Plant Quarantine Laboratory

Nanjing, China

Plant Quarantine Laboratory

Nanjing, China
SEARCH FILTERS
Time filter
Source Type

Wu C.P.,Nanjing University | Wu C.P.,Plant Quarantine Laboratory | Chen G.Y.,Nanjing University | Li B.,Plant Quarantine Laboratory | And 4 more authors.
Forest Pathology | Year: 2011

A nested and real-time PCR assay was developed for the rapid and accurate detection of Ceratocystis fagacearum, which is the causal agent of oak wilt in stained wood and soil. Based on the differences of the internal transcribed spacer (ITS) sequences of Ceratocystis spp., one pair of species-specific primers, CF01/CF02, was designed. Whereas a 280-bp product was amplified using the purified DNA from three isolates of C. fagacearum as the template, no PCR product was obtained from template of other 18 fungi. The detection sensitivity was 10 pg genomic DNA per 25-μl PCR reaction volume. To increase detection sensitivity, a nested PCR was developed by using ITS1/ITS4 as the first-round primers and CF01/CF02 in the second round, as it can detect 1 pg genomic DNA per 25-μl PCR reaction volume. More importantly, CF01/CF02 primers were successfully adapted to real-time PCR with a detection limit of 0.1 pg genomic DNA per 20-μl PCR reaction volume. Using these two methods, we could rapidly and accurately detect the pathogen in artificially infected wood and soil. © 2009 Blackwell Verlag GmbH.


Chen G.Y.,Nanjing University | Wu C.P.,Nanjing University | Wu C.P.,Plant Quarantine Laboratory | Li B.,Plant Quarantine Laboratory | And 3 more authors.
Journal of Plant Diseases and Protection | Year: 2010

Leptosphaeria maculans, the causal agent of blackleg on oilseeds in many regions around the world, has not been reported in China. In this study, L. maculans was detected from imported canola seeds by polymerase chain reaction (PCR) using a species-specific primer pair. The isolated L. maculans exhibited morphology identical to the known L. maculans strain on PDA culture plates and produced typical blackleg symptom when inoculated on oilseed leaves. It is the first report on the detection and identification of L. maculans from Canadian oilseed in China. © Eugen Ulmer KG, Stuttgart.

Loading Plant Quarantine Laboratory collaborators
Loading Plant Quarantine Laboratory collaborators