Molecular Identification of Ladybird Beetles (Coccinella: Coccinellidae) Using DNA Barcodes

Hanan Abdalla, Yosur Gamal Fiteha, Mohamed Abdel-Salam Rashed, Ramadan Mohamed Ali, Aya Mohamed Abdel-Salam Hammad, Ahmed Saber Mohamed Bream, Mahmoud Magdy
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Abstract

DNA barcoding is a molecular genetics technique commonly applied for species identification based on the cytochrome c oxidase subunit I (COI) gene located on the mitochondrial DNA genome. Based on our analysis, the DNA barcodes were generated from 16 specimen of Coccinella (family Coccinellidae). All the samples were collected from diverse sites in Egypt and Libya. Morphological traits based on the number of dorsal spots, DNA extraction, PCR amplification of COI, sequencing, nucleotide BLAST, and phylogenetics analyses were used to classify the specimens. A phylogenetic tree was constructed using the maximum likelihood method, including 46 COI sequences (the study and NCBI sequences). Three Coccinella clusters were defined and classified as Coccinella septempunctata, Coccinella novemnotata, and Coccinella undecimpunctata each species clustered into a unique branch. DNA barcodes discriminated clearly against the analyzed species. This study validated the COI efficiency as a marker for DNA barcoding of insects (Coccinella, family Coccinellidae).
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基于DNA条形码的瓢虫分子鉴定(瓢虫科
DNA条形码是一种常用的分子遗传学技术,基于线粒体DNA基因组上的细胞色素c氧化酶亚基I (COI)基因进行物种鉴定。基于我们的分析,生成了16份球菌属(Coccinella)标本的DNA条形码。所有的样本都是从埃及和利比亚的不同地点收集的。根据背斑数量、DNA提取、COI PCR扩增、测序、核苷酸BLAST和系统发育分析等形态学特征对标本进行分类。采用最大似然法构建系统发育树,包括46条COI序列(本研究序列和NCBI序列)。定义了3个瓢虫群,分别为七星瓢虫、新星瓢虫和非影响瓢虫,每一种都聚集在一个独特的分支中。DNA条形码对所分析的物种有明显的区别。本研究验证了COI作为昆虫(Coccinella, coccinellae科)DNA条形码标记的有效性。
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