%A ZHANG CHUNLI, LIN MULAN, YANG JIHONG, SHI XIAOYAN %T Detection of the Paulownia Witches' Broom Mycoplasmalike Organism by Polymerase Chain Reaction %0 Journal Article %D 1994 %J Biodiv Sci %R 10.17520/biods.1994047 %P 55-60 %V 02 %N Suppl. %U {https://www.biodiversity-science.net/CN/abstract/article_8584.shtml} %8 1994-12-20 %X

A polymerase chain reaction (PCR) method with high sensitivity and specificity has been developed with a pair of oligonucleotide primers designed and synthesized according to the DNA sequence specific to paulownia witches' broom mycoplasmalike organism (PWB-MLO). Under optimum conditions, i. e. 22. 5mM of Mg2+, 3.0 units of Taq DNA polymerase in a 100μl PCR reaction mixture and annealing at 55℃, a unique 377-bp DNA fragment was amplified aftter 30 cycles from all field-grown, MLO-infected paulownia tissues with or without PWB symptom. No amplified DNA was detected under the same conditions in the samples from healthy paulowina, healthy and MLO-infected jujube and rhododendron plants. With the established PCR system, the PWB-MLO DNA from approximately 2pg of crude DNA from diseased paulownia petiole tissues could be detected. This method is suitable for practical diagnosis and customs quarantine.