Detection and Practical Differentiation of Phytoplasmas from Several Host Plants Using PCR-RFLP

Abstract

Phytoplasma as a phytopathogenic prokaryote with a wide host range is a pathogen that needs more attention in Indonesia. This pathogen is relatively difficult to detect and identify due to its complicated biological properties. This study involved detection of phytoplasmas by polymerase chain reaction (PCR) technique with P1/P7 primers from seven symptomatic plants, i.e. Bermuda grass white leaf, bamboo yellows, witches’ broom of peanut, soybean, yard long bean, and cactus, and sweet potato little leaf. The phytoplasma DNA of the 16S rRNA gene resulting from PCR amplification was examined by digestion reaction using three endonuclease enzymes AluI, RSaI, and MSeI to generate restriction fragment length polymorphism (RFLP) profile. The seven diseased plants were confirmed positive to be associated with phytoplasma as indicated by the PCR product of 1800 bp. Based on the RFLP profiles of the three enzymes, the phytoplasmas were divided into two groups, namely group I (Bermuda grass and bamboo) and group II (peanuts, soybeans, yard long beans, cactus, and sweet potatoes). Cactus phytoplasma is a sub-group (strain) because it has a slightly different fragment of MSeI RFLP profile.

Keywords

• 16SrRNA gene
• fastidious prokaryotes
• restriction endonucleases,
• yellows disease
• witches’ broom