欢迎您！东篱公司
退出
-
申报数据库
-
立项数据库
-
成果数据库
-
项目获奖数据库
中文摘要：
【目的】粉蚧是一类重要的世界性检疫性害虫,对果蔬产业以及水果的进出口贸易造成了巨大威胁。通常,口岸截获的粉蚧多为若虫或残体,加之隐存种的存在和较小的近缘种间差异,严重影响了基于形态学特征的粉蚧类害虫识别鉴定的准确性和及时性。本研究旨在明确DNA条形码技术对重大潜在入侵害虫大洋臀纹粉蚧Planococcus minor（Maskell）的鉴定有效性。【方法】以旅检截获的36头大洋臀纹粉蚧为对象、其近缘种柑橘臀纹粉蚧Pl.citri（Risso）为参照,以线粒体COI基因5＇端和3＇端序列以及核糖体28S r DNA D2-D3区段序列为分子标记进行比对分析,以K-2-P模型计算种内种间遗传距离,以最大似然法（maximum likelihood,ML）构建进化树并进行系统发育分析,同时利用Species Identifier物种识别软件评价3种基因片段对大洋臀纹粉蚧的鉴定效果。【结果】当分别以COI基因5＇端和3＇端序列为分子标记时,截获大洋臀纹粉蚧的碱基序列与NCBI中大洋臀纹粉蚧的序列一致性分别为100%和99%-100%,而与近缘种柑橘臀纹粉蚧COI基因5＇端和3＇端的核苷酸序列一致性分别为97%-98%和96%-98%;且大洋臀纹粉蚧和柑橘臀纹粉蚧分别存在5个和11个稳定的物种特异性识别位点;系统发育分析显示,截获的大洋臀纹粉蚧均与数据库中的大洋臀纹粉蚧聚为一支。当以28S r DNA D2-D3区段序列为分子标记时,臀纹粉蚧属各物种间高度保守,无法区分大洋臀纹粉蚧与其近缘种柑橘臀纹粉蚧;种间遗传距离仅为0.004。此外,物种识别软件评价结果显示,基于COI基因5＇端和3＇端序列的鉴定结果完全正确,而基于28S r DNA D2-D3区段序列的鉴定结果却存在45.2%-61.9%的模糊鉴定。【结论】基于COI基因5＇端和3＇端的DNA条形码技术完全可用于大洋臀纹粉蚧的快速准确鉴定及检测,对有效阻截其入侵和进一步扩散蔓延意义重大。
英文摘要：
【Aim 】Mealybugs are one of the most important quarantine pests that injury a variety of tropical fruits and vegetables worldwide. Usually,mealybugs intercepted in the quarantine inspection include eggs,nymphs or debris,and there exist several cryptic species and small difference in related species,which make them difficult to be identified by traditional morphological identification methods correctly and timely. In this study,the validity of DNA barcoding in identification of Planococcus minor（ Maskell）,a potential major alien invasive species to China,was evaluated. 【Methods】The 5＇-and 3＇-end of the mitochondrial cytochrome c oxidase subunit I（ COI） gene and 28 S ribosomal DNA（ r DNA） of36 individuals of Pl. minor intercepted during passenger inspection were amplified using universal primers,and Pl. citri（ Risso）,a related species of Pl. minor,from Langfang,Hebei used as a reference. The obtained partial fragment of 5＇- and 3＇-end of COI gene and D2-D3 fragment of 28 S r DNA were sequenced. The phylogenetic tree was established by using a maximum likelihood（ ML） method.The intra- and inter-species genetic distances were calculated using the Kimura-2-Parameter model.Meanwhile,the validity of Pl. minor identification based on the above three gene fragments was tested with Species Identifier software. 【Results 】When the 5＇- and 3＇-end of COI gene of Pl. minor were sequenced and BLAST, respectively, the nucleotide sequence identity between the fragments from Gen Bank and our study is 100% and 99%- 100%,respectively. The nucleotide sequence identity of the 5＇- and 3＇-end of COI gene between Pl. minor and Pl. citri is 97%- 98% and 96%- 98%,respectively. There are 5 and 11 stable species-specific identification sites in the fragments of the 5＇- and3＇-end of COI gene for Pl. minor and Pl. citri,respectively. Phylogenetic analysis indicated that Pl.minor intercepted during passenger inspection and those from the NCBI clustered in a clade. The sequencing results showed t
同期刊论文项目
同项目期刊论文
期刊信息
位置：