• Journal of Photoscience
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• 제7권2호
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• pp.53-58
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• 2000

We performed the cloning of a chalcone synthase (CHS) gene, the key enzyme in the anthocyanin biosynthesis, from the cDNA library constructed with grape suspension cells irradiated UV-B. The PCR fragment was used to cloning the CHS gene. One CHS cDNA clone containing an open reading frame and a partial stilbene synthase (STS)cDNA, the stilbene-type phytoalexin, were isolated. The CHS cDNA clone (VCHS) showed 87% sequence homology with VvCHS (V.vinifea) and 72.3% identity with VSTSY(V.vinifea). its amino acid sequences were longer than any other CHS genes as 454 residues. Two genes were weakly expressed in white light irradiated cells, but highly induced in UV-B irradiated condition during 32 hours. Interestingly, the STS was quickly and abundantly expressed from 2 hours when supplemented with jasmonic acid (JA) and the maximum expression was observed at 4 hours and then gradually decreased. But, the additional UV-B or white light quickly degraded the STS expression than only JA treated grape suspension cells. The CHS also was rapidly induced with JA and the synergistical effect was observed at the addigional light treatment of UV-B or white light. These results are indicated that CHS and STS have different response mechanisms against the environmental stresses.

• 한국육종학회지
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• 제40권4호
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• pp.422-429
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• 2008

본 실험에서 Lilium longiflorum과 11품종에 대한 biochemical, enzymology 등의 분석방법을 적용하여, 화색생합성조절유전자 chalcone synthase(CHS), flavanone 3-hydroxylase(FHT)의 활성을 분석한 결과는 다음과 같다. 1. Flavonoid의 기본골격형성을 위한 첫 효소인 CHS가 malonyl-CoA와 p-coumaroyl-CoA 기질을 촉매하여 naringenin(NAR) 산물을 생성하였고, caffeoyl-CoA와 p-coumaroyl-CoA 기질에는 eridictiol(ERI)를 생성하지 않아, 백합의 CHS는 기질특이성을 나타냈다. 2. L. longiflorum에서 FHT 효소는 NAR를 기질로써 사용하여, dihydrokaempferol(DHK) 산물을 생성하였고, 그 외 11백합품종에서도 FHT 효소의 뚜렷한 활성이 확인되었다.

• Journal of Plant Biotechnology
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• 제46권4호
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• pp.247-254
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• 2019

Flavonoids are widely distributed secondary metabolites in plants that have a variety biological functions, as well as beneficial biological and pharmacological activities. In barley (Hordeum vulgare L.), for example, high levels of saponarin accumulate during primary leaf development. However, the effect of saponarin biosynthetic pathway genes on the accumulation of saponarin in barley is poorly understood. Accordingly, the aim of the present study was to examine the saponarin contents and expression levels of saponarin biosynthetic pathway genes [chalcone synthase (CHS), chalcone isomerase (CHI), and UDP-Glc:isovitexin 7-O-glucosyltransferase (OGT)] during early seedling developmental and under several abiotic stress conditions. Interestingly, the upregulation of HvCHS, HvCHI, and HvOGT during early development was associated with saponarin accumulation during later stages. In addition, exposure to abiotic stress conditions (e.g., light/dark transition, drought, and low or high temperature) significantly affected the expression of HvCHS and HvCHI but failed to affect either HvOGT expression or saponarin accumulation. These findings suggested that the expression of HvOGT, which encodes an enzyme that catalyzes the final step of saponarin biosynthesis, is required for saponarin accumulation. Taken together, the results of the present study provide a basis for metabolic engineering in barley plants, especially in regards to enhancing the contents of useful secondary metabolites, such as saponarin.

• Journal of Plant Biotechnology
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• 제48권1호
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• pp.12-17
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• 2021

Saponarin found in young barley sprouts has a variety of beneficial biological and pharmacological properties, including antioxidant, hypoglycemic, antimicrobial, and hepatoprotective activities. Our previous work demonstrated that saponarin content was correlated with the expression levels of three biosynthetic pathway genes [chalcone synthase (HvCHS1), chalcone isomerase (HvCHI), and UDP-Glc:isovitexin 7-O-glucosyltransferase (HvOGT1)] in young barley seedlings under various abiotic stress conditions. In this study, we investigated the saponarin content and expression levels of three saponarin biosynthetic pathway genes in hulled and hulless domestic barley cultivars. In the early developmental stages, some hulled barley cultivars (Kunalbori1 and Heukdahyang) had much higher saponarin contents than did the hulless barley cultivars. An RNA expression analysis showed that in most barley cultivars, decreased saponarin content correlated with reduced expression of HvCHS1 and HvCHI, but not HvOGT1. Heat map analysis revealed both specific increases in HvCHS1 expression in certain hulled and hulless barley cultivars, as well as general changes that occurred during the different developmental stages of each barley cultivar. In summary, our results provide a molecular genetic basis for the metabolic engineering of barley plants to enhance their saponarin content.

• Journal of Microbiology
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• 제43권6호
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• pp.475-486
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• 2005

Fungal secondary metabolites constitute a wide variety of compounds which either playa vital role in agricultural, pharmaceutical and industrial contexts, or have devastating effects on agriculture, animal and human affairs by virtue of their toxigenicity. Owing to their beneficial and deleterious characteristics, these complex compounds and the genes responsible for their synthesis have been the subjects of extensive investigation by microbiologists and pharmacologists. A majority of the fungal secondary metabolic genes are classified as type I polyketide synthases (PKS) which are often clustered with other secondary metabolism related genes. In this review we discuss on the significance of our recent discovery of chalcone synthase (CHS) genes belonging to the type III PKS superfamily in an industrially important fungus, Aspergillus oryzae. CHS genes are known to playa vital role in the biosynthesis of flavonoids in plants. A comparative genome analyses revealed the unique character of A. oryzae with four CHS-like genes (csyA, csyB, csyC and csyD) amongst other Aspergilli (Aspergillus nidulans and Aspergillus fumigatus) which contained none of the CHS-like genes. Some other fungi such as Neurospora crassa, Fusarium graminearum, Magnaporthe grisea, Podospora anserina and Phanerochaete chrysosporium also contained putative type III PKSs, with a phylogenic distinction from bacteria and plants. The enzymatically active nature of these newly discovered homologues is expected owing to the conservation in the catalytic residues across the different species of plants and fungi, and also by the fact that a majority of these genes (csyA, csyB and csyD) were expressed in A. oryzae. While this finding brings filamentous fungi closer to plants and bacteria which until recently were the only ones considered to possess the type III PKSs, the presence of putative genes encoding other principal enzymes involved in the phenylpropanoid and flavonoid biosynthesis (viz., phenylalanine ammonia-lyase, cinnamic acid hydroxylase and p-coumarate CoA ligase) in the A. oryzae genome undoubtedly prove the extent of its metabolic diversity. Since many of these genes have not been identified earlier, knowledge on their corresponding products or activities remain undeciphered. In future, it is anticipated that these enzymes may be reasonable targets for metabolic engineering in fungi to produce agriculturally and nutritionally important metabolites.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1998년도 The 12th Symposium on Plant Biotechnology Vol.12
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• pp.79-82
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• 1998

White and blue/white varieties of Torenia hybrida cv. Summerwave (SWB) were successfully obtained from the blue variety of by cosuppressing gene expression of two of the enzymes involved in anthocyanin biosynthesis; chalcone synthase (CHS) and dihydroflavonol 4-reductase (DFR). Such molecular brceding is the only precise and efficient way to widen the flower color variation of SWB due to its male and female sterility. Flower color and the degree of suppression varies depending on the transgenic lines. The dorsal and ventral petal lobes and corolla tube consistently lose anthocyanins prior to lateral petal lobes. A pink variety was also obtained by cosuppressing the flavonoid 3`5`-hydroxylase (F3`5`H) gene. Yellow torenia was obtained from T-33, an in-house cultivar that contained both carotenoids and anthocyanins, by blockage of anthocyanin biosynthesis with cosuppressing CHS or DFR genes.

• Journal of Microbiology and Biotechnology
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• 제24권11호
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• pp.1536-1541
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• 2014

Pinocembrin is a flavonoid that exhibits diverse biological properties. Although the major source of pinocembrin is propolis, it can be synthesized biologically using microorganisms such as Escherichia coli, which has been used to synthesize diverse natural compounds. Pinocembrin is synthesized from phenylalanine by the action of three enzymes; phenylalanine ammonia lyase (PAL), 4-coumarate:CoA ligase (4CL), and chalcone synthase (CHS). In order to synthesize pinocembrin from glucose in Escherichia coli, the PAL, 4CL, and CHS genes from three different plants were introduced into an E. coli strain. Next, we tested the different constructs containing 4CL and CHS. In addition, the malonyl-CoA level was increased by overexpressing acetyl-CoA carboxylase. Through these strategies, a high production yield (97 mg/l) of pinocembrin was achieved.

• Journal of Plant Biotechnology
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• 제36권4호
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• pp.415-422
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• 2009

This study aimed to develop carnation cultivars with new coloring system. We used four genes of Petunia hybrida - chalcone synthase (CHS), flavanone 3-hydroxylase (FHT), dihydroflavonol 4-reductase (DFR), and anthocyanidin synthase (ANS) - as probes, in order to isolate four genes from carnations (Dianthus Caryophyllus). The isolated genes were used as probes in order to select mutants out of collected carnations, using Northern blot analysis. The Northern blot analysis revealed 10 DFR mutants - Gumbyul, Eunbyul, Ballatyne, Crystal, Eugenia, Koreno, Imp. White Sim, West Crystal, White Alpine, and White Charotte. Six among the selected 10 cultivarswere excluded from the target cultivars, because Eugenia, Imp. White Sim, and White Alpine were proved to be double mutants of DFR and ANS, Koreno was considered to be a double mutant of DFR and CHS, and Gumbyul and Ballatyne were proved to be double mutants of DFR and CHI (Chalcone isomerase). Consequently, we selected five DFR mutants, including Virginie, which was already selected as a DFR mutant. Finally, we measured DFR activities in order to confirm the selection, and the results showed that all of the five cultivars - Eunbyul, Crystal, West Crystal, White Charotte, and Virginie - had got no DFR activity.

• 생명과학회지
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• 제29권10호
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• pp.1055-1061
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• 2019

식충식물(carnivorous plant)인 Sarracenia purpurea는 생육에 있어 매우 높은 수준의 광량을 필요로 하며 색, 과즙 등으로 벌레를 유인하여 영양분을 흡수하는 것으로 알려져 있다. S. purpurea는 약한 빛 조건에서 녹색을 띄었으며 강한 빛 조건에서는 잎을 포함한 포충낭 전체가 검붉은 색으로 변화하였다. 이와 같은 색의 변화는 질소비료를 첨가하였을 때에 관찰되지 않았다. S. purpurea를 붉게 물들이는 색소는 안토시아닌(anthocyanin)인 것으로 보고된 바가 있다. 본 연구에서는 빛의 세기와 질소 첨가에 따른 안토시아닌의 함량과 생합성 관련 유전자의 발현 특성을 분석하였다. $240{\mu}mol\;m^{-2}s^{-1}$의 강한 빛 조건에서의 안토시아닌 함량은 $40{\mu}mol\;m^{-2}s^{-1}$의 약한 빛에서 보다 6.15배 높았으며, 0.8% 요소 비료로 질소를 첨가하였을 때에는 약한 빛 조건과 큰 차이가 없었다. 안토시아닌의 초기 생합성 과정에 관여하는 CHALCONE SYNTHASE (CHS) 유전자는 강한 빛에서 발현이 증가하였고 질소 양분을 첨가하였을 때에 감소하였다. 이상의 결과는 빛과 토양의 환경 변화가 S. purpurea의 안토시아닌의 함량을 조절한다는 것을 보여주고 있다.

• 생명과학회지
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• 제18권2호
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• pp.234-240
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• 2008

매향' 딸기의 안토시아닌 생합성은 개화 후 26일째 시작되어 과실의 성숙기 동안 계속된다. 딸기로부터 안토시아닌의 생합성에 관여하는 주요 유전자를 분리하였다. 각각의 유전자에 대해, 다양한 식물체의 유사 유전자의 염기서열을 비교하여 PCR (polymerase chain reaciton) primer를 제작하였다. 숙기의 딸기에서 분리된 total RNA로부터 합성된 CDNA와 각 primer를 이용하여 RT (reverse transcriptase)-PCR을 수행하였다. 각 CDNA clone의 염기서열을 작성하여 분석한 결과, 이들은 안토시아닌 생합성에 관여하는 phenylalanine ammonia lyase (PAL), 4-cummarate CoA ligase (4CL), chalcone synthase (CHS), chalcone isomerase (CHI), flavanone-3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), anthocyanidine synthase (ANS) 그리고 UDP-glucose:flavonoid-3-O-glucosyltransferase (UFGT) 효소에 해당되었다. Northern blot 분석 결과, 이들 유전자는 과실 발달과정에서 시기적으로 조절되었다. 특히 PAL을 제외한 모든 유전자는 과실에서만 주로 발현되었다. PAL, DFR 그리고 ANS유전자는 과실 초기 발달 단계인 개화 후 10일에 검출된 후 감소하다가, 22일에 다시 증가하기 시작하여 34일에 최대가 되었다. 한편, 다른 유전자들은 초기에는 발현되지 않다가, 안토시아닌이 축적되기 시작하는 개화 후 $22{\sim}30$일에 처음으로 검출되었다. 본 연구를 통해, 딸기 과실 발달과정에서 안토시아닌 생합성 과정에 관여하는 여러 유전자가 과실 숙기에 함께 조절되는 현상을 알 수 있다. 이러한 연구 결과는 안토시아닌 합성과정을 제어하는 조절 유전자가 존재한다는 것을 시사한다. 그리고 딸기의 안토시아닌 생합성 유전자의 발현패턴을 크게 두 가지로 나눌 수 있는 것으로 보아, 딸기의 안토시아닌 생합성에는 적어도 두 가지 서로 다른 조절 기작이 관여하여 색소 발달 과정을 제어할 것으로 보인다.