Korean Journal of Medicinal Crop Science (한국약용작물학회지)

The Korean Society of Medicinal Crop Science (한국약용작물학회)
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Development of SCAR Marker for Discriminating between Violet Flowered Lines and White Flowered Lines in Chinese Bellflower (Platycodon grandiflorum A.)

청도라지와 백도라지의 구분을 위한 SCAR 마커 개발

  • Published : 2007.02.28
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Abstract

To develop a convenient method for discriminating between violet flowered lines and white flowered lines in Chinese bellflower, RAPD analysis was carried out and SCAR markers were generated. Eighteen specific RAPD bands were obtained from 6 OPERON primer sets. Two out of eighteen RAPD bands were cloned into pGEM-T-Easy vectors and then subjected to the nucleotide sequence analysis. PgR1 and PgR2 DNA fragment, each specific for violet and white flowered lines, consist of 887 bp and 863 bp sequences, respectively. Two SCAR markers were developed from RAPD clones: SPgR1 (355 bp) from PgR1 and SPgR2 (493 bp) from PgR2. One (SPgR2) of these two markers was useful to differentiate between violet flowered lines and white flowered lines in Chinese bellflower.

청도라지와 백도라지를 DNA 수준에서 구분하기 위하여, RAPD 분석을 바탕으로 SCAR primer (SPgR1, SPgR2)를 제작하여 이를 적용한 실험 결과는 다음과 같다. 총 24개의 임의 프라이머를 적용하여 6개의 청도라지와 백도라지 특이적인 프라이머를 선발하였고, 이들 프라이머를 이용하여 PCR을 수행한 결과 총 18개의 다형성을 나타내는 DNA fragment를 확보하였다. 확보된 DNA fragment 중 2개에 대한 염기서열 분석을 수행한 결과, 청도라지 특이적인 DNA 밴드는 887 bp의 염기서열로 구성되어 있었고 백도라지는 863 bp의 염기서열로 구성되어 있었다. 이들 염기서열을 바탕으로 두개의 SCAR primer를 제작하였고, 이중 백도라지 특이적인 SPgR2를 이용하여 PCR 한 결과, 청도라지에서는 약 500 bp 크기에서 두 개의 특이적인 밴드가 형성되었으며, 백도라지의 경우 한 개의 특이적인 밴드만 관찰되어 청도라지와 백도라지를 구분할 수 있었다. 결론적으로 본 실험을 통하여 개발된 SCAR 마커는 청도라지와 백도라지를 구분하기에 유용함을 확인하였다.

Keywords

References

  1. Adam-Blondon AF, Sevignac M, Bannerot H, Dron M (1994) SCAR, RAPD and RFLP markers linked to a dominant gene (Are) conferring resistance to anthracnose in common bean. Theor. Appl. Genet. 88:865-870 https://doi.org/10.1007/BF01253998
  2. Altschul FS, Thomas L, Madden AA, Schaffer, Jinghui Z, Zheng Z, Web M, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25:3389-3402 https://doi.org/10.1093/nar/25.17.3389
  3. Bang KH, Park CG, Jin DC, Kim HS, Park HW, Park CH, Seong NS (2003) Discrimination of species specific DNA markers using RAPD and AFLP analysis between Atractylodes japonica Koidz. and Atractylodes macrocephala Koidz. Korean J. Medicinal Crop Sci. 11:268-273
  4. Bang KH, Sung JS, Park CH, Jin DC, Park CG, Yu HS, Park HW, Seong NS (2004) Discrimination of Atractylodes rhizome white using anatomical characteristics and SCAR markers. Korean J. Medicinal Crop Sci. 12:53-59
  5. Das M, Bhattacharya S, Pal A (2005) Generation and characterization of SCARs by cloning and sequencing of RAPD products: A strategy for species-specific development in Bamboo. Ann. Bot. 95:835-841 https://doi.org/10.1093/aob/mci088
  6. Hernandez P, Martin A, Doraco G (1999) Development of SCARs by direct sequencing of RAPD products: A practical tool for the introgression and marker assisted selection of wheat. Mol. Breeding 5:245-253 https://doi.org/10.1023/A:1009637928471
  7. Hu J, van Eysden J, Quiros CF (1995) Generation of DNAbased markers in specific genome regions by two-primer RAPD reactions. PCR Methods Appl. 4:346-351 https://doi.org/10.1101/gr.4.6.346
  8. In DS, Kim YC, Bang KH, Chung JW, Kim OT, Hyun DY, Cha SW, Kim TS, Seong NS (2005) Genetic relationships of Panax species by RAPD and ISSR analyses. Korean J. Medicinal Crop Sci. 13:249-253
  9. Lee WB, Choi SY, Kim YS (1993) An application of random amplified polymorphic DNA (RAPD) to systematics of some species Lilium in Korea. Korean J. Plant Tax. 23:35-42 https://doi.org/10.11110/kjpt.1993.23.2.035
  10. Mabberley DJ (1987) The Plant Book. p. 461
  11. Murray YHG, Thomson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res. 8:4321-4326 https://doi.org/10.1093/nar/8.19.4321
  12. Naqvi NI, Chattoo BB (1996) Development of a sequence characterized amplified region (SCAR) based indirect selection method for a dominant blast resistance gene in rice. Genome 39:26-30 https://doi.org/10.1139/g96-004
  13. Paran I, Michelmore RW (1993) Development of reliable PCR based markers linked to downy mildew resistance genes in lettuce. Theor. Appl. Genet. 85:985-993
  14. 이창복 (1980) 대한식물도감, 향문사., p. 725
  15. 중국약재공사 (1995) 중국상용중약재, 과학출판사., p. 420-428