Improved Nucleic Acid Spot Hybridization Technique for Detection of Potato Spindle Tuber Viroid (PSTVd)

• 作者: Qiu Cai-ling Lu Dian-qiu Liu De-fu Shi Jiao-xu Bao Liu-yuan Ma Zhong-lian Liu Li Feng Zhen-yue Huang Xian-min Jiang Rui Chen Yue Wang Shi-min
• 来源期刊: 东北农业大学学报（英文版）
• 学科: 农学
• 年，卷（期）: 2022,（1）
• 页码范围: 26-32
• 页数: 7页
• 分类号: S532|S435.32
• 语种: 英文

摘要

Potato spindle tuber viroid (PSTVd) disease is one of the major diseases that threatens potato production. Therefore, an advanced, rapid and sensitive detection technology is needed to detect the disease for better control. In order to establish an easier nucleic acid spot hybridization (NASH) method, some studies were tried as the followings: (1) the pre-hybridization step of nucleic acid spot hybridization (NASH) was omitted compared with ordinary way; (2) RNA extraction (phenol extraction and Ames buffer extraction) methods were compared; (3) fixed RNA by UV lamp and oven compared with UV cross-linker; (4) hybridized the RNA in shaking incubator and so on. The results showed that RNA extracted by Ames buffer was more effective than by the phenol extraction method. Besides, the result of hybridization without pre-hybridization step was better than that with 1.5 h of pre-hybridization. The more important discovery was that the shaking incubator could replace the hybridization oven and the ordinary UV lamp could replace the UV cross-linker. After a long term repeated research and testing, a new hybridization system that could rapidly detect the PSTVd by improved NASH technique merely using common instruments and equipment was established.