• 제목/요약/키워드: Secondary metabolite accumulation

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Growth, secondary metabolite production and antioxidant enzyme response of Morinda citrifolia adventitious root as affected by auxin and cytokinin

  • Baque, Md. Abdullahil;Hahn, Eun-Joo;Paek, Kee-Yoeup
    • Plant Biotechnology Reports
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    • 제4권2호
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    • pp.109-116
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    • 2010
  • Morinda citrifolia adventitious roots were cultured in shake flasks using Murashige and Skoog medium with different types and concentrations of auxin and cytokinin. Root (fresh weight and dry weight) accumulation was enhanced at 5 $mg\;l^{-1}$ indole butyric acid (IBA) and at 7 and 9 $mg\;l^{-1}$ naphthalene acetic acid (NAA). On the other hand, 9 $mg\;l^{-1}$ NAA decreased the anthraquinone, phenolic and flavonoid contents more severely than 9 $mg\;l^{-1}$ IBA. When adventitious roots were treated with kinetin (0.1, 0.3 and 0.5 $mg\;l^{-1}$) and thidiazuron (TDZ; 0.1, 0.3 and 0.5 $mg\;l^{-1}$) in combination with 5 $mg\;l^{-1}$ IBA, fresh weight and dry weight decreased but secondary metabolite content increased. The secondary metabolite content (including 1,1-diphenyl-2-picrylhydrazyl activity) increased more in TDZ-treated than in kinetin-treated roots. Antioxidative enzymes such as catalase (CAT) and guaiacol peroxidase (G-POD), which play important roles in plant defense, also increased. A strong decrease in ascorbate peroxidase activity resulted in a high accumulation of hydrogen peroxide. This indicates that adventitious roots can grow under stress conditions with induced CAT and G-POD activities and higher accumulations of secondary metabolites. These results suggest that 5 $mg\;l^{-1}$ IBA supplementation is useful for growth and secondary metabolite production in adventitious roots of M. citrifolia.

Influence of Hormones and Selection of Stable Cell Lines of Plumbago rosea for Accumulation of Plumbagin

  • Komaraiah P.;Jogeswar G.;Naga Amrutha R.;Sri Laxmi P.;Lavanya B.;Rama Krishna S.V.;Kavi Kishor P.B.
    • Journal of Plant Biotechnology
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    • 제5권3호
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    • pp.181-185
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    • 2003
  • Callus and suspension cultures derived from leaf explants of Plumbago rosea were established on Murashige and Skoog's medium containing 1 mg/L IAA, 0.5 mg/L NAA and 0.3 mg/L BAP. Callus cultures were tested for their growth and accumulation of plumbagin, a naphthoquinone and was identified by $^1H$ NMR and electron ionization mass spectroscopy. While auxins (not 2,4-D) influenced growth and plumbagin accumulation, cytokinins did not influence them much. Increasing concentrations of IAA in presence of NAA and BAP increased plumbagin in suspensions only up to 1 mg/L. Growth of callus was optimum (8.3 g DCW/I) at a hormonal combination of 1.5 mg/L IAA, 0.5 mg/L NAA and 0.3 mg/L BAP, but high plumbagin accumulation (4.9 mg/g DCW) was recorded at 1.0 mg/L IAA plus 0.3 mg/L BAP. Since instability in growth and secondary metabolite accumulation was noticed, several cell lines/clumps of callus were screened for plumbagin accumulation by visual and analytical methods. Biomass and accumulation of plumbagin showed a negative correlation in several cell lines. But one cell line showed stability both in terms of biomass and plumbagin accumulation over a period of 6 months.

C-4 Euphorbia maculata 엽육조직 내 탄닌물질의 축적 양상 (Patterns of Tannin Accumulation in Leaves of C-4 Euphorbia maculata)

  • 김인선
    • Applied Microscopy
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    • 제33권3호
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    • pp.233-241
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    • 2003
  • 유액을 형성하는 대극과 C-4 식물 Euphorbia maculata의 엽육조직은 유관속초와 엽육세포 두 유형의 유세포로 분화되는데, 본 연구에서는 이들 세포 내에 축적되는 2차 대사물질인 탄닌의 발달양상을 형태 구조적으로 연구하였다. 탄닌의 축적은 엽육조직 발달초기 상태에서부터 시작되어 유관속초세포에서 뚜렷하게 진행되었고, 분화를 마친 성숙한 잎의 유관속초세포와 인접한 엽육세포에는 현저하게 다른 양상으로 축적되었다. 일부 미분화 유관속초세포는 발달초기에 일어난 액포융합현상으로 세포용적의 대부분을 차지하는 거대한 탄닌액포를 형성하여 세포질에는 영향을 미치지 않으나, 이후 엽육조직이 발달하면 유관속초세포 내 액포에는 소과립상에서부터 소구체, 구체 및 비교적 커다란 탄닌액포에 이르기까지 다양한 형태로 집적되었다. 특히, 탄닌이 세포소기관들 사이로 침투하여 소기관들의 경계를 따라 분포되거나, 침착이 더욱 진행되어 세포질 내 거의 모든 세포소기관에 영향을 미친 경우에는 세포질의 전자밀도를 매우 높게 나타났다. 반면, 엽육세포층 내에서는 탄닌의 축적현상이 드물게 일어나 대부분의 세포들은 거의 영향을 받지 않는 것으로 나타났다. 그러나, 유관속초세포 내 탄닌의 축적현상이 할발하게 일어난 경우에는 이들과 바로 인접한 엽육세포의 세포질에도 탄닌의 축적이 야기되어 일부 세포소기관의 전자밀도는 높게 나타났다. Euphorbia maculata는 식물체 전체에 망상으로 분포하는 유관에 다량의 알카로이드성 유액이 분비되는데, 액포 및 세포질에 축적되는 이러한 탄닌은 이들 알카로이드성 물질과 밀접한 관계를 유지하며 병충해 및 초식동물로부터의 보호 등 식물체 방어기작에 중요한 기능을 수행할 것으로 추정된다.

Instability of Anthocyanin Accumulation in Vitis vinifera L. var. Gamay Freaux Suspension Cultures

  • Qu Junge;Zhang Wei;Yu Xingju;Jin Meifang
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권2호
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    • pp.155-161
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    • 2005
  • The inherent instability of metabolite production in plant cell culture-based bioprocessing is a major problem hindering its commercialization. To understand the extent and causes of this instability, this study was aimed at understanding the variability of anthocyanin accumulation during long-term subcultures, as well as within subculture batches, in Vitis vinifera cell cultures. Therefore, four cell line suspensions of Vitis vinifera L. var. Gamay Freaux, A, B, C and D, originated from the same callus by cell-aggregate cloning, were established with starting anthocyanin contents of $2.73\;\pm\;0.15,\;1.45\;\pm\;0.04,\;0.7\;\pm\;0.024\;and\;0.27\;\pm\;0.04$CV (Color Value)/g-FCW (fresh cell weight), respectively. During weekly subculturing of 33 batches over 8 months, the anthocyanin biosynthetic capacity was gradually lost at various rates, for all four cell lines, regardless of the significant difference in the starting anthocyanin content. Contrary to this general trend, a significant fluctuation in the anthocyanin content was observed, but with an irregular cyclic pattern. The variabilities in the anthocyanin content between the subcultures for the 33 batches, as represented by the variation coefficient (VC), were 58, 57, 54, and $84\%$ for V. vinifera cell lines A, B, C and D, respectively. Within one subculture, the VCs from 12 replicate flasks for each of 12 independent subcultures were averaged, and found to be $9.7\%$, ranging from 4 to $17\%$. High- and low-producing cell lines, VV05 and VV06, with 1.8-fold differences in their basal anthocyanin contents, exhibited different inducibilities to L-phenylalanine feeding, methyl jasmonate and light irradiation. The low-producing cell line showed greater potential in enhanced the anthocyanin production.

Differential Induction of Protein Expression and Benzophenanthridine Alkaloid Accumulation in Eschscholtzia californica Suspension Cultures by Methyl Jasmonate and Yeast Extract

  • Cho, Hwa-Young;Rhee, Hong-Soon;H. Yoon, Sung-Yong;Park, Jong-Moon
    • Journal of Microbiology and Biotechnology
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    • 제18권2호
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    • pp.255-262
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    • 2008
  • Methyl jasmonate (MJ) and yeast extract (YE) induce protein expression and benzophenanthridine alkaloid accumulation in Eschscholtzia californica suspension cell cultures. One hundred ${\mu}M$ MJ primarily induced dihydrosanguinarine $(509.0{\pm}7.4mg/l)$ ; 0.2g/l YE induced sanguinarine $(146.8{\pm}3.8mg/l)$ and an unknown compound. These results occur because dihydrobenzophenanthridine oxidase (DHBO) is induced by YE and not by MJ. YE and chitin (CHI) had similar effects on sanguinarine production and DHBO expression. Differential induction of secondary metabolites was shown in E. californica suspension cultures and the expression of proteins confirmed the metabolite results. Furthermore, treatment by various oligosaccharides helped us to understand the elicitation effect of YE in signal transduction pathways.

Evaluation on Anticancer Effect Against HL-60 Cells and Toxicity in vitro and in vivo of the Phenethyl Acetate Isolated from a Marine Bacterium Streptomyces griseus

  • Lee, Ji-Hyeok;Zhang, Chao;Ko, Ju-Young;Lee, Jung-Suck;Jeon, You-Jin
    • Fisheries and Aquatic Sciences
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    • 제18권1호
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    • pp.35-44
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    • 2015
  • We previously identified Streptomyces griseus as an anti-cancer agent (Kim et al., 2014). In this study, we isolated compounds from S. griseus and evaluated their anticancer effect and toxicity in vitro and in vivo. Preparative centrifugal partition chromatography (CPC) was used to obtain three compounds, cyclo($_{\small{L}}$-[4-hydroxyprolinyl]-$_{\small{L}}$-leucine], cyclo($_{\small{L}}$-Phe-trans-4-hydroxy-$_{\small{L}}$-Pro) and phenethyl acetate (PA). We chose PA, which had the highest anticancer activity, as a target compound for further experiments. PA induced the formation of apoptotic bodies, DNA fragmentation, DNA accumulation in $G_0/G_1$ phase, and reactive oxygen species (ROS) formation. Furthermore, PA treatment increased Bax/Bcl-xL expression, activated caspase-3, and cleaved poly-ADP-ribose polymerase (PARP) in HL-60 cells. Simultaneous evaluation in vitro and in vivo, revealed that PA exhibited no toxicity in Vero cells and zebrafish embryos. We revealed, for the first time, that PA generates ROS, and that this ROS accumulation induced the Bcl signaling pathway.

Optimization of shoot cultures and bioactive compound accumulation in Rosa rugosa during acclimatization

  • Jang, Hae-Rim;Park, Byung-Jun;Park, Seung-A;Pee, Ok-Ja;Park, So-Young;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • 제43권1호
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    • pp.104-109
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    • 2016
  • Rosa rugosa is a medicinal, ornamental, and edible plant native to Eastern Asian countries, including Korea, Japan, and China. The aim of this study was to establish a system for biomass production and secondary metabolite accumulation during in vitro culture and acclimatization of Rosa rugosa. The highest rate of multiple shoot proliferation was achieved with $8.8{\mu}M$ benzyladenine (BA) (83.3%). However, the number of shoots (14.4 per explant) at $4.4{\mu}M$ BA was higher than that at $8.8{\mu}M$ BA. Compared to BA, a combination of thidiazuron (TDZ) and indole butyric acid (IBA) exhibited significantly lower shoot induction, with only 50.0~79.2% and 4.2~16.7% relative shoot formation, respectively. During acclimatization, shoots were sampled every week and their total phenolic contents were analyzed. Among various growth factors, fresh weight showed the most dramatic increase from the 3rd week (88.0 mg/plant) to 4th week (132.7 mg/plant). Total phenolics and flavonoids contents were the highest at $1^{st}$ week of acclimatization. Depending on developmental stages, total phenolics and flavonoids contents were higher in 1-yr-old shoots grown ex vitro than in those of older field-grown or in vitro-grown plants. Amongst different ages of field grown plants, 6-year-old plants, the oldest in this study, showed the lowest content in total phenolics.

Light/Dark Responsiveness of Kinetin-Inducible Secondary Metabolites and Stress Proteins in Rice Leaf

  • Cho, Kyoung-Won;Kim, Dea-Wook;Jung, Young-Ho;Shibato, Junko;Tamogami, Shigeru;Yonekura, Masami;Jwa, Nam-Soo;Kubo, Akihiro;Agrawal, Ganesh Kumar;Rakwal, Randeep
    • Journal of Crop Science and Biotechnology
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    • 제10권2호
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    • pp.112-116
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    • 2007
  • Kinetin(KN) is an inducer of rice(Oryza sativa L.) defense/stress responses, as evidenced by the induction of inducible secondary metabolite and defense/stress protein markers in leaf. We show a novel light-dependent effect of KN-triggered defense stress responses in rice leaf. Leaf segments treated with KN(100 ${\mu}M$) show hypersensitive-like necrotic lesion formation only under continuous light illumination. Potent accumulation of two phytoalexins, sakuranetin and momilactone A(MoA) by KN that peaks at 48 h after treatment under continuous light is completely suppressed by incubation under continuous dark. Using two-dimensional gel electrophoresis we identified KN-induced changes in ribulose-1, 5-bisphosphate carboxylase/oxygenase, energy- and pathogenesis-related proteins(OsPR class 5 and 10 members) by N-terminal amino acid sequencing and mass spectrometry. These changes were light-inducible and could not be observed in the dark(and control). Present results provide a new dimension(light modulation/regulation) to our finding that KN has a potential role in the rice plant self-defense mechanism.

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당근 Phytoalexin 6-Methoxymellein 생합성에 미치는 Polychlorinated Biphenyls 및 Phenol의 영향 (Effects of Polychlorinated Biphenyls and Phenols on the Biosynthesis of 6-Methoxymellein, a Phytoalexin)

  • 임도형;임다솜;금영수
    • 한국환경농학회지
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    • 제35권3호
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    • pp.216-222
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    • 2016
  • Polychlorinated biphenyl은 잔류성 환경 오염물질로서, 다양한 생리 독성 때문에 인축에 대한 세부적 연구가 수행된 바 있다. 한편 포유동물을 대상으로 진행된 연구에서 hydroxy PCBs, PCB quinones, 및 methylsulfonyl PCBs 등의 대사물이 발견되었으며, 이들은 모화합물과는 상이한 독성을 나타낸다고 보고되었다. 그러나 이와 같은 동물에 대한 연구와 비교하여, 식물 및 토양 미생물에 대한 영향, 독성 등에 대한 연구는 매우 제한적이다. 일반적으로 식물의 경우, 농약 등과 같은 외래 물질 또는 자외선, 병원균 등의 외래 스트레스원의 작용을 받게되면, 특이 생물활성을 지닌 파이토알렉신 등의 이차 대사산물을 생합성하는 것이 알려져 있다. 본 연구에서는 수 종의 PCB 및 hydroxy PCB, phenol류 등을 화학적으로 합성하고, 이들의 처리에 따른 당근 중, 파이토알렉신인 6-메톡시멜레인 생합성에 미치는 영향을 검토하였다. 증류수를 처리한 대조구의 경우, 파이토알렉신의 농도는 처리 후 2일에 최대치까지 증가하였으며, 이후 지속적으로 감소하였다. 한편 2-hydroxy/4-hydroxybiphenyl의 경우, 처리 후 3일까지 증대한 반면, 비교적 지용성이 높은 PCB-126 및 3,5-dichloro-2-hydroxybiphenyl 처리구에서는 처리 1일 후, 최대치를 나타내어 화합물의 구조 또는 지용성 등에 따라 다른 유도효과를 나타내었다. 이상의 결과에 따라, 환경 오염물질의 성상에 따라, 토양 중, 재배 농작물의 생리 활성 2차 대사물질의 생합성이 다양하게 변화됨을 알 수 있었다.