Multiplex PCR for simultaneous identification of Ralstonia solanacearum and Xanthomonas perforans

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農業知識庫

作者：S. Umesha;P. Avinash; 公佈日期：105 年 12 月 30 日

研究報告

995

Ralstonia solanacearum is a causative agent of

bacterial wilt in many economically important crops, and

Xanthomonas perforans is the causal organism of bacterial

spot, one of the most important diseases of vegetables. A

multiplex PCR protocol has been developed for the

simultaneous, specific and rapid identification of R. solanacearum

and X. perforans in plant materials. Speciesspecific

primers RS-F-759 and RS-R-760 for R. solanacearum,

RST2 and RST3 for X. perforans were used for

identification of both pathogens at primer concentrations of

1:4 by optimization of multiplex PCR at annealing temperature

of about 61 ± 1 C. With these primer sets, specific

amplification of 281- and 840-bp PCR products was

obtained for R. solanacearum and X. perforans, respectively.

The multiplex PCR assay was validated with susceptible

plants mechanically inoculated with both the

pathogens; specific PCR products confirmed the presence

of R. solanacearum and X. perforans. The multiplex PCR

is valuable in identification as well as primary screening of

cultivars of both pathogens. The present study is a rapid

and easy method for early identification of pathogens from

asymptomatic and symptomatic plant materials.

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