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ARS Home » Research » Publications at this Location » Publication #113385
Title: DESIGN OF A POLYMERASE CHAIN REACTION FOR SPECIFIC DETECTION OF CORN STUNT SPIROPLASMA

Author
item BARROS, THEREZA - UNIV DE BRASILIA BRASILIA
item Davis, Robert
item RESENDE, RENATO - UNIV DE BRASILIA BRASILIA
item Dally, Ellen

Submitted to: Plant Disease
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/16/2001
Publication Date: N/A
Citation: N/A

Interpretive Summary: Corn stunt disease is a major limiting factor in production of corn (Zea mays L.) in the Americas. The pathogen that causes this disease is a minute, helical, cell wall-less bacterium known as the corn stunt spiroplasma, Spiroplasma kunkelii. The overall objective of our work was to aid the development of corn varieties that are resistant to the disease and to enhance research concerning the geographical distribution and spread of the corn stunt spiroplasma. In this paper, we report the design of oligonucleotide primers for use in the polymerase chain reaction (PCR). We designed primers on the basis of nucleotide sequences of the gene encoding a membrane protein, spiralin, in different species of Spiroplasma. By using these primers, we achieved sensitive detection of spiroplasma DNA, and specific detection of corn stunt spiroplasma DNA. The availability of this sensitive and specific PCR for detection and identification of S. kunkelii should facilitate studies of the ecology of this pathogen, as well as its influence on the incidence, spread, and severity of corn stunting disorders.

Technical Abstract: Corn stunt disease is a major limiting factor in production of corn (Zea mays L.) in the Americas. To develop a polymerase chain reaction (PCR) assay specific for detection of the causal agent, Spiroplasma kunkelii, PCR primers were designed on the basis of unique regions of the nucleotide sequence of the S. kunkelii spiralin gene. DNA was amplified in PCRs containing template DNAs derived from laboratory strains of S. kunkelii and from naturally diseased corn plants collected in the field. No DNA amplification was observed in PCRs containing template DNAs derived from other Spiroplasma species tested or from healthy corn or corn infected by maize bushy stunt phytoplasma. The availability of a sensitive and specific PCR for detection and identification of S. kunkelii should facilitate studies of the ecology of this pathogen, as well as its influence in the incidence, spread, and severity of corn stunting diseases.