Development of Rapid Molecular Detection Marker for Colletotrichum spp. in Leaf and Fruit Tissues of Sweet Persimmon

  • Iee, Sang-Pyo (Plant Molecular Biology & Microbiol Biotechnology Lab, Div. Of Applid Plant Science, College of Agriculture and Life Sciences, Kangwon National University) ;
  • Lee, Youn-Su (Plant Molecular Biology & Microbiol Biotechnology Lab, Div. Of Applid Plant Science, College of Agriculture and Life Sciences, Kangwon National University)
  • Published : 2002.12.01

Abstract

Sweet persimmon (Diospyros kaki Thunb.) is widely cultivated in the southern part of Korea and its cultivation is increasing. However, anthracnose disease caused by Colletotricuhum species is one of the major hinderances to the cultivation and production of sweet persimmon. Therefore, in the current study, PCR was used to specifically detect Colletotrichum spp., based on the sequences of the ITS II regions in the rDNA. Using the sequence data, CO-1 was designated to detect Colletotrichum together the with ITS 4 primer. The result showed that a single segment of ca. 500 bp was observed only in Colletotrichum, but not in any other fungal and bacterial isolates. The annealing temperatures and template DNA quantites were also investigated to identify optimal conditions for detection. Using these species-specific primers, a unique band was obtained at annealing temperatures ranging from $55^{\circ}C\;and\;61^{\circ}C$ and template DNA levels from 10 pg- $10{\mu}g$.

Keywords

References

  1. Plant Pathology Agrios,G.N.
  2. Phytopathology v.82 Morphological and pathological characteristics of strains of Colletotrichum gloeosporioides from citrus Agostini,J.P.;L.W.Timmer;D.J.Mitchell https://doi.org/10.1094/Phyto-82-1377
  3. Mycol. Res. v.94 Restriction fragment length polymorphism in Colletotrichum gloeosporioides infecting Stylosanthes spp. in Austria. Breithwaite,K.S.;J.A.G.Irwin;J.M.Manners https://doi.org/10.1016/S0953-7562(09)81345-2
  4. Curr. Genet. v.22 Cloning and genetic analysis of two highly polymorphic moderately repetitive nuclear DNAs from Phytophthora infestans. Goodwin,S.B.;A.Drenth;W.E.Fry https://doi.org/10.1007/BF00351469
  5. Mycologia v.84 Taxonomy and morphology of Colletotrichum species pathogenic to strawberry Gunnell,P.S.;W.D.Gubler https://doi.org/10.2307/3760246
  6. PCR Protocols. A guide to methods and applications Optimization of PCRs Innis,M.A.;D.H.Gelfand;M.A.Innis;D.H.Gelfand;J.J.Sninsky;T.J.White(eds.)
  7. J. Microbiol. Biotechnol. v.10 Rapid and accurate species-specific detection of Phytophthora infestans through analysis of ITS regions in its rDNA Kim,K.S.;Y.S.Lee
  8. J. Microbiol. Biotechnol. v.10 Rapid identification of Lactobacillus planuarum in Kimchi using polymerase chain reaction Kim,T.W.;S.G.Min;D.H.Choi;J.S.Jo;H.Y.Kim
  9. J. Microbiol. Biotechnol. v.11 Inhibitory effects of acetic acid and temperature on growth of Campylobacter jejuni ATCC 33291 Kim,W.J.;S.Y.Shin;H.J.Hwang
  10. J. Microbiol. Biotechnol. v.11 Detection of Aspergillus and Penicillium genera by enzyme-linked immunosorbent assay using a monoclonal antibody Kwak,B.Y.;D.H.Shon;B.J.Kwon;C.H.Kweon;K.H.Lee
  11. J. Microbiol. Biotechnol. v.11 16S/23S intergenic spacer region as a genetic marker for thiobacillus thiooxidans and T. ferrooxidans. Lee,H.Y.;W.Y.Choi;K.S.Cho;W.J.Choi
  12. Plant Disease v.82 Identification of races of Colletotrichum lindemuthianum with the aid of PCR-based molecular markers Mesquita,A.G.G.;T.J.Paula;M.A.Moreira Jr;E.G.de Barros https://doi.org/10.1094/PDIS.1998.82.10.1084
  13. J. Microbiol. Biotechnol. v.11 Estimation of distribution of a commensal thermophile in soil by competitive PCR and terminal restriction fragment length polymorphism analysis Rhee,S.K.;S.P.Hong;J.W.Bae;C.O.Jeon;S.G.Lee;J.J.Song;H.Poo;M.H.Sung
  14. Exp. Mycol. v.18 Ribosomal new species grouping in the genus Colletotrichum Sherriff,C.;M.J.Whelan;G.M.Amold;J.M.Lafay;J.Brygoo;J.A.Bailey https://doi.org/10.1006/emyc.1994.1014
  15. sublineolum. Mycological Research v.99 rDNA sequence analysis confirms the distinction between Colletotrichum graminicola and C. Sherriff,C.;M.J.Whelan;G.M.Arnold;J.A.Bailey https://doi.org/10.1016/S0953-7562(09)80649-7
  16. J. Microbiol. Biotechnol. v.9 Specific detection of enteropathogen Camphlobacter jejuni in food using a polymerase chain reaction Shin,S.S.;J.H.Park;W.J.Kim
  17. Genome v.39 Phylogeny and systematics of 18 Colletotrichum species based on ribosomal DNA spacer sequences Sreenivasaparasad,S.;P.R.Mills;B.M.Meehan;A.E.Brown https://doi.org/10.1139/g96-064
  18. Physiol. Mol. Plant Pathol. v.41 DNA sequence variation and interrelationships among Colletotrichum species causing strawgerry anthracnose Sreenivasaprasad,S.;A.E.Brown;P.P.Mills https://doi.org/10.1016/0885-5765(92)90026-R
  19. Exp. Mycol. v.16 Genetic and morphological comparisons of Glomerella (Colletotrichum) isolates from maize and from sorghum Vailancourt,L.J.;R.M.Hanau https://doi.org/10.1016/0147-5975(92)90030-U
  20. Gene v.23 Molecular cloning of the rDNA of Saccharomyces rosei and comparison of its transcription initiation region with that of Saccharomyces carlsbergensis Verbeet,M.P.;J.Klootwijk;H.Herrikhuizen;R.Fontijin;E.Vrengdenhil;R.J.Planta https://doi.org/10.1016/0378-1119(83)90216-0
  21. Braz. Phytopathol. v.20 Characterization of races of Colletotrichum lindermuthianum by the random amplified polymorphic DNA technique Vilarinhos,A.D.;T.T.Paula Jr.;E.G.Barros;M.A.Moreia
  22. Nucleic Acids Res. v.21 A simple method of preparing plant samples for PCR. Wang,H.;M.Qi;A.J.Cutler https://doi.org/10.1093/nar/21.17.4153
  23. PCR Protocols Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics White,T.J.;T.Burns;S.Lee;J.Taylor;M.A.Innis;D.H.Gelfand;J.J.Sninsky;T.J.White(eds.)
  24. Nucleic Acids Res. v.18 DNA polymorphisms amplified by arbitrary primers are useful as genetic markers Williams,J.G.K.;A.R.Kubelik;K.J.Livak;J.A.Rafalski;S.V.Tingey https://doi.org/10.1093/nar/18.22.6531
  25. J. Microbiol. Biotechnol. v.9 A molecular marker specific to Metarhizium anisophiae var. majus. Youn,C.S.;G.H.Sung;J.H.Sung;J.O.Lee