Thrips Identification on Imported Crops Through Multiplex PCR and Morphological Identification: Detection of the Quarantine Thrips Frankliniella occidentalis and Other Related Species (Thysanoptera: Thripidae)

應用複合性PCR及形態特徵鑑定輸入農作上之薊馬：檢疫薊馬西方花薊馬及其他相關薊馬（纓翅目：薊馬科）之檢測

Watchalawalee Boonmee ； 王子政(Tzu-Cheng Wang) ； 陳素琴(Su-Chin Chen) ； 葉文斌(Wen-Bin Yeh)

《台灣昆蟲》 41卷2期 (2021/05) Pp. 37-47+ap1-3

https://doi.org/10.6662/TESFE.202105_41(2).001

摘要

Thrips cause severe damage to many crops and are frequently found on imported crops. Thus, many thrips species are categorized as quarantine pests. In this study, thrips species, intercepted in imported agricultural products, were identified based on morphological characteristics and through multiplex polymerase chain reaction (PCR). Five specific primers for the three quarantine thrips species Anaphothrips obscurus (Müller), F. occidentalis Pergande, and Thrips fuscipennis Haliday and two other common species, Thrips tabaci Lindeman and Frankliniella intonsa (Trybom), were designed based on the internal transcribed spacer (ITS) region, a well-known marker for species identification. Five species-specific primer pairs and one universal primer pair based on conserved 28S ribosomal DNA were pooled in each multiplex PCR. Identification results were consistent, whether based on morphological or molecular characteristics. Perhaps due to the sample quality, the molecular identification efficiency was approximately 87%. The most common thrips species on imported crops was T. tabaci, followed by F. occidentalis, which was observed primarily on strawberry and artichoke crops from the United States; some F. occidentalis specimens were also found on leucadendron and eryngium plants from Australia and the Netherlands. The other thrips species found on imported crops were F. intonsa, Scirtothrips dorsalis Hood, several species of the Tubulifera suborder, and one other quarantine species, namely Thrips subnudula Karny, which was identified morphologically on imported leacospemum plants from Australia. This study demonstrated that multiplex PCR based on specific primers of the ITS region is a rapid and reliable method for identifying quarantine F. occidentalis, which was identified with 93% accuracy. In addition, morphological identification is crucial when nontarget quarantine thrips species are observed.

關鍵字

quarantine pest ； molecular identification ； internal transcribed spacer

並列摘要

嚴重危害眾多農作的薊馬常於進口作物上發現，有些種類遂被列為檢疫對象。本研究以進口農作為對象，針對西方花薊馬（Frankliniella occidentalis）及數種檢疫薊馬進行檢測；結合形態特徵，並依據常用於物種鑑定的核醣體區間（internal transcribed spacer, ITS），設計5對專一性引子，應用複合性PCR（multiplex PCR）檢測黃呆薊馬（Anaphothrips obscurus）、西方花薊馬、玫瑰花薊馬（Thrips fuscipennis）3種檢疫薊馬及2種常見的蔥薊馬（Thrips tabaci）及歐洲薊馬（Frankliniella intonsa）；每一複合性的PCR檢測中均含這5組專一性引子及一組28S rDNA區段的廣效性引子。分子鑑定的結果與形態特徵相互一致。進口作物上的薊馬多為蔥薊馬及美國草莓及薊類作物上的西方花薊馬，部分西方花薊馬也可在澳洲及紐西蘭的百合及刺芹上發現，其它進口作物上檢測到的薊馬有歐洲薊馬、小黃薊馬（Scirtothrips dorsalis）及管尾亞目薊馬物種（Tubulifera），並含有一種經形態鑑定，來自澳洲針墊花的Thrips subnadula。本研究證實應用ITS區段種專一性的複合性PCR是一個快速有效鑑定檢疫的西方花薊馬的方法，準確率可達93%；此外，對於非標的性的檢疫薊馬種類，仍須依賴形態特徵的鑑定。

並列關鍵字

檢疫有害生物 ； 分子鑑定 ； 核醣體區間