异色瓢虫低温胁迫下过冷却点变化及抗寒基因表达分析

doi:10.3864/j.issn.0578-1752.2016.04.007

Abstract

Abstract: 【Objective】Harmonia axyridis is an important predatory lady beetle as natural enemies attacking agricultural and forestry pests. H. axyridis has strong cold-hardiness as it can overwinter as disapausing adults in Northeast population of China. The objective of this study is to use the super cooing point and the changes of cold resistance gene mRNA level as the index of cold-resistance ability, and to define the effect of low temperature stress on improving cold resistance of H. axyridis.【Method】The wild H. axyridis adults were collected and treated for 2, 12 and 24 h by three different low temperatures of 0, 5 and 10℃ to get the best low temperature stress. The changes of super cooling point were detected all the time during the period of low temperature processing. The H. axyridis adults of experimental populations and wild adults were used as the control groups and experimental groups, respectively. All of these adults were treated under the condition of 5℃ low temperature and stored for 10, 20 and 30 d, and the changes of super cooling point and survival rate were measured after the treatment. Six important cold hardiness genes were sure by transcriptome and digital gene expression tag profiling (DGE). The relative expressions of six important cold hardiness genes at mRNA level were detected by real-time fluorescent quantitative PCR. 【Result】The super cooling point was decreased significantly after acclimation for 12 h at 5℃. Acclimation for 12 h at 5℃ could effectively decrease the super cooling point and other indexes of cold resistance in H. axyridis. Setting treatment group which was acclimated for 12 h at 5℃ and control group deposited for 10 , 20, and 30 d at 5℃, the survival rate and super cooling point of control group were all higher than the treatment group. In the same time, the 6 genes’ relative expression of mRNA level results showed HSP21.4, trehalase, transketolase, ATP-grasp fold domain protein and dopa decarboxylase were up-regulated when cold acclimation, but erythrocyte binding protein was down-regulated. HSP21.4 was always highly expressed in cryopreservation, and the treatment group was strikingly higher than the control group. Others were on low expression in cryopreservation, and strikingly lower than the control group. 【Conclusion】The low temperature stress can reduce the super cooing point of the H. axyridis, but it can’t improve the survival ability of summer high temperature resistant population. HSP21.4 and erythrocyte binding protein can help H. axyridis to improve the cold-resistance ability respectively by high expression and low expression. Trehalase, transketolase, ATP-grasp fold domain protein and dopa decarboxylase are highly expressed in cold acclimation to resist cold environment.

Key words: Harmonia axyridis, low temperature stress, cold-resistant gene, quantitative expression, preservation in low temperature

WU Meng-jing, XU Qing-ye, LIU Ya, SHI Xing-rong, SHEN Qi-da, YANG Meng-meng, WANG Shi-gui, TANG Bin. The Super Cooling Point Change of Harmonia axyridis Under Low Temperature Stress and Its Cold-Resistance Genes’ Expression Analysis[J].Scientia Agricultura Sinica, 2016, 49(4): 677-685.

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