Molecular Identifying Liriomyza Trifolii (Burgess) Based On Real-time PCR And Genetic Relationship Between Newly Established Population In China And Population From Other World

Liriomyza trifolii (Burgess) (Diptera: Agromyzidae) is an important pest of vegetable andcut-flower crops around the world. Liriomyza trifolii in China mainland was first reported fromZhongshan of Guangdong province in 2006. We report here that this pest has recently expandedits distribution in China, along with a host plant range extension and population explosion.The molecular identifying Liriomyza trifolii (Burgess) based on real-time PCR was established inthis paper. Using partial DNA sequences of mitochondrial COI of L. trifolii occurred inGuangdong and Taiwan provinces in China, Japan, Philippines, Israel, Germany, USA, Mexicoand Honduras, and related species from Genbank, the L. trifolii-specific probe distinguishingfrom L. sativae Blanchard is designed. It obtained the same results in individuals of differentstages. It is a sentitive, fast and less costly assay system compared to other molecular method,such as random amplified polymorphic DNA-PCR, PCR-restriction fragment-lengthpolymorphism, DNA sequencing and multiplex polymerase chain reaction. The partial COIsequence of mtDNA from 14 leafminers populations were analyzed. The probes and primers forL. trifolii were designed basis on the align sequence of the amplified COI mtDNA of Liriomyzaspp. by using the CLUSTSAL 1.81 of the sequence software. The probe was labeled twofluorescent dyes, that is a reporter of 6-carboxy-fluorescein (FAM) at the 5' end and a quencherof 6-carboxy- tetramethhylrhodamine (TAMRA) at the 3'end. The real-time PCR methoddeveloped in identitifying L. trifolii in this study was in 4hrs. It is a stable and preciseidentifying method.The results showed that the mitochondrial cytochrome oxidaseâ… gene sequenced from 14populations collected from Zhuhai, Ningbo, Taiwan, Zhongshan in China and Japan, Israel,Germany and USA were identical. No genetic variation was observed between population eitherfrom different geographical localities or from different host plants. The phylogenetic analysisindicated that China population was grouped into the trifolii-W clade, belonging to a nonpepperlineage, which also includes other recently introduced Asian populations.