Characterization and Pathogenicity of Rhizoctonia Species Causing Root Rot and Damping-off on Sugar Beet in Turkey.

During 2015-2017 growing seasons, seventy-one isolates of Rhizoctonia spp. were obtained from the samples collected from the sugar beet fields of Konya province, which is one of the most important sugar beet growing areas of Turkey. Anastomosis groups (AGs) of Rhizoctonia spp. isolates were determined by hyphal conjugation with the tester strains and sequence analysis of the rDNA ITS region. The obtained data confirmed the species identity of 61 isolates out of 71 as R. solani (AG-2-2-IIIB, AG-4-(HGI, HGII, HGIII), AG-5, AG-11) and the remaining 10 isolates as binucleate Rhizoctonia (AG-K, AG-A). Pathogenicity tests revealed that AG-2-2-IIIB, AG-4-(HGI, HGII, HGIII) and AG-K isolates were highly virulent on sugar beet. The disease severity of 71 isolates varied between 13 and 100%. Based on the virulence, the isolates formed four categories; (i) 11 isolates: non-pathogenic, (ii) 15 isolates: low virulent, (iii) 6 isolates: moderately virulent and (iv) 39 isolates: highly virulent. To our knowledge, the AGs of AG-4-(HGI, HGII, HGIII), AG-11, AG-A are first reports on sugar beet in Turkey and the AG-5, AG-11 and AG-K are first AG groups for Konya region.

A field based detection method for Rose rosette virus using isothermal probe-based Reverse transcription-recombinase polymerase amplification assay.

Rose rosette disease, caused by Rose rosette virus (RRV; genus Emaravirus) is a major threat to the rose industry in the U.S. The only strategy currently available for disease management is early detection and eradication of the infected plants, thereby limiting its potential spread. Current RT-PCR based diagnostic methods for RRV are time consuming and are inconsistent in detecting the virus from symptomatic plants. Real-time RT-qPCR assay is highly sensitive for detection of RRV, but it is expensive and requires well-equipped laboratories. Both the RT-PCR and RT-qPCR cannot be used in a field-based testing for RRV. Hence a novel probe based, isothermal reverse transcription-recombinase polymerase amplification (RT-exoRPA) assay, using primer/probe designed based on the nucleocapsid gene of the RRV has been developed. The assay is highly specific and did not give a positive reaction to other viruses infecting roses belonging to both inclusive and exclusive genus. Dilution assays using the in vitro transcript showed that the primer/probe set is highly sensitive, with a detection limit of 1 fg/µl. In addition, a rapid technique for the extraction of viral RNA (<5min) has been standardized from RRV infected tissue sources, using PBS-T buffer (pH 7.4), which facilitates the virus adsorption onto the PCR tubes at 4°C for 2min, followed by denaturation to release the RNA. RT-exoRPA analysis of the infected plants using the primer/probe indicated that the virus could be detected from leaves, stems, petals, pollen, primary roots and secondary roots. In addition, the assay was efficiently used in the diagnosis of RRV from different rose varieties, collected from different states in the U.S. The entire process, including the extraction can be completed in 25min, with less sophisticated equipments. The developed assay can be used with high efficiency in large scale field testing for rapid detection of RRV in commercial nurseries and landscapes.