Amplification of 16S rRNA genes from culturable and nonculturable Mollicutes

First report of a ‘Candidatus Phytoplasma aurantifolia’ isolate associated with a yellowing disease of Barleria prionitis

“…Barleria prionitis (Acanthaceae), known as porcupine flower, is a medicinal plant species native to India and commonly found across Africa, Sri Lanka, and tropical Asia (Chavana et al, 2010) (Deng & Hiruki, 1991), followed by fU5/rU3 (Lorenz et al, 1995 (Arocha et al, 2008). Therefore, these results have a significant phytosanitary impact for the epidemiology of 16SrII phytoplasma diseases in the country.…”

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Barleria prionitis (Acanthaceae), known as porcupine flower, is a medicinal plant species native to India and commonly found across Africa, Sri Lanka, and tropical Asia (Chavana et al, 2010) (Deng & Hiruki, 1991), followed by fU5/rU3 (Lorenz et al, 1995 [doi:10.1111/j.1365-3059.2008.01969.x] Chavan CB, Hogade MG, Bhinge S, Kumbhar M, Tamboli A, 2010

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First report of a phytoplasma associated with sapodilla flattened stem disease in Iran

“…Total DNA was extracted separately from two symptomatic and eight symptomless trees using the CTAB method. Samples were analysed for presence of phytoplasma DNA by direct-PCR using the phytoplasma universal primer pair P1/P7 (Deng and Hiruki 1991;Schneider et al 1995) and nested-PCR using primer pairs P1/P7 (first round) and R16F2n/R16R2 (second round) (Gundersen and Lee 1996). The PCR was performed in 20 μl of reaction mixture containing 10 μl PCR Master Mix (Amplicon), 1 μl of each primer (10 μM), 2 μl of template DNA and 6 μl sterile distilled water.…”

First report of Candidatus Phytoplasma aurantifolia (16SrII group) associated with Conocarpus erectus disease in Iran

“…The lysates obtained from each sample were extracted two times by phenol: chloroform:isoamyl alcohol and chloroform:isoamyl alcohol, respectively. Total DNA was analyzed by the nested polymerase chain reaction assay, in which the partial regions of 16S rDNA were amplified using two primer pairs of P1/P7 (Deng and Hiruki, 1991) and R16F2n/R16R2 (Gundersen and Lee, 1996). The nested PCR products were separated by electrophoresis in 1% agarose gels and the fragments of the expected size were excised and purified by GF-1 AmbiClean Kit (Vivantis, Malaysia) according to the manufacturer's instructions.…”