China is the largest potato producing country worldwide,with this crop representing the fourth largest staple food crop in China.However,the steady presence of Potato spindle tuber viroid（PSTVd） over the past five d...China is the largest potato producing country worldwide,with this crop representing the fourth largest staple food crop in China.However,the steady presence of Potato spindle tuber viroid（PSTVd） over the past five decades has a significant economic impact on potato production.To determine why PSTVd control measures have been ineffective in China,more than 1 000 seed potatoes collected between 2009 and 2014 were subjected to PSTVd detection at the Supervision and Testing Center for Virus-free Seed Potatoes Quality,Ministry of Agriculture,China.A high PSTVd infection rate（6.5%） was detected among these commercial seed potatoes.Some breeding lines of potato collected from 2012 to 2015 were also tested for PSTVd infection,revealing a high rate of PSTVd contamination in these potato propagation materials.Furthermore,comparison of the full-length sequences of 71 different Chinese PSTVd isolates revealed a total of 74 predominant PSTVd variants,which represented 42 different sequence variants of PSTVd.Comparative sequence analysis revealed 30 novel PSTVd sequence variants specific to China.Comprehensive phylogenetic analysis uncovered a close relationship between the Chinese PSTVd sequence variants and those isolated from Russia.It is worth noting that three intermediate strains and six mild strains were identified among these variants.These results have important implications for explaining the ineffective control of PSTVd in China and thus could serve as a basic reference for designing more effective measures to eliminate PSTVd from China in the future.展开更多
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...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.展开更多
基金supported by the China Agriculture Research System(CARS-10-P14)the National Science Foundation for Post-doctoral Scientists of China(20110491125)+2 种基金the Heilongjiang Funds for Distinguished Young Scientist,China (JC 201018)the Harbin Application Technology Research and Development Projects,China(2013AE6AW059)the Young Scientists Science Foundation of Heilongjiang Province,China(QC2015026)
文摘China is the largest potato producing country worldwide,with this crop representing the fourth largest staple food crop in China.However,the steady presence of Potato spindle tuber viroid（PSTVd） over the past five decades has a significant economic impact on potato production.To determine why PSTVd control measures have been ineffective in China,more than 1 000 seed potatoes collected between 2009 and 2014 were subjected to PSTVd detection at the Supervision and Testing Center for Virus-free Seed Potatoes Quality,Ministry of Agriculture,China.A high PSTVd infection rate（6.5%） was detected among these commercial seed potatoes.Some breeding lines of potato collected from 2012 to 2015 were also tested for PSTVd infection,revealing a high rate of PSTVd contamination in these potato propagation materials.Furthermore,comparison of the full-length sequences of 71 different Chinese PSTVd isolates revealed a total of 74 predominant PSTVd variants,which represented 42 different sequence variants of PSTVd.Comparative sequence analysis revealed 30 novel PSTVd sequence variants specific to China.Comprehensive phylogenetic analysis uncovered a close relationship between the Chinese PSTVd sequence variants and those isolated from Russia.It is worth noting that three intermediate strains and six mild strains were identified among these variants.These results have important implications for explaining the ineffective control of PSTVd in China and thus could serve as a basic reference for designing more effective measures to eliminate PSTVd from China in the future.
基金Supported by Doctoral Research Startup Project of Zhaotong University。
文摘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.
