Submitted to: American Phytopathological Society Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: 5/8/2015
Publication Date: 8/1/2015
Citation: Ling, K., Li, R. 2015. Genome sequencing, genetic diversity and field detection of Cucumber green mottle mosaic virus using LAMP technology. American Phytopathological Society Abstracts. P. 570.
Technical Abstract: The recent outbreaks of Cucumber green mottle mosaic virus on cucumber, melon and watermelon in Australia, Canada, and the U.S. highlight the importance in implementing a cleaned seed program to manage this seed-borne virus from introduction. Both Canadian and Australian isolates were closely related to the Asian genotype with 99% sequence identity. However, the California isolate was more divergent, which was more closely resembled those from Europe (95% sequence identity) than from Asia (90-92% sequence identity). In addition to the genetic diversity, recent discovery of a strong serological specificity between the two genotypes causes concerns on the reliability of a serological test (i.e., ELISA), particularly for seed health test, where virus titer is typically lower. To improve reliability for a sensitive detection of CGMMV isolates in all genotypes, a loop-mediated isothermal amplification (LAMP) was developed with six oligonucleotides derived from a highly conserved region in a multi-sequence alignment of available 31 genome sequences. This LAMP system was confirmed to provide a sensitive detection to all genotypes of CGMMV. In combination with a simple crude tissue RNA preparation, this LAMP detection system was used to monitor CGMMV infection on greenhouse cucumber. The availability of this simple and sensitive detection would greatly enhance our ability to provide an early and reliable detection of CGMMV-infected plants as well as for seed health test.