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免疫组织化学和发育学研究,以阐明雪花莲凝集素对褐飞虱(Nilaparvata lugens (Stal))的作用机制。

Immunohistochemical and developmental studies to elucidate the mechanism of action of the snowdrop lectin on the rice brown planthopper, Nilaparvata lugens (Stal).

作者信息

Gatehouse A M.R., Gatehouse J A., Bharathi M, Spence J, Powell K S.

机构信息

University of Durham, Department of Biological Sciences, South Road, Durham, UK

出版信息

J Insect Physiol. 1998 Jul;44(7-8):529-539. doi: 10.1016/s0022-1910(98)00054-7.

Abstract

Rice brown planthoppers (Nilaparvata lugens) were fed on artificial diet containing snowdrop lectin (Galanthus nivalis agglutinin; GNA), which has been shown to be toxic towards this insect pest. In addition to decreasing survival, the lectin affected development, reducing the growth rate of nymphs by approximately 50% when present at a concentration of 5.3&mgr;M. Immunolocalisation studies showed that lectin binding was concentrated on the luminal surface of the midgut epithelial cells within the planthopper, suggesting that GNA binds to cell surface carbohydrate moieties in the gut. Immunolabelling at a lower level was also observed in the fat bodies, the ovarioles, and throughout the haemolymph. These observations suggest that GNA is able to cross the midgut epithelial barrier, and pass into the insect's circulatory system, resulting in a systemic toxic effect. Electron microscope studies showed morphological changes in the midgut region of planthoppers fed on a toxic dose of GNA, with disruption of the microvilli brush border region. No significant proteolytic degradation of GNA was observed either in the gut or honeydew of planthoppers fed on lectin-containing diet. The presence of glycoproteins which bind GNA in the gut of the brown planthopper was confirmed using digoxigen-labeled lectins to probe blots of extracted gut polypeptides.

摘要

用含有雪花莲凝集素(Galanthus nivalis agglutinin;GNA)的人工饲料喂养稻褐飞虱(Nilaparvata lugens),已证明这种凝集素对这种害虫有毒性。除了降低存活率外,凝集素还影响发育,当浓度为5.3μM时,若虫的生长速率降低约50%。免疫定位研究表明,凝集素结合集中在褐飞虱中肠上皮细胞的管腔表面,这表明GNA与肠道中的细胞表面碳水化合物部分结合。在脂肪体、卵巢小管以及整个血淋巴中也观察到较低水平的免疫标记。这些观察结果表明,GNA能够穿过中肠上皮屏障,进入昆虫的循环系统,从而产生全身毒性作用。电子显微镜研究显示,以有毒剂量的GNA喂养的褐飞虱中肠区域出现形态变化,微绒毛刷状缘区域遭到破坏。在以含凝集素饲料喂养的褐飞虱的肠道或蜜露中,均未观察到GNA有明显的蛋白水解降解。使用地高辛标记的凝集素探测提取的肠道多肽印迹,证实了褐飞虱肠道中存在与GNA结合的糖蛋白。

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