Development of Dipstick for the Rapid Detection of Three Important Monilinia Species on Fruits

doi:10.13560/j.cnki.biotech.bull.1985.2018-1006

Abstract

Abstract: A rapid detection dipstick for fruit quarantine in port was studied and developed using three important brown rot pathogens（Monilinia fructicola,Monilinia laxa and Monilinia fructigena）on fruits. Using nitrocellulose membrane as the solid phase carrier,the translation elongation factor 1-alpha（EF-1α）of above 3 Monilinia species as detecting targets,and PCR amplification products of biotin marker as detecting markers,DNA-DNA hybridization was applied in the rapid detection of 3 Monilinia species with the developed dipstick. The specificity and sensitivity of the Monilinia dipstick（abbr. as MON dipstick below）were evaluated by using DNA of brown rot strains and the constructed recombinant plasmids of EF-1α as positive standard samples,respectively. The results showed that the MON dipstick specifically detected 3 Monilinia species;the whole detection process（including DNA extraction and amplification process）was completed in 2 h,and the lowest detection limit reached 10 fg/µL. In sum,the dipstick developed in this study is of high specificity,sensitivity and stability,and the detection result is visible by human eyes,thus it does not rely on expensive equipment and can be expanded and used in local quarantine laboratory.

Key words: Monilinia fructicola, Monilinia laxa, Monilinia fructigena, dipstick, rapid detection

LIN Hui-jiao, YANG Hua-wei, GU Heng-sen, JIANG Xiang, ZHANG Hai-lei, LIU Yu-chen, ZHOU Er-xun. Development of Dipstick for the Rapid Detection of Three Important Monilinia Species on Fruits[J]. Biotechnology Bulletin, 2019, 35(6): 205-212.

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