Proteomic analysis of the salivary glands of the rice brown planthopper, Nilaparvata lugens (Stal) (Homoptera: Delphacidae)
Proteomic analysis of the salivary glands of the rice brown planthopper, Nilaparvata lugens (Stal) (Homoptera: Delphacidae)
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DOI：
10.1303/aez.2009.525
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发表时间：
2009-11-01
影响因子：
1.3
通讯作者：
Hattori, Makoto
中科院分区：
文献类型：
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作者：
Konishi, Hirosato;Noda, Hiroaki;Hattori, Makoto
稻褐飞虱 (Nilaparvata lugens) 是一种以韧皮部为食的水稻害虫。为了了解唾液在进食过程中的作用，我们使用二维聚丙烯酰胺凝胶电泳分析唾液腺蛋白质，然后对蛋白质进行埃德曼降解。在分析的 52 个主要点中，确定了 25 个 N 端序列。其余27个蛋白质的内部氨基酸序列通过克利夫兰肽图谱方法确定。在对照褐飞虱表达序列标签 (EST) 数据库中的条目进行搜索后，我们鉴定了 25 种蛋白质中的 22 种，其 N 末端区域已测序，以及 27 种蛋白质中的 13 种进行了内部测序。通过使用公共非冗余蛋白质数据库的同源性搜索，我们鉴定了 EST 不匹配的蛋白质。结果，鉴定出许多负责能量代谢、蛋白质合成、折叠和修饰的管家蛋白，其中肌动蛋白、微管蛋白、热休克蛋白、蛋白质二硫键异构酶和ATP合酶与从冈比亚按蚊和植食性瘿蚊Mayetiola destructor等食血节肢动物的唾液腺中鉴定出的蛋白质有共同之处。然而，我们无法识别先前在褐飞虱唾液腺中检测到的葡萄糖苷酶和酚氧化酶。相比之下，在来自褐飞虱的唾液腺匀浆中发现了相对较高丰度的具有 EF-hand 结构域的新型蛋白质；它们相应的 EST 专门来自唾液腺。它们的钙结合特性表明它们对飞虱取食韧皮部的重要性。
The rice brown planthopper, Nilaparvata lugens is a phloem-feeding insect pest of the rice plant Oryza sativa. To understand the role of saliva in its feeding process, we analyzed salivary gland proteins using two-dimensional polyacrylamide gel electrophoresis followed by Edman degradation of proteins. Of the 52 major spots analyzed, 25 N-terminal sequences were determined. The internal amino acid sequences of the remaining 27 proteins were determined by the Cleveland peptide mapping method. We identified 22 of the 25 proteins whose N-terminal regions were sequenced and 13 of the 27 proteins that were internally sequenced after searching them against the entries in the expressed sequence tag (EST) database of N. lugens. Through a homology search using public nonredundant protein databases, we identified EST-unmatched proteins. As a result, many housekeeping proteins responsible for energy metabolism, protein synthesis, folding and modification were identified, of which actins, tubulins, heat-shock proteins, protein disulfide isomerases and ATP synthases were in common with proteins identified from the salivary glands of hematophagous arthropods such as Anopheles gambiae, and a phytophagous gall midge Mayetiola destructor. However, we could not identify the glucosidases and phenoloxidases previously detected in the salivary glands of N. lugens. In contrast, novel proteins with an EF-hand domain were found in relatively high abundance in salivary gland homogenates derived from N. lugens; their corresponding ESTs were specifically derived from the salivary glands. Their calcium-binding properties suggest their importance of phloem feeding by the planthopper.