Abstract:

Invasive alien organisms pose a severe threat to China’s ecological security and food security. Invasive agricultural pests (mites) are often mixed with native allied pests (mites) with feature diverse morphologies and complex groups. Accurate differentiation between them requires professional taxonomic expertise, which remains a major challenge for grassroots agricultural technicians. This study established a standardized protocol combining morphological identification and DNA barcoding technology as follows: field technicians preserve suspected invasive pests (mites) in 75% ethanol and submit samples to a laboratory capable of molecular biology analysis, and the application of DNA barcoding technology enables precise identification of invasive agricultural pest (mites) species. Using this protocol, the samples of suspected invasive agricultural pests (mites) collected in Shaanxi Province were identified, and the following species were accurately distinguished: the invasive species Frankliniella occidentalis from the native species F. intonsa, the invasive species Liriomyza sativae, L. huidobrensis from the native species Chromatomyia horticola, and L. bryoniae, the invasive Mediterranean cryptic species of Bemisia tabaci from the native Trialeurodes vaporariorum, and the invasive Tetranychus urticae from the native green-type Tetranychus truncatus and Tetranychus viennensis. Furthermore, the protocol confirmed the distribution of the invasive pests Tuta absoluta and the Bruchophagus roddi in northern Shaanxi. This standardized protocol allows for accurate identification of alien invasive insect (mite) species within one week, and the cost of single sample can be controlled at around 30 RMB. It holds significant importance for the rapid and accurate identification, monitoring, early warning, and control of invasive pests.

Key words: DNA barcoding technology, alien invasion, agricultural pests, identification, procedure and standards