利用新PCR引子檢測小綠葉蟬（Empoasca spp.）攜帶花生簇葉病菌質體

Peanut Witches' Broom Phytoplasma Detection in Empoasca spp. Using New PCR Primers

陳覽繽(Lan-Pin Tan) ； 蔡正隆(Cheng-Lung Tsai) ； 蔡孟旅(Meng-Lu Tsai) ； 陳昇寬(Sheng-Kuan Chen) ； 陳美娥(Mei-Er Chen)

《台灣昆蟲》 41卷1期 (2021/02) Pp. 10-16

https://doi.org/10.6662/TESFE.202102_41(1).002

摘要

植物菌質體（Candidatus Phytoplasma）為一植物病原細菌，可透過吸食韌皮部汁液之昆蟲傳播，感染多種植物造成不同之疾病。花生受菌質體感染引發簇葉病（peanut witches' broom disease），造成經濟損失。目前已知南斑葉蟬（Orosius orientalis）為傳播花生簇葉病的媒介昆蟲。然而，在花生田中亦可發現小綠葉蟬（Empoasca spp.）的蹤跡，小綠葉蟬與南斑葉蟬皆為吸食韌皮部汁液之昆蟲，因此本研究檢測小綠葉蟬是否也攜帶花生簇葉病菌質體，以探究其為花生簇葉病之媒介昆蟲的可能性。利用PCR檢測小綠葉蟬全蟲的DNA，透過P1／P7及f1／r1通用性引子對增幅植物菌質體之16S rRNA基因，初步檢測小綠葉蟬是否攜帶花生簇葉病菌質體，結果顯示以f1／r1引子對檢測採集樣本中有76％的小綠葉蟬及52％的南斑葉蟬可增幅出產物。進一步設計專一性引子對f3／r3，降低通用性引子對f1／r1增幅非目標產物之情形，結果於採集樣本中41％的小綠葉蟬及53％的南斑葉蟬可增幅出菌質體標的產物。本研究證實小綠葉蟬也可攜帶花生簇葉病菌質體，是否能藉以傳播則有待進一步檢測；此成果可作為花生簇葉病菌質體病害防治中防除媒介昆蟲之基礎資訊。同時，本研究研發之f3／r3引子對可應用於防治花生簇葉病菌質體媒介昆蟲之檢測工具。

關鍵字

花生簇葉病 ； 植物菌質體 ； 小綠葉蟬 ； 南斑葉蟬

並列摘要

Phytoplasmas are plant pathogens transmitted by phloem-feeding insects and cause many diseases. Peanut witches' broom (PnWB) disease is induced by phytoplasmas and causes substantial economic loss for farmers. Until now, Orosius orientalis has been identified as the vector of PnWB diseases. However, Empoasca spp. are also found in peanut fields. Both Orosius orientalis and Empoasca spp. are phloem-feeding insects. Therefore, phytoplasma detection in Empoasca spp. was performed to determine the possibility of Empoasca spp. being a vector. The first step was to detect if the phytoplasma 16S rRNA gene was present in total Empoasca spp. DNA through a polymerase chain reaction (PCR) with P1/P7 and f1/r1 universal primers. Further, f3/r3 primers were designed to avoid the untargeted PCR products from universal f1/r1 primers. Phytoplasmas were detected in 76% of Empoasca spp. and 52% of Orosius orientalis samples when the f1/r1 primers were used, and in 41% of Empoasca spp. and 53% of O. orientalis samples when the f3/r3 primers were used. The results of this study can be used as fundamental information in developing preventative measures against the insects that transmit PnWB diseases. Additionally, the f3/r3 primers developed in this study can serve as a tool for the detection of PnWB phytoplasma in insects.

並列關鍵字

peanut witches' broom disease ； phytoplasma ； Empoasca sp. ； Orosius orientalis