Detection of "Candidatus Phytoplasma Asteris" Associated with Black Locust Witches' Broom in Korea

"Candidatus phytoplasma asteris" Group에 속하는 아까시나무 빗자루병 검출

  • Han, Sangsub (Division of Forest Sciences, College of Agriculture & Life science, Chonbuk National University)
  • 한상섭 (전북대학교 농업생명과학대학 산림과학부)
  • Received : 2007.10.04
  • Accepted : 2007.11.20
  • Published : 2007.12.31

Abstract

Typical phytoplasma witches' broom symptoms were observed in black locust (Robinia pseudoacacia L.) in Korea. The symptoms of the disease were showing abnormally small leaves, shortened intemodes and proliferation of shoots. The phytoplasmas were detected consistently in all the symptomatic samples by the amplification with phytoplasma universal primer pairs P1/P7 and R16F2n/R2, and the expected size was 1.8 kb and 1.2 kb. However, the phytoplasma DNA was not detected in healthy seedling. Based on sequence analysis of amplified region, this phytoplasma has close homologies with aster yellow, mulberry dwarf, maize bushy stunt, ash witches' broom and sumac witches' broom phytoplasmas, more than 99.2% but showed homologies with black locust witches' broom (GeneBank Accession No. AF 244363), and jujube witches' broom, 88.6% and 87.7%, respectively. This phylogetic analysis indicates that the black locust witches' broom phytoplasma founded in korea should be classified in the Candidatus phytoplasma asteris (16Sr I) group and clearly distinct from the black locust witches' broom group 16Sr III (peach X-disease phytoplasma group).

전형적인 파이토플라스마성 빗자루증상을 보이는 아까시나무가 전북지방에서 발견되었으며, 일반적으로 작은 잎들이 총생하였고, 마디와 마디 사이가 좁고, 가지가 위축되어 있는 증상을 보였다. 파이토플라스마 검출 primer인 P1/P7(약 1.8 kb)과 R16F2n/R2(약 1.2 kb)를 이용하여 증폭한 결과 빗지루증상을 보인 아까시나무에서 파이토플라스마 검출되었으며, 건전한 아카시에서는 PCR산물이 증폭되지 않았다. 염기서열 분석에 의한 유의성 검정결과 아까시나무 빗자루병 파이토플라스마는 aster yellow, 뽕나무오갈병, maize bushy stunt, 물푸레나무 빗자루병, 붉나무 빗자루병 파이토플라스마와 99.2% 이상의 유의성을 보였으며, 아까시나무 빗자루병 (GeneBank Accession No. AF 244363) 및 대추나무 빗자루병 파이토플라스마와는 각 각 88.6% 및 87.7%의 유의성을 보여 본 연구에서 분석한 아까시나무 빗자루병 파이토플라스마는 Candidatus phytoplasma asteris(16Sr I) group에 속하였으며, 16Sr III(peach X-disease phytoplasma group) group에 속하는 아카시나무 빗자루병 파이토플라스마와는 다른 계통으로 분류되었다.

Keywords

Acknowledgement

Supported by : 전북대학교

References

  1. Ciferri, R and Corte, A. 1960. Virus witches' broom of Robinia pseudoacacia, a new disease for Italy. Atti. 1st. Bot. Univ. Pavia Ser. 5(17): 122-128
  2. Davis, R.E and Dal1y, E.L. 2000. Black locust witches' broom phytoplasma is related to Virginia grapevine yellows phytoplasma. Phytopathology 90(6S): S18
  3. Deng, SJ. and Hiruki, C. 1991. Genetic relatedness between two nonculturable mycoplasmalike organisms revealed by nucleic acid hybridization and polymerase chain reaction. Phytopathology 81: 1475-1479 https://doi.org/10.1094/Phyto-81-1475
  4. Grant, T. and Hartley, C. 1938. A witches' broom on black locust and similar disease on honey locust. Plant Dis. Reptr. 22: 28-3l
  5. Gundersen, D.E., Lee, l.-M. 1996. Ultrasensitive detection of phytoplasma by nested-PCR assays using two universal primer pairs. Phytopathology. Mediterr.35: 144-151
  6. Hartley, C and Haasis, F.w. 1929. Brooming disease of black locust (Robinia pseudoacacia). Phytopathology 19: 163-166
  7. Kim, C.J. 1964. Mosaic disease of black locust (Robinia pseudoacacia L.) I. Symptom and transmission by grafting. Plant Protection 3: 1-5
  8. Kim, C.J and Hwang, I.C. 1989. Witches'-broom symptom of locust (Robinia pseudoacacia L.). Korean K. Plant Pathol. 5: 413
  9. Kollar, A., Seemuller, E., Bonet, F., Saillaird, S. and Bove, J.M. 1990. Isolation of the DNA of various plant pathogenic mycoplasmalike organisms from infected plants. Phytopathology 80: 3233-3237
  10. Lee, I-M., Gundersen, D.E., Hammond, R.W. and Davis, R.E. 1994. Use of mycoplasmalike organism (MLO) grouppspecific oligonucleotide primer for nested-PCR assays to detect mixed-MLO infections in a single host plant. Phytopathology 84: 559-566 https://doi.org/10.1094/Phyto-84-559
  11. Schneider, B., Seemuller, E., Smart, C.D. and Kirkpatrick, B.C. 1995. Phylogenetic classification of plant pathogenic mycoplasmalike organisms or phytoplasmas. In: Morecular and Diagnostic Procedures in Mycoplasmology, ed. by R. Razin and J.G. Tully, pp. 1:369-380. San Diego, CA: Academic
  12. Seliskar, C.E., Wilson, C.L and Bouene, C.E. 1973. Mycoplasmalike bodies found in phloem of black locust affected with witches' broom. Phytopathology 63: 30-34 https://doi.org/10.1094/Phyto-63-30
  13. Waters, C.L. 1898. Witches' broom on the locust. Plant World 1: 83-84
  14. Watwerworth, H.E. and Mock, R. 1999. An assessment of nested PCR to detect phytoplasma in imported dormant buds and internodal tissues of quarantined fruit tree germ plasm. Plant Dis. 83: 1047-1050 https://doi.org/10.1094/PDIS.1999.83.11.1047