摘要
把柑桔黄龙病(Citrus Huanglongbing,HLB)阳性核酸样品进行梯度稀释后,分别采用HLBp、CQULAP两组TaqMan水解探针法和以rpl基因为目的片段的SYBR Green染料法进行HLB实时荧光定量PCR(Quantitative Real-time PCR,qPCR)检测,结果发现,HLBp探针法可以从稀释105倍的阳性叶片核酸样品(病菌拷贝数约为60个)中检测到病原菌,高于CQULAP探针法和SYBR Green染料法。将3种qPCR检测体系用于赣州安远县、寻乌县和赣县送检的60份田间样品检测,结果发现,HLBp探针法的阳性率为86.7%,CQULAP探针的阳性率为60.0%,SYBR Green染料法的阳性率为68.3%。
Three Quantitative Real-time PCR（qPCR）methods were compared using TaqMan probes（HLBp and CQULAP）and SYBR Green targeted rpl gene to diagnose citrus Huanglongbing（HLB）disease.Results showed that TaqMan probe HLBp could detect the pathogen in 105 fold of dilution from total DNA extracts of the samples（about 60 copy of pathogen）.It was more sensitive than TaqMan probe CQULAP and SYBR Green methods.To further confirm sensitivity of three qPCR protocols,60 leaf samples from orchards in Anyuan county,Xunwu county,Ganxian county were used for detection of HLB by TaqMan probe HLBp,TaqMan CQULAP and SYBR Green targeted rpl gene,respectively.Assays showed that TaqMan probe HLBp has the highest positive rate of 86.66%,TaqMan probe CQULAP and SYBR Green methods showed positive rate of 60%and 68.33%,respectively.
出处
《中国南方果树》
北大核心
2016年第2期33-37,共5页
South China Fruits
基金
国家自然科学基金(31560602)
江西省科技计划重大项目(20152ACF60003)
江西省科技计划重大项目(20143ACF60012)
江西省科技厅青年科学基金(20131542040011)
江西省教育厅重点项目(GJJ12564)
江西省教育厅科技项目(GJJ14661)资助
江西省自然科学基金(20122BAB204024)
江西省自然科学基金(20131542040011)
