First report of Cotton leafroll dwarf virus in Thailand using a species-specific PCR validated with isolates from Brazil

Abstract

Partial virus genome sequence with high nucleotide identity to Cotton leafroll dwarf virus (CLRDV) was identified from two cotton (Gossypium hirsutum) samples from Thailand displaying typical cotton leaf roll disease symptoms. We developed and validated a PCR assay for the detection of CLRDV isolates from Thailand and Brazil.

Two Complete Genome Sequences of Phasey Bean Mild Yellows Virus, a Novel Member of the Luteoviridae from Australia

“…From this, total RNA was extracted using a BioSprint 15 plant DNA kit (Qiagen), as per the manufacturer’s instructions, except that RNase A was not used. PCR products were amplified using previously described conditions and cycling parameters ( 5 ), using the overlapping primer pairs PLF/PLR, Pol3167F/Pol3982R, and Pol3870F/AS3 ( 5 – 7 ), and were sequenced directly using an Applied Biosystems automated sequencing system (AGRF, Brisbane, Australia). After the removal of the primer sequences, the resulting partial genome fragment from isolate 2805 was 1,360 nucleotides (nt) in length.…”

Cotton leafroll dwarf virus detected in Timor-Leste

“…Unless further sequence information becomes available for CpSDaV which proves otherwise, the virus detected in Timor-Leste should be considered a strain of CLRDV. This detection of a CLRDV-like sample from TimorLeste represents a significant geographic jump in the distribution of this virus, with the next nearest reported location being Thailand (Sharman et al 2015). Limited host range data is available for CLRDV but is reported to include several Malvaceae species such as G. hirsutum, G. barbardense, Hibiscus sabdariffa and Sida acuta (Kaowsiri 1982).…”

“…Total nucleic acids were extracted and partial virus genome amplified from the sample by reverse transcription PCR as previously described by Sharman et al (2015) . The CLRDV-specific PCR (primers CLRDV3675F / Pol3982R) produced an amplicon of expected size of approximately 310 bp from the Timor-Leste sample (BRIP no.…”

“…Sequence alignments and phylogenetic analyses were done as previously described (Sharman et al 2015). The almostcomplete coat protein gene sequence of 591 nucleotide (nt) from the Timor-Leste sample was analysed to determine phylogenetic relationships to published sequences.…”

Faba bean polerovirus 1 (FBPV-1); a new polerovirus infecting legume crops in Australia

“…Synthesis of cDNA was done using SuperScript III reverse transcriptase (Thermo Fisher) as per instructions using primer AS3 [2]. PCR conditions were as described by Sharman et al [30] using primers BWYV3969F (GTC TCC GAR GCC TCT TCC CAA) and AS3 under the following cycling conditions: initial denaturation at 95 °C for 1 min, then 35 cycles of 95 °C for 15 s, 62 °C for 20 s, 56 °C for 10 s and 72 °C for 40 s; followed by a final extension step at 72 °C for 3 min. Out of the 74 plants tested, 19 were positive.…”