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Cloning, expression and chitin-binding activity of two peritrophin-like protein genes in the common cutworm, Spodoptera fitura

引用

Insect Science 2014年第4期21卷 449-458页

作者： Wei-Jun Chen Li-Xia Huang Dan Hu Li-Yan Liu Jun Gu Li-Hua Huang Qi-Li Feng Guangdong Provincial Key Laboratory of Biotechnology for Plant Development School of Life Sciences South China Normal UniversityGuangzhou China

Insect midgut secretes a semi-permeable peritrophic membrane （PM）, which plays important roles in protecting the midgut and helping with food digestion. The lep- idopteran larvae produce type 1 PM, which is degraded when insects develop into the metamorphic stages. However, the underlying mechanism is unclear. In the present study, two peritrophin-like proteins （peritrophin-57 and 37） were identified from the midgut expression sequence tag library and transcriptome of the common cutworm, Spodoptera litura. The temporal and spatial expression patterns and responses to the induction of 20- hydroxyecdysone （20E） and starvation were examined by real-time quantitative polymerase chain reaction according to their common sequence region. The chitin-binding activity was also studied using a competitor, calcofluor. The open reading frames are 1 554 and 1 020 bp, respectively. They shared four highly conserved peritrophin-A domains and were ex- pressed only in the midgut rather than in the other tissues, including fat body, epidermis, Malpighian tube and hemolymph. Their transcriptional expression could only be detected at the larval stages rather than in eggs, prepupae, pupae and adults. The purified protein of peritrophin-37 bound to chitin in a dose-dependent manner. These results indicate that the two proteins are peritrophins, the structural components of PM. In addition, the messenger RNA levels of the two peritrophins were significantly down-regulated by 20E injection, whereas feeding/starvation had no effect on the expression. These findings suggest that the increase of20E titer may be an important factor which controls the degradation of PM during metamorphosis.

关键词： metamorphosis peritrophic membrane peritrophin starvation 20-hydroxyecdysone

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