稻田金背鲤尾柄肌纤维特征及相关代谢酶与基因表达研究

张文争; 杨立; 姚俊杰; 蒙庆米; 纪达

doi:10.12131/20220320

稻田金背鲤尾柄肌纤维特征及相关代谢酶与基因表达研究

doi: 10.12131/20220320

张文争^{1, 2,},
杨立^{1, 2},
姚俊杰^{1, 2, ,},
蒙庆米^{1, 2},
纪达^{1, 2}

1.
贵州大学渔业资源与环境研究中心，贵州贵阳 550025
2.
贵州大学高原山地动物遗传育种与繁殖教育部重点实验室，贵州贵阳 550025

基金项目: 贵州省农业重大产业科学研究攻关项目 (黔教合KY字〔2019〕013)

详细信息

作者简介:
张文争 (1995—)，女，硕士研究生，研究方向为水生生物发育与繁殖生物学。E-mail: 2682792491@qq.com

通讯作者:
姚俊杰 (1968—)，男，教授，博士，研究方向为水生生物发育与繁殖生物学。E-mail: junjieyao@163.com

中图分类号: S 917.4
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出版历程
- 收稿日期: 2022-12-22
- 修回日期: 2023-02-12
- 录用日期: 2023-02-23
- 网络出版日期: 2023-03-01
- 刊出日期: 2023-08-05

Study on muscle fiber characteristics, metabolic enzymes and gene expression of caudal stalk of golden-backed carp (Cyprinus carpio var. Jinbei)

ZHANG Wenzheng^{1, 2
,},
YANG Li^{1, 2},
YAO Junjie^{1, 2
, ,},
MENG Qingmi^{1, 2},
JI Da^{1, 2}

1.
Research Center of Fishery Resources and Environment, Guizhou University, Guiyang 550025, China
2.
Key Laboratory of Animal Genetics and Breeding and Reproduction of Plateau and Mountain Animals of Guizhou University, Guiyang 550025, China

摘要

摘要: 稻田金背鲤 (Cyprinus carpio var. Jinbei) 是经过多个世代自然选择和人工选择生活于稻田的鲤鱼。为推广金背鲤-稻田养殖模式，为该模式下的良种选育提供数据支撑，以池塘养殖的金背鲤为对照组，采用组织学、酶学及分子生物学方法分析了其尾柄肌纤维及相关代谢酶与基因表达，探究了稻田金背鲤尾柄肌肉在稻田浅水生态条件下生长的适应性特征。结果显示：在水稻扬花期和渔获期，尾柄肌纤维直径稻田组均大于池塘组，并存在显著性差异 (P<0.05)；在水稻扬花期和渔获期，尾柄肌纤维横截面积稻田组大于池塘组，在渔获期存在显著性差异 (P<0.05)；尾柄肌纤维密度稻田组均小于池塘组，在渔获期存在显著性差异 (P<0.05)；在水稻扬花期和渔获期，代谢酶中的乳酸脱氢酶 (LDH)、过氧化氢酶 (CAT) 和Ca²⁺-ATP酶活性稻田组均高于池塘组；在水稻扬花期，稻田组的过氧化物酶体增殖物激活受体γ辅助激活因子α (PGC-1α) 基因表达量显著高于池塘组 (P<0.05)，而稻田组的腺苷酸活化蛋白激酶 (AMPK)、沉默信息调节因子1 (SIRT1) 基因表达量在水稻扬花期和渔获期均显著低于池塘组 (P<0.05)。研究显示，稻田金背鲤尾柄肌肉有更大的肌纤维直径和横截面积，尾柄肌肉呈现较高的LDH和Ca²⁺-ATP酶活性，这是其适应稻田浅水生活的特点；稻田金背鲤的尾柄抗阻运动会引起肌肉肥大，与其激活蛋白质合成通路有关，而池塘金背鲤通常进行长时间的耐力运动，与其线粒体生物合成有关；在水稻扬花期，稻田金背鲤PGC-1α基因表达量较高，可能与此阶段水稻开花、稻田中昆虫增多、生物多样性增加以及生物量增加有关。
- 稻田金背鲤 /
- 尾柄肌纤维 /
- 代谢酶 /
- 基因表达
Abstract: Golden-backed carp (Cyprinus carpio var. Jinbei) is a common carp that has lived in rice fields for many generations through long-term natural and artificial selection. In order to promote its paddy field breeding model and provide data supports for the breeding of improved varieties by this model, we analyzed the caudal stalk muscle fibers, related metabolic enzymes and gene expressions by using histology, enzymology and molecular biology methods, and explored the adaptive characteristics of caudal stalk muscle growth under the shallow ecological conditions of rice field. The results show that the diameter of caudal stalk muscle fiber in paddy field group was larger than that in pond group at flowering stage and harvest stage with significant differences (P<0.05). The cross-section area of caudal stalk muscle fiber in paddy field group was larger than that in pond group at flowering stage and harvest stage, with significant differences at harvest stage (P<0.05), while the density of caudal stalk muscle fiber in paddy field group was smaller than that in pond group. There were significant differences at harvest stage (P<0.05). The activities of lactate dehydrogenase (LDH), catalase (CAT) and Ca²⁺-ATPase in metabolic enzymes were higher in paddy field group than in pond group at flowering stage and harvest stage. The expression level of peroxisome proliferation-activated receptor γ-coactivator α (PGC-1α) gene in paddy field group was significantly higher than that in pond group (P<0.05). The results reveal that the expression levels of adenylate activated protein kinase (AMPK) and silence-regulating factor 1 (SIRT1) gene in paddy field group were significantly lower than those in pond group at flowering stage and harvest stage (P<0.05). The resistance movement of the caudal stalk of golden-backed carp in rice field caused muscle hypertrophy, which was related with the activation of protein synthesis pathway, while its endurance exercise, was related with the mitochondrial biosynthesis. The higher PGC-1α expression level of golden-backed carp in rice flowering stage may be related with the increase of rice flowering, insects, biodiversity and biomass.
- Golden-backed carp (Cyprinus carpio var. Jinbei) /
- Caudal stalk muscle fiber /
- Metabolic enzymes /
- Gene expression

HTML全文

图 1 耐力运动激活能量感应网络 AMPK/SIRT1/PGC-1α 信号通路

Figure 1. Endurance exercise activates AMPK/SIRT1/PGC-1α signaling pathway in energy sensing network

下载: 全尺寸图片幻灯片

图 2 稻田和池塘金背鲤尾柄肌纤维 HE 染色图

注：a. 稻花前稻田金背鲤尾柄肌纤维 (200×)；b. 水稻扬花期稻田金背鲤尾柄肌纤维 (200×)；c. 水稻扬花期稻田金背鲤尾柄肌纤维 (400×)； d. 渔获期稻田金背鲤尾柄肌纤维 (200×)；e. 水稻扬花同时期池塘金背鲤尾柄肌纤维 (200×)；f. 渔获期池塘金背鲤尾柄肌纤维 (200×)。

Figure 2. HE staining of muscle fibers of caudal stalk of golden-backed carps in paddy fields and ponds

Note: a. Caudal stalk muscle fibers of golden-backed carp before rice flower (200×); b. Caudal stalk muscle fibers (200×) of golden-backed carp in rice field during flowering period; c. Caudal stalk muscle fibers (400×) of golden-backed carp in rice field during flowering period; d. Caudal stalk muscle fibers (200×) of golden-backed carp in paddy field; e. Caudal stalk muscle fibers of pond golden-backed carp (200×) at the same period of rice flowering; f. Caudal stalk muscle fibers of pond golden-backed carp (200×) at the harvest stage.

下载: 全尺寸图片幻灯片

图 3 稻田和池塘金背鲤生长过程中主要代谢酶活性变化

注：1. 稻花前期；2. 水稻扬花期；3. 渔获期；每个数据表示3个重复；不同的小写字母表示相同组别 (稻田组) 不同采样时间点的酶活性差异显著 (P<0.05)，****表示不同处理组在相同采样点上有极显著性差异 (P<0.01)。

Figure 3. Changes of main metabolic enzymes during growth of golden-backed carp in paddy fields and ponds

Notes: 1. Before rice flowering period; 2. Flowering period of rice; 3. Harvest period; Values are $ \overline { X}\pm { \rm {SD}}$1 (n=3). Different letters indicate a significant difference (P<0.05) among different growth periods in rice field; ****. Extremely significant difference (P<0.01) between the two groups at the same sampling site.

下载: 全尺寸图片幻灯片

图 4 稻田和池塘养殖的金背鲤不同生长时期 AMPK、SIRT1 和 PGC-1α 基因的相对表达量

注：1. 稻花前期；2. 水稻扬花期；3. 渔获期；每个数据表示 3 个重复；不同的小写字母表示稻田组在不同生长时间点基因表达量差异显著 (P<0.05)，而*表示不同处理组在相同采样点上有显著差异 (P<0.05)。

Figure 4. Relative expression of AMPK, SIRT1 and PGC-1α gene in different growth periods of golden-backed carps in paddy field and pond

Note: 1. Before flowering period; 2. Flowering period of rice; 3. Harvest period. Values are Mean±SD (n=3). Different letters indicate significant differences in paddy field in each growth period (P<0.05).*. Significant differences between the two groups at the same sampling time (P<0.05).

下载: 全尺寸图片幻灯片

表 1 本实验所需引物

Table 1. Primers for this study

引物 Primer	序列 (5'—3') Sequence (5'–3')	长度 Length/bp	温度 Temperature/℃	应用 Application
AMPK-F	CGGAGACACACTAGGAGTGG	141	60	qPCR
AMPK-R	ATCTCGCGTCGAATCTTCCC	141	60	qPCR
PGC-1α-F	TCTCCATACTCCCGCTCCG	230	60	qPCR
PGC-1α-R	TCAGCCCTCTCACATTCTCGTT	230	60	qPCR
SIRT1-F	GGCAGCTATGGTCAGCCTAC	135	60	qPCR
SIRT1-R	TGCTTGAGGACAGGACTTACAC	135	60	qPCR
β-actin-F	AAAACCAACCATGTGCGACG	238	60	qPCR
β-actin-R	CCGTGCTCAATGGGGTACTT	238	60	qPCR

下载: 导出CSV

表 2 金背鲤尾柄肌纤维直径

Table 2. Muscle fiber diameter of caudal stalk of 　　　　　　　　　golden-backed carps　　　　　　μm

采样时期 Sampling period	稻田实验组 Paddy field experimental group	池塘对照组 Pond control group
稻花前期 Before flowering period	31.88±4.41^a	31.88±4.41
水稻扬花期 Flowering period	54.88±10.31^b*	41.68±4.42
渔获期 Harvest period	62.98±4.92^bc*	46.00±2.69
注：不同上标字母表示数据间有显著性差异 (P<0.05)；表示同一行数据间有显著性差异 (P<0.05)。稻花前期指同一批幼鱼未投放到稻田和池塘阶段。后表同此。 Note: Different superscript letters indicate significant differences between the data (P<0.05); . Significant difference between the data within the same row (P<0.05). Before flowering period indicates the stage when the same batch of juvenile fish had not been released into paddy fields and ponds yet. The same case in the following tables.

下载: 导出CSV

表 3 金背鲤尾柄肌纤维横截面积

Table 3. Cross-sectional area of muscle fibers of caudal　　　　　　 stalk of golden-backed carps　　　　μm²

采样时期 Sampling period	稻田实验组 Paddy field experimental group	池塘对照组 Pond control group
稻花前期 Before flowering period	1 065.99±549.36^a	1 065.99±549.36
水稻扬花期 Flowering period	1 711.76±748.25^a	1 327.79±600.17
渔获期 Harvest period	2 845.43±70.82^b^*	1 360.62±393.08

下载: 导出CSV

表 4 金背鲤尾柄肌纤维密度

Table 4. Muscle fiber density of caudal stalk of 　　　　　　　　　golden-backed carps　　　根·mm⁻²

采样时期 Sampling period	稻田实验组 Paddy field experimental group	池塘对照组 Pond control group
稻花前期 Before flowering period	1252.56±916.11	1252.56±916.11
水稻扬花期 Flowering period	702.45±406.78	845.10±306.00
渔获期 Harvest period	351.59±8.88^*	781.40±244.04

下载: 导出CSV

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