[논문]콩 발아기간 중 isoflavone 생합성 유전자 발현 변이

임진수; 김서영; 김용호

doi:10.7740/kjcs.2021.66.4.365

콩 발아기간 중 isoflavone 생합성 유전자 발현 변이
Differential Expression of Isoflavone Biosynthetic Genes in Soybean During Germination 원문보기

Korean journal of crop science = 韓國作物學會誌, v.66 no.4, 2021년, pp.365 - 374

임진수 (순천향대학교 의료생명공학과) , 김서영 (순천향대학교 의료과학과) , 김용호 (순천향대학교 의료생명공학과)

초록
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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.

Abstract ▼ AI-Helper

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.

주제어

표/그림 (7)

표 Table 1. Primers used for quantitative real-time PCR.
표 Table 2. Mean content (μg g^-1, DW) of individual isoflavones during soybean germination.
그림 Fig. 1. Schematic representation of the phenylpropanoid pathway for isoflavone biosynthesis (adapted from Yuk et al., 2016). Abbreviations: PAL (phenylalanine ammonia lyase), C4H (cinnamic acid 4-hydroxylase), 4CL (4-coumarate: CoA ligase), CHS (chalcone synthase), CHR (chalcone reductase), CHI (chalcone isomerase), IFS (isoflavone synthase), HID (2-hydroxyisoflavone dehydratase), IF7GT (isoflavone 7-O-uridine diphosphate glycosyltransferase), IF7MaT (isoflavone 7-O-glucoside-6-O-malonyltransferase).
그림 Fig. 2. Relative expression levels of isoflavone biosynthetic genes during soybean germination. Data are presented as the means of measurements from three replicates. Bars represent SE. Gene expression levels were normalized to the expression of the internal control SKIP16.
그림 Fig. 2. Relative expression levels of isoflavone biosynthetic genes during soybean germination. Data are presented as the means of measurements from three replicates. Bars represent SE. Gene expression levels were normalized to the expression of the internal control SKIP16 (Continued).
표 Table 3. Relative expression levels of isoflavone biosynthetic genes compared with the CHS6 gene during soybean germination.
그림 Fig. 3. Relative expression levels of isoflavone biosynthetic genes in the cotyledon and hypocotyl at 5 days after germination in soybeans cultivars 'Daepung2ho' (A) and 'Pungsannamulkong' (B). Relative transcript levels were analyzed using qRT-PCR. Gene expression levels were normalized to the expression of the internal control SKIP16 and calculated based on the CHS6 value.

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