Biodiversity Science ›› 1994, Vol. 02 ›› Issue (Suppl.): 55-60.doi: 10.17520/biods.1994047

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Detection of the Paulownia Witches' Broom Mycoplasmalike Organism by Polymerase Chain Reaction

ZHANG CHUNLI, LIN MULAN, YANG JIHONG, SHI XIAOYAN   

  1. Wuhan Institute of Virology, Academia Sinica, Wuhan 430071, China
  • Received:1994-04-04 Revised:1994-05-18 Online:1994-12-20

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.

Key words: microbial diversity, resource conservation

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