Lee, Jung-Han
(Department of Applied Biology & Environmental Sciences, Gyeongsang National Univesity)
,
Han, Ki-Soo
(Department of Applied Biology & Environmental Sciences, Gyeongsang National Univesity)
,
Lee, Sun-Cheol
(Department of Applied Biology & Environmental Sciences, Gyeongsang National Univesity)
,
Shim, Chang-Ki
(Department of Applied Biology & Environmental Sciences, Gyeongsang National Univesity)
,
Bae, Dong-Won
(Central Laboratory, Gyeongsang National Univesity)
,
Kim, Dong-Kil
(Research Institute of Life Science, Gyeongsang National Univesity)
,
Kim, Hee-Kyu
(Research Institute of Life Science, Gyeongsang National Univesity)
We developed a polyclonal antibody (PAh) based- ELISA system to accurately and rapidly monitor inocula on plant surface before onset of anthracnose. Titer of mouse antisera against conidia of Colletotrichum gloeosporioides was determined by using indirect ELISA. It was high enough to be detectable u...
We developed a polyclonal antibody (PAh) based- ELISA system to accurately and rapidly monitor inocula on plant surface before onset of anthracnose. Titer of mouse antisera against conidia of Colletotrichum gloeosporioides was determined by using indirect ELISA. It was high enough to be detectable up to ${\times}$ 12,800 dilutions. Absorbance readings exceeded (1.5even at a 10$^{-5}$ dilution. Sensitivity of PAb was precise enough to detect spore concentration as low as 50 conidia/well by indirect ELISA. PAb1 and PAb2 proved to be very sensitive and highly specific to the target pathogen, C. gloeosporioides, apparently discriminating other unrelated pathogens, or epiphytes. Absorbance values for original isolate exceeded 1.0, but no reaction was detected with other isolates, except three other anthracnose fungi: C. gloeosporioides (pepper strain), Glomerella cingulata (apple strain) and C. lagenarium. Our data suggest that PAb1 and PAb2 bind with the protein epitope that partially contains residues of amino acid, arginine, and Iysine. This kit fulfills the require-ments for detecting inoculums before infection and during onset of anthracnose on sweet persimmon.
We developed a polyclonal antibody (PAh) based- ELISA system to accurately and rapidly monitor inocula on plant surface before onset of anthracnose. Titer of mouse antisera against conidia of Colletotrichum gloeosporioides was determined by using indirect ELISA. It was high enough to be detectable up to ${\times}$ 12,800 dilutions. Absorbance readings exceeded (1.5even at a 10$^{-5}$ dilution. Sensitivity of PAb was precise enough to detect spore concentration as low as 50 conidia/well by indirect ELISA. PAb1 and PAb2 proved to be very sensitive and highly specific to the target pathogen, C. gloeosporioides, apparently discriminating other unrelated pathogens, or epiphytes. Absorbance values for original isolate exceeded 1.0, but no reaction was detected with other isolates, except three other anthracnose fungi: C. gloeosporioides (pepper strain), Glomerella cingulata (apple strain) and C. lagenarium. Our data suggest that PAb1 and PAb2 bind with the protein epitope that partially contains residues of amino acid, arginine, and Iysine. This kit fulfills the require-ments for detecting inoculums before infection and during onset of anthracnose on sweet persimmon.
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성능/효과
In conclusion, PAbl and PAb2 proved to be very sensi tive and highly specific to the target pathogen, Colletotri- chum gloeosporioides, apparently discriminating other unrelated pathogens, or epiphytes. This kit fiilfills the requirements for detecting inoculums before infection and onset of anthracnose, and would provide us with a more highly accurate information than that obtained with PCR method or classical identification method (Singh et aL, 2000).
for Colletotrichum lagenarium. Taxonomy of Gloeo sporium kaki was revised to C. gloeosporioides by von Arx J. A., and G cingulata is a telemorph of C. gloeosporioides. Still, it is likely that persimmon strains, formerly Gloeo sporium kaki and other two C.
참고문헌 (10)
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Singh, U., Trevors, C. M., Boer, S. H. and Janse, J. D. 2000. Fimbrial- specific monoclonal antibody-based ELISA for European potato strains of Envinia chrysanthemi and comparison to PCR. Plant Dis. 84:443-448
Somai, B. M. and Keinath, A. P. 2002. Development of PCRELISA for detection and differentiation of Didymella bryoniae from related phoma species. Plant Dis. 86:710-716
Sundaram, S., Plasencia, J. and Banttari, E. E. 1991. Enzymelinked immunosorbent assay for detection of verticillium spp. Using antisera produced to V. dahliae from potato. Phytopathology 81: 1485-1489
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