低温与无水胁迫下凡纳滨对虾肌肉品质劣变标记蛋白识别

Identification of Marker Proteins Indicating Potential Flesh Quality Deterioration of Litopenaeus vannamei Under Low Temperature Stress andWaterless Stress

【目的】探明凡纳滨对虾(Litopenaeus vannamei)保活流通过程中低温和无水双重胁迫诱导肌肉品质劣变与差异表达蛋白之间的相关性,识别品质劣变的潜在标记蛋白。【方法】在模拟的海水环境中[溶解氧质量浓度为7.2 mg/L,温度控制在(12±1)℃]观察对虾在低温和无水胁迫下的生理反应,通过串联质谱标签蛋白质组学方法,分析环境胁迫导致的肝胰腺蛋白质组显著差异性变化,并将其与肌肉品质进行相关性分析。【结果】与正常对照组相比,低温组和低温+无水组分别鉴定了33和44个显著差异蛋白质(DEPs),这些蛋白质大多在溶酶体、糖酵解或糖异生、黏附等通路显著富集(P<0.05),同时胰蛋白酶-1、二肽基肽酶显著上调(P<0.05),而磷酸丙糖异构酶和醛脱氢酶显著下调(P<0.05)。此外,颜色和质构指标的变化与微管蛋白、凝溶胶蛋白、层黏连蛋白、内源性蛋白酶和代谢酶表达水平显著相关。【结论】本研究初次探明肝胰腺DEPs与肌肉品质间的内在联系,识别的典型DEPs可作为品质劣变的潜在标志蛋白,为监测无水运输期间凡纳滨对虾肌肉品质劣变提供分子靶标。

【Objective】The aim of this study is to investigate the correlation between muscle quality deterioration and differently expressed protein in hepatopancreas of Litopenaeus vannamei under low temperature(LT)stress and waterless duration(WD)stress during the anaerobic transportation,and to identify the potentially proteomic markers indicating the quality deterioration.【Method】The physiological responses of shrimp under LT stress and WD stress were observed in a simulated marine environment with a dissolved oxygen content of 7.2 mg/L and a temperature control of(12±1)℃.The significant difference of hepatopancreas proteome induced by the dual stress was analyzed by tandem mass spectrometry,and its correlation with muscle quality was analyzed accordingly.【Results】Compared with the control,33 and 44 differently expressed proteins were identified in LT and LT+WD groups,respectively,among which the most DEPs were significantly enriched in lysosomal,glycolytic or gluconeogenic,and adhesion pathways(P<0.05),while trypsin-1 and dipeptidyl peptidase underwent significant up-regulatied(P<0.05)and phosphoglycan isomerase and aldehyde dehydrogenase were significantly down-regulated(P<0.05).Moreover,the deterioration in colour and texture attributes was closely associated with the expression of tubulin,gelsolin,laminin,endogenous protease and metabolic enzymes.【Conclusion】This research found out that the intrinsic relationship between hepatopancreas DEPs and muscle quality for the first time.The identified typical DEPs could be used as potential marker proteins indicating quality deterioration,and thus providing a molecular target for monitoring muscle quality deterioration of L.vannamei during waterless live transport.