Physiological and Metabolic Mechanisms of Procambarus clarkii in Response to High Temperature Stress

doi:10.13560/j.cnki.biotech.bull.1985.2024-0184

Abstract

Abstract:

【Objective】 To understand the physiological and metabolic mechanisms of Procambarus clarkii in response to high temperature stress, and to provide references for improving P. clarkii culture management and variety breeding. 【Method】 The P. clarkii were exposed to 32℃ and 37℃, and 26℃ serving as the control condition. Gill samples were collected at 24 and 72 h post-exposure. The study employed a comprehensive analysis of histological observation, biochemical indicators（malondialdehyde, reactive oxygen species, total antioxidant capacity, pyruvate kinase, hexokinase, alkaline phosphatase, phosphoenolpyruvate carboxykinase, lysozyme, and acid phosphatase）, and transcriptome to comprehensively explore the physiological and metabolic responses of P. clarkii to high temperature stress. 【Result】 High temperature damaged the gill tissues and the biochemical indicators overall decreased with the increasing of temperature. When compared to the control group, a total of 226 and 6 937 differentially expressed genes（DEGs）were identified at 24 and 72 h in 32℃ group, and a total of 2 928 and 9 449 DEGs were identified at 24 and 72 h in 37℃ group. GO and KEGG enrichment analyses indicate that DEGs were significantly enriched in O-glycan biosynthesis, extracellular matrix-receptor interaction, arachidonic acid metabolism, sphingolipid metabolism, as well as starch and sucrose metabolism processes. Additionally, the DEGs were found to cluster in modules pertinent to immune and energy metabolism based on weighted gene co-expression network analysis（WGCNA）, hub genes like histones（H2A）and mitochondrial components（ND6）were identified. Ten DEGs were randomly selected to validate the RNA-seq results using reverse transcription-quantitative polymerase chain reaction（RT-qPCR）, and the results and the expression profiles of these DEGs were of a well consistent with the high-throughput data, which confirmed the reliability of transcriptome results. 【Conclusion】 At temperatures of 32℃ and 37℃, the gills of P. clarkii are damaged. The glucose utilization rate of P. clarkii is reduced in high temperature environments, causing cellular function to be maintained through the adjustments of heat shock proteins, antioxidant capacity, and metabolism.

Key words: Procambarus clarkii, high temperature, gill, physiological metabolism, mechanism

ZOU Yong-feng, BAO Zhi-ming, CAO Pan-hui, ZHANG Jia-yuan, GUO Jie-yu, SU Xian-bin, XU Yu, XU Zhi-qiang, GUO Hui. Physiological and Metabolic Mechanisms of Procambarus clarkii in Response to High Temperature Stress[J]. Biotechnology Bulletin, 2024, 40(11): 321-334.

Figures/Tables 12

References 37

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基因 Gene	序列 Sequence（5'-3'）	基因ID Gene ID
KP106	F: ACAACACTGCTGGCGGACTG	LOC123770190
KP106	R: CTTCGTCTTCTCCTTCGTCTCCTC	LOC123770190
TRET1	F: CGACGGAGAGAAGAGGATGATGAC	LOC123770879
TRET1	R: CCTAAGTTGATATTGCTGGCGTTGG	LOC123770879
HRCP3	F: TCCGCCGCTCCCAAGAAATC	LOC123759871
HRCP3	R: GCTGAATACCTGGAGGGCTCTG	LOC123759871
CAH2	F: CCTTGGTGTCTCTGGCGATGG	LOC123772758
CAH2	R:AAGTGGAACTGAGTGAAGGTGTAGG	LOC123772758
SAMH1	F: TCAGCGAGAAGGACGTGTTGTG	LOC123760680
SAMH1	R: TGATGAAAGCCTCCCAGCAGTG	LOC123760680
HSP71D	F: CCTCCTTGATGTGGCACCTCTG	LOC123759425
HSP71D	R: AGTGACCGCTGGTTGGTTATCTG	LOC123759425
HSP70	F: TGGCGGCTCCAGTAATTGGTATC	LOC123774884
HSP70	R: TGAGTCGTTCTGTGTCGGTGAAG	LOC123774884
HSP71	F: CCTACGGTGCGGCTGTTCAG	LOC123759427
HSP71	R: AAGACAGAGGTGCCACATCAAGG	LOC123759427
HSP83	F: GTGACCTCTCCGTGCTGTATCG	LOC123770521
HSP83	R: CTCGCAGTGCCTGAGCCTTC	LOC123770521
HSP7D	F: CAGTCTGAGGGCGTGAAGGATG	LOC123762431
HSP7D	R: TGAATACTTGCTGTTGCTTGGTTGG	LOC123762431
β-actin	F: CACTGCCGCCTCATCCTCTTC R: GAACCTCTCGTTGCCAATGGTAATG	LOC123745344

基因 Gene	序列 Sequence（5'-3'）	基因ID Gene ID
KP106	F: ACAACACTGCTGGCGGACTG	LOC123770190
KP106	R: CTTCGTCTTCTCCTTCGTCTCCTC	LOC123770190
TRET1	F: CGACGGAGAGAAGAGGATGATGAC	LOC123770879
TRET1	R: CCTAAGTTGATATTGCTGGCGTTGG	LOC123770879
HRCP3	F: TCCGCCGCTCCCAAGAAATC	LOC123759871
HRCP3	R: GCTGAATACCTGGAGGGCTCTG	LOC123759871
CAH2	F: CCTTGGTGTCTCTGGCGATGG	LOC123772758
CAH2	R:AAGTGGAACTGAGTGAAGGTGTAGG	LOC123772758
SAMH1	F: TCAGCGAGAAGGACGTGTTGTG	LOC123760680
SAMH1	R: TGATGAAAGCCTCCCAGCAGTG	LOC123760680
HSP71D	F: CCTCCTTGATGTGGCACCTCTG	LOC123759425
HSP71D	R: AGTGACCGCTGGTTGGTTATCTG	LOC123759425
HSP70	F: TGGCGGCTCCAGTAATTGGTATC	LOC123774884
HSP70	R: TGAGTCGTTCTGTGTCGGTGAAG	LOC123774884
HSP71	F: CCTACGGTGCGGCTGTTCAG	LOC123759427
HSP71	R: AAGACAGAGGTGCCACATCAAGG	LOC123759427
HSP83	F: GTGACCTCTCCGTGCTGTATCG	LOC123770521
HSP83	R: CTCGCAGTGCCTGAGCCTTC	LOC123770521
HSP7D	F: CAGTCTGAGGGCGTGAAGGATG	LOC123762431
HSP7D	R: TGAATACTTGCTGTTGCTTGGTTGG	LOC123762431
β-actin	F: CACTGCCGCCTCATCCTCTTC R: GAACCTCTCGTTGCCAATGGTAATG	LOC123745344

模块 Model	基因ID Gene ID	基因描述 Gene description
Red	LOC123769279	未鉴定蛋白质 Uncharacterized protein
Red	LOC123769505	锚蛋白-3样异构体X1 Ankyrin-3-like isoform X1
Red	LOC123762606	未鉴定蛋白质 Uncharacterized protein
Red	LOC123749095	真核翻译起始因子3亚基c样 Eukaryotic translation initiation factor 3 subunit C-like
Red	LOC123767106	菱形相关蛋白3样异构体X1 Rhomboid-related protein 3-like isoform X1
Red	LOC123767084	肌苷-尿苷偏好核苷水解酶样异构体X1 Inosine-uridine preferring nucleoside hydrolase-like isoform X1
Red	LOC123768178	唾液蛋白样 Sialin-like
Red	LOC123775008	毛发角蛋白样 Trichohyalin-like
Yellow	ND6	ND6基因产物 ND6 gene product
Yellow	COX2	COX2基因产物 COX2 gene product
Yellow	CYTB	CYTB基因产物 CYTB gene product
Yellow	LOC123750758	1-样凝血反应蛋白亚型X1的崩解素和金属蛋白酶 A disintegrin and metalloproteinase with thrombospondin motifs 1-like isoform X1
Yellow	LOC123754039	UDP葡萄糖转移酶2样 UDP-glucosyltransferase 2-like
Yellow	LOC123746798	富含精氨酸/丝氨酸的卷曲蛋白2样异构体X6 Arginine/serine-rich coiled-coil protein 2-like isoform X6
Yellow	LOC123760103	整合酶/重组酶xerD同源物 Integrase/recombinase xerD homolog
Yellow	LOC123763597	脑畸形1同源物 Llissencephaly-1 homolog
Yellow	LOC123765558	转录终止因子Rho样 Transcription termination factor Rho-like
Yellow	LOC123775192	胞质Fe-S簇组装因子NUBP2同源异构体X1 Cytosolic Fe-S cluster assembly factor NUBP2 homolog isoform X1
Magenta	LOC123751850	未鉴定蛋白质 Uncharacterized protein
Magenta	LOC123745689	未鉴定蛋白质 Uncharacterized protein
Magenta	LOC123752918	组蛋白H2A样 Histone H2A-like
Magenta	LOC123749390	组蛋白H2A Histone H2A
Magenta	LOC123750872	组蛋白H2A Histone H2A-like
Magenta	LOC123750985	未鉴定蛋白异构体X1 Uncharacterized protein isoform X1
Purple	LOC123768029	未鉴定蛋白质 Uncharacterized protein
Purple	LOC123768775	神经细胞黏附分子2样异构体X1 Neural cell adhesion molecule 2-like isoform X1
Purple	LOC123769508	未鉴定蛋白质 Uncharacterized protein
Purple	LOC123769760	未表征的蛋白异构体X1 Uncharacterized protein isoform X1
Purple	LOC123772183	钠/葡萄糖共转运蛋白4样 Sodium/glucose cotransporter 4-like
Purple	LOC123747964	部分未鉴定蛋白 Uncharacterized protein, partial
Purple	LOC123759325	三胸蛋白osa样 Trithorax group protein osa-like
Purple	LOC123760348	肌醇多磷酸4-磷酸酶I型a样 Inositol polyphosphate-4-phosphatase type I A-like

模块 Model	基因ID Gene ID	基因描述 Gene description
Red	LOC123769279	未鉴定蛋白质 Uncharacterized protein
Red	LOC123769505	锚蛋白-3样异构体X1 Ankyrin-3-like isoform X1
Red	LOC123762606	未鉴定蛋白质 Uncharacterized protein
Red	LOC123749095	真核翻译起始因子3亚基c样 Eukaryotic translation initiation factor 3 subunit C-like
Red	LOC123767106	菱形相关蛋白3样异构体X1 Rhomboid-related protein 3-like isoform X1
Red	LOC123767084	肌苷-尿苷偏好核苷水解酶样异构体X1 Inosine-uridine preferring nucleoside hydrolase-like isoform X1
Red	LOC123768178	唾液蛋白样 Sialin-like
Red	LOC123775008	毛发角蛋白样 Trichohyalin-like
Yellow	ND6	ND6基因产物 ND6 gene product
Yellow	COX2	COX2基因产物 COX2 gene product
Yellow	CYTB	CYTB基因产物 CYTB gene product
Yellow	LOC123750758	1-样凝血反应蛋白亚型X1的崩解素和金属蛋白酶 A disintegrin and metalloproteinase with thrombospondin motifs 1-like isoform X1
Yellow	LOC123754039	UDP葡萄糖转移酶2样 UDP-glucosyltransferase 2-like
Yellow	LOC123746798	富含精氨酸/丝氨酸的卷曲蛋白2样异构体X6 Arginine/serine-rich coiled-coil protein 2-like isoform X6
Yellow	LOC123760103	整合酶/重组酶xerD同源物 Integrase/recombinase xerD homolog
Yellow	LOC123763597	脑畸形1同源物 Llissencephaly-1 homolog
Yellow	LOC123765558	转录终止因子Rho样 Transcription termination factor Rho-like
Yellow	LOC123775192	胞质Fe-S簇组装因子NUBP2同源异构体X1 Cytosolic Fe-S cluster assembly factor NUBP2 homolog isoform X1
Magenta	LOC123751850	未鉴定蛋白质 Uncharacterized protein
Magenta	LOC123745689	未鉴定蛋白质 Uncharacterized protein
Magenta	LOC123752918	组蛋白H2A样 Histone H2A-like
Magenta	LOC123749390	组蛋白H2A Histone H2A
Magenta	LOC123750872	组蛋白H2A Histone H2A-like
Magenta	LOC123750985	未鉴定蛋白异构体X1 Uncharacterized protein isoform X1
Purple	LOC123768029	未鉴定蛋白质 Uncharacterized protein
Purple	LOC123768775	神经细胞黏附分子2样异构体X1 Neural cell adhesion molecule 2-like isoform X1
Purple	LOC123769508	未鉴定蛋白质 Uncharacterized protein
Purple	LOC123769760	未表征的蛋白异构体X1 Uncharacterized protein isoform X1
Purple	LOC123772183	钠/葡萄糖共转运蛋白4样 Sodium/glucose cotransporter 4-like
Purple	LOC123747964	部分未鉴定蛋白 Uncharacterized protein, partial
Purple	LOC123759325	三胸蛋白osa样 Trithorax group protein osa-like
Purple	LOC123760348	肌醇多磷酸4-磷酸酶I型a样 Inositol polyphosphate-4-phosphatase type I A-like