Pub Date : 2024-02-01Epub Date: 2024-01-15DOI: 10.1007/s10126-024-10286-z
Hua Yu, Zhao-Xia Zou, Wei Wei, Ying Li
The relationship between conjugated linoleic acid (CLA) and lipogenesis has been extensively studied in mammals and some cell lines, but it is relatively rare in fish, and the potential mechanism of action of CLA reducing fat mass remains unclear. The established primary culture model for studying lipogenesis in grass carp (Ctenopharyngodon idella) preadipocytes was used in the present study, and the objective was to explore the effects of CLA on intracellular lipid and TG content, fatty acid composition, and mRNA levels of adipogenesis transcription factors, lipase, and apoptosis genes in grass carp adipocytes in vitro. The results showed that CLA reduced the size of adipocyte and lipid droplet and decreased the content of intracellular lipid and TG, which was accompanied by a significant down-regulation of mRNA abundance in transcriptional regulators including peroxisome proliferator-activated receptor (PPAR) γ, CCAAT/enhancer-binding protein (C/EBP) α, sterol regulatory element-binding protein (SREBP) 1c, lipase genes including fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), lipoprotein lipase (LPL). Meanwhile, it decreased the content of saturated fatty acids (SFAs) and n - 6 polyunsaturated fatty acid (n-6 PUFA) and increased the content of monounsaturated fatty acid (MUFA) and n - 3 polyunsaturated fatty acid (n-3 PUFA) in primary grass carp adipocyte. In addition, CLA induced adipocyte apoptosis through downregulated anti-apoptotic gene B-cell CLL/lymphoma 2 (Bcl-2) mRNA level and up-regulated pro-apoptotic genes tumor necrosis factor-α (TNF-α), Bcl-2-associated X protein (Bax), Caspase-3, and Caspase-9 mRNA level in a dose-dependent manner. These findings suggest that CLA can act on grass carp adipocytes through various pathways, including decreasing adipocyte size, altering fatty acid composition, inhibiting adipocyte differentiation, promoting adipocyte apoptosis, and ultimately decreasing lipid accumulation.
共轭亚油酸(CLA)与脂肪生成之间的关系已在哺乳动物和一些细胞系中得到广泛研究,但在鱼类中却相对罕见,CLA减少脂肪量的潜在作用机制仍不清楚。本研究采用已建立的草鱼(Ctenopharyngodon idella)前脂肪细胞脂肪生成原代培养模型,目的是探讨CLA对体外草鱼脂肪细胞内脂质和TG含量、脂肪酸组成以及脂肪生成转录因子、脂肪酶和凋亡基因mRNA水平的影响。结果表明,CLA能缩小脂肪细胞和脂滴的体积,降低细胞内脂质和TG的含量,同时还能显著下调包括过氧化物酶体增殖激活受体(PPAR)γ在内的转录调节因子的mRNA丰度、CCAAT/增强子结合蛋白(C/EBP)α、固醇调节元件结合蛋白(SREBP)1c,以及脂肪酸合成酶(FAS)、乙酰-CoA 羧化酶(ACC)、脂蛋白脂肪酶(LPL)等脂肪酶基因的 mRNA 丰度。同时,它降低了原代草鱼脂肪细胞中饱和脂肪酸(SFAs)和 n - 6 多不饱和脂肪酸(n-6 PUFA)的含量,增加了单不饱和脂肪酸(MUFA)和 n - 3 多不饱和脂肪酸(n-3 PUFA)的含量。此外,CLA 通过下调抗凋亡基因 B 细胞 CLL/淋巴瘤 2(Bcl-2)mRNA 水平,上调促凋亡基因肿瘤坏死因子-α(TNF-α)、Bcl-2 相关 X 蛋白(Bax)、Caspase-3 和 Caspase-9 mRNA 水平,以剂量依赖性方式诱导脂肪细胞凋亡。这些研究结果表明,CLA 可通过多种途径作用于草鱼脂肪细胞,包括缩小脂肪细胞体积、改变脂肪酸组成、抑制脂肪细胞分化、促进脂肪细胞凋亡以及最终减少脂质积累。
{"title":"Conjugated Linoleic Acid Reduces Lipid Accumulation via Down-regulation Expression of Lipogenic Genes and Up-regulation of Apoptotic Genes in Grass Carp (Ctenopharyngodon idella) Adipocyte In Vitro.","authors":"Hua Yu, Zhao-Xia Zou, Wei Wei, Ying Li","doi":"10.1007/s10126-024-10286-z","DOIUrl":"10.1007/s10126-024-10286-z","url":null,"abstract":"<p><p>The relationship between conjugated linoleic acid (CLA) and lipogenesis has been extensively studied in mammals and some cell lines, but it is relatively rare in fish, and the potential mechanism of action of CLA reducing fat mass remains unclear. The established primary culture model for studying lipogenesis in grass carp (Ctenopharyngodon idella) preadipocytes was used in the present study, and the objective was to explore the effects of CLA on intracellular lipid and TG content, fatty acid composition, and mRNA levels of adipogenesis transcription factors, lipase, and apoptosis genes in grass carp adipocytes in vitro. The results showed that CLA reduced the size of adipocyte and lipid droplet and decreased the content of intracellular lipid and TG, which was accompanied by a significant down-regulation of mRNA abundance in transcriptional regulators including peroxisome proliferator-activated receptor (PPAR) γ, CCAAT/enhancer-binding protein (C/EBP) α, sterol regulatory element-binding protein (SREBP) 1c, lipase genes including fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), lipoprotein lipase (LPL). Meanwhile, it decreased the content of saturated fatty acids (SFAs) and n - 6 polyunsaturated fatty acid (n-6 PUFA) and increased the content of monounsaturated fatty acid (MUFA) and n - 3 polyunsaturated fatty acid (n-3 PUFA) in primary grass carp adipocyte. In addition, CLA induced adipocyte apoptosis through downregulated anti-apoptotic gene B-cell CLL/lymphoma 2 (Bcl-2) mRNA level and up-regulated pro-apoptotic genes tumor necrosis factor-α (TNF-α), Bcl-2-associated X protein (Bax), Caspase-3, and Caspase-9 mRNA level in a dose-dependent manner. These findings suggest that CLA can act on grass carp adipocytes through various pathways, including decreasing adipocyte size, altering fatty acid composition, inhibiting adipocyte differentiation, promoting adipocyte apoptosis, and ultimately decreasing lipid accumulation.</p>","PeriodicalId":690,"journal":{"name":"Marine Biotechnology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139465750","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The study aimed to compare the effects of crystalline L-lysine and L-glutamate (CAA), Lys-Glu dipeptide (KE) on the growth and muscle development of grass carp (Ctenopharyngodon idellus), and related molecular mechanisms. Five experimental diets (CR, 0.5% CAA, 1.5% CAA, 0.5% KE, 1.5% KE) containing Lys and Glu as free (Lys and Glu, CAA) dipeptide (Lys-Glu, KE) forms were prepared, respectively. A total of 450 juvenile grass carp with an initial weight of 10.69 ± 0.07 g were randomly assigned to 15 cages, and 5 treatments with 3 replicates of 30 fish each for 61 days of feeding. The results showed that the group of 0.5% KE exhibited the best growth performances according to the indicator's weight gain rate (WGR) and specific growth rate (SGR), although no statistically significant occurred among all groups; diet supplemented with 0.5% CAA significantly elevated the condition factor (CF) and viscerasomatic index (VSI) of juvenile grass carp. Diet supplemented with different Lys and Glu co-forms at different levels promoted the muscle amino acid content compared with those of CR group. Comparing with the CR group and other groups, the hardness of 0.5% CAA group significantly increased, and the springiness of 0.5% KE group excelled. Both the muscle fiber diameter and density of 0.5% KE group showed significant difference with those of the CR group, and a negative correlation between them was also observed. To uncover the related molecular mechanism of the differences caused by the different co-forms of Lys and Glu, the effect of different diets on the expressions of protein absorption, muscle quality, and antioxidation-related genes was analyzed. The results suggested that comparing with those of CR group, the dipeptide KE inhibited the expressions of genes associated with protein metabolism, such as AKT, S6K1, and FoxO1a but promoted PCNA expression, while the free style of CAA would improve the FoxO1a expression. Additionally, the muscle development-related genes (MyoD, MyOG, and Myf5) were significantly boosted in CAA co-form groups, and the expressions of fMYHCs were blocked but fMYHCs30 significantly promoted in 0.5% KE group. Finally, the effect of different co-forms of Lys and Glu on muscle antioxidant was examined. The 0.5% CAA diet was verified to increase GPX1a but obstruct Keap1 and GSTP1 expressions, resulting in enhanced SOD activity and reduced MDA levels in plasma. Collectively, the different co-forms of Lys and Glu influenced the growth of juvenile grass carp, and also the muscle development and quality through their different regulation on the protein metabolism, muscle development- and antioxidative-related genes.
{"title":"Insights into Dietary Different Co-Forms of Lysine and Glutamate on Growth Performance, Muscle Development, Antioxidation and Related Gene Expressions in Juvenile Grass Carp (Ctenopharyngodon idellus).","authors":"Yuyang Cai, Li He, Shenping Cao, Peng Zeng, Linhan Xu, Yanan Luo, Xiang Tang, Qixiang Wang, Zhen Liu, Zhimin He, Suchun Liu","doi":"10.1007/s10126-023-10278-5","DOIUrl":"10.1007/s10126-023-10278-5","url":null,"abstract":"<p><p>The study aimed to compare the effects of crystalline L-lysine and L-glutamate (CAA), Lys-Glu dipeptide (KE) on the growth and muscle development of grass carp (Ctenopharyngodon idellus), and related molecular mechanisms. Five experimental diets (CR, 0.5% CAA, 1.5% CAA, 0.5% KE, 1.5% KE) containing Lys and Glu as free (Lys and Glu, CAA) dipeptide (Lys-Glu, KE) forms were prepared, respectively. A total of 450 juvenile grass carp with an initial weight of 10.69 ± 0.07 g were randomly assigned to 15 cages, and 5 treatments with 3 replicates of 30 fish each for 61 days of feeding. The results showed that the group of 0.5% KE exhibited the best growth performances according to the indicator's weight gain rate (WGR) and specific growth rate (SGR), although no statistically significant occurred among all groups; diet supplemented with 0.5% CAA significantly elevated the condition factor (CF) and viscerasomatic index (VSI) of juvenile grass carp. Diet supplemented with different Lys and Glu co-forms at different levels promoted the muscle amino acid content compared with those of CR group. Comparing with the CR group and other groups, the hardness of 0.5% CAA group significantly increased, and the springiness of 0.5% KE group excelled. Both the muscle fiber diameter and density of 0.5% KE group showed significant difference with those of the CR group, and a negative correlation between them was also observed. To uncover the related molecular mechanism of the differences caused by the different co-forms of Lys and Glu, the effect of different diets on the expressions of protein absorption, muscle quality, and antioxidation-related genes was analyzed. The results suggested that comparing with those of CR group, the dipeptide KE inhibited the expressions of genes associated with protein metabolism, such as AKT, S6K1, and FoxO1a but promoted PCNA expression, while the free style of CAA would improve the FoxO1a expression. Additionally, the muscle development-related genes (MyoD, MyOG, and Myf5) were significantly boosted in CAA co-form groups, and the expressions of fMYHCs were blocked but fMYHCs30 significantly promoted in 0.5% KE group. Finally, the effect of different co-forms of Lys and Glu on muscle antioxidant was examined. The 0.5% CAA diet was verified to increase GPX1a but obstruct Keap1 and GSTP1 expressions, resulting in enhanced SOD activity and reduced MDA levels in plasma. Collectively, the different co-forms of Lys and Glu influenced the growth of juvenile grass carp, and also the muscle development and quality through their different regulation on the protein metabolism, muscle development- and antioxidative-related genes.</p>","PeriodicalId":690,"journal":{"name":"Marine Biotechnology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139047979","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-01Epub Date: 2024-01-26DOI: 10.1007/s10126-024-10287-y
Maíra Barbosa E Reis, Açucena Imparato Maximo, Jessica Maria Magno, Daniel de Lima Bellan, João Luiz Aldinucci Buzzo, Fernanda Fogagnoli Simas, Hugo Alexandre Oliveira Rocha, Edvaldo da Silva Trindade, Carolina Camargo de Oliveira
Natural substances are strategic candidates for drug development in cancer research. Marine-derived molecules are of special interest due to their wide range of biological activities and sustainable large-scale production. Melanoma is a type of skin cancer that originates from genetic mutations in melanocytes. BRAF, RAS, and NF1 mutations are described as the major melanoma drivers, but approximately 20% of patients lack these mutations and are included in the triple wild-type (tripleWT) classification. Recent advances in targeted therapy directed at driver mutations along with immunotherapy have only partially improved patients' overall survival, and consequently, melanoma remains deadly when in advanced stages. Fucose-containing sulfated polysaccharides (FCSP) are potential candidates to treat melanoma; therefore, we investigated Fucan A, a FCSP from Spatoglossum schröederi brown seaweed, in vitro in human melanoma cell lines presenting different mutations. Up to 72 h Fucan A treatment was not cytotoxic either to normal melanocytes or melanoma cell lines. Interestingly, it was able to impair the tripleWT CHL-1 cell proliferation (57%), comparable to the chemotherapeutic cytotoxic drug cisplatin results, with the advantage of not causing cytotoxicity. Fucan A increased CHL-1 doubling time, an effect attributed to cell cycle arrest. Vascular mimicry, a close related angiogenesis process, was also impaired (73%). Fucan A mode of action could be related to gene expression modulation, in special β-catenin downregulation, a molecule with protagonist roles in important signaling pathways. Taken together, results indicate that Fucan A is a potential anticancer molecule and, therefore, deserves further investigation.
天然物质是癌症研究药物开发的战略候选者。源自海洋的分子因其广泛的生物活性和可持续的大规模生产而备受关注。黑色素瘤是一种皮肤癌,源于黑色素细胞的基因突变。BRAF、RAS 和 NF1 基因突变被认为是黑色素瘤的主要诱因,但大约 20% 的患者缺乏这些基因突变,被归入三重野生型(tripleWT)分类。针对驱动基因突变的靶向治疗和免疫疗法的最新进展仅部分改善了患者的总体生存率,因此,黑色素瘤晚期患者仍然是致命的。含岩藻糖硫酸化多糖(FCSP)是治疗黑色素瘤的潜在候选物质;因此,我们在体外研究了来自Spatoglossum schröederi褐藻的Fucan A(一种FCSP)在出现不同突变的人类黑色素瘤细胞系中的作用。Fucan A处理72小时后,对正常黑色素细胞或黑色素瘤细胞系均无细胞毒性。有趣的是,它能损害三重WT CHL-1细胞的增殖(57%),与化疗细胞毒性药物顺铂的结果相当,但优点是不会产生细胞毒性。Fucan A 增加了 CHL-1 的倍增时间,这种效应归因于细胞周期停滞。与血管生成过程密切相关的血管模拟也受到了影响(73%)。Fucan A 的作用模式可能与基因表达调节有关,特别是β-catenin 的下调,β-catenin 是一种在重要信号通路中起主角作用的分子。综上所述,研究结果表明 Fucan A 是一种潜在的抗癌分子,因此值得进一步研究。
{"title":"A Fucose-Containing Sulfated Polysaccharide from Spatoglossum schröederi Potentially Targets Tumor Growth Rather Than Cytotoxicity: Distinguishing Action on Human Melanoma Cell Lines.","authors":"Maíra Barbosa E Reis, Açucena Imparato Maximo, Jessica Maria Magno, Daniel de Lima Bellan, João Luiz Aldinucci Buzzo, Fernanda Fogagnoli Simas, Hugo Alexandre Oliveira Rocha, Edvaldo da Silva Trindade, Carolina Camargo de Oliveira","doi":"10.1007/s10126-024-10287-y","DOIUrl":"10.1007/s10126-024-10287-y","url":null,"abstract":"<p><p>Natural substances are strategic candidates for drug development in cancer research. Marine-derived molecules are of special interest due to their wide range of biological activities and sustainable large-scale production. Melanoma is a type of skin cancer that originates from genetic mutations in melanocytes. BRAF, RAS, and NF1 mutations are described as the major melanoma drivers, but approximately 20% of patients lack these mutations and are included in the triple wild-type (tripleWT) classification. Recent advances in targeted therapy directed at driver mutations along with immunotherapy have only partially improved patients' overall survival, and consequently, melanoma remains deadly when in advanced stages. Fucose-containing sulfated polysaccharides (FCSP) are potential candidates to treat melanoma; therefore, we investigated Fucan A, a FCSP from Spatoglossum schröederi brown seaweed, in vitro in human melanoma cell lines presenting different mutations. Up to 72 h Fucan A treatment was not cytotoxic either to normal melanocytes or melanoma cell lines. Interestingly, it was able to impair the tripleWT CHL-1 cell proliferation (57%), comparable to the chemotherapeutic cytotoxic drug cisplatin results, with the advantage of not causing cytotoxicity. Fucan A increased CHL-1 doubling time, an effect attributed to cell cycle arrest. Vascular mimicry, a close related angiogenesis process, was also impaired (73%). Fucan A mode of action could be related to gene expression modulation, in special β-catenin downregulation, a molecule with protagonist roles in important signaling pathways. Taken together, results indicate that Fucan A is a potential anticancer molecule and, therefore, deserves further investigation.</p>","PeriodicalId":690,"journal":{"name":"Marine Biotechnology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139562583","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Understanding the genetic composition and regional adaptation of marine species under environmental heterogeneity and fishing pressure is crucial for responsible management. In order to understand the genetic diversity and adaptability of yellowfin seabream (Acanthopagrus latus) along southern China coast, this study was conducted a seascape genome analysis on yellowfin seabream from the ecologically diverse coast, spanning over 1600 km. A total of 92 yellowfin seabream individuals from 15 sites were performed whole-genome resequencing, and 4,383,564 high-quality single nucleotide polymorphisms (SNPs) were called. By conducting a genotype-environment association analysis, 29,951 adaptive and 4,328,299 neutral SNPs were identified. The yellowfin seabream exhibited two distinct population structures, despite high gene flow between sites. The seascape genome analysis revealed that genetic structure was influenced by a variety of factors including salinity gradients, habitat distance, and ocean currents. The frequency of allelic variation at the candidate loci changed with the salinity gradient. Annotation of these loci revealed that most of the genes are associated with osmoregulation, such as kcnab2a, kcnk5a, and slc47a1. These genes are significantly enriched in pathways associated with ion transport including G protein-coupled receptor activity, transmembrane signaling receptor activity, and transporter activity. Overall, our findings provide insights into how seascape heterogeneity affects adaptive evolution, while providing important information for regional management in yellowfin seabream populations.
了解海洋物种在环境异质性和捕捞压力下的遗传组成和区域适应性对于负责任的管理至关重要。为了了解中国南方沿海黄鳍鲷的遗传多样性和适应性,本研究对黄鳍鲷进行了海景基因组分析。共对来自15个地点的92条黄鳍鲷进行了全基因组重测序,调用了4,383,564个高质量的单核苷酸多态性(SNPs)。通过基因型-环境关联分析,共鉴定出29,951个适应性SNP和4,328,299个中性SNP。尽管基因在不同地点之间流动频繁,黄鳍鲷仍表现出两种不同的种群结构。海景基因组分析表明,遗传结构受到多种因素的影响,包括盐度梯度、栖息地距离和洋流。候选位点的等位基因变异频率随盐度梯度而变化。对这些基因座的注释显示,大多数基因与渗透调节有关,如 kcnab2a、kcnk5a 和 slc47a1。这些基因在与离子转运相关的通路中明显富集,包括 G 蛋白偶联受体活性、跨膜信号受体活性和转运体活性。总之,我们的研究结果为海景异质性如何影响适应性进化提供了见解,同时也为黄鳍鲷种群的区域管理提供了重要信息。
{"title":"Seascapes Shaped the Local Adaptation and Population Structure of South China Coast Yellowfin Seabream (Acanthopagrus latus).","authors":"Wenhao Wang, Junrou Huang, Yan Hu, Jianxiang Feng, Dong Gao, Wenyu Fang, Meng Xu, Chunlei Ma, Zhenqiang Fu, Qinglong Chen, Xuanguang Liang, Jianguo Lu","doi":"10.1007/s10126-023-10277-6","DOIUrl":"10.1007/s10126-023-10277-6","url":null,"abstract":"<p><p>Understanding the genetic composition and regional adaptation of marine species under environmental heterogeneity and fishing pressure is crucial for responsible management. In order to understand the genetic diversity and adaptability of yellowfin seabream (Acanthopagrus latus) along southern China coast, this study was conducted a seascape genome analysis on yellowfin seabream from the ecologically diverse coast, spanning over 1600 km. A total of 92 yellowfin seabream individuals from 15 sites were performed whole-genome resequencing, and 4,383,564 high-quality single nucleotide polymorphisms (SNPs) were called. By conducting a genotype-environment association analysis, 29,951 adaptive and 4,328,299 neutral SNPs were identified. The yellowfin seabream exhibited two distinct population structures, despite high gene flow between sites. The seascape genome analysis revealed that genetic structure was influenced by a variety of factors including salinity gradients, habitat distance, and ocean currents. The frequency of allelic variation at the candidate loci changed with the salinity gradient. Annotation of these loci revealed that most of the genes are associated with osmoregulation, such as kcnab2a, kcnk5a, and slc47a1. These genes are significantly enriched in pathways associated with ion transport including G protein-coupled receptor activity, transmembrane signaling receptor activity, and transporter activity. Overall, our findings provide insights into how seascape heterogeneity affects adaptive evolution, while providing important information for regional management in yellowfin seabream populations.</p>","PeriodicalId":690,"journal":{"name":"Marine Biotechnology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139037277","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alkalinity is regarded as one of the primary stressors for aquatic animals in saline-alkaline water. Alternative splicing (AS) can significantly increase the diversity of transcripts and play key roles in stress response; however, the studies on AS under alkalinity stress of crustaceans are still limited. In the present study, we devoted ourselves to the study of AS under acute alkalinity stress at control (50 mg/L) and treatment groups (350 mg/L) by RNA-seq in pacific white shrimp (Litopenaeus vannamei). We identified a total of 10,556 AS events from 4865 genes and 619 differential AS (DAS) events from 519 DAS genes in pacific white shrimp. Functional annotation showed that the DAS genes primarily involved in spliceosome. Five splicing factors (SFs), U2AF1, PUF60, CHERP, SR140 and SRSF2 were significantly up-regulated and promoted AS. Furthermore, alkalinity activated the Leukocyte transendothelial migration, mTOR signaling pathway and AMPK signaling pathway, which regulated MAPK1, EIF3B and IGFP-RP1 associated with these pathways. We also studied three SFs (HSFP1, SRSF2 and NHE-RF1), which underwent AS to form different transcript isoforms. The above results demonstrated that AS was a regulatory mechanism in pacific white shrimp in response to acute alkalinity stress. SFs played vital roles in AS of pacific white shrimp, such as HSFP1, SRSF2 and NHE-RF1. DAS genes were significantly modified in immunity of pacific white shrimp to cope with alkalinity stress. This is the first study on the response of AS to acute alkalinity stress, which provided scientific basis for AS mechanism of crustaceans response to alkalinity stress.
{"title":"Alternative Splicing Reveals Acute Stress Response of Litopenaeus vannamei at High Alkalinity.","authors":"Xiang Shi, Ruiqi Zhang, Zhe Liu, Guiyan Zhao, Jintao Guo, Xue Mao, Baoyi Fan","doi":"10.1007/s10126-023-10281-w","DOIUrl":"10.1007/s10126-023-10281-w","url":null,"abstract":"<p><p>Alkalinity is regarded as one of the primary stressors for aquatic animals in saline-alkaline water. Alternative splicing (AS) can significantly increase the diversity of transcripts and play key roles in stress response; however, the studies on AS under alkalinity stress of crustaceans are still limited. In the present study, we devoted ourselves to the study of AS under acute alkalinity stress at control (50 mg/L) and treatment groups (350 mg/L) by RNA-seq in pacific white shrimp (Litopenaeus vannamei). We identified a total of 10,556 AS events from 4865 genes and 619 differential AS (DAS) events from 519 DAS genes in pacific white shrimp. Functional annotation showed that the DAS genes primarily involved in spliceosome. Five splicing factors (SFs), U2AF1, PUF60, CHERP, SR140 and SRSF2 were significantly up-regulated and promoted AS. Furthermore, alkalinity activated the Leukocyte transendothelial migration, mTOR signaling pathway and AMPK signaling pathway, which regulated MAPK1, EIF3B and IGFP-RP1 associated with these pathways. We also studied three SFs (HSFP1, SRSF2 and NHE-RF1), which underwent AS to form different transcript isoforms. The above results demonstrated that AS was a regulatory mechanism in pacific white shrimp in response to acute alkalinity stress. SFs played vital roles in AS of pacific white shrimp, such as HSFP1, SRSF2 and NHE-RF1. DAS genes were significantly modified in immunity of pacific white shrimp to cope with alkalinity stress. This is the first study on the response of AS to acute alkalinity stress, which provided scientific basis for AS mechanism of crustaceans response to alkalinity stress.</p>","PeriodicalId":690,"journal":{"name":"Marine Biotechnology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139416049","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-01Epub Date: 2024-01-16DOI: 10.1007/s10126-024-10290-3
Ju Li, Xiaohong Li, Simiao Fu, Yuxuan Meng, Xiaoyan Lv, Xin Zhang, Guozheng Liu, Jinsheng Sun
Limb autotomy and regeneration represent distinctive responses of crustaceans to environmental stress. Glucose metabolism plays a pivotal role in energy generation for tissue development and regeneration across various species. However, the relationship between glucose metabolism and tissue regeneration in crustaceans remains elusive. Therefore, this study is aimed at analyzing the alterations of glucose metabolic profile during limb autotomy and regeneration in Eriocheir sinensis, while also evaluating the effects of carbohydrate supplementation on limb regeneration. The results demonstrated that limb autotomy triggered a metabolic profile adaption at the early stage of regeneration. Hemolymph glucose levels were elevated, and multiple glucose catabolic pathways were enhanced in the hepatopancreas. Additionally, glucose and ATP levels in the regenerative limb were upregulated, along with increased expression of glucose transporters. Furthermore, the gene expression and activity of enzymes involved in gluconeogenesis were repressed in the hepatopancreas. These findings indicate that limb regeneration triggers metabolic profile adaptations to meet the elevated energy requirements. Moreover, the study observed that supplementation with corn starch enhanced limb regeneration capacity by promoting wound healing and blastema growth. Interestingly, dietary carbohydrate addition influenced limb regeneration by stimulating gluconeogenesis rather than glycolysis in the regenerative limb. Thus, these results underscore the adaptation of glucose metabolism during limb autotomy and regeneration, highlighting its essential role in the limb regeneration process of E. sinensis.
肢体自切和再生是甲壳动物对环境压力的独特反应。葡萄糖代谢在不同物种的组织发育和再生的能量生成过程中发挥着关键作用。然而,甲壳类动物的葡萄糖代谢与组织再生之间的关系仍然难以捉摸。因此,本研究旨在分析中华绒螯虾肢体自切和再生过程中葡萄糖代谢谱的变化,同时评估补充碳水化合物对肢体再生的影响。结果表明,肢体自体切除会在再生早期引发代谢特征适应。血淋巴葡萄糖水平升高,肝胰脏中多种葡萄糖分解途径增强。此外,再生肢体中的葡萄糖和 ATP 水平上调,葡萄糖转运体的表达也增加了。此外,肝胰腺中参与葡萄糖生成的酶的基因表达和活性受到抑制。这些发现表明,肢体再生会引发新陈代谢特征的适应,以满足能量需求的增加。此外,研究还观察到,补充玉米淀粉可促进伤口愈合和胚芽生长,从而提高肢体再生能力。有趣的是,膳食中碳水化合物的添加通过刺激再生肢体的糖元生成而不是糖酵解来影响肢体再生。因此,这些结果强调了葡萄糖代谢在肢体自切和再生过程中的适应性,突出了其在中华鳖肢体再生过程中的重要作用。
{"title":"Adaptation of Glucose Metabolism to Limb Autotomy and Regeneration in the Chinese Mitten Crab.","authors":"Ju Li, Xiaohong Li, Simiao Fu, Yuxuan Meng, Xiaoyan Lv, Xin Zhang, Guozheng Liu, Jinsheng Sun","doi":"10.1007/s10126-024-10290-3","DOIUrl":"10.1007/s10126-024-10290-3","url":null,"abstract":"<p><p>Limb autotomy and regeneration represent distinctive responses of crustaceans to environmental stress. Glucose metabolism plays a pivotal role in energy generation for tissue development and regeneration across various species. However, the relationship between glucose metabolism and tissue regeneration in crustaceans remains elusive. Therefore, this study is aimed at analyzing the alterations of glucose metabolic profile during limb autotomy and regeneration in Eriocheir sinensis, while also evaluating the effects of carbohydrate supplementation on limb regeneration. The results demonstrated that limb autotomy triggered a metabolic profile adaption at the early stage of regeneration. Hemolymph glucose levels were elevated, and multiple glucose catabolic pathways were enhanced in the hepatopancreas. Additionally, glucose and ATP levels in the regenerative limb were upregulated, along with increased expression of glucose transporters. Furthermore, the gene expression and activity of enzymes involved in gluconeogenesis were repressed in the hepatopancreas. These findings indicate that limb regeneration triggers metabolic profile adaptations to meet the elevated energy requirements. Moreover, the study observed that supplementation with corn starch enhanced limb regeneration capacity by promoting wound healing and blastema growth. Interestingly, dietary carbohydrate addition influenced limb regeneration by stimulating gluconeogenesis rather than glycolysis in the regenerative limb. Thus, these results underscore the adaptation of glucose metabolism during limb autotomy and regeneration, highlighting its essential role in the limb regeneration process of E. sinensis.</p>","PeriodicalId":690,"journal":{"name":"Marine Biotechnology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139471342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-01Epub Date: 2024-01-19DOI: 10.1007/s10126-024-10285-0
Michael Acquafredda, Ximing Guo, Daphne Munroe
There is clear evidence that the oceans are warming due to anthropogenic climate change, and the northeastern coast of USA contains some of the fastest warming areas. This warming is projected to continue with serious biological and social ramifications for fisheries and aquaculture. One species particularly vulnerable to warming is the Atlantic surfclam (Spisula solidissima). The surfclam is a critically important species, linking marine food webs and supporting a productive, lucrative, and sustainable fishery. The surfclam is also emerging as an attractive candidate for aquaculture diversification, but the warming of shallow coastal farms threatens the expansion of surfclam aquaculture. Little is known about the adaptive potential of surfclams to cope with ocean warming. In this study, the surfclam transcriptome under heat stress was examined. Two groups of surfclams were subjected to heat stress to assess how artificial selection may alter gene expression. One group of clams had been selected for greater heat tolerance (HS) and the other was composed of random control clams (RC). After a 6-h exposure to 16 or 29 °C, gill transcriptome expression profiles of the four temperature/group combinations were determined by RNA sequencing and compared. When surfclams experienced heat stress, they exhibited upregulation of heat shock proteins (HSPs), inhibitors of apoptosis (IAPs), and other stress-response related genes. RC clams differentially expressed 1.7 times more genes than HS clams, yet HS clams had a stronger response of key stress response genes, including HSPs, IAPs, and genes involved with mitigating oxidative stress. The findings imply that the HS clams have a more effective response to heat stress after undergoing the initial selection event due to genetic differences created by the selection, epigenetic memory of the first heat shock, or both. This work provides insights into how surfclams adapt to heat stress and should inform future breeding programs that attempt to breed surfclam for greater heat tolerance, and ultimately bring greater resiliency to shellfish farms.
{"title":"Transcriptomic Response of the Atlantic Surfclam (Spisula solidissima) to Acute Heat Stress.","authors":"Michael Acquafredda, Ximing Guo, Daphne Munroe","doi":"10.1007/s10126-024-10285-0","DOIUrl":"10.1007/s10126-024-10285-0","url":null,"abstract":"<p><p>There is clear evidence that the oceans are warming due to anthropogenic climate change, and the northeastern coast of USA contains some of the fastest warming areas. This warming is projected to continue with serious biological and social ramifications for fisheries and aquaculture. One species particularly vulnerable to warming is the Atlantic surfclam (Spisula solidissima). The surfclam is a critically important species, linking marine food webs and supporting a productive, lucrative, and sustainable fishery. The surfclam is also emerging as an attractive candidate for aquaculture diversification, but the warming of shallow coastal farms threatens the expansion of surfclam aquaculture. Little is known about the adaptive potential of surfclams to cope with ocean warming. In this study, the surfclam transcriptome under heat stress was examined. Two groups of surfclams were subjected to heat stress to assess how artificial selection may alter gene expression. One group of clams had been selected for greater heat tolerance (HS) and the other was composed of random control clams (RC). After a 6-h exposure to 16 or 29 °C, gill transcriptome expression profiles of the four temperature/group combinations were determined by RNA sequencing and compared. When surfclams experienced heat stress, they exhibited upregulation of heat shock proteins (HSPs), inhibitors of apoptosis (IAPs), and other stress-response related genes. RC clams differentially expressed 1.7 times more genes than HS clams, yet HS clams had a stronger response of key stress response genes, including HSPs, IAPs, and genes involved with mitigating oxidative stress. The findings imply that the HS clams have a more effective response to heat stress after undergoing the initial selection event due to genetic differences created by the selection, epigenetic memory of the first heat shock, or both. This work provides insights into how surfclams adapt to heat stress and should inform future breeding programs that attempt to breed surfclam for greater heat tolerance, and ultimately bring greater resiliency to shellfish farms.</p>","PeriodicalId":690,"journal":{"name":"Marine Biotechnology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10869415/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139490511","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-13DOI: 10.1007/s10126-024-10283-2
Qiong Yang, Hong Yu, Qi Li
The fecundity of triploid female Crassostrea gigas exhibited significant variation and was lower compared to diploid individuals. Previous studies categorized mature stage triploid female C. gigas into two groups: female α, characterized by a high number of oocytes, and female β, displaying few or no oocytes. To investigate the molecular mechanisms underlying irregular oogenesis and fecundity differences in triploid C. gigas, we performed a comparative analysis of gonad transcriptomes at different stages of gonadal development, including female α, female β, and diploids. During early oogenesis, functional enrichment analysis between female diploids and putative female β triploids revealed differently expressed genes (DEGs) in the ribosome and ribosome biogenesis pathways. Expression levels of DEGs in these pathways were significantly decreased in the putative female β triploid, suggesting a potential role of reduced ribosome levels in obstructing triploid oogenesis. Moreover, to identify regulatory pathways in gonad development, female oysters at the early and mature stages were compared. The DNA repair and recombination proteins pathways were enriched in female diploids and female α triploids but absent in female β triploids. Overall, we propose that decreased ribosome biogenesis in female triploids hinders the differentiation of germ stem cells, leading to the formation of a large number of abnormal germ cells and ultimately resulting in reduced fecundity. The variation in fertility among triploids appeared to be related to the degree of DNA damage repair during female gonad development. This study offers valuable insights into the oogenesis process in female triploid C. gigas.
{"title":"Comparative Transcriptome Analysis Reveals the Role of Ribosome Reduction in Impeding Oogenesis in Female Triploid Crassostrea Gigas","authors":"Qiong Yang, Hong Yu, Qi Li","doi":"10.1007/s10126-024-10283-2","DOIUrl":"https://doi.org/10.1007/s10126-024-10283-2","url":null,"abstract":"<p>The fecundity of triploid female <i>Crassostrea gigas</i> exhibited significant variation and was lower compared to diploid individuals. Previous studies categorized mature stage triploid female <i>C. gigas</i> into two groups: female α, characterized by a high number of oocytes, and female β, displaying few or no oocytes. To investigate the molecular mechanisms underlying irregular oogenesis and fecundity differences in triploid <i>C. gigas</i>, we performed a comparative analysis of gonad transcriptomes at different stages of gonadal development, including female α, female β, and diploids. During early oogenesis, functional enrichment analysis between female diploids and putative female β triploids revealed differently expressed genes (DEGs) in the ribosome and ribosome biogenesis pathways. Expression levels of DEGs in these pathways were significantly decreased in the putative female β triploid, suggesting a potential role of reduced ribosome levels in obstructing triploid oogenesis. Moreover, to identify regulatory pathways in gonad development, female oysters at the early and mature stages were compared. The DNA repair and recombination proteins pathways were enriched in female diploids and female α triploids but absent in female β triploids. Overall, we propose that decreased ribosome biogenesis in female triploids hinders the differentiation of germ stem cells, leading to the formation of a large number of abnormal germ cells and ultimately resulting in reduced fecundity. The variation in fertility among triploids appeared to be related to the degree of DNA damage repair during female gonad development. This study offers valuable insights into the oogenesis process in female triploid <i>C. gigas</i>.</p>","PeriodicalId":690,"journal":{"name":"Marine Biotechnology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139462514","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-05DOI: 10.1007/s10126-023-10280-x
Kohei Yoneda, Chun Hung Man, Yoshiaki Maeda, Iwane Suzuki
A marine thraustochytrid, Aurantiochytrium, is a promising organism to produce docosahexaenoic acid (DHA) and squalene. Utilization of inexpensive substances such as proteins in wastes and by-products from the food industry for cultivation is a considerable option to reduce production cost; however, the proteolytic ability of Aurantiochytrium spp. is low compared to taxonomically close Shizochytrium aggregatum. We previously identified extracellular protease (extracellular protease 1, EP1) in S. aggregatum ATCC 28209 from the supernatant of the culture and found that a similar protease gene (EP2) was located downstream of the EP1 gene. In the present study, we created the transformants expressing SaEP1 and/or SaEP2 to enhance the proteolytic ability of Aurantiochytrium sp. 18W-13a strain and cultivated them in the medium containing casein as a test protein substrate. Through SDS-PAGE analysis, we confirmed that casein in the supernatant was more efficiently degraded by the transformants than the wild type, suggesting that the expressed protease(s) were properly expressed and excreted. After 4-day cultivation in the casein medium, the value of optical density at 660 nm and the cell number in the culture of the transformant that expressed both SaEP1 and SaEP2 (designated as EP12 strain) showed 1.48- and 1.38-fold higher than those of wild type, respectively. The DHA and squalene yield of the EP12 strain were respectively 158.3 and 0.23 mg L-1, and these values were 1.42- and 2.01-fold higher than those of wild type, respectively, suggesting that the EP12 created in the present study is a favorable strain for the cultivation using protein-containing medium.
{"title":"Genetic Modification of Aurantiochytrium sp. 18W-13a for Enhancement of Proteolytic Activity by Heterologous Expression of Extracellular Proteases.","authors":"Kohei Yoneda, Chun Hung Man, Yoshiaki Maeda, Iwane Suzuki","doi":"10.1007/s10126-023-10280-x","DOIUrl":"https://doi.org/10.1007/s10126-023-10280-x","url":null,"abstract":"<p><p>A marine thraustochytrid, Aurantiochytrium, is a promising organism to produce docosahexaenoic acid (DHA) and squalene. Utilization of inexpensive substances such as proteins in wastes and by-products from the food industry for cultivation is a considerable option to reduce production cost; however, the proteolytic ability of Aurantiochytrium spp. is low compared to taxonomically close Shizochytrium aggregatum. We previously identified extracellular protease (extracellular protease 1, EP1) in S. aggregatum ATCC 28209 from the supernatant of the culture and found that a similar protease gene (EP2) was located downstream of the EP1 gene. In the present study, we created the transformants expressing SaEP1 and/or SaEP2 to enhance the proteolytic ability of Aurantiochytrium sp. 18W-13a strain and cultivated them in the medium containing casein as a test protein substrate. Through SDS-PAGE analysis, we confirmed that casein in the supernatant was more efficiently degraded by the transformants than the wild type, suggesting that the expressed protease(s) were properly expressed and excreted. After 4-day cultivation in the casein medium, the value of optical density at 660 nm and the cell number in the culture of the transformant that expressed both SaEP1 and SaEP2 (designated as EP12 strain) showed 1.48- and 1.38-fold higher than those of wild type, respectively. The DHA and squalene yield of the EP12 strain were respectively 158.3 and 0.23 mg L<sup>-1</sup>, and these values were 1.42- and 2.01-fold higher than those of wild type, respectively, suggesting that the EP12 created in the present study is a favorable strain for the cultivation using protein-containing medium.</p>","PeriodicalId":690,"journal":{"name":"Marine Biotechnology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139097100","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
In recent years, a new type of Spiroplasma has been found that can cause “tremor disease” of the Chinese mitten crab Eriocheir sinensis. The outbreak of epidemic tremor disease has caused a serious setback in the Chinese mitten crab farming industry, with an incidence rate of more than 30% and mortality rates of 80–100%. Therefore, finding a sensitive method to detect tremor disease in E. sinensis has become a current research focus. In this research, a loop-mediated isothermal amplification detection method coupled with hydroxynaphthol blue dye (LAMP-HNB) was developed and used to rapidly detect Spiroplasma eriocheiris. First, we designed and synthesized specific outer primers, inner primers and loop primers based on the 16S ribosomal RNA gene of S. eriocheiris. Second, the LAMP-HNB detection method for S. eriocheiris was successfully established by screening the primers, adjusting the temperature and time of the reaction, and optimizing the concentrations of Mg2+ and dNTPs. In the specific tests, only samples infected with S. eriocheiris showed positive results, and other infections caused by bacteria and parasites tested negative, proving that the test has high specificity. Moreover, the detection limit was 2.5 × 10–6 ng/µL, indicating high sensitivity. This method for detecting S. eriocheiris provides rapid visual output based on LAMP-HNB detection and is a simple, fast, sensitive, and inexpensive method that can be applied to a wide range of field investigations.
{"title":"Rapid Visual Detection of Spiroplasma eriocheiris by Loop-Mediated Isothermal Amplification with Hydroxynaphthol Blue Dye","authors":"Shun Zhou, Zongrui Yang, Baoshan Guo, Jingyuan Yi, Yunfei Pang, Ruixin Feng, Jiaxue Song, Yunji Xiu","doi":"10.1007/s10126-023-10282-9","DOIUrl":"https://doi.org/10.1007/s10126-023-10282-9","url":null,"abstract":"<p>In recent years, a new type of <i>Spiroplasma</i> has been found that can cause “tremor disease” of the Chinese mitten crab <i>Eriocheir sinensis</i>. The outbreak of epidemic tremor disease has caused a serious setback in the Chinese mitten crab farming industry, with an incidence rate of more than 30% and mortality rates of 80–100%. Therefore, finding a sensitive method to detect tremor disease in <i>E. sinensis</i> has become a current research focus. In this research, a loop-mediated isothermal amplification detection method coupled with hydroxynaphthol blue dye (LAMP-HNB) was developed and used to rapidly detect <i>Spiroplasma eriocheiris</i>. First, we designed and synthesized specific outer primers, inner primers and loop primers based on the <i>16S ribosomal RNA</i> gene of <i>S. eriocheiris</i>. Second, the LAMP-HNB detection method for <i>S. eriocheiris</i> was successfully established by screening the primers, adjusting the temperature and time of the reaction, and optimizing the concentrations of Mg<sup>2+</sup> and dNTPs. In the specific tests, only samples infected with <i>S. eriocheiris</i> showed positive results, and other infections caused by bacteria and parasites tested negative, proving that the test has high specificity. Moreover, the detection limit was 2.5 × 10<sup>–6</sup> ng/µL, indicating high sensitivity. This method for detecting <i>S. eriocheiris</i> provides rapid visual output based on LAMP-HNB detection and is a simple, fast, sensitive, and inexpensive method that can be applied to a wide range of field investigations.</p>","PeriodicalId":690,"journal":{"name":"Marine Biotechnology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139082102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}