橡胶树白粉菌分生孢子DNA的微量提取方法建立OA北大核心CSTPCD

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.