Department of Entomology, College of Agriculture, Food and Environment, University of Kentucky, Lexington, KY, 40546, United States of America.
Key Laboratory of Insect Developmental and Evolutionary Biology, Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai, 200032, China.
Sci Rep. 2020 Feb 11;10(1):2363. doi: 10.1038/s41598-020-59197-8.
The functional information on heat-shock proteins (Hsp) and heat-shock promoters from an important agricultural insect pest, Spodoptera frugiperda, is still lacking. We conducted a genome-wide identification of Hsp genes and identified a total of 21 genes belonging to four major insect Hsp families (small heat-shock proteins, Hsp60, Hsp70, and Hsp90) in S. frugiperda. Expression of most of S. frugiperda (SfHsp) genes could be detected in Sf9 cells, embryos and larval tissues of S. frugiperda. The heat-inducible activity of heat-shock promoters from several SfHsp genes was tested in Sf9 cells and embryos. The promoter of SfHsp70D showed the high constitutive activity in cell line and embryos, while the activity of SfHsp20.15 and SfHsp20.71 promoters was most dramatically induced in Sf9 cells and embryos. In embryos, the heat-induced activity of SfHsp20.71 and SfHsp70D promoters outperformed commercially used ie1 and ie2 promoters. The heat-induced activity of SfHsp70D and SfHsp19.07 promoters were more robust than ie2 promoter in Sf9 cells. These SfHsp promoters with high basal activity or with heat-induced activity from low basal activity, could be used in S. frugiperda or other lepidopteran insects for many applications including transgenesis and genome editing.
关于农业重要害虫夜蛾（Spodoptera frugiperda）的热休克蛋白（Hsp）和热休克启动子的功能信息仍然缺乏。我们对 Hsp 基因进行了全基因组鉴定，在 S. frugiperda 中鉴定了总共 21 个属于四个主要昆虫 Hsp 家族（小热休克蛋白、Hsp60、Hsp70 和 Hsp90）的基因。Sf9 细胞、S. frugiperda 胚胎和幼虫组织中均可检测到大多数 S. frugiperda（SfHsp）基因的表达。在 Sf9 细胞和胚胎中测试了几个 SfHsp 基因的热休克启动子的热诱导活性。SfHsp70D 的启动子在细胞系和胚胎中表现出高组成活性，而 SfHsp20.15 和 SfHsp20.71 启动子的活性在 Sf9 细胞和胚胎中诱导最为显著。在胚胎中，SfHsp20.71 和 SfHsp70D 启动子的热诱导活性优于商用的 ie1 和 ie2 启动子。SfHsp70D 和 SfHsp19.07 启动子在 Sf9 细胞中的热诱导活性比 ie2 启动子更稳健。这些具有高基础活性或从低基础活性诱导热诱导活性的 SfHsp 启动子可用于 S. frugiperda 或其他鳞翅目昆虫的许多应用，包括转基因和基因组编辑。
