한국작물학회지 (KOREAN JOURNAL OF CROP SCIENCE)

  • 제66권4호
  • /
  • Pages.365-374
  • /
  • 2021
  • /
  • 0252-9777(pISSN)
  • /
  • 2287-8432(eISSN)
한국작물학회 (The Korean Society of Crop Science)
권호 표지
DOI QR코드

DOI QR Code

콩 발아기간 중 isoflavone 생합성 유전자 발현 변이

Differential Expression of Isoflavone Biosynthetic Genes in Soybean During Germination

  • 임진수 (순천향대학교 의료생명공학과) ;
  • 김서영 (순천향대학교 의료과학과) ;
  • 김용호 (순천향대학교 의료생명공학과)
  • Lim, Jin-Su (Department of Medical Biotechnology, SoonChunHyang University) ;
  • Kim, Seo-Young (Major in Medical Science, SoonChunHyang University) ;
  • Kim, Yong-Ho (Department of Medical Biotechnology, SoonChunHyang University)
  • 투고 : 2021.10.01
  • 심사 : 2021.10.07
  • 발행 : 2021.12.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

이소플라본 함량이 높으며 장류콩인 대풍2호와 이소플라본 함량이 낮으며 나물콩인 풍산나물콩을 재료로 하여 콩 발아기간 동안 이소플라본 생합성 관련 유전자 6종(CHS6, HID, IF7GT, IF7MaT, GmIMaT1 및 GmIMaT3)의 발현량을 qRT-PCR로 분석하였다. 1. 공시재료 모두 발아기간 경과에 따라 이소플라본 함량이 높아졌으며, 총 이소플라본 함량 중 malonyl-glucosides 함량이 80% 이상을 차지하여 제일 높았으며 acetyl-glucosides는 거의 분석되지 않았다. 한편, 자엽과 배축에서 이소플라본 축적 정도가 각각 다르게 나타났으며 개별 이소플라본 함량에서도 차이가 있었다. 2. 이소플라본 생합성 관련 유전자들은 콩 발아시기 경과에 따라 발현량이 높아져 이소플라본 축적 정도와 상관이 있는 것으로 판단되나 유전자들의 발현량이 시기별로 각각 달라 개별 이소플라본 함량과 유전자간 뚜렷한 상관은 찾을 수 없었다. 3. HID 유전자는 대풍2호와 풍산나물콩 모두 발아 3일차를 제외하고는 발아시간 경과에 따라 유전자의 발현량이 높아졌으나 다른 유전자들의 상대적 발현량은 품종 간 차이가 있었으며, 또한 발아시간 경과에 따른 유전자의 발현 양상은 유전자별로 각각 달랐다. 4. 발아시간 별로 유전자들의 상대적 발현량을 비교한 결과 품종 간 차이가 있었으며 자엽과 배축에서도 상대적 발현량이 다르게 나타났다. 자엽에서는 GmIMaT1을 제외한 다른 유전자들의 발현량이 대풍2호와 풍산나물콩에서 비슷하게 나타난 반면, 배축에서는 HID 발현량이 2품종 모두 높게 나타났으나 다른 유전자들은 유전자 발현량에 일정한 경향이 없었다.

Soybean isoflavones are essential secondary metabolites synthesized through the phenylpropanoid pathway, and they play vital roles in human health. Isoflavone content is a complex quantitative trait controlled by multiple genes, and the genetic mechanisms underlying isoflavone biosynthesis remain largely unknown. Therefore, the present study analyzed the content of isoflavone and expression of six key genes involved in its biosynthesis (i.e., CHS6, HID, IF7GT, IF7MaT, GmIMaT1, and GmIMaT3) during soybean seed germination. Isoflavone content was quantified using high-performance liquid chromatography, and isoflavone biosynthetic gene expression was analyzed using quantitative real-time PCR. Two cultivars, namely 'Daepung2ho' and 'Pungsannamulkong', which are high- and low-isoflavone cultivars, respectively, were used. Isoflavone accumulation gradually increased with the progression of the germination period. As such, malonyl glucosides accounted for over 80% of the total content, whereas acetyl glucosides were present at trace amounts. Transcriptional analysis of isoflavone biosynthetic genes demonstrated expression patterns parallel to isoflavone content; however, there was no clear correlation between isoflavone content and gene expression. Moreover, most isoflavone biosynthetic genes showed different expression patterns depending on the individual gene or genotypes. Among the tested genes, HID showed consistently higher expression, except at 3 days after germination, and its expression was upregulated in 'Daepung2ho' but downregulated in 'Pungsannamulkong'. In addition, all tested genes exhibited different expression patterns between cotyledons and hypocotyls and responded differently to the germination period. These findings suggest that the expression levels of isoflavone biosynthetic genes are not consistent with the germination period and appear to be genotype-dependent.

키워드

과제정보

이 논문은 2021학년도 순천향대학교 교수 연구년제에 의하여 연구하였음.

참고문헌

  1. Ahmad, M. Z., P. Li, J. Wang, N. U. Rehman, and J. Zhao. 2017. Isoflavone Malonyltransferases GmIMaT1 and GmIMaT3 differently modify isoflavone glucosides in soybean (Glycine max) under various stresses. Front. Plant Sci. https://doi.org/10.3389/fpls.2017.00735
  2. Berger, M., C. A. Rasolohery, R. Cazalis, and J. Dayde. 2008. Isoflavone accumulation kinetics in soybean seed cotyledons and hypocotyls: distinct pathways and genetic controls. Crop Sci. 48 : 700-708. https://doi.org/10.2135/cropsci2007.08.0431
  3. Chen, H., P. Seguin, S. Jabaji, and W. Liu. 2011. Spatial distribution of isoflavones and isoflavone-related gene expression in high-and low-isoflavone soybean cultivars. Can. J. Plant Sci. 91 : 697-705. https://doi.org/10.4141/cjps10192
  4. Chennupati, P., P. Seguin, R. Chamoun, and S. Jabaji. 2012. Effects of high-temperature stress on soybean isoflavone concentration and expression of key genes involved in isoflavone synthesis. J. Agric. Food Chem. 60 : 12421-12427. https://doi.org/10.1021/jf3036319
  5. Chu, S., J. Wang, Y. Zhu, S. Liu, X. Zhou, H. Zhang, C. Wang, W. Yang, Z. Tian, H. Cheng, and D. Yu. 2017. An R2R3-type MYB transcription factor, GmMYB29, regulates isoflavone biosynthesis in soybean. PLOS Genetics.https://doi.org/10.1371/journal.pgen.1006770
  6. Devi, M. K. A, G. Kumar, and P. Giridhar. 2020. Effect of biotic and abiotic elicitors on isoflavone biosynthesis during seed development and in suspension cultures of soybean (Glycine max L.). 3 Biotech. https://doi.org/10.1007/s13205-020-2065-1
  7. Gupta, O. P., A. Dahuja, A. Sachdev, P. K. Jain, S. Kumari, T. Vinutha, and S. Praveen. 2018. Cytosine methylation of isoflavone synthase gene in the genic region positively regulates its expression and isoflavone biosynthesis in soybean seeds. DNA and Cell Biology. https://doi.org/10.1089/dna.2018.4584
  8. Gupta, O. P., D. Nigam, A. Dahuja, S. Kumar, T. Vinutha1, A. Sachdev, and S. Praveen. 2017. Regulation of isoflavone biosynthesis by miRNAs in two contrasting soybean genotypes at different seed developmental stages. Front. Plant Sci. https://doi.org/10.3389/fpls.2017.00567
  9. Gutierrez-Gonzalez, J. J., S. K. Guttikonda, L. Tran, D. L. Aldrich, R. Zhong, O. Yu, H.T. Nguyen, and D. A. Sleper. 2010a. Differential expression of isoflavone biosynthetic genes in goybean during water deficits. Plant and Cell Physiology 51(6) : 936-948. https://doi.org/10.1093/pcp/pcq065
  10. Gutierrez-Gonzalez, J. J., X. Wu, J. D. Gillman, J. D. Lee, R. Zhong, O. Yu, G. Shannon, M. Ellersieck, H. T. Nguyen, and D. A. Sleper. 2010b. Research article intricate environment-modulated genetic networks control isoflavone accumulation in soybean seeds. BMC Plant Biology. http://www.biomedcentral.com/1471-2229/10/105
  11. Hu, R., C. Fan, H. Li, Q. Zhang, and Y. Fu. 2009. Evaluation of putative reference genes for gene expression normalization in soybean by quantitative real-time RT-PCR. BMC Molecular Biology. https://doi.org/10.1186/471-2199-10-93
  12. Jia, Y., Y. Ma, P. Zou, G. Cheng, J. Zhou, and S. Cai. 2019. Effects of different oligochitosans on isoflavone metabolites, antioxidant activity, and isoflavone biosynthetic genes in soybean (Glycine max) seeds during germination. J. Agric. Food Chem. 67 : 4652-4661. https://doi.org/10.1021/acs.jafc.8b07300
  13. Kim, D. G., J. I. Lyu, Y. J. Lim, J. M. Kim, N. N. Hung, S. H. Eom, S. H. Kim, J. B. Kim, C. H. Bae, and S. J. Kwon. 2021. Differential gene expression associated with altered isoflavone and fatty acid contents in soybean mutant diversity pool. Plants. https://doi.org/10.3390/plants10061037
  14. Lee, J. W., Y. J. Yi, J. H. Lee, M. S. Jo, D. J. Choi, M. H. Ma, H. S. Kim, D. O. Kim, H. T. Yun, and Y. H. Kim. 2018. Quantification of isoflavone malonylglucosides in soybean seed during germination. Korean J. Crop Sci. 63(3) : 239-247. https://doi.org/10.7740/KJCS.2018.63.3.239
  15. Lim, Y. J., H. Y. Jeong, C. S. Gil, S. J. Kwon, J. K. Na, C. H. Lee, and S. H. Eom. 2020. Isoflavone accumulation and the metabolic gene expression in response to persistent UV-B irradiation in soybean sprouts. Food Chemistry. https://doi.org/10.1016/j.foodchem.2019.125376
  16. Livak, K. J. and T. D. Schmittgen. 2001. Analysis of relative gene expression data using real-time quantitative PCR and 2-△△Ct method. Methods 25 : 402-408. https://doi.org/10.1006/meth.2001.1262
  17. Miladinovic, J., V. Dordevic, S. Balesevic-Tubic, K. Petrovic, M. Ceran, J. Cvejic, M. Bursac, and D. Miladinovic. 2019. Increase of isoflavones in the aglycone form in soybeans by targeted crossings of cultivated breeding material. Scientific Reports. https://doi.org/10.1038/s41598-019-46817-1.
  18. Pei, R., J. Zhang, L. Tian, S. Zhang, F. Han, S. Yan, L. Wang, B. Li, and J. Sun. 2018. Identification of novel QTL associated with soybean isoflavone content. The Crop Journal 6(3) : 244-252. https://doi.org/10.1016/j.cj.2017.10.004
  19. Wan, Q., S. Chen, Z. Shan, Z. Yang, L. Chen, C. Zhang, S. Yuan, Q. Hap, X. Zhang, D. Qiu, H. Chen, and X. Zhou. 2017. Stability evaluation of reference genes for gene expression analysis by RT-qPCR in soybean under different conditions. PLOS ONE. https://doi.org/10.1371/journal.pone.0189405
  20. Wu, D., D. Li, X. Zhao, Y. Zhan, W. Teng, L. Qiu, H. Zheng, W. Li, and Y. Han. 2020. Identification of a candidate gene associated with isoflavone content in soybean seeds using genome-wide association and linkage mapping. The Plant Journal 104 : 950-963. https://doi.org/10.1111/tpj.14972
  21. Yuk, H. J., Y. H. Song, M. J. Curtis-Long, D. W. Kim, S. G. Woo, Y. B. Lee, Z. Uddin, C. Y. Kim, and K. H. Park. 2016. Ethylene induced a high accumulation of dietary isoflavones and expression of isoflavonoid biosynthetic genes in soybean (Glycine max) leaves. J. Agric. Food Chem. 64 : 7315-7324. https://doi.org/10.1021/acs.jafc.6b02543