热胁迫对不同类胡萝卜素总含量的贵贝 Chlamys nobilis 的死亡率和 HSP90 表达水平的影响。

Cell Stress and Chaperones Pub Date : 2020-01-01 Epub Date: 2019-11-25 DOI:10.1007/s12192-019-01052-5
Dewei Cheng, Hongxing Liu, Hongkuan Zhang, Karsoon Tan, Ting Ye, Hongyu Ma, Shengkang Li, Huaiping Zheng
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摘要

贵贝(Chlamys nobilis)是一种具有重要经济价值的海洋双壳贝类,自 20 世纪 80 年代以来一直在中国南部海域养殖。遗憾的是,该扇贝品种在夏季经常出现大量死亡。本研究旨在探讨热休克蛋白 90(HSP90)的表达和类胡萝卜素的水平能否增强扇贝的高温抗逆性。首先,鉴定并克隆了扇贝的 HSP90 同源物(命名为 CnHSP90)。CnHSP90 的完整 cDNA 序列为 2631 bp,包括一个 2181 bp 的开放阅读框(ORF),编码 726 个氨基酸的多肽,具有五个 HSP90 家族特征,与 HSP90 家族成员具有高度同源性。CnHSP90 在包括肠、肾、内脏、套膜、鳃和性腺在内的所有受检组织中均普遍表达,其中以性腺的表达量最高。对总类胡萝卜素含量(TCC)明显不同的金色扇贝和棕色扇贝进行急性热挑战,测定其 LTE50(36 h 热休克时的半致死温度)和 LTI50(热休克后的半致死时间)以及 CnHSP90 基因表达与 TCC 的相关性。金色扇贝的LTE50(32.14 °C)高于棕色扇贝(31.19 °C),在所有测试温度下的LTI50都更长,表明金色扇贝比棕色扇贝更能抵抗热应激。同样,金色扇贝鳃中 CnHSP90 的 mRNA 表达水平也明显高于棕色扇贝(P<0.05)。
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Effects of thermal stress on mortality and HSP90 expression levels in the noble scallops Chlamys nobilis with different total carotenoid content.

The noble scallop Chlamys nobilis is an economically important marine bivalve cultivated in the southern sea of China since the 1980s. Unfortunately, mass mortality of this scallop species often occurs in summer. The present study was conducted to investigate whether the expression of heat shock protein 90 (HSP90) and level of carotenoids could enhance high-temperature stress resistance in scallop. First, the HSP90 homolog of C. nobilis (designated CnHSP90) was identified and cloned. The complete cDNA sequence of CnHSP90 was 2631 bp, including a 2181-bp open reading frame (ORF) encoding a 726 amino acid polypeptide with five HSP90 family signatures, and sharing high homology with members of the HSP90 family. CnHSP90 was ubiquitously expressed in all examined tissues including the intestine, kidney, adductor, mantle, gill, and gonad, with the highest in the gonad. Golden and brown scallops, which contain significantly different total carotenoid content (TCC), were subjected to acute thermal challenge, and the LTE50 (semi-lethal temperature at 36 h heat shock) and LTI50 (semi-lethal time after heat shock) as well as the correlation between CnHSP90 gene expression and TCC were determined. The LTE50 of golden scallop (32.14 °C) was higher than that of brown scallops (31.19 °C), with longer LTI50 at all tested temperatures, indicating that golden scallops were more resistant to thermal stress than brown scallops. Similarly, the mRNA expression levels of CnHSP90 in gill of golden scallops were significantly higher (P < 0.05) than that of brown scallops at 6, 12, 24, and 36 h, with a strong positive correlation between CnHSP90 expression level and TCC. This suggests that both carotenoids and HSP90 levels could improve thermal resistance in the noble scallops.

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