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Phromnoi S.,Kasetsart University | Phromnoi S.,Center for Agricultural Biotechnology | Sirinarumitr K.,Kasetsart University | Sirinarumitr K.,Center for Agricultural Biotechnology | And 2 more authors.
Virus Genes | Year: 2010

Canine parvovirus (CPV) causes a very severe enteric disease especially in puppies. Twenty-six isolates of CPV were obtained from dogs at the Animal Hospital, Kasetsart University, Thailand. Whole VP2 gene of 26 isolates was amplified using polymerase chain reaction (PCR) and its sequences were analyzed. Nineteen out of 26 isolates were characterized as CPV type 2a variants and the rest of the isolates were characterized as CPV type 2b. These results indicated that both types are currently prevalent field CPV circulating in Thailand and type 2a is the predominant genotype. Neither CPV type 2 nor type 2c was observed in this study. © 2010 Springer Science+Business Media, LLC.

Temiyakul P.,Kasetsart University | Temiyakul P.,Center for Agricultural Biotechnology | Taylor P.W.J.,University of Melbourne | Mongkolporn O.,Kasetsart University
Journal of Phytopathology | Year: 2010

Chilli anthracnose is caused by a complex of Colletotrichum species. Breeding for resistance to anthracnose has been focused on introgressing resistance from Capsicum chinense and C. baccatum into commercial C. annuum cultivars. Trispecies hybrids of C. annuum cv. Bangchang, C. chinense PBC932 and C. baccatum PBC80 were successfully produced. Assessments for resistance in F1 progeny to Colletotrichum capsici isolate 158ci (Cc158ci) and C. acutatum isolate MJ5 (CaMJ5) were carried out by inoculating fruit using a laboratory microinjection method. Due to the poor fruit set of the F1 hybrid, a double-inoculation method was developed to inoculate the same chilli fruit with two isolates of two Colletotrichum species. The positions (apex, centre, end) at which the fruit were inoculated with either isolate did not affect disease development. At 7 days after inoculation, Cc158ci produced larger lesions on chilli fruit than CaMJ5; and lesions from single inoculation were larger than those from double inoculation. The double-inoculation technique was applied to the trispecies F1 hybrid to select individual F1 plants that contained resistance to both Colletotrichum species. Of the nine F1 plants that produced fruits for inoculation, all were resistant to Cc158ci at both mature green and ripe fruit stages. Two plants were also resistant to CaMJ5 at both fruit maturity stages, and one plant was resistant at the ripe fruit stage but susceptible at the green fruit stage. © 2010 Blackwell Verlag GmbH.

Phromnoi S.,Kasetsart University | Phromnoi S.,Center for Agricultural Biotechnology | Sinsiri R.,Kasetsart University | Sirinarumitr T.,Kasetsart University | Sirinarumitr T.,Center for Agricultural Biotechnology
Kasetsart Journal - Natural Science | Year: 2010

Canine parvovirus (CPV) appears to be endemic in almost all populations of wild and domesticated dogs. It causes serious contagious enteric disease. The VP2 protein of CPV is a major capsid protein and plays an important role in the host immune response. In the present study, the recombinant VP2 was expressed in Escherichia coli (E. coli) using the pBAD expression system. Virus DNA from the infected feces was extracted and used to amplify the whole VP2 gene by using specific primers. Subsequently, the whole VP2 gene was ligated with plasmid pBAD202/D-TOPO and used to transform into the E. coli strain TOP10. The SDS-PAGE analysis revealed a specific band approximately 80 kDa and was found mainly in the pellet of the bacterial lysate. The optimum time and concentration of arabinose for expression of recombinant VP2 protein was 8 h and 0.002%, respectively. By dot blot and Western blot analysis, the recombinant VP2 protein showed specific interaction with mouse antihistidine monoclonal antibody and rabbit anti-CPV hyperimmune serum. The recombinant protein VP2 might be a useful tool for the development of a diagnostic test for the detection of CPV and a vaccine against CPV.

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