热带作物学报2024，Vol.45Issue(7)：1314-1322,9.DOI:10.3969/j.issn.1000-2561.2024.07.002
橡胶树白粉菌分生孢子DNA的微量提取方法建立
Establishment of a Microscale DNA Extraction Method for Rubber Tree Powdery Mildew Spores
摘要
Abstract
Powdery mildew caused by Erysiphe quercicola is a significant disease in China's natural rubber industry,posing a severe threat to latex production.With the continuous advancement of predictive and forecasting technologies,the combination of qPCR detection and spore capture techniques holds promise as an efficient method for predicting the prevalence of powdery mildew.In this study,we focused on the microscale extraction of DNA from rubber tree powdery mildew spores.We compared various pre-treatment methods,different types of grinding beads with varying materials,and different types of grinding machines to identify the most suitable spore sample grinding system.Furthermore,we compared the efficiency of the CTAB method with several commercial extraction kits.Ultimately,we established a fea-sible microscale DNA extraction method for spores:first,the powdery mildew spore water solution subjected to 4 ℃ ultrasonication for 10 min,subsequently,2 mm zirconia beads(0.2 g),glass beads(0.4-0.6 mm,0.2 g),and 0.2 mm zirconia beads(0.2 g)added,tissue grinder 70 Hz at 4 ℃,grind 40 s,stop 10 s,20 cycles,finally,DNA extracted using the CTAB method.This method effectively and stably extracts DNA from 101 spores/mL rubber tree powdery mildew spores,without being affected by complex field conditions,and demonstrates high sensitivity and specificity.It provides a reliable theoretical basis and an effective technical approach for the prediction and forecasting of rubber tree powdery mildew.关键词
橡胶树/白粉菌/分生孢子/DNA/微量提取Key words
rubber tree/Erysiphe quercicola/spores/DNA/microscale extraction分类
农业科技引用本文复制引用
冯德宇,梁晓宇,王晗茉,黄曦泽,周绍尧,王萌,张宇..橡胶树白粉菌分生孢子DNA的微量提取方法建立[J].热带作物学报,2024,45(7):1314-1322,9.基金项目
国家自然科学基金项目(No.32160370) （No.32160370）
国家天然橡胶产业技术体系病害岗位专家经费项目(No.CARS-33-BC1) （No.CARS-33-BC1）
海南省研究生创新科研课题(No.Qhys2022-107). （No.Qhys2022-107）
