摘要
CTAB法是一种广泛用于从染病植物中提取总核酸,并用于后续相关病原物PCR检测的核酸提取法,已普遍运用于DNA病毒、植原体、细菌、真菌侵染植物的总DNA提取和病原检测。本文尝试将CTAB法用于一种由RNA病毒(烟草丛顶病毒)引起的烟草丛顶病染病组织总核酸的提取,以总核酸进行烟草丛顶病毒的RT-PCR检测,并与常规RNA病毒核酸提取法——Trizol法作比较。结果表明CTAB法和Trizol法提取的核酸用于烟草丛顶病毒RT-PCR检测时,均能扩增到目的条带,检测结果一致。在对复合侵染材料进行检测时发现,CTAB法提取的染病植物组织总核酸既包括DNA也包含RNA,有利于对DNA和RNA病原物同时进行检测;CTAB法所用试剂较Trizol法便宜,毒性小,且提取的核酸量较多、保存时间较长。因此CTAB法是一种高效、简便、经济的从烟草丛顶病染病组织中提取烟草丛顶病毒RNA的方法。
CTAB method is one of the conventional pathogenic total nucleic acids extraction ways from diseased plants,which can then be used for pathogen detection in PCR-based techniques.This method has been widely used in total nucleic acids extraction and pathogen detection from the plants infected by DNA virus,phytoplasma,bacterium and fungus.In this study,the CTAB method was tried to extract the total nucleic acids from infected tobacco plants by a RNA virus of Tobacco bushy top virus（TBTV）,and compared with Trizol method,a routine RNA virus nucleic acids extraction method,for TBTV RT-PCR detection.The RT-PCR detection results showed that TBTV expected cDNA band could be amplified from the RNA templates extracted from both two methods.The extracts include both DNA and RNA when CTAB is used to extract the total nucleic acids from the co-infected plants,and thus are beneficial to detect both DNA and RNA pathogens.The chemicals used in CTAB method are much cheaper and low toxic than Trizol reagent.Additionally,the total nucleic acids extracted with CTAB method are high quantity and have long-term storage compared with Trizol method.Therefore,CTAB method is a more efficient,simple and cheap method for RNA extraction from TBTV infected tobacco plants.
出处
《云南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2011年第1期26-29,共4页
Journal of Yunnan Agricultural University:Natural Science
基金
国家自然科学基金项目(30660101)
