表皮蛋白是昆虫表皮关联器官的主要组成部分，如体壁和翅，然而其在渐变态昆虫翅发育中的功能尚不清楚。本文从飞蝗中鉴定出一种属于CPR 家族RR-2亚家族的翅表皮蛋白LmACP8。LmACP8主要在翅芽中表达，且在三龄、四龄和五龄若虫蜕皮前表达量较高，其编码的蛋白定位于翅芽和成虫翅的原表皮层。利用RNA干扰，LmACP8的表达缺失显著降低了其蛋白质含量，从而导致飞蝗若虫向成虫转变过程中翅形态发生异常。进一步研究证实这种异常形态发生是由于翅内皮层严重受损所致。个体实验结果发现，蜕皮激素（20-hydroxyecdysone，20E）能够抑制LmACP8的表达，而在干扰激素受体基因LmHR39（Hormone receptor 39）后，LmACP8表达显著上调。由此可见，LmACP8参与飞蝗若虫向成虫转变过程中翅的发育，且其表达受LmHR39介导的20E信号通路负调控。

突变效率和孵化率是影响基因编辑昆虫构建的两个关键因素。在CRISPR/Cas9介导的dsLmRNase2^-/-突变体蝗虫构建过程中，我们发现注射与卵囊接触20 min的鞣化卵比未鞣化的新鲜卵获得突变体飞蝗的效率更高。然而，鞣化和未鞣化卵产生的dsLmRNase2突变遗传到G₁代的效率相似。此外，发育正常的鞣化和未鞣化的G₀代卵和成虫的有效突变率没有显著差异，表明鞣化并不影响CRISPR/Cas9介导的突变效率。同时，我们发现飞蝗合胞体分裂期比鞣化时间长，为显微注射的鞣化卵和未鞣化卵提供了足够的时间窗口以完成有效地基因编辑。我们进一步发现，鞣化卵显微注射后感染率较低进而表现出更高的孵化率。抗压和超微结构分析表明，鞣化卵具有压缩的卵壳，能够承受较高的外部压力。综上所述，鞣化卵具有更强的防御能力以提高孵化率，并保持了更高的基因组突变效率，为开发CRISPR/Cas9介导的飞蝗突变体构建提供了一种优化的技术方法。

5-Aminolevulinic acid (ALA), a major photosensitivity insecticide, has attracted increasing attention as a new type of highly efficient, environmental friendly pesticide to be used to control the pest. To examine whether or not ALA acts effectively to grasshopper, Oxya chinensis and elucidate the detoxification mechanism of ALA, the susceptibility to ALA was assessed in O. chinensis and two major metabolic detoxification enzymes including glutathione S-transferases (GSTs) and general esterases (ESTs)-specific activities were compared in different development stages and different body sections
of O. chinensis treated by ALA and the control. The results showed that the ALA exhibited obvious toxicity to the grasshopper in different development stages. In the low-dose treatment (0.0597 mmol L-1), the mortalities of O. chinensis reached a significant level (55.5% in the 1st instar nymphs, 61.4% in the 2nd instar nymphs, 71.4% in the 3rd instar nymphs, and 64.4% in the 4th instar nymphs. But, there was no dose-dependent toxic effect. Thereby, we proposed that ALA has the potential for acting as photosensitivity insecticide for controlling O. chinensis. GSTs activity assays using CDNB and DCNB as substrates indicated that the thorax and abdomen of the different instar nymphs treated by ALA showed 1.52-5.56 fold significantly increased GSTs activities compared with the control. However, for the ESTs-specific activity assay, there was no significant difference between O. chinensis treated by ALA and the control within different instar nymphs, when α-NA, α-NB and β-NA were used as substrates. Therefore, GSTs-mediated metabolic detoxification
as evidenced by significantly increased GSTs activities might contribute to protect against oxidative damage and oxidative
stress by ALA in O. chinensis.

A cDNA encoding a sigma-class glutathione S-transferase of the locust, Locusta migratoria manilensis (LmGSTs1), was cloned by reverse transcriptase-polymerase chain reaction. The 830 bp-long cDNA encoded a 615 bp open reading frame (204 amino acid polypeptide), which exhibited the structural motif and domain organization characteristic of GST sigmaclass. It revealed 59, 57, 57, and 56% identities to sigma-class GSTs from Blattella germanica, Gryllotalpa orientalis, Nasonia vitripennis, and Pediculus humanus corporis, respectively. A recombinant protein (LmGSTs1) was functionally expressed in Escherichia coli cells in a soluble form and purified to homogeneity. LmGSTs1 was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a model substrate for GSTs, as well as with p-nitro-benzyl chloride. Its optimal activity was observed at pH 8.0 and at 30°C. Incubation for 30 min at temperatures below 50°C scarcely affected the activity. The I50 of reactive blue (RB) was 18.5 μmol L-1. In the presence of 0.05 mmol L-1 ethacrynic acid (ECA), LmGSTs1 showed (81±3)% of the original activities.