异色瓢虫3个小分子热激蛋白序列及低温诱导表达分析

doi:10.3864/j.issn.0578-1752.2017.16.009

Abstract

Abstract: 【Objective】 Harmonia axyridis is an important predator insect, and it is widely used in agricultural and forestry production. In this study, three small heat shock proteins (sHSP) were screened out and cloned using high throughput sequencing techniques. Also the expression levels of these three sHSP (sHPSs/sHSP genes) were detected under cold stress in order to explore the improvement of H. axyridis cold resistance role under low temperature stress conditions. 【Method】 Based on H. axyridis gene sequences in the transcriptome, several specific primers were designed to obtain the full-length open reading frame (ORF) sequence of the three sHSP genes. The expression of these sHSP genes was detected by quantitative real-time PCR (qRT-PCR) under short-time cooling or short-time cooling recover conditions, and between the black and yellow females of experimental and overwintering populations under low-temperature storage.【Result】 HaHSP47.74 and HaHSP21.53 were highly expressed at the pupal and adult stages during different developmental stages. And the expression of HaHSP21.52 in 4th instar larvae on the 4th day was significantly higher than that in control group. In short-term cooling treatment, the expression of HaHSP47.74 was significantly higher than that of the control group when the temperature was reduced to 0℃ and -5℃. The expression of HaHSP21.53 and HaHSP21.52 decreased significantly during cooling. There was no significant difference in the expression of the three sHSP genes in short-term heating treatment. The HaHSP47.74 expression of black female was significantly higher on the 5th day, while the yellow female was significantly higher from the 5th to 15th day in the experimental population under the condition of low temperature storage. The expression of HaHSP21.53 in black females was significantly higher from the 5th day to 20th day, while in the yellow females was significantly higher on the 15th day. Moreover, there was no significant expression of HaHSP21.52 in black or yellow female, and HaHSP47.74, HaHSP21.53, HaHSP21.52 in black females and yellow females were not significantly higher in the low temperature storage treatment in overwintering population. 【Conclusion】 sHSP may play a role in developmental stages of H. axyridis. HaHSP47.74 expression increased significantly under short-term cooling conditions, suggesting that sHSP in H. axyridis may play a key role under low temperature stress. While higher expression of HaHSP47.74 and HaHSP21.53 was found in the experimental population under low temperature storage, which indicated that these two genes played an important role in the cold acclimation. In addition, the expression level of black and yellow females of H. axyridis was different, it was speculated that the resistance and mechanism of different colors of H. axyridis are different.

Key words: Harmonia axyridis, small heat shock protein, cold-resistance gene, preservation in low temperature, qRT-PCR, gene expression

WANG HuiJuan, ZHAO Jing, SHI ZuoKun, QIU LingYu, WANG Su, ZHANG Fan, WANG ShiGui, TANG Bin. Sequence Analysis and Induced Expression of Three Novel Small Heat Shock Proteins Mediating Cold-Hardiness in Harmonia axyridis[J].Scientia Agricultura Sinica, 2017, 50(16): 3145-3154.

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Sequence Analysis and Induced Expression of Three Novel Small Heat Shock Proteins Mediating Cold-Hardiness in Harmonia axyridis

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