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热休克蛋白(heat shock protein,HSP)在昆虫应对外界胁迫刺激时起着重要作用。为了系统研究小菜蛾Plutella xylostella HSP基因家族,根据家蚕的HSP蛋白序列,采用本地Blast程序对小菜蛾全基因组数据库进行同源序列检索,从小菜蛾基因组数据库中鉴定了25个HSP基因,包括2个HSP90、8个HSP70和15个sHSP(small heat shock protein,sHSP)基因。小菜蛾、家蚕Bombyx mori、黑腹果蝇Drosophila melanogaster和赤拟谷盗Tribolium castaneum的HSP系统进化分析显示,昆虫的小分子量热休克蛋白sHSP具有很强的种属特异性,HSP70家族的保守性比sHSP强。小菜蛾HSP基因表达模式分析显示,与敏感品系对比,抗性品系(抗毒死蜱和抗氟虫氰品系)中HSP基因具有不同的表达模式。小菜蛾1,2和3龄幼虫HSP基因表达模式较为接近,而与4龄幼虫中的表达模式相差较大;4龄幼虫和蛹中的表达模式相近;雌成虫和雄成虫中的表达模式显著不同,与果蝇精子形成有关的两个热休克蛋白HSP23和HSP27基因[分别为CCG003980.1(Px23.5)和CCG005412.2(Px27.5)],在小菜蛾雄成虫中的表达量显著高于雌成虫。研究结果表明小菜蛾HSP基因不仅在杀虫剂抗性、发育分化,甚至在生殖上均可能起着重要的作用。本研究为深入研究小菜蛾HSP与生长发育、抗逆行为的相互关系奠定了基础。
Heat shock protein (HSP) plays an important role in insect response to external stress. In order to systematically study the HSP gene family of Plutella xylostella in Plutella xylostella, homologous sequences were searched using the Blast program based on the HSP protein sequence of the silkworm, and 25 HSP genes were identified from the diamondback moth genome database HSP90, 8 HSP70 and 15 sHSP (sHSP) genes. The phylogenetic analysis of the HSPs of diamondback moth, Bombyx mori, Drosophila melanogaster and Tribolium castaneum showed that insect small-molecular-weight heat-shock protein sHSP has strong species-specificity and the conserved ratio of HSP70 family Strong sHSP. The analysis of HSP gene expression pattern in Plutella xylostella shows that the HSP genes have different expression patterns in the resistant lines (anti-chlorpyrifos and fipronil) compared with the susceptible lines. The expression patterns of HSPs in 1st, 2nd and 3rd instar larvae of Plutella xylostella were similar, but different from those in 4th instar larvae. The expression patterns were similar in 4th instar larvae and pupae, while the expression patterns were significantly different between female and male adults , Two heat shock proteins HSP23 and HSP27 [related to CCG003980.1 (Px23.5) and CCG005412.2 (Px27.5)], respectively, associated with sperm formation in Drosophila melanogaster, were significantly higher in male diamondback moth Female adults. The results showed that the HSPs of Plutella xylostella may play an important role not only in the resistance, development and differentiation of insecticides, but also in reproduction. This study lays the foundation for further study on the relationship between HSP and growth and development and anti-retrograde behaviors in Plutella xylostella.