热带作物学报2024，Vol.45Issue(6)：1120-1126,7.DOI:10.3969/j.issn.1000-2561.2024.06.004
槟榔黄化植原体TaqMan探针实时荧光定量PCR检测方法的建立
Establishment of TaqMan Probe Real-time Fluorescent Quantitative PCR Detection Method for Areca Palm Yellow Leaf Phytoplasma
摘要
Abstract
Areca palm is an important tropical economic crop in Hainan Province.Areca palm yellow leaf disease(YLD)caused by phytoplasma infection is a devastating disease in the production of areca in China.In order to estab-lish an accurate and efficient detection method for areca palm yellow leaf phytoplasma,this study designed and syn-thesized specific primers AMf/AMr and AM-Prode based on the 16S rDNA gene of areca palm yellow leaf phyto-plasma.The method was used to test the accuracy,sensitivity,specificity and repeatability,and to detect phytoplasma diseases in other plants.The method could accurately detect the positive samples,and the healthy samples had no am-plification curve.In the sensitivity test,the method could detect the sample concentration level of 1.16×101 copies/μL,and the standard curve equation was y=-3.4185x+43.624,the amplification efficiency was 96.12％,and the correlation coefficient R2=0.9833.In the specificity test,the method had good specificity for the detection of YLD,and the ge-nomes of areca,other disease pathogens and the endophytes did not interfere with the method.In the repeatability test,the method had good repeatability for the detection of YLD.The detection method could be used to detect 8 phyto-plasma diseases such as chinaberry yellow leaf disease,'Pericampylus glaucus'witches'broom disease and pepper yellow leaf disease and the detection of phytoplasma has certain universality.The establishment of detection method is conducive to providing reliable technical means for the accurate diagnosis of YLD,pathogen monitoring and vector insect detection.关键词
槟榔/槟榔黄化病/植原体/TaqMan实时荧光定量PCR/病害检测Key words
areca/areca palm yellow leaf disease/phytoplasma/TaqMan qPCR/disease detection分类
农业科技引用本文复制引用
林兆威,孟秀利,唐庆华,牛晓庆,宋薇薇..槟榔黄化植原体TaqMan探针实时荧光定量PCR检测方法的建立[J].热带作物学报,2024,45(6):1120-1126,7.基金项目
海南省自然科学基金项目(No.321QN345) （No.321QN345）
海南省院士创新平台科研专项(No.YSPTZX202138,No.YSPTZX202151)o （No.YSPTZX202138,No.YSPTZX202151）
