摘要
本研究合成了苹果茎沟病毒(ASGV)、苹果褪绿叶斑病毒(ACLSV)和苹果茎痘病毒(ASPV)的生物素标记cDNA探针,对斑点杂交和免疫印迹杂交检测这3种病毒的效果进行了分析。以含ACLSV和ASGV克隆片段的大肠杆菌菌液为样品时,斑点杂交检测灵敏度分别为80 cfu/μL和50 cfu/μL。以离体培养砂梨植株为材料,采用斑点杂交法检测砂梨离体培养植株粗提液中的3种病毒,均产生强的杂交信号,且特异性好。比较斑点杂交和ELISA检测病毒含量相对较低的热处理再生植株中ASGV和ACLSV,结果表明斑点杂交具有较高的灵敏度。组织印迹杂交检测砂梨离体植株ASGV和ACLSV的结果显示,这2种病毒在离体植株的各部位均有分布,自基部至茎尖各部位印迹均产生很强的杂交信号。
Biotinylated cDNA probes specific to Apple stem grooving virus （ ASGV）, Apple cholortotic leaf spot virus （ACLSV） and Apple stem pitting virus （ASPV） were prepared from cloned fragments. Efficiency of dot-blot and tissue printing hybridization for detection of these three viruses was evaluated. The detection sensitivities of dot-blot hybridization for E. coli containing cloned fragments were 80 cfu/μL and 50 cfu/μL for ACLSV and ASGV respectively. The prepared biotinylated cDNA probes could specifically recognize ASGV, ACLSV and ASPV in crude extract of in vitro-cultured pear plants and give strong hybridization signals. Comparison of detection results for ASGV and ACLSV in regenerated in vitro-cultured pear plants from thermotherapy revealed a higher sensitivity of dot-blot hybridization than ELISA. Tissue printing hybridization was also successfully used for detection of ASGV and ACLSV in in vitro-cultured pears. The results showed that these two viruses distributed in the whole plants.
出处
《植物病理学报》
CAS
CSCD
北大核心
2006年第6期488-493,共6页
Acta Phytopathologica Sinica
基金
国家自然科学基金资助项目(30370997)
科技部"八六三"资助项目(2001AA241142)
关键词
斑点杂交
梨
潜隐病毒
检测
组织印迹
dot-blot hybridization
pear
latent viruses
detection
tissue printing
