Molecular detection and identification of phytoplasma affecting smooth bromegrass (Bromopsis inermis) [Fitoplazmos, pažeidžiančios beginklės dirsuolės (Bromopsis inermis) augalus, nustatymas ir apibūdinimas molekuliniais metodais]. – Botanica Lithuanica, 11(3): 191–194
Smooth bromegrass (Bromopsis inermis (Leyss.) Holub) plants exhibiting symptoms of general yellowing, stunting, bushiness and sterility of spikes were observed in the fields and waysides in the Vilnius region of Lithuania. The detected phytoplasma was identified and classified using restriction fragment length polymorphism (RFLP) analysis of PCR amplified 16S rDNA. A phytoplasma-characteristic fragment of 16S rDNA was amplified in nested polymerase chain reactions which were primed by phytoplasma universal primers (R16F2n/R2) and template DNA extracted from diseased plants, confirming phytoplasma infection in the symptomatic plants. The 1.2 kbp 16 rDNA product was subjected to single enzyme digestions with 10 different restriction endonucleases. RFLP analysis of amplified 16S rDNA indicated that the diseased smooth bromegrass was infected by a phytoplasma strain belonging to group 16SrI (aster yellows phytoplasma group) and subgroup 16SrI-B. The phytoplasma strain found in smooth bromegrass was named bromegrass stunt (BrS) phytoplasma.
Keywords: Bromopsis inermis, PCR, RFLP, 16S rRNA gene.