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为构建白背飞虱、褐飞虱、灰飞虱3种稻飞虱DNA条形码,准确鉴定各虫态稻飞虱种类,随机选取35个供试虫体提取全基因组DNA,PCR扩增其中线粒体细胞色素氧化酶Ⅰ(mtCOⅠ)基因,分析所获得的基因组成特点,计算P距离值,构建系统发育树,并对稻飞虱若虫、卵进行种类鉴定。PCR扩增获得了长度为706 bp的稻飞虱mtCOⅠ基因片段,碱基组成特点种内、种间不同;3种稻飞虱的种间P距离值为25.53%~34.21%,种内为0~0.51%,种间值明显高于种内;系统发育树显示同种个体聚集在同一分支;同种卵、若虫的mtCOⅠ基因与已获得的成虫基因序列同源性最高值为100%,P距离值为0。研究表明,3种稻飞虱的DNA条形码可基于mtCOⅠ基因构建,且可准确鉴定各虫态。
To construct the DNA barcodes of three planthoppers of BPPH, BPH and SBPH and accurately identify the species of the planthopper, 35 samples of the tested parasites were randomly selected to extract the whole genome DNA and PCR amplification of mitochondrial cytochromes (MtCOⅠ) gene was analyzed. The characteristics of the obtained genes were analyzed. The P distance value was calculated and the phylogenetic tree was constructed. The species identification of rice planthopper nymphs and eggs was also carried out. The mtCOⅠ gene fragment of planthopper was obtained by PCR amplification. The base composition was different between species and species. The interspecific P distance of three planthopper species was 25.53% -34.21%, and the species was 0 ~ 0.51%, and interspecific values were significantly higher than those in the species. Phylogenetic trees showed that the same individuals clustered in the same branch. The highest homology of the mtCOⅠ gene of the same egg and nymph with the obtained adult was 100% The value is 0. Studies have shown that the DNA barcodes of the three planthopper species can be constructed based on the mtCO I gene, and each of the insect states can be accurately identified.