PDF(1919 KB)
Physiological Responses of Different Apostichopus japonicus Breeding Populations to Heat Stress
GAOJijun, XUEMan, WANGQinglin, ZHANGJian, RENHai, YUShanshan, ZHAOChunlong, HEZhenping
Journal of Agriculture ›› 2026, Vol. 16 ›› Issue (8) : 56-66.
PDF(1919 KB)
PDF(1919 KB)
Physiological Responses of Different Apostichopus japonicus Breeding Populations to Heat Stress
This study aims to elucidate the physiological mechanisms underlying heat tolerance,a key selective trait in the sea cucumber Apostichopusjaponicas, by examining dynamic changes in antioxidant, immune-related, and metabolic enzyme activities under varying heat shock temperatures, and by evaluating the thermal stability of the critical heat-tolerant enzyme malate dehydrogenase. Using a graded temperature elevation protocol for heat shock treatmentwith four geographical hybrid populations(WFD♀×RZ♂, WFD♀×YT♂, WFD♀×TS♂, WFD♀×WFD♂) and two early heat shock populations (blastocyst stage, big ear larvae stage) as materials, the enzymatic activities and thermal stability of superoxide dismutase (SOD), catalase (CAT), lactate dehydrogenase (LDH), pyruvate kinase (PK), and cytoplasmic malate dehydrogenase (cMDH) were systematically measured and analyzed in the body walls of juvenile sea cucumbers from different breeding populations. The results revealed that: (1) the WFD♀×RZ♂ population exhibited significantly higher SOD, CAT, PK, and cMDH activities compared to other hybrid combinations (P<0.05); (2) under heat stress conditions (34℃ and 36℃), individuals heat-shocked at the late auricularia stage showed significantly greater SOD, CAT, LDH, and cMDH activities than those treated at the blastula stage (P<0.05); (3) starting from the 5-minute time point post-heat treatment, the residual cMDH activity remained significantly higher in the late auricularia group than in the blastula group across all subsequent time points; (4) among the hybrid populations, the cMDH thermal stability in the WFD♀×YT♂ and WFD♀×RZ♂ groups was markedly superior to that in the other two populations. In summary, these findings indicate that the WFD♀×RZ♂ population and individuals exposed to heat shock at the late auricularia stage possess enhanced potential for heat tolerance. This study provides valuable theoretical insights and empirical data to support the screening of stress-resistant sea cucumber germplasm and the advancement of sustainable aquaculture practices.
Apostichopusjaponicus / high temperature stress / enzyme activity / breeding group / thermal stability / malate dehydrogenase
| [1] |
朱志煌, 林琪, 吴建绍, 等. 仿刺参南移养殖温度耐受性相关基因的差异表达分析[J]. 渔业研究, 2024, 46(1):1-10.
本研究将仿刺参幼苗从北方南移至福建进行养殖，在自然水温下饲养1年，并作为南移福建养殖的实验组，同时以北方相同苗龄的仿刺参作为对照组，比较两组间的抗热性能；采用荧光定量法分析经过热诱导1、2、3 h后的实验组和对照组仿刺参的HSP90a、gp96、HSP70、HSP26和proeintl(2)efl温度耐受性相关基因的转录表达变化情况。研究发现：1）实验组仿刺参的亚致死温度为32 ℃，明显高于对照组的30 ℃，实验组和对照组仿刺参的半致死温度分别为33.1、31.9 ℃，表明高温驯化能够提高仿刺参的耐热能力。2）实验组与对照组仿刺参的5种温度耐受性相关基因的表达存在差异。在30 ℃高温刺激下，对照组仿刺参的HSP90a、gp96、HSP70、HSP26基因表达在温度刺激1、2 h后没有明显增加，而刺激3 h后才出现上调，表达量分别达到（9.801±1.303）、（2.508±0.910）、（8.649±1.936）、（34.787±4.978）；实验组仿刺参的HSP90a、gp96、HSP70、HSP26表达在温度刺激1 h后即出现显著上调，表达量分别达到（42.000±8.798）、（20.019±6.224）、（218.750±78.701）、（93.710±5.674）。实验组仿刺参的4种温度耐受性相关基因的上调时间早于对照组，表明实验组仿刺参对外界高温的适应能力强于对照组。因此，高温驯化后的仿刺参能够更早地对外界高温变化做出反应。
|
| [2] |
李倩, 夏光丽, 曹荣, 等. 仿刺参性腺酶解过程风味变化[J]. 渔业科学进展, 2022, 43(6):239-248.
|
| [3] |
李尚俊, 孙国华, 李雪燕, 等. 高温胁迫下仿刺参表观遗传调控相关基因的表达特征[J]. 中国水产科学, 2017, 24(3):470-476.
|
| [4] |
|
| [5] |
徐冬雪. 仿刺参热胁迫响应的分子调控特征研究与SNP标记筛选[D]. 青岛: 中国科学院研究生院(海洋研究所), 2016.
|
| [6] |
|
| [7] |
王青林, 董云伟, 董双林, 等. 高温刺激对刺参浮游期幼体存活率及Hsp70表达量的影响[J]. 中国海洋大学学报(自然科学版), 2011, 41(3):47-51.
|
| [8] |
于姗姗, 王青林, 董云伟. 亲本高温驯化和囊胚期热应激对刺参幼体生长、发育影响的初步研究[J]. 南方水产科学, 2015, 11(4):46-52.
|
| [9] |
|
| [10] |
|
| [11] |
谢兆文, 王寿昆, 林旋, 等. 高温胁迫对仿刺参抗氧化酶等指标的影响[J]. 水产科学, 2016, 35(4):327-333.
|
| [12] |
李晓英, 张冬冬, 徐梦雪, 等. 5个不同地理群体仿刺参的高温耐受力及养殖性能评估[J]. 海洋渔业, 2020, 42(6):722-731.
|
| [13] |
江慧琼, 刘雅婷, 陈清华. 超氧化物歧化酶的生理功能及其在动物生产中的应用前景[J]. 中国畜牧兽医, 2024, 51(3):945-954.
氧化应激会导致机体内的氧化与抗氧化的失衡,从而产生过多的活性氧(reactive oxygen species,ROS)基团造成细胞和组织的损伤、机体炎症等疾病,影响机体的生长发育。超氧化物歧化酶(superoxide dismutase,SOD)是存在于生物体内的一种抗氧化金属酶,它能够通过歧化反应催化超氧阴离子自由基(superoxide anionradical,O<sub>2</sub><sup>-</sup>),从而清除动物机体内过多的ROS。研究发现,SOD不仅在抗氧化功能上具有专一性、高效性,能够减缓机体衰老,并且还能在一定程度上提高动物肠道健康、生产性能和调节机体免疫功能。在集约化畜禽养殖、生产机器的大规模使用情况下,辐射对于动物所造成的损伤需要引起关注,有效地利用SOD的抗辐射功能以缓解或改善辐射对于机体的影响,值得进一步探索。目前,针对缓解机体氧化应激的添加剂主要有维生素、微量元素、植物提取物等,而利用SOD来缓解动物氧化应激、提高生产性能的研究相对较少,主要源于天然SOD提取成本高、分子质量大、易失活等。但随着人工合成SOD的出现,降低了制备SOD的成本,且SOD具备高活性、小分子易吸收的特点,这使得SOD在畜禽生产中的应用逐渐增多,未来可以挖掘更多SOD的应用潜力,将SOD更广泛地应用于动物生产中。笔者对SOD抗氧化、抗衰老、抗炎症、抗辐射、调节机体免疫等生理功能进行了总结,以期为SOD在动物生产中的应用提供更多理论依据。
|
| [14] |
Death-associated protein kinase (DAPk), a multi-domain serine/threonine kinase, regulates numerous cell death mechanisms and harbors tumor suppressor functions. In this study, we report that DAPk directly binds and functionally activates pyruvate kinase M2 (PKM2), a key glycolytic enzyme, which contributes to the regulation of cancer cell metabolism. PKM2 was identified as a novel binding partner of DAPk by a yeast two-hybrid screen. This interaction was validated in vitro by enzyme-linked immunosorbent assay using purified proteins and in vivo by co-immunoprecipitation of the two endogenous proteins from cells. In vitro interaction with full-length DAPk resulted in a significant increase in the activity of PKM2. Conversely, a fragment of DAPk harboring only the functional kinase domain (KD) could neither bind PKM2 in cells nor activate it in vitro. Indeed, DAPk failed to phosphorylate PKM2. Notably, transfection of cells, with a truncated DAPk lacking the KD, elevated endogenous PKM2 activity, suggesting that PKM2 activation by DAPk occurs independently of its kinase activity. DAPk-transfected cells displayed changes in glycolytic activity, as reflected by elevated lactate production, whereas glucose uptake remained unaltered. A mild reduction in cell proliferation was detected as well in these transfected cells. Altogether, this work identifies a new role for DAPk as a metabolic regulator, suggesting the concept of direct interactions between a tumor suppressor and a key glycolytic enzyme to limit cell growth. Moreover, the work documents a unique function of DAPk that is independent of its catalytic activity and a novel mechanism to activate PKM2 by protein-protein interaction.
|
| [15] |
|
| [16] |
|
| [17] |
|
| [18] |
|
| [19] |
|
| [20] |
冯其明. 仿刺参个体生长差异的行为及生理机制研究[D]. 青岛: 中国科学院大学(中国科学院海洋研究所), 2023.
|
| [21] |
|
| [22] |
|
| [23] |
王青林. 温度驯化对刺参(Apostichopus japonicus)生长及耐热性的影响及生理生态学机制[D]. 青岛: 中国海洋大学, 2012.
|
| [24] |
|
| [25] |
|
| [26] |
|
| [27] |
崔静, 李太武, 苏秀榕, 等. 南移养殖的刺参(Apostichopus japonicus)cDNA文库的构建及原肌球蛋白基因的研究[J]. 海洋与湖沼, 2010, 41(6):850-856.
|
| [28] |
To test predictions of biochemical symmorphosis, we measured the activity of seven consecutive glycolytic enzymes at three positions along the heterothermic white muscle of the bluefin tuna. Biochemical symmorphosis predicts that adjustments in sequential enzyme concentrations along a thermal gradient should occur as a function of the thermal sensitivity of the enzymes to ensure that no one enzyme in the pathway is in excess at any point along the gradient. We found no evidence for adjustments in enzyme quantity or quality along the thermal gradient, as well as no evidence for the prediction that the more temperature-sensitive enzymes would exhibit more dramatic compensation. Conservation of glycolytic flux in the cold exterior and warm interior muscle may be achieved by the near insensitivity of glyceraldehyde-3-phosphate dehydrogenase to temperature in this tissue. This may have the added benefit of moderating flux during seasonal or transient changes in the thermal gradient. According to the strictest application of biochemical symmorphosis, such a mechanism represents adequate, yet suboptimal design.
|
| [29] |
\n This study aims to establish a quantitative and qualitative evaluation model of sea cucumber\n Apostichopus japonicus\n research. Data from 2000 to 2021 were obtained from the Web of Science Core Collection (WoSCC) of Thomson Reuters. Bibliometrics and CiteSpace software were used to analyze authors, exporting countries, journals, influential articles, research areas, institutions, research hot spots, and trends. A total of 1,358 research papers on\n A. japonicus\n research were identified from 2000 to 2021. The number of papers published in this field is rapidly increasing, and the research phase can be divided into initial, developmental, and stabilization phases. Research on\n A. japonicus\n is mostly conducted in China, followed by Japan and the United States. Hongsheng Yang, Chenghua Li, and Shuanglin Dong are the lead authors. Research activities are focused on genetics and breeding, growth and development, immunology and disease, aestivation, regeneration, and food processing. Gut microbiota, activation, and collagen are potential research hot spots. The project highlights differences in the level of research between countries and teams, and regions with more developed industries or richer resources need further support. Governments or organizations are encouraged to 1) promote the development of the\n A. japonicus\n industry through the development or implementation of policies; 2) further participate in the research, production, and processing of\n A. japonicus\n ; and 3) strengthen international exchange and cooperation to bring economic benefits to farmers in suitable breeding areas through technology sharing.\n
|
| [30] |
|
| [31] |
韩莎, 胡炜, 李成林, 等. pH胁迫对刺参存活、生长及抗氧化酶活性的影响[J]. 渔业科学进展, 2018, 39(5):91-98.
|
| [32] |
Malate dehydrogenase (MDH) (EC 1.1.1.37) catalyzes the conversion of oxaloacetate and malate. This reaction is important in cellular metabolism, and it is coupled with easily detectable cofactor oxidation/reduction. It is a rather ubiquitous enzyme, for which several isoforms have been identified, differing in their subcellular localization and their specificity for the cofactor NAD or NADP. The nucleotide binding characteristics can be altered by a single amino acid change. Multiple amino acid sequence alignments of MDH show that there is a low degree of primary structural similarity, apart from several positions crucial for catalysis, cofactor binding and the subunit interface. Despite the low amino acids sequence identity their 3-dimensional structures are very similar. MDH is a group of multimeric enzymes consisting of identical subunits usually organized as either dimer or tetramers with subunit molecular weights of 30-35 kDa. MDH has been isolated from different sources including archaea, eubacteria, fungi, plant and mammals.
|
| [33] |
\n Orthologous proteins of species adapted to different temperatures exhibit differences in stability and function that are interpreted to reflect adaptive variation in structural “flexibility.” However, quantifying flexibility and comparing flexibility across proteins has remained a challenge. To address this issue, we examined temperature effects on cytosolic malate dehydrogenase (cMDH) orthologs from differently thermally adapted congeners of five genera of marine molluscs whose field body temperatures span a range of ∼60 °C. We describe consistent patterns of convergent evolution in adaptation of function [temperature effects on\n K\n M\n of cofactor (NADH)] and structural stability (rate of heat denaturation of activity). To determine how these differences depend on flexibilities of overall structure and of regions known to be important in binding and catalysis, we performed molecular dynamics simulation (MDS) analyses. MDS analyses revealed a significant negative correlation between adaptation temperature and heat-induced increase of backbone atom movements [root mean square deviation (rmsd) of main-chain atoms]. Root mean square fluctuations (RMSFs) of movement by individual amino acid residues varied across the sequence in a qualitatively similar pattern among orthologs. Regions of sequence involved in ligand binding and catalysis—termed mobile regions 1 and 2 (MR1 and MR2), respectively—showed the largest values for RMSF. Heat-induced changes in RMSF values across the sequence and, importantly, in MR1 and MR2 were greatest in cold-adapted species. MDS methods are shown to provide powerful tools for examining adaptation of enzymes by providing a quantitative index of protein flexibility and identifying sequence regions where adaptive change in flexibility occurs.\n
|
| [34] |
钱圆. 刺参对不同溶氧水平和干露的生理生态学响应及其机理研究[D]. 青岛: 中国海洋大学, 2011.
|
| [35] |
于明志, 常亚青. 低温对不同群体仿刺参幼参某些生理现象的影响[J]. 大连水产学院学报, 2008, 23(1):31-36.
|
| [36] |
孟思远, 常亚青, 李文东, 等. 仿刺参幼参阶段4个生长性状遗传力的估计[J]. 大连水产学院学报, 2010, 25(6):475-479.
|
| [37] |
|
| [38] |
We characterized functional and structural properties of cytoplasmic malate dehydrogenases (cMDHs) from six limpets of the genus Lottia that have different vertical and latitudinal distributions. Particular attention was given to the cryptic species pair Lottia digitalis (northern occurring) and L. austrodigitalis (southern occurring) because of recent contraction in the southern range of L. digitalis and a northward range extension of L. austrodigitalis. As an index of adaptation of function, we measured the effects of temperature on the apparent Michaelis–Menten constant (Km) of the cofactor NADH(KmNADH). KmNADHvalues of cMDHs from the mid- to high-intertidal, low-latitude species L. scabra and L. gigantea were less sensitive to high temperature than those of cMDHs from the low- and mid-intertidal, high-latitude species L. scutum and L. pelta. cMDH of L. digitalis was more sensitive to high temperatures than the cMDH ortholog of L. austrodigitalis. Thermal stability (rate of loss of activity at 42.5°C) showed a similar pattern of interspecific variation. Comparison of the deduced amino acid sequences showed that interspecific differences ranged from one to as many as 17 residues. Differences in KmNADH and thermal stability between orthologs of L. digitalis and L. austrodigitalis result from a single amino acid substitution. At position 291, the glycine residue in cMDH of L. digitalis is replaced by a serine in cMDH of L. austrodigitalis, a change that favors additional hydrogen bonding and reduced conformational entropy. This difference between closely related congeners demonstrates the role of minor alterations in protein sequence in temperature adaptation and suggests that such variation is important in governing shifts in biogeographic range in response to climate change.
|
| [39] |
宋志东, 王际英, 王世信, 等. 不同生长发育阶段刺参体壁营养成分及氨基酸组成比较分析[J]. 水产科技情报, 2009, 36(1):11-13.
|
| [40] |
|
| [41] |
|
/
| 〈 |
|
〉 |
