摘要
本研究以苹果牛眼果腐病菌Neofabraea perennans的3个菌株为供试材料,通过对10种活性染料设置5种不同浓度和5种不同染色时间的实验条件,筛选到染料PI能够区别孢子死活,其最佳的染色条件为0.05 mg/mL,室温避光染色4 min。再通过设置5种不同水浴温度(范围)和5种不同水浴时间(范围)对孢子进行处理,获得孢子的最佳热处理条件为55湿热处理10 min,并以传统萌发实验作为对照,结果显示,不同条件处理后的孢子染色率不能与孢子萌发率完全相符,其孢子不萌发数高于被染色比例,极有可能是孢子在所试温度下被钝化而无法萌发。本研究初步建立了基于PI染色法和荧光显微镜的苹果牛眼果腐病菌Neofabraea perennans的活性检测方法,能够对其分生孢子进行活性检测,从制样到检测完成仅需约30 min,可用于口岸对该病菌的活性检测。
In this study,three strains of Neofabraea perennans were used as test materials.The experimental conditions were set with 5 different concentrations and 5 different staining times for 10 active dyes and it was screened that PI dyes could distinguish between dead and live spores.Next the spores were treated by setting 5 different water bath temperatures(ranges)and 5 different water bath times(ranges).The optimal heat treatment condition for obtaining spores was 55 humid heat treatment for 10 minutes,and the traditional germination experiment was used as a control.The statistical results showed that the staining rate of spores treated with different conditions could not be completely consistent with the germination rate of spores,and the number of spores that did not germinate was higher than the proportion of stained spores.This study preliminarily established a method for detecting the viability test of conidia activity of Neofabraea perennans species based on PI staining and fluorescence microscopy 30 minutes,and the method can be used for the quarantine viability detection of the pathogen at the port.
作者
李娜
黄河清
章桂明
LI Na;HUANG He-Qing;ZHANG Gui-Ming(Shenzhen Customs Inspection and Quarantine Technology Center,Shenzhen 518000)
出处
《中国口岸科学技术》
2022年第4期31-37,共7页
China Port Science and Technology
基金
国家重点研发计划(2016YFF0203204)。
关键词
苹果牛眼果腐病菌
荧光显微镜技术
活性检测
孢子萌发
Neofabraea perennans
fungus fluorescence microscope
viability detection
spore germination
