中国水产科学
中國水產科學
중국수산과학
Journal of Fishery Sciences of China
2014年
6期
1172-1180
,共9页
于坤%叶海辉%黄陈翠%巩杰%黄辉洋
于坤%葉海輝%黃陳翠%鞏傑%黃輝洋
우곤%협해휘%황진취%공걸%황휘양
拟穴青蟹%ANT2%温度%盐度%应激%实时定量PCR
擬穴青蟹%ANT2%溫度%鹽度%應激%實時定量PCR
의혈청해%ANT2%온도%염도%응격%실시정량PCR
Scylla paramamosain%ANT2%temperature%salinity%stress%real time-PCR
采用qRT-PCR技术,对不同温度(10℃,15℃,20℃,25℃)以及低温(10℃,15℃)和盐度(10,35)联合作用下拟穴青蟹(Scylla paramamosain)(甲壳长3.7~5.8 cm,体质量80~100 g) ANT2基因的表达进行了检测。结果表明:在4个温度下肝胰腺、肌肉、鳃中 ANT2基因表达量依次为低、中、高,存在组织表达特异性。随着温度降低, ANT2基因总体上趋低表达,但在15℃时ANT2基因在肌肉(1~12 h)和鳃中均显著高于20℃( P<0.05)。该结果提示, ANT2基因与能量代谢相关,且在一定低温下被诱导表达,可能参与低温胁迫的应答。相同低温下,3种组织中ANT2基因在盐度10的表达量通常高于盐度35的(P<0.05),说明低温下低盐环境有利于提高青蟹体内代谢,增强耐寒能力。
採用qRT-PCR技術,對不同溫度(10℃,15℃,20℃,25℃)以及低溫(10℃,15℃)和鹽度(10,35)聯閤作用下擬穴青蟹(Scylla paramamosain)(甲殼長3.7~5.8 cm,體質量80~100 g) ANT2基因的錶達進行瞭檢測。結果錶明:在4箇溫度下肝胰腺、肌肉、鰓中 ANT2基因錶達量依次為低、中、高,存在組織錶達特異性。隨著溫度降低, ANT2基因總體上趨低錶達,但在15℃時ANT2基因在肌肉(1~12 h)和鰓中均顯著高于20℃( P<0.05)。該結果提示, ANT2基因與能量代謝相關,且在一定低溫下被誘導錶達,可能參與低溫脅迫的應答。相同低溫下,3種組織中ANT2基因在鹽度10的錶達量通常高于鹽度35的(P<0.05),說明低溫下低鹽環境有利于提高青蟹體內代謝,增彊耐寒能力。
채용qRT-PCR기술,대불동온도(10℃,15℃,20℃,25℃)이급저온(10℃,15℃)화염도(10,35)연합작용하의혈청해(Scylla paramamosain)(갑각장3.7~5.8 cm,체질량80~100 g) ANT2기인적표체진행료검측。결과표명:재4개온도하간이선、기육、새중 ANT2기인표체량의차위저、중、고,존재조직표체특이성。수착온도강저, ANT2기인총체상추저표체,단재15℃시ANT2기인재기육(1~12 h)화새중균현저고우20℃( P<0.05)。해결과제시, ANT2기인여능량대사상관,차재일정저온하피유도표체,가능삼여저온협박적응답。상동저온하,3충조직중ANT2기인재염도10적표체량통상고우염도35적(P<0.05),설명저온하저염배경유리우제고청해체내대사,증강내한능력。
The mud crab Scylla paramamosain, a eurythermal and euryhaline species, is a commercially exploited crab inhabiting the southeastern coast of China. In recent years, mud crab aquaculture has suffered as a result of unsettled weather, which has led to a seasonal temperature and salinity changes in the sea water. This study aims to explore the molecular mechanism of cold tolerance in S. paramamosain by examining the expression of adenine nucleotide trans-locase 2 (ANT2) at different temperatures and salinities. ANT belongs to the mitochondrial carrier superfamily (MCF). It is the most abundant inner membrane protein of the mitochondrion and is responsible for the transport of ADP and ATP in the cytoplasm and mitochondrial matrix through the inner mitochondrial membrane. In this study, the relative expression of ANT2 mRNA was detected using real-time quantitative polymerase chain reaction at different tempera-tures (10℃, 15℃, 20℃, 25℃), and low temperatures (10℃, 15℃) combined with salinity (10, 35) in mud crabs (cara-pace width 3.7–5.8 cm, body weight 80–100 g). Real-time qPCR indicated that, at different temperatures, ANT2 was expressed in the hepatopancreas, muscle and gill. The relative expression of ANT2 mRNA was lowest in the hepatopan-creas, at a medium level in muscle, and highest in the gill, and the results indicated that the expression of ANT2 was tissue-specific. With declining temperature, the expression of ANT2 mRNA decreased generally, but at 15℃in the mus-cle (1–12 h) and gill, the expression of ANT2 was significantly higher than that at 20℃(P<0.05). The expression pattern of ANT2 at different temperatures demonstrated that it might be associated with energy metabolism, and participate in stress responses at a certain range of low temperatures, stimulating the expression of ANT2 in the mud crab. At the same temperature, the expression of ANT2 at a salinity of 10 was significantly higher than at the higher salinity of 35 (P<0.05), which indicated low salinity might be helpful for metabolism in mud crabs, enhancing cold resistance. This research might provide theoretical guidance for crab aquaculture and management to some extent.