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http://www.globalsciencebooks.info/Online/GSBOnline/images/2011/AmJPSB_5%28SI1%29/AmJPSB_5%28SI1%29107-114o.pdf
Context in source publication
Context 1
... occurrence and spread of the potato tuber necrosis strain of PVY, i.e., PVY NTN could be considered another example. Unlike the common strain (PVY O ) and the tobacco veinal necrosis strain (PVY N ), PVY NTN induces potato tuber necrotic ringspot disease (PTNRD) in sensitive potato varieties and veinal necrosis in tobacco plants (Fig. 1), thus posing a significant threat to the potato industry (Singh et al. 2008). PVY NTN was first found in Hungary in the 1980s (Beczner et al. 1984) and subsequently in many other parts of the world (Singh et al. 2008), suggesting that the spread might be a recent event. Capable of causing deformed- tubers, Potato mop-top virus (PMTV), ...
Citations
... Various techniques including biological index, serological assay, and molecular approaches have been developed and used to detect viruses, viroids included, in potato plants, tubers, and vectors (DeHaan 1994;Singh and Nie 2003). These diagnostic methods not only play a pivotal role in seed certification programs worldwide but also contribute to the development of more effective management regimes for controlling or managing various viral diseases (Frost et al. 2013;Nie 2011). For instance, RT-PCR-based detection has been used to monitor the spread of potato virus Y (PVY) in growing plants and developing tubers during the growing season (Fageria et al. 2013) and to identify viruliferous aphids and potential PVY-vectoring aphid species (Boquel et al. 2013;Pelletier et al. 2012), thus enabling the development of improved management practices for managing PVY in seed potato crops in New Brunswick, Canada (MacKenzie et al. 2016. ...
In this study, a set of duplex reverse transcription (RT)-PCR-mediated high resolution DNA melting (HRM) analyses for simultaneous detection of potato mop-virus (PMTV) and its protist vector, Spongospora subterranea f.sp. subterranea (Sss), was developed. The infestation of soil by PMTV was detected by using a tobacco-based baiting system. Total RNA extracted from the soil led to successful RT-PCR gel-electrophoresis detection of both PMTV and Sss. To facilitate more efficient detection, newly designed primer pairs for PMTV RNA species (i.e., RNA-Rep, -CP, and -TGB) were analyzed together with the existing Sss primers using real-time RT-PCR. The resulting amplicons exhibited melting profiles that could be readily differentiated. Under duplex RT-PCR format, all PMTV and Sss primer combinations led to successful detection of respective PMTV RNA species and Sss in the samples by high resolution DNA melting (HRM) analyses. When the duplex HRM assay was applied to soil samples collected from six fields at four different sites in New Brunswick, Canada, positive detection of PMTV and/or Sss was found in 63-100% samples collected from fields in which PMTV-infected tubers had been observed. In contrast, the samples from fields where neither PMTV- nor Sss-infected tubers had been observed resulted in negative detection by the assay. Bait tobacco bioassay for PMTV and Sss produced similar results. Between 63%-83% and 100% of the soil samples collected from PMTV-infested fields led to PMTV and Sss infections in the bait tobacco plants, respectively; whereas no PMTV or Sss infected plants were obtained from soil samples collected from PMTV/Sss-free fields.
