAO S G, LEI Z M, et al.. Analysis on polymorphism of CAST gene in Gansu local cattle and its relationships with meat quality trait [J]. Chin. Agric. Sci. Bull., 2013, 29(35):17-22.
[38]	潘兴云, 薛妍君, 蒋林惠, 等. 肉多汁性的研究及核磁共振技术在多汁性检测中的前景[J]. 科技信息, 2010(24): 6-7.
[39]	PUNIA BANGAR S, SINGH SANDHU K, TRIF M, et al.. Enrichment in different health components of barley flour using twin-screw extrusion technology to support nutritionally balanced diets [J/OL]. Front. Nutr., 2021,8:823148 [2024-02-20]. .
[40]	PALMA-ACEVEDO A, PÉREZ-WON M, TABILO-MUNIZAGA G, et al.. Effects of PEF-assisted freeze-drying on protein quality,microstructure,and digestibility in Chilean abalone “loco” (Concholepas concholepas) mollusk [J/OL].Front.Nutr., 2022, 9:810827 [2024-02-20]. .
[41]	李晓丽, 朱宇旌, 李方方, 等. 钙蛋白酶抑制蛋白在动物营养中的研究进展[J]. 饲料与畜牧, 2014(12): 43-45.
[42]	张淑静.肉质候选基因在不同品种猪中的表达差异及与肉质的关系[D].合肥:安徽农业大学,2016.
	ZHANG S J. The difference analysis of the gene expression of meat quality candidate genes and it’s the relation of meat quality in different porcine breeds [D]. Hefei: Anhui Agricultural University, 2016
[43]	刘孟君,扎西央宗,尼珍,等.天然放牧条件下河谷型藏绵羊及其与萨福克羊杂交后代肉品质的比较[J].动物营养学报,2022,34(5):3077-3095.
	LIU M J, Zhaxiyangzong, NI Z, et al.. Comparison of meat quality of valley-type Tibetan sheep and its hybrid progeny with suffolk sheep under natural grazing condition [J]. J. Anim. Nutr., 2022, 34(5): 3077-3095.
[44]	周雨.西藏苏格绵羊不同部位肉品质特征与主要营养成分分析[J].黑龙江畜牧兽医,2019(5):55-58.
[45]	周雨,罗章,张文会,等.西藏不同品种藏系绵羊育肥屠宰后肉品质的分析[J].畜牧与饲料科学,2019,40(3):21-27.
	ZHOU Y, LUO Z, ZHANG W H, et al.. Meat quality analysis of different breeds of fattening Tibetan sheep after slaughtering [J].Anim. Husb. Feed. Sci., 2019, 40(3):21-27.
[46]	德庆卓嘎, 央金, 扎西, 等. 西藏多玛绵羊羊肉品质研究[J]. 家畜生态学报, 2014, 35(9): 78-82.
	Deqingzhuoga, Yangjin, Zhaxi, et al.. Research progress on mutton quality of Duoma sheep in Tibet [J]. J. Domestic Anim. Ecol., 2014, 35(9): 78-82.
[47]	姚亮伟, 沙玉柱, 郭新羽, 等. 不同海拔藏绵羊肉品质、营养成分及肉质相关基因表达特征分析[J]. 中国农业科技导报, 2024, 26(3): 66-75.
	YAO L W, SHA Y Z, GUO X Y, et al.. Analysis of meat quality, nutritional components and expression characteristics of meat quality-related genes in Tibetan sheep at different altitudes [J]. J. Agric. Sci. Technol., 2024, 26(3): 66-75.
[48]	田佳. MyoG、H-FABP、LPL基因对新疆两个地方绵羊品种产肉量及肉质的影响[D].乌鲁木齐:新疆农业大学,2014.
	TIAN J. Effects of MyoG, H-FABP, LPL genes on meat production and meat quality in two Xinjiang sheep breeds [D]. Urumqi: Xinjiang Agricultural University, 2014.

基因 Gene	引物序列 Primers sequence （5’-3’）	扩增长度 Amplification length/bp	基因索引号 Gene accession No.
β-actin	F：AGCCTTCCTTCCTGGGCATGGA R：GGACAGCACCGTGTTGGCGTAA	113	NM_001009428.3
MSTN	F：AACAGCGAGCAGAAGGAAAA R：GGTTAAATGCCAACCATTGC	145	NM_001009788.1
CAST	F：CGAGATTTCCGGTGGTGGAA R：GCTTGGATTCAACTGGCACG	122	NM_001009394.1
LPL	F：TGGAGTGACGGAATCTGTGG R：ACGTTGGAGTCTGGTTCCCT	150	NM_001267884.2
FABP3	F：GTCTCTTTCCCGACCTAGCC R：TAGCAAAACCGACACCGAGT	118	XM_004007004.5
ADSL	F：CGCTTGCTTCCCGTTATGC R：CGCCAGGTCCGGAATTTGTA	73	AJ_001048
SCD	F：CCCAGCTGTCAGAGAAAAGG R：GATGAAGCACAACAGCTTGT	115	NM_001009428.3

基因 Gene	引物序列 Primers sequence （5’-3’）	扩增长度 Amplification length/bp	基因索引号 Gene accession No.
β-actin	F：AGCCTTCCTTCCTGGGCATGGA R：GGACAGCACCGTGTTGGCGTAA	113	NM_001009428.3
MSTN	F：AACAGCGAGCAGAAGGAAAA R：GGTTAAATGCCAACCATTGC	145	NM_001009788.1
CAST	F：CGAGATTTCCGGTGGTGGAA R：GCTTGGATTCAACTGGCACG	122	NM_001009394.1
LPL	F：TGGAGTGACGGAATCTGTGG R：ACGTTGGAGTCTGGTTCCCT	150	NM_001267884.2
FABP3	F：GTCTCTTTCCCGACCTAGCC R：TAGCAAAACCGACACCGAGT	118	XM_004007004.5
ADSL	F：CGCTTGCTTCCCGTTATGC R：CGCCAGGTCCGGAATTTGTA	73	AJ_001048
SCD	F：CCCAGCTGTCAGAGAAAAGG R：GATGAAGCACAACAGCTTGT	115	NM_001009428.3

指标Index	返青期Regreen stage	青草期Grassy stage	Ｐ值P value
干物质DM	94.567±0.135	86.698±0.095	0.001
粗蛋白质CP	6.586±0.102	19.084±0.340	<0.001
粗脂肪EE	0.886±0.006	1.792±0.046	<0.001
酸性洗涤纤维ADF	31.640±0.669	30.820±0.382	<0.001
中性洗涤纤维NDF	48.432±0.382	52.623±0.709	0.001

指标Index	返青期Regreen stage	青草期Grassy stage	Ｐ值P value
干物质DM	94.567±0.135	86.698±0.095	0.001
粗蛋白质CP	6.586±0.102	19.084±0.340	<0.001
粗脂肪EE	0.886±0.006	1.792±0.046	<0.001
酸性洗涤纤维ADF	31.640±0.669	30.820±0.382	<0.001
中性洗涤纤维NDF	48.432±0.382	52.623±0.709	0.001

指标 Index	部位 Part	返青期 Regreen stage	青草期 Grassy stage	P值 P value
失水率 Water loss capacity/%	背最长肌 Longissimus dorsi muscle	38.7±0.000	23.3±0.010	<0.001
	前腿肌 Muscle of fore leg	27.2±0.000	24.8±0.012	0.001
	后腿肌 Muscle of hind leg	36.3±0.001	23.7±0.003	<0.001
剪切力 Shear force/N	背最长肌 Longissimus dorsi muscle	37.360±0.208	55.810±0.985	<0.001
	前腿肌 Muscle of fore leg	60.293±0.305	59.045±2.954	0.691
	后腿肌 Muscle of hind leg	46.512±0.28	75.207±0.557	<0.001
熟肉率 Cooked meat rate/%	背最长肌 Longissimus dorsi muscle	48.1±0.032	52.7±0.026	0.308
	前腿肌 Muscle of fore leg	55.8±0.028	65.5±0.038	0.066
	后腿肌 Muscle of hind leg	58.5±0.30	50.1±0.001	0.037
pH_{1 h}	背最长肌 Longissimus dorsi muscle	7.193±0.189	6.202±0.086	0.020
	前腿肌 Muscle of fore leg	7.798±0.197	6.197±0.067	<0.001
	后腿肌 Muscle of hind leg	7.435±0.179	6.488±0.060	0.010
pH_{24 h}	背最长肌 Longissimus dorsi muscle	6.935±0.092	6.248±0.039	<0.001
	前腿肌 Muscle of fore leg	7.022±0.432	6.280±0.018	0.117
	后腿肌 Muscle of hind leg	7.145±0.326	6.352±0.062	0.038