Physiological responses of juvenile Apostichopus japonicus (Selenka) to collaborative stress of high temperature and low salinity: Growth and induced heat shock protein gene expression
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摘要: 全球气候变化背景下的极端高温和强降雨频发天气导致的养殖池塘持续性的高温和低盐环境给刺参池塘养殖带来了严峻的挑战。为了研究刺参对高温和低盐环境的生理响应,本实验分析了高温和低盐协同胁迫对刺参幼参生长及诱导型hsp70和hsp90基因表达的影响。实验设置4个温度梯度(16℃、20℃、24℃和28℃)和3个盐度梯度(22、27和32),共12个不同的胁迫组。经过40 d的长期胁迫,研究发现,随着温度的升高和盐度的降低,幼参的体重增加量减少,并在高温、低盐组出现体重负增长。长期胁迫提高了刺参幼参的诱导型hsp70和hsp90基因表达量,从而在一定程度上增强了对极端天气的抵抗能力。同时,高温下盐度22处理组刺参幼参诱导型hsp70和hsp90基因的表达量较盐度27条件下低。双因子方差分析结果显示,高温和低盐对幼参特定生长率和诱导型hsp70和hsp90基因表达量的交互作用均不显著,并且盐度对诱导型hsp70和hsp90基因表达量没有显著的影响。因此,相较低盐,高温对刺参幼参的影响更大,可作为刺参育种的选择压力。在高温期应该采取有效措施降低海水温度,并防止极端天气造成的养殖水体温度分层。该研究丰富了刺参生理生态学理论,可为刺参良种选育工作提供思路,并为指导极端天气下刺参生产实践活动提供一定的理论依据。Abstract: Under the background of global climate change, the continuous high temperature and low salinity caused by extreme short-time heavy rainfall in summer brings severe challenges to pond culture of Apostichopus japonicus. In order to study the physiological responses of A. japonicus to high temperature and low salinity, the effects of collaborative stress of high temperature and low salt on growth and induced hsp70 and hsp90 expression of juvenile A. japonicus were analyzed in this paper. Four temperature gradients (16℃, 20℃, 24℃, and 28℃) and three salinity gradients (22, 27, and 32) were set, and there were a total of 12 different stress groups. After a period of 40 days stress, the weight gain of juvenile A. japonicus decreased with the increase of temperature and the decrease of salinity, there was negative growth in the high temperature and low salinity group. The expression of induced hsp70 and hsp90 was increased after a long period of stress, thus enhancing the resistance of juveniles to extreme weather. Meanwhile, the expression of induced hsps of juveniles treated with salinity 22 was lower than that in the condition of salinity 27. The results of two-factor ANOVA showed that there was no significant interaction between high temperature and low salinity on specific growth rate and induced hsp70 and hsp90 expression, and salinity had no significant effect on induced hsp70 and hsp90 expression. Therefore, compared with low salinity, high temperature had a greater impact on juvenile A. japonicus, which could be used as the selection pressure in A. japonicus breeding. During the high temperature period, effective measures should be taken to reduce the sea temperature and prevent the stratification of aquaculture water caused by extreme weather. This study enriched the physiological and ecological theories of A. japonicus, provided ideas for the breeding of A. japonicus, and provided certain theoretical basis for guiding the production and practice of A. japonicus culture under extreme weather.
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Key words:
- high temperature /
- low salinity /
- collaborative stress /
- Apostichopus japonicus /
- growth /
- heat shock protein
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图 1 高温和低盐协同胁迫对刺参幼参特定生长率的影响
小写字母不同表示同一盐度下不同温度处理组差异显著(p<0.05);大写字母表示同一温度下不同盐度处理组差异显著(p<0.05)
Fig. 1 Effects of high temperature and low salinity collaborative stress on specific growth rate of juvenile Apostichopus japonicus
Data in the same row having different lower case letters are significantly different (p<0.05) among different temperature levels; data in the same column having different capital letters are significantly different (p<0.05) among different salinity levels
表 1 用于本实验的引物序列
Tab. 1 Primers used in this study
引物名称 引物序列 Hsp70-F 5’-ATGCCTAGAACCAGTAGAGAAAG-3’ Hsp70-R 5’-TGTCGTTCGTGATGGTGATT-3’ Hsp90-F 5’-TTGTTGAAAGGGAGGAGG-3’ Hsp90-R 5’-GGCATCAGAGGCGTTAGA-3’ β-actin-F 5’-ACACGGTATCGTCACAAACTGG-3’ β-actin-R 5’-AGGATAGCGTGAGGAAGAGCAT-3’ 表 2 高温和低盐协同胁迫对刺参幼参生长的影响
Tab. 2 Effects of high temperature and low salinity collaborative stress on growth of juvenile Apostichopus japonicus
温度/℃ 盐度 初体重/g 末体重/g 个体数 16 22 3.574±0.024a 5.264±0.263c 18 16 27 3.543±0.029a 7.717±0.413b 18 16 32 3.544±0.033a 10.618±0.782a 18 20 22 3.549±0.025a 4.278±0.274c 18 20 27 3.530±0.015a 5.239±0.261c 18 20 32 3.558±0.029a 7.290±0.582b 18 24 22 3.507±0.012a 2.765±0.149de 18 24 27 3.534±0.034a 3.963±0.147c 18 24 32 3.555±0.049a 5.077±0.166c 18 28 22 3.525±0.036a 2.089±0.047e 18 28 27 3.553±0.007a 2.429±0.075e 18 28 32 3.520±0.032a 3.898±0.094cd 18 注:同一列上标字母不同表示差异显著(p<0.05)。 表 3 温度和盐度对刺参幼参生长的双因子方差分析
Tab. 3 Two-way ANOVA of growth for the juvenile Apostichopus japonicus maintained in different temperature and salinity
影响因子 自由度 均方 F值 p 末体重/g 温度/℃ 3 2.094 172.761 < 0.001 盐度 2 1.389 114.546 < 0.001 交互作用 6 0.035 2.917 < 0.05 特定生长率 温度/℃ 3 10.894 220.109 < 0.001 盐度 2 7.100 143.439 < 0.001 交互作用 6 0.093 1.874 > 0.05 表 4 温度和盐度对刺参幼参诱导型hsp基因表达量的双因子方差分析
Tab. 4 Two-way ANOVA of hsp gene expression for the juvenile Apostichopus japonicus maintained in different temperature and salinity levels
影响因子 自由度 均方 F值 p hsp70 温度/℃ 3 0.694 7.207 < 0.05 盐度 2 0.241 2.502 > 0.05 交互作用 6 0.015 0.159 > 0.05 hsp90 温度/℃ 3 1.472 8.541 < 0.001 盐度 2 0.321 1.861 > 0.05 交互作用 6 0.027 0.157 > 0.05 -
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