Analysis of High Temperature Tolerance in Early Development of Esox lucius

doi:10.13560/j.cnki.biotech.bull.1985.2020-1335

Abstract

Abstract:

Analyzing the high temperature tolerance in early development stage of Esox lucius is aimed to lay a theoretical foundation for the breeding and ecological adaptation of E. lucius. High temperature stress method was used to study the high temperature tolerance of the fertilized eggs and juvenile E. lucius,which was evaluated by 24hUILT₅₀and CT_max. Real-time fluorescence quantitative PCR（qRT-PCR）and relative quantitative 2^-ΔΔCt method were used to compare the expressions of HSP70 gene in the brain,gill,heart and liver of juvenile E. lucius under different temperatures and high temperature stressed times. The results showed that the breaking time of the membranes of the fertilized eggs in E. lucius was shortened gradually as the incubation temperature increased. However,the floating rate of the fry showed the tendency of rising slowly then falling sharply. With the increase of domestication temperature,the 24hUILT₅₀ value of the juveniles increased and the occurrence time of CT_max delayed under high temperature stress. Under the stress of different high temperature,the expressions of HSP70 gene in the brain,gill and heart tissues of juvenile E. lucius increased first and then decreased,while that in the liver tissue continued to increase. When juvenile E. lucius were subjected to 30℃ high temperature stress for 0-24 h,the HSP70 gene expressions in their brains gill and heart tissues increased first and then decreased,while that in the liver tissue tended to be decreasing first and then increasing and then decreasing. The expressions of HSP70 gene in the brain,gill,heart and liver of juvenile E. lucius varied at different degrees under different temperatures and different high temperature stressed times,but the variations were time-specific and tissue-specific,with different response ranges. After high temp-erature domestication,the temperature tolerance of E. lucius increased,which may be involved in important regulation through HSP70.

Key words: Esox lucius, high temperature tolerance, critical temperature, heat shock protein 70

ZHANG Yu, HAYSA· Ayelhan, RABIGUL· Sawut, SHI Chun-ming, ZHANG Ren-ming. Analysis of High Temperature Tolerance in Early Development of Esox lucius[J]. Biotechnology Bulletin, 2021, 37(5): 76-83.

Figures/Tables 8

Table 1 Fluorescent quantitative PCR primers

基因 Gene	引物序列 Primer sequence（5'-3'）	片段长度Frag-ment length/bp
HSP70	F：ATCAGCCAGGGGTGCTCATT	200
HSP70	R：CTTGTTCTCCTTCCCAGTGC	200
管家基因β-actin Housekeeping genes	F：AAGATGAAATCGCCGCACTG	215
管家基因β-actin Housekeeping genes	R：TGCCAGATCTTCTCCATG	215

Table 2 Effects of different incubation water temperature on floatation rate of E. lucius fry

温度 Temperature/℃	受精卵数量/粒 The number of fertilized eggs/Grain	50%破膜时间 50% film breaking time/h	正常上浮鱼苗数量/尾 Number of floating fry/Tail	上浮率 Floating ratio/%
15±0.5	959	188	526	73.47
17±0.5	1096	139	602	73.58
19±0.5	1254	123	653	69.76
21±0.5	950	102	372	52.46

Fig.1 24hUILT50 of E. lucius at different acclimation temperatures

Table 3 CTmax of E. lucius fry at different acclimation temperatures

驯化温度Acclimation temperature/℃	升温速率Heating rate/（℃/d）	CT_max/℃	CT_max出现时间CT_max appearance time/min
18±0.5	1±0.5	32.90	1152
21±0.5	1±0.5	33.06	1259
24±0.5	1±0.5	33.36	1418
27±0.5	1±0.5	33.27	1735

Fig.2 Expression of HSP70 gene in E. lucius different tissues under different high temperature stress Different letters refer to there is significant difference within the relative expression of the HSP70 gene in the same tissue under different high temperature stress(P<0.05)

Fig.3 Expression of HSP70 gene in E. lucius different tissues under the same high temperature stress Different letters indicate there is significant difference within the relative expression of the HSP70 gene in different tissues under the same high temperature stress(P<0.05)

Fig.4 Expression of HSP70 gene in E. lucius different tis-sues under different times of high temperature stress Different letters indicate there is significant difference within the relative expression of the HSP70 gene in the same tissue under different times of high temperature stress(P<0.05)

Fig.5 Expression of HSP70 gene in E. lucius different tissues under the same time of high temperature stress Different letters indicate there was significant difference within the relative expression of the HSP70 gene in the different tissues under the same time of high temperature stress(P<0.05)

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