利用 PCR-RFLP 技術快速鑑定台灣進口農產品中常見的介殼蟲 (同翅目：介殼蟲總科)

邱一中; 吳文哲; 翁振宇; 陳淑佩; 石正人; Yi-Chung Chiu; Wen-Jer Wu; Cheng-Yu Wong; Shu-Pei Chen; Cheng-Jen Shih

利用 PCR-RFLP 技術快速鑑定台灣進口農產品中常見的介殼蟲 (同翅目：介殼蟲總科)

Other Title

Application of the PCR-RFLP Technique for the Rapid Diagnosis of Scale Insects (Homoptera: Coccoidea) on Imported Agricultural Products in Taiwan

Journal

台灣昆蟲

Journal Volume

24

Journal Issue

2

Pages

159-171

Date Issued

2004-06

Date

2004-06

Author(s)

邱一中

吳文哲

翁振宇

陳淑佩

石正人

Yi-Chung Chiu

Wen-Jer Wu

Cheng-Yu Wong

Shu-Pei Chen

Cheng-Jen Shih

URI

https://scholars.tari.gov.tw/handle/123456789/5627

Abstract

依據海關人員截獲農產品上附有的介殼蟲，經鑑定共計有 30 種，本研究針對其中 16 種經常截獲的介殼蟲，建立以核糖體形成基因為基礎的聚合連鎖反應-限制片段長度多態型技術 (ribosomal DNA/polymerase chain reaction-restriction fragment length polymorphism, rDNA/PCR-RFLP)，做為介殼蟲分子快速診斷鑑定的操作標準流程，提供海關、機場的檢疫人員依操作標準進行常見介殼蟲的診斷鑑定。試驗篩選出 NS7 和 ITS6 引子對，可供增幅供試的 16 種介殼蟲之 rDNA/ITS1 特定區域。並進一步篩選出 MspI、TaqI 和 HaeIII 核酸內切的切圖譜具有較高的種類特異性，可作為種類鑑定依據。此外，利用分類學慣用的檢索表概念，以 RFLP 切圖譜差異之分生特徵，建立試驗 16 種介殼蟲的分子檢索表，提供快速、簡單且準確的診斷鑑定方式。 There are 30 scale insect species which have been intercepted from agricultural products coming into Taiwan by quarantine workers at customs. This study attempted to develop a molecular diagnostic technique (polymerase chain reaction-restriction fragment length polymorphism, PCR- RFLP) using genomic DNA markers (ribosomal DNA, rDNA) for identifying 16 scale insect species. NS7 and ITS6 primer sets were used to amplify ITS1 and its flanking regions of rDNA from genomic DNA as a template extracted from a single specimen. DNA extracted from these 16 scale insect species yielded a single fragment after PCR amplification. These PCR products were then cut with various restriction endonucleases in order to compare the results of restriction fragment length polymorphism. Our results also reveal that it is possible to discriminate these 16 scale insect species based on the species-specific patterns acquired from digesting the PCR products with the endonucleases, TaqI, MspI, and HaeIII. Futhermore, we also a set up a molecular key for these 16 scale insect species based on their RFLP digestion patterns.

Subjects

介殼蟲

快速鑑定

限制片段長度多態型 (RFLP)

分子檢索表

scale insect

rapid identification

restriction fragment length polymorphism (RFLP)

molecular key

Publisher

臺灣昆蟲學會

Type

journal article

File(s)

Name

24-2-07.pdf

Size

1.68 MB

Format

Adobe PDF

Checksum

(MD5):f19679891e87b948501d9ccef9f9693b