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      Mitochondrial COI and 16sRNA Evidence for a Single Species Hypothesis of E. vitis, J. formosana and E. onukii in East Asia

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          Abstract

          Tea green leafhopper is one of the most damaging tea pests in main tea production regions of East Asia. For lack of recognized morphological characters, the dominant species of tea green leafhoppers in Mainland China, Taiwan and Japan have always been named as Empoasca vitis Göthe, Jacobiasca formosana Paoli and Empoasca onukii MATSUDA, respectively. Furthermore, nothing is known about the genetic relationships among them. In this study, we collected six populations from Mainland China, four populations from Japan and one population from Taiwan, and examined the genetic distances in the COI and 16sRNA regions of mtDNA among them. The results showed that the genetic distances based on single gene or the combined sequences among eleven leafhopper populations were 0.3–1.2%, which were all less than the species boundary of 2%. Moreover, there were at least two haplotypes shared by two distinct populations from different regions. The phylogenetic analysis based on single gene or combined sets also supported that tea green leafhoppers from Mainland China, Taiwan and Japan were closely related to each other, and there were at least two specimens from different regions clustered ahead of those from the same region. Therefore, we propose that the view of recognizing the dominant species of tea green leafhoppers in three adjacent tea production regions of East Asia as different species is unreliable or questionable and suggest that they are a single species.

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          DNA barcodes affirm that 16 species of apparently generalist tropical parasitoid flies (Diptera, Tachinidae) are not all generalists.

          Many species of tachinid flies are viewed as generalist parasitoids because what is apparently a single species of fly has been reared from many species of caterpillars. However, an ongoing inventory of the tachinid flies parasitizing thousands of species of caterpillars in Area de Conservación Guanacaste, northwestern Costa Rica, has encountered >400 species of specialist tachinids with only a few generalists. We DNA-barcoded 2,134 flies belonging to what appeared to be the 16 most generalist of the reared tachinid morphospecies and encountered 73 mitochondrial lineages separated by an average of 4% sequence divergence. These lineages are supported by collateral ecological information and, where tested, by independent nuclear markers (28S and ITS1), and we therefore view these lineages as provisional species. Each of the 16 apparently generalist species dissolved into one of four patterns: (i) a single generalist species, (ii) a pair of morphologically cryptic generalist species, (iii) a complex of specialist species plus a generalist, or (iv) a complex of specialists with no remaining generalist. In sum, there remained 9 generalist species among the 73 mitochondrial lineages we analyzed, demonstrating that a generalist lifestyle is possible for a tropical caterpillar parasitoid fly. These results reinforce the emerging suspicion that estimates of global species richness are likely underestimates for parasitoids (which may constitute as much as 20% of all animal life) and that the strategy of being a tropical generalist parasitic fly may be yet more unusual than has been envisioned for tachinids.
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            Global relationships of Bemisia tabaci (Hemiptera: Aleyrodidae) revealed using Bayesian analysis of mitochondrial COI DNA sequences.

            Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a species complex that is one of the most devastating agricultural pests worldwide and affects a broad range of food, fiber and ornamental crops. Unfortunately, using parsimony and neighbor joining methods, global phylogenetic relationships of the major races/biotypes of B. tabaci remain unresolved. Aside from the limitations of these methods, phylogenetic analyses have been limited to only small subsets of the global collection of B. tabaci, and thus limited taxon sampling has confounded the analyses. To improve our understanding of global B. tabaci phylogenetic relationships, a Bayesian phylogenetic technique was utilized to elucidate the relationships among all COI DNA sequence data available in GenBank for B. tabaci worldwide (366 specimens). As a result, the first well-resolved phylogeny for the B. tabaci species complex was produced showing 12 major well-resolved (0.70 posterior probability or above) genetic groups: B. tabaci (Mediterranean/Asia Minor/Africa), B. tabaci (Mediterranean), B. tabaci (Indian Ocean), B. tabaci (sub-Saharan Africa silverleafing), B. tabaci (Asia I), B. tabaci (Australia), B. tabaci (China), B. tabaci (Asia II), B. tabaci (Italy), B. tabaci (New World), B. tabaci (sub-Saharan Africa non-silverleafing) and B. tabaci (Uganda sweet potato). Further analysis of this phylogeny shows a close relationship of the New World B. tabaci with Asian biotypes, and characteristics of the major sub-Saharan Africa non-silverleafing clade strongly supports an African origin of B. tabaci due to its position at the base of the global phylogeny, and the diversity of well-resolved sub-clades within this group. Bayesian re-analyses of B. tabaci ITS, COI, and a combined dataset from a previous study resulted in seven major well-resolved races with high posterior probabilities, also showing the utility of the Bayesian method. Relationships of the 12 major B. tabaci genetic groups are discussed herein.
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              Insect pests of tea and their management.

              Globally, 1031 species of arthropods are associated with the intensively managed tea Camellia sinensis (L.) O. Kuntze monoculture. All parts of the plant, leaf, stem, root, flower, and seed, are fed upon by at least one pest species, resulting in an 11%-55% loss in yield if left unchecked. There has been heavy use of organosynthetic pesticides since the 1950s to defend the plant against these pests, leading to rapid conversion of innocuous species into pests, development of resistance, and undesirable pesticide residues in made tea. As a result of importer and consumer concerns, pesticide residues have become a major problem for the tea industry. Integrated pest management (IPM) may help to overcome the overuse of pesticides and subsequent residues. We review the advances made in our understanding of the biology and ecology of major insect and mite pests of tea, host plant resistance, cultural practices, biocontrol measures, and need-based application of botanicals and safer pesticides to understand the present status of IPM and to identify future challenges to improvement.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2014
                15 December 2014
                : 9
                : 12
                : e115259
                Affiliations
                [1 ]Key Laboratory of Tea Plants Biology and Resources Utilization of Agriculture Ministry, Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310008, PR China
                [2 ]Graduate School of Chinese Academy of Agricultural Sciences, Beijing, 100081, PR China
                [3 ]Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, College of Life Sciences of China Jiliang University, Hangzhou, 310018, PR China
                St. Petersburg Pasteur Institute, Russian Federation
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: JYF BYH QX. Performed the experiments: JYF. Analyzed the data: JYF. Contributed reagents/materials/analysis tools: JYF. Wrote the paper: JYF BYH QX. Cloned COI and 16sRNA genes, did the phylogenetic analysis and wrote the manuscript: JYF.

                Article
                PONE-D-14-30254
                10.1371/journal.pone.0115259
                4266653
                25506929
                ba96457b-3034-49b7-9c71-3ab58d3d5603

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 10 July 2014
                : 20 November 2014
                Page count
                Pages: 15
                Funding
                This work was supported by the National Natural Science Foundation of China (31470693, 31100503), the public technology application research of Zhejiang Province (2013C32086), the Fundamental Research Funds of National Nonprofit Research Institute for Tea Research Institute, CAAS, (0032014014), the Natural Science Foundation of Zhejiang province, China (Y3080150), the Fundamental Research Funds of National Science and Technology (2013FY113200), and the Key Innovation Team of Zhejiang Province (Grant No. 2011R09027-13). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and life sciences
                Evolutionary biology
                Evolutionary systematics
                Molecular systematics
                DNA barcoding
                Phylogenetics
                Animal Phylogenetics
                Zoology
                Custom metadata
                The authors confirm that all data underlying the findings are fully available without restriction. All relevant data are within the paper and its Supporting Information files.

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