辽宁地区柞园新记录害虫多斑柞跳象的初步鉴定

Preliminary Identification of Ｒhynchaenus (Orchestes) maculosus, a Newly-recorded Pest Species Infesting Oak in Liaoning Tussah Silkworm Rearing Area

近期在辽宁柞蚕区发现了一种新的柞树潜叶性害虫。将昆虫线粒体细胞色素C氧化酶亚基Ⅰ基因（mtD NA COⅠ）序列作为DNA条形码,结合形态特征对害虫进行分类鉴定。以单头试虫的基因组DNA为模板,用mtD NA COⅠ通用引物PCR扩增得到了试虫的mtD NA COⅠ基因片段（GenB ank登录号：KX657707~KX657709）。将获得的基因片段序列在NCBI数据库进行BLAST同源性搜索,在GenB ank中未搜索到该试虫的相关序列信息,但与鞘翅目Coleoptera象甲科Curculionidae象甲亚科Curculioninae的Orchestes jota mtD NA COⅠ同源片段序列（GenB ank登录号：KJ963227.1）的相似度最高,为86%。室内观察其成虫、幼虫和蛹的形态特征：成虫体长3.7~4.0 mm,喙长0.8~1.0 mm,鞘翅,后足发达,腿节粗壮;老熟幼虫5~7 mm,体节12节,无足,具步泡突;蛹为裸蛹。基于试虫的mtD NA COⅠ分子鉴定标记,结合试虫与原有记录形态特征的比较结果,初步鉴定新发现的柞树潜叶性害虫为跳象亚科Rhynchaeninae跳象属Rhynchaenus的多斑柞跳象Rhynchaenus（Orchestes）maculosus。

A new leaf-mining insect pest,which causes damage to oak leaves,was found in Liaoning tussah silkworm rearing area. The pest species was identified and classified based on its morphological characteristics combined with mitochondrial cytochrome C oxidase subunitⅠ（ mtD NA COⅠ） gene sequence as DNA barcode. Using genomic DNA of the individual pest larva as template,various segments of mtD NA COⅠ gene were amplified and sequenced（ GenB ank accession No. KX657707 to KX657709）. Sequence similarity search was carried out using the basic local alignment search tool（ BLAST） in NCBI database. The results showed no sequence information related to this leaf-miner in NCBI database. The highest sequence similarity of 86% was found between mtD NA COⅠ gene sequence of this pest and that of Orchestes jota（ GenB ank accession No.KJ963227. 1） which belongs to subfamily of Curculioninae（ Coleoptera： Curculionidae）. Observations of morphological characteristics showed that the adult of this pest is3. 7 to 4. 0 mm long,has a snout of 0. 8 and 1. 0 mm long,with elytra wings,strong meta-legs and thick tibiae.The mature larva is 5 to 7 mm long,has 12 segments,has no legs but with leg-like bulges. The pupa is in bare form.Based on the mtD NA COⅠ molecular identification label of this pest and the comparative result of morphological characteristics between the collected leaf-miners and the original records,this newly founded leaf-mining insect pest on oak was preliminarily classified into subfamily Curculioninae, genus Rhynchaenus and named as Rhynchaenus（ Orchestes） maculosus.