Abstract
BackgroundTea green leafhopper is one of the most dominant pests in Chinese tea plantations. Recent evidence, including morphological and molecular data, revealed that tea green leafhopper in China is the same species as in Japan, Empoasca (Matsumurasca) onukii Matsuda. Previous morphological study that revealed variation in the structure of the male genitalia within and among populations of this species suggested that there may be significant population-level genetic variation. To provide powerful molecular markers to explore the population genetic diversity and population genetic structure of this pest in China, microsatellite markers were obtained by AFLP of sequences containing repeats (FIASCO).ResultsEighteen polymorphic markers were evaluated for five populations of E. (M.) onukii, Two related empoascine leafhopper species were selected to test the transferability of the markers. Population genetic structure of E. (M.) onukii was detected using Structure analysis, principal coordinate analysis (PCoA) and variance analysis. The identified markers were polymorphic with total number of alleles ranging from 6 to 24 per locus, observed and expected heterozygosity ranged from 0.133 to 0.9 and 0.183 to 0.926, respectively, and the polymorphic information content value over all populations varied from 0.429 to 0.911.ConclusionsThis is the first study to demonstrate that microsatellite markers provide valuable information for genetic structure of E. (M.) onukii in Chinese tea plantations. There is obvious genetic differentiation between the two populations in the Southwest tea area. These microsatellite markers will be the powerful tools for genetic studies of E. (M.) onukii and improve understanding of the microevolution of this species.Electronic supplementary materialThe online version of this article (doi:10.1186/s12863-016-0420-3) contains supplementary material, which is available to authorized users.
Highlights
Tea green leafhopper is one of the most dominant pests in Chinese tea plantations
The (AG)12, (AC)12 and (AAC)8-enriched partial genomic libraries were constructed by FIASCO. 100 ng of DNA was digested with MseI (BioLabs, Beijing, China) and ligated to prepared MseI AFLP adaptors (5′-TACTCAGGACTC AT-3′/5′-GACGATGAGTCCTGAG-3′) using T4 DNA ligase (TaKaRa, Dalian, China) The linkeradaptor-PCR was performed in a final volume of 50 μL containing: 2 mM MgCl2, 250 uM dNTP, 0.8 uM MseI AFLP adaptors and 0.32 U of Taq DNA polymerase (TaKaRa, Dalian, China)
Evaluation of microsatellite markers One hundred and seventy-three clones were selected to be sequenced from three microsatellite-enriched libraries, (AC)n, (AG)n and (AAC)n
Summary
Tea green leafhopper is one of the most dominant pests in Chinese tea plantations. Recent evidence, including morphological and molecular data, revealed that tea green leafhopper in China is the same species as in Japan, Empoasca (Matsumurasca) onukii Matsuda. Zhang et al BMC Genetics (2016) 17:112 the genetic polymorphism and relationships among seven populations of this species [9] or sequencing of mtDNA from the COI and 16S rRNA gene regions from different populations [10,11,12] These studies were conducted without first confirming the species identifications of the leafhoppers included, using morphology. Because of the high species diversity (>800 species worldwide and >200 in China) and morphological similarity of the Chinese fauna of Empoasca it is important to examine the male genitalia of specimens used in such studies to ensure accurate identification In these previous studies only a few genetic markers were used for a single individual, and their variation was too low for studying population structure. The population structure of E. (M.) onukii is still poorly understood
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