Enhanced Production of Phytoestrogenic Isoflavones from Hairy Root Cultures of Psoralea corylifolia L. Using Elicitation and Precursor Feeding

Amit N. Shinde,Nutan Malpathak,Devanand P. Fulzele 한국생물공학회 2009 Biotechnology and Bioprocess Engineering Vol.14 No.3

The effect of biotic elicitors (yeast extract, chitosan), signaling molecule (salicylic acid), and polyamines (putrescine and spermidine) was studied with respect to isoflavones accumulation in hairy root cultures of Psoralea corylifolia L. Untreated hairy roots (control) accumulated 1.55% dry wt of daidzein and 0.19% dry wt of genistein. In precursor feeding experiment, phenylalanine at 2 mM concentration led to 1.3 fold higher production of daidzein (1.91% dry wt) and genistein (0.27% dry wt). In biotic elicitors, chitosan (2 mg/L) was found to be the most efficient elicitor to induce daidzein (2.78% dry wt) and genistein (0.279% dry wt) levels in hairy roots. Salicylic acid at 1 mM concentration stimulated the maximum accumulation of daidzein (2.2% dry wt) and genistein (0.228% dry wt) 2 days after elicitation. In case of polyamines, putrescine (50 mM) resulted in highest accumulation of daidzein (3.01% dry wt) and genistein (0.227% dry wt) after 5 days of addition. Present results indicated the effectiveness of elicitation and precursor feeding on isoflavones accumulation in hairy roots of p.corylifolia. This is the first report of elicitation on isoflavones production by hairy roots of P.corylifolia. The effect of biotic elicitors (yeast extract, chitosan), signaling molecule (salicylic acid), and polyamines (putrescine and spermidine) was studied with respect to isoflavones accumulation in hairy root cultures of Psoralea corylifolia L. Untreated hairy roots (control) accumulated 1.55% dry wt of daidzein and 0.19% dry wt of genistein. In precursor feeding experiment, phenylalanine at 2 mM concentration led to 1.3 fold higher production of daidzein (1.91% dry wt) and genistein (0.27% dry wt). In biotic elicitors, chitosan (2 mg/L) was found to be the most efficient elicitor to induce daidzein (2.78% dry wt) and genistein (0.279% dry wt) levels in hairy roots. Salicylic acid at 1 mM concentration stimulated the maximum accumulation of daidzein (2.2% dry wt) and genistein (0.228% dry wt) 2 days after elicitation. In case of polyamines, putrescine (50 mM) resulted in highest accumulation of daidzein (3.01% dry wt) and genistein (0.227% dry wt) after 5 days of addition. Present results indicated the effectiveness of elicitation and precursor feeding on isoflavones accumulation in hairy roots of p.corylifolia. This is the first report of elicitation on isoflavones production by hairy roots of P.corylifolia.

Identification of elicitors enhances the polyphenolic compounds and pharmacological potential in hairy root cultures of <i>Aster scaber</i>

Ghimire, B.K.,Thiruvengadam, M.,Chung, I.-M. Elsevier 2019 South African journal of botany : official journal Vol.125 No.-

<P><B>Abstract</B></P> <P> <I>Aster scaber</I> Thunb is an important Korean mountainous vegetable and also a medicinal herb, which produces valuable bioactive compounds. This study showed that hairy root cultures can be established to improve the accumulation of biomass and phenolic compounds in <I>A. scaber.</I> Hairy roots were molecularly confirmed by polymerase chain reaction (PCR). We examined the influences of methyl jasmonate (MeJA) and yeast extract (YE) elicitors for the enhancement of phenolic compounds and pharmaceutical activities on hairy root cultures of <I>A. scaber</I>. Application of 100 mg/L of YE resulted in significantly higher fresh and dry root biomass (97.91 and 9.82 g/L) than MeJA and non-elicited control root cultures. MeJA (100 μM) and YE (100 mg/L) elicitation dramatically induced the accumulation of total phenolics (244.5 ± 2.5; 215.10 ± 2.0 GAE) and flavonoids (6.7 ± 0.3; 6.0 ± 0.2 QE) than non-elicited root cultures (181.65 ± 1.5 mg/g GAE and 4.65 ± 0.2 mg/g QE). Phenolic compounds including six hydroxycinnamic acids, seven flavonols, seven hydroxybenzoic acids, vanillin, homogentisic acid, and resveratrol were identified via UHPLC analysis in elicited (MeJA 100 μM and YE 100 mg/L) and control root cultures. Because of these phenolic compounds enhancement, the antioxidant, antimicrobial, antidiabetic, anti-inflammatory and anticancer activities were high in the in the MeJA- and YE-elicited hairy roots than in the control cultures. MeJA was more effective in the elicitation of phenolic compounds and pharmaceutical potential than YE. The identification of elicitors can lead the large quantity of phenolic compounds to encounter extensive range demand through marketable production without endangering of <I>A. scaber</I>. This elicitation approach could be large scale production of the commercially possible amount of these valuable bioactive compounds by hairy root cultures in <I>A. scaber.</I> </P> <P><B>Highlights</B></P> <P> <UL> <LI> Hairy root biomass was enhanced in methyl jasmonate (MeJA) or yeast extract (YE) elicitation. </LI> <LI> Phenolic compounds were significantly raised in MeJA or YE elicited hairy roots. </LI> <LI> Biological activities were exhibited higher in MeJA or YE than non-elicited hairy roots. </LI> </UL> </P>

Re-Elicitation with Methyl Jasmonate in Eschscholtzia californica Cell Suspension Cultures

BYUN, SANG YO 한국미생물 · 생명공학회 2000 Journal of microbiology and biotechnology Vol.10 No.1

Elicited cells with methyl jasmonate continued to produce benzophenanthridine alkaloids throughout medium changes in suspension cultures of Eschscholtzia californica. Large increases in alkaloid production were observed by re-elicitations with medium changes. The total alkaloid production increased during the successive elicitation steps reaching a maximum level on the 4th elicitation. The highest total alkaloid produced was 250㎎/l, which was 2-fold higher than that of the single elicitation and 4-fold higher than that of the normal culture without elicitation. The large increases in alkaloid production in successive re-elicitations with medium changes are believed to be caused by the accumulation of the signal transduction compound, jasmonate.

A Practical Guideline of Selecting Stakeholders for Requirements Elicitation – An Empirical Study

Fares Anwar,Rozilawati Razali 보안공학연구지원센터 2015 International Journal of Software Engineering and Vol.9 No.2

Requirements elicitation is widely regarded as the most challenging activity within the scope of requirements engineering. The activity concerns the understanding of problems that a system should address, which later leads to the identification of system requirements. As an abstract process, its effectiveness is greatly influenced by the credibility and suitability of the stakeholders involved. High-quality requirements could only be achieved if they are gathered from the right stakeholders. There are several methods proposed by earlier studies that could guide the selection of stakeholders for requirements elicitation. The methods however are theoretical rather than pragmatic. Therefore, there is a need of practical method that could help practitioners in identifying and selecting the appropriate stakeholders. Prior to having such a method, the contributing factors that influence the selection process should be firstly identified and understood. This study addressed this issue by collating the important factors that contribute to an effective selection of stakeholders for requirement elicitation purposes. The factors were identified through a series of empirical study conducted in two countries: seven in-depth individual interviews and a focus group session with five experienced practitioners. The results were analysed by using content analysis through Nvivo 9. The factors were then merged as a practical guideline that consists of a step-by-step procedure. This guide can be used by practitioners to execute the process of identifying and selecting the appropriate stakeholders for requirements elicitation.

A Collaborative Requirements Elicitation Model For Crowdsourcing Platforms

Chantal Mukundwa,Seok-Won Lee(이석원) 한국컴퓨터정보학회 2019 韓國컴퓨터情報學會論文誌 Vol.24 No.3

Crowdsourcing is growing its interests in designing where various designers work independently to a given design task. Recent researchers discovered that collaboration by sharing designs among designers helps to produce high-quality designs. However, design task may still be hard even with that collaboration in case the requirements are not well-defined. Most customers sometimes do not know what they really want and do not know how to clearly define the requirements. Consequently, the lack of requirements creates issues on designers, such as spending much time and effort on collecting requirements alone or from the customers. The designers even end up missing important necessities to complete their tasks. To address this issue, we proposed a collaborative requirements elicitation method that supports designers who are working on the same task. We developed CREFD (Collaborative Requirements Elicitation For Designers and Developers) tool to enable designers collaboratively provide requirements, identify dependencies, add annotations and votes to the provided requirements. We performed the hypothetical and empirical evaluations to test and compare the proposed method with one of the existing elicitation methods, the results show that the proposed method helps in collecting accepted and well-organized requirements better than individual requirements elicitation.

Optimized Production of Isoflavones in Cell Cultures of Psoralea corylifolia L. Using Elicitation and Precursor Feeding

Amit N. Shinde,Nutan Malpathak,Devanand P. Fulzele 한국생물공학회 2009 Biotechnology and Bioprocess Engineering Vol.14 No.5

The effect of biotic elicitors (yeast extract, chitosan), signaling molecule (salicylic acid), and polyamines (putrescine and spermidine) was studied with respect to isoflavones accumulation in cell suspension cultures of Psoralea corylifolia L.Untreated cell suspension (control) accumulated 1.66% dry wt of daidzein and 0.165% dry wt of genistein. In precursor feeding experiment, phenylalanine at 0.5 mM concentration led to 1.3 fold higher production of daidzein (1.99% dry wt) and genistein (0.22% dry wt). In biotic elicitors, yeast extract (100 mg/L) was found to be the most efficient elicitor to induce higher production levels of daidzein (2.21% dry wt) and genistein (0.293% dry wt) in suspension cultures. Salicylic acid (signaling molecule) at 1 mM concentration stimulated the maximum accumulation of daidzein (3.4% dry wt) and genistein (0.41% dry wt) 2 days after elicitation. In case of polyamines, spermidine (100 mM) resulted in highest accumulation of daidzein (3.2% dry wt) and genistein (0.475% dry wt) after 7 days of addition, which was 2.4 fold of that in control. This is the first report on kinetics of isoflavone production in response to elicitation in cell suspension of P.corylifolia. The effect of biotic elicitors (yeast extract, chitosan), signaling molecule (salicylic acid), and polyamines (putrescine and spermidine) was studied with respect to isoflavones accumulation in cell suspension cultures of Psoralea corylifolia L.Untreated cell suspension (control) accumulated 1.66% dry wt of daidzein and 0.165% dry wt of genistein. In precursor feeding experiment, phenylalanine at 0.5 mM concentration led to 1.3 fold higher production of daidzein (1.99% dry wt) and genistein (0.22% dry wt). In biotic elicitors, yeast extract (100 mg/L) was found to be the most efficient elicitor to induce higher production levels of daidzein (2.21% dry wt) and genistein (0.293% dry wt) in suspension cultures. Salicylic acid (signaling molecule) at 1 mM concentration stimulated the maximum accumulation of daidzein (3.4% dry wt) and genistein (0.41% dry wt) 2 days after elicitation. In case of polyamines, spermidine (100 mM) resulted in highest accumulation of daidzein (3.2% dry wt) and genistein (0.475% dry wt) after 7 days of addition, which was 2.4 fold of that in control. This is the first report on kinetics of isoflavone production in response to elicitation in cell suspension of P.corylifolia.

Development of Origanum vulgare Cell Suspension Culture to Produce Polyphenols and the Stimulation Effect of Salicylic Acid Elicitation and Phenylalanine Feeding

Yan-Ping Li,Dao-Bang Tang,Xiao-Qiang Wang,Meng Wang,Qing-Feng Zhang,Yuan Liu,Bei-Yun Shen,Jiguang Chen,Zhongping Yin 한국생물공학회 2021 Biotechnology and Bioprocess Engineering Vol.26 No.3

Three types of calli were induced from Origanum vulgare (O. vulgare) aseptic seedlings, and the friable calli with white appearance and high growth rate were further screened and used to develop cell suspension culture to produce polyphenols. Murashige and Skoog (MS) medium with 3.0 mg/L Kinetin (KT) and 0.5 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) was suitable for both O. vulgare cells growth and polyphenols accumulation. To further enhance the polyphenols accumulation, O. vulgare cells were treated by phenylalanine (Phe) feeding and salicylic acid (SA) elicitation. Compared with the individual Phe feeding and SA elicitation, SA elicitation combined with Phe feeding showed a much better promotion effect on the polyphenols synthesis in O. vulgare cells, especially rosmarinic acid (RosA) accumulation. With the combined treatment of 200 μM SA and 100 μM Phe, total polyphenols content and yield were 41.36 mg/g and 752.93 mg/L, respectively. RosA content and yield reached 31.25 mg/g and 570.37 mg/L, which were 5.44 and 5.47 times that of the control. Furthermore, the total polyphenols extracted from the cultured cells treated by SA elicitation combined with Phe feeding displayed a much higher antioxidant capacity than that of untreated cells, meanwhile its 1,1-diphenyl-2- trinitrophenyl hydrazine (DPPH) and superoxide anion radical-scavenging activity were much stronger than that of vitamin C. What’s more, our results also showed that RosA was the principal contributor to the fine antioxidant capacity of the total polyphenols extracted from the SA and Phe treated cells. Our research indicated that SA elicitation combined with Phe feeding significantly improved the polyphenols yield and antioxidant capacity of the cultured O. vulgare cells, and therefore has a promising application prospect in natural polyphenols production.

가중치 추출 기법의 비교: AHP, JA, Swing기법을 중심으로

김윤주 ( Yoon Ju Kim ),심준섭 ( Jun Seop Shim ) 중앙대학교 국가정책연구소 2007 국가정책연구 Vol.21 No.1

기존의 정책분야와 관련한 의사결정 연구는 집단을 분석단위로 하여 종합적인 가중치를 추출하는데 초점이 맞추어졌으며, 매우 한정된 가중치 추출 기법만이 사용 되었다. 본 연구의 궁극적인 목적은 의사결정 분석기법들 간의 가중치 추출방법의 차이를 고찰하는 것이다. 본 연구를 위해 다양한 가중치 추출기법들 중 폭넓게 사용되고 있으며, 효용을 인정받고 있는 AHP(Analytical Hierarchy Process)와 JA(Judgement Analysis), Swing(Swing weight method)기법을 논의 및 비교 하였다. 본 연구에서는 각 기법들의 이론적 측면과 방법적 측면, 적용의 측면에서 접근하였다. 연구의 결과로 각 기법들 간의 차이가 존재함을 확인하였다. 따라서 향후의 연구에서는 적용 목적, 대상, 또는 방법 등을 고려하여 상황에 가장 적합한 가중치 추출 기법이 선택되어야 할 것이다. 본 연구는 앞으로의 가중치 추출 기법의 비교 연구와 개인을 분석 단위로 하는 가중치 추출 기법의 적용연구를 제시하는데 의의를 갖는다. It has long been discussed how to make a rational decision with uncertainty and various information. This study aims to compare methods of decision making process focusing on the weight elicitation. To accomplish the research goal, we compared three methods: Analytical Hierarchy Process(AHP), Judgment Analysis(JA), and Swing weigh method(SW). Those methods have been widely used and showed their practical applicability. This paper starts with a comparison of theoretical facets of each methodology. It also considers both methodological standpoint and practical application of each method. From this study, these three methods show somehowdifferences which stem from theoretical and technological diversity of the weight elicitation. Therefore, this study concludes that every weight extracting methods should be accurately used with moreconsideration about subject, object, and method of study.

Effects of Elicitors on Seedling Growth, Total Polyphenol and Chlorophyll Content and Antioxidant Activity of Barley (Hordeum vulgare L.)

Timnoy Salitxay,Phonesavanh Phouthaxay,Yeoun-Gyu Pang,Yu-Chi Yeong,Prakash Babu Adhikari,박철호 한국작물학회 2016 Korean journal of crop science Vol.61 No.1

This study is focused on the evaluation of growth parameters, total polyphenol content (TPC), chlorophyll content as well as the DPPH (1,1-diphenyl-2-picryhydrazyl) free radical scavenging activity of young barley seedling (YBS) affected by elicitation. Salicylic acid (SA), methyl jasmonate (MJ), amino acid liquid fertilizer (ALF) and microbial metabolism activator (MMA) were used. Elicitation was conducted for two times and various concentrations were used in this study. The result revealed that, MJ 1 ml/L treated-YBS gave the longest seedling length of 1.33 cm, followed by the ones treated with SA 1.38 mg/L and ALF 2 ml/L, respectively. ALF 3 ml/L treatment gave the highest fresh weight of 10 seedlings, followed by MJ 5 ml/L and SA 13.8 mg/L treatment with 1.56 g, 1.55 g and 1.53 g respectively. SA 138.12 mg/L elicitor treated-YBS gave the highest Chl a, Chl b content of 8.57 μg/mg and 3.83 μg/mg, respectively while the highest carotenoid content was found in MJ ml/L treatment with 1.62 μg/mg. Among elicitor treated-YBS, SA showed better TPC. The highest TPC was found in SA 1.38 mg/L treatment with 18.82 mg/g TAE. Likewise, SA 1.38 mg/L showed the highest DPPH free radical scavenging activity among all the treatments. However, the lowest TPC was found in ALF 1ml/L treated-YBS with 9.46 mg/g TAE, which was even lower than the control (14.31 mg/g TAE). Keywords : Bayley (Hordeum vulgare L), Chlorophyll, DPPH (1,1-diphenyl-2-picryhydrazyl) free radical scavenging activity, Elicitation, Growth parameters, Total polyphenol

Effects of Elicitors on Seedling Growth, Total Polyphenol and Chlorophyll Content and Antioxidant Activity of Barley (Hordeum vulgare L.)

Salitxay, Timnoy,Phouthaxay, Phonesavanh,Pang, Yeoun-Gyu,Yeong, Yu-Chi,Adhikari, Prakash Babu,Park, Cheol-Ho The Korean Society of Crop Science 2016 한국작물학회지 Vol.61 No.1

This study is focused on the evaluation of growth parameters, total polyphenol content (TPC), chlorophyll content as well as the DPPH (1,1-diphenyl-2-picryhydrazyl) free radical scavenging activity of young barley seedling (YBS) affected by elicitation. Salicylic acid (SA), methyl jasmonate (MJ), amino acid liquid fertilizer (ALF) and microbial metabolism activator (MMA) were used. Elicitation was conducted for two times and various concentrations were used in this study. The result revealed that, MJ 1 ml/L treated-YBS gave the longest seedling length of 1.33 cm, followed by the ones treated with SA 1.38 mg/L and ALF 2 ml/L, respectively. ALF 3 ml/L treatment gave the highest fresh weight of 10 seedlings, followed by MJ 5 ml/L and SA 13.8 mg/L treatment with 1.56 g, 1.55 g and 1.53 g respectively. SA 138.12 mg/L elicitor treated-YBS gave the highest Chl a, Chl b content of $8.57{\mu}g/mg$ and $3.83{\mu}g/mg$, respectively while the highest carotenoid content was found in MJ ml/L treatment with $1.62{\mu}g/mg$. Among elicitor treated-YBS, SA showed better TPC. The highest TPC was found in SA 1.38 mg/L treatment with 18.82 mg/g TAE. Likewise, SA 1.38 mg/L showed the highest DPPH free radical scavenging activity among all the treatments. However, the lowest TPC was found in ALF 1ml/L treated-YBS with 9.46 mg/g TAE, which was even lower than the control (14.31 mg/g TAE).