고려인삼(Panax Ginseng C.A. Meyer) 모상근으로부터 Ginsenosides 생산에 미치는 Jasmonic acid와 Methyl jasmonate의 영향

박효진,오승용,최경화,맹성주,윤의수,양덕춘 고려인삼학회 2000 Journal of Ginseng Research Vol.24 No.2

To elucidate the effects of jasmonic acid and methyl jasmonate on the production of ginsenosides and growth, ginseng hairy root KGHR-8 clone was cultured on the 1/2 MS medium without growth regulators, which was supplemented with of various concentrations jasmonic acid and methyl jasmonate and culture period. The highest growth rate was obtained when 1$\mu\textrm{m}$ jasmonic acid and methyl jasmorlate were treated. However, the growth was inhibited at more than 30$\mu\textrm{m}$ of concentration. Treatment with high concen Dation of jasmonic acid (10$\mu\textrm{m}$) and methyl jasmonate (50$\mu\textrm{m}$) increased the contents and productivity of ginsenosides reversion of the growth inhibition. The highest contents and productivity of ginsenosides were appeared at 4 weeks after onset of the treatment of jasmonic acid and at 3 weeks in the case of methyl jasmonate. 인삼모상근의 생장과 ginsenosides의 함량을 높이기 위하여 생장조절제가 첨가되지 않은 1/2 MS 배지에 jasmonic acid와 methyl jasmonate의 농도와 처리시기를 달리하여 인삼모상근 KGHR울 세포주를 배양하였다. 인삼모상근 생장은jasmonic acid와 methyl jasmonate 모두 1$\mu\textrm{m}$ 농도에서 가장 양호하였으며 30$\mu\textrm{m}$ 이상 농도가 증가할수록 모상근생장이 감소하였다. 그러나 생장이 낮았던 jasmonic acid 10$\mu\textrm{m}$ 처리구와 methyl jasmonate 50$\mu\textrm{m}$에서 ginsenosides 함량과 생산성이 더 높았다. 배양시기별로 jasmonic acid와 methyl jasmonate의 처리 효과는 jasmonic acid는 배양 후4주, methyl jasmonate는 3주에 처리하는 것이 ginsenosides의 함량과 생산성을 높이는데 효과적이었다.

대사유도물질 처리에 의한 발아녹두의 아이소플라본 생합성 양상

이지현,정일민,박세준,김욱한,김소연,김진애,정우식 韓國作物學會 2004 Korean journal of crop science Vol.49 No.6

발아 녹두에 세 가지 스트레스 관련 화합물 salicylic acid, methyl jasmonic acid, acetyl salicylic acid를 처리하여 isoflavone의 생합성양상을 관찰한 결과를 요약하면 다음과 같다. 1. 숙주나물의 자엽에서는 isoflavone총량이 건조중 1g당 832.5ug 인 무처리구와 비교하여 10mM salicylic acid를 처리한 경우 169~% , 12mM acetyl salicylic acid로 처리한 경우 165~% 의 isoflavone 총량이 증가한 반면 0.5~% methyl jasmonic acid를 처리한 경우는 오히려 무처리구보다 47~% 수준으로 감소하였다. 2. 숙주나물의 자엽하부(hypocotyl and root)의 isoflavone 생성량에서는 1g당 284.8ug 이 생성된 무처리구와 비교하여 세 가지 처리 모두에서 유의성이 있는 차이를 보였다. 10mM salicylic acid 처리구의 경우 419~% , 12mM acetyl salicylic acid 처리구의 경우 401~% 의 isoflavone 총량의 증가를 보였고, 0.5~% methyl jasmonic acid처리구의 경우에는 121~% 증가하였다. 3.숙주나물의 자엽부위와 자엽하부에서 검출된 isoflavone의 합을 각 처리별 isoflavone생산총량으로 하여 무처리구의 건조중 1g당 1117.3ug 을 기준으로 비교하여보면 건조중 1g당 10mM salicylic acid 처리구에서는 2601.02ug 으로 233~% 증가하였고, 12mM acetyl salicylic acid 2514.4ug 으로 225~% 증가한 반면, 0.5~% methyl jasmonic acid 처리구에서는 738.8ug 으로 66~% 수준으로 감소하였다. 4. 숙주나물 자엽부위의 경우 무처리구와 비교하여 증가를 보였던 10mM salicylic acid처리구와 12mM acetyl salicylic acid 처리구에서는 malonyldaidzine과 malonylglycitin이 증가가 두드러지게 나타났다. 5. 숙주나물 자엽하부의 경우 무처리구와 비교하여 증가를 보였던 10mM salicylic acid 처리구와 12mM acetyl salicylic acid 처리구에서는 malonylglycitin의 증가가 두드러지게 나타났다 We have studied physiological responses of mung bean sprout to the treatment of elicitors. Chemicals such as salicylic acid and methyl jasmonic acid are not only the intermediates found in plant defense system but also could affect plant secondary metabolism. We found that mild treatment of salicylic acid and acetyl salicylic acid (aspirin) increase isoflavone production dramatically in mung bean sprout which has very low level of isoflavones compared with soybean sprout. The isoflavone content in salicylic acid treated- and acetyl salicylic acid treated-mung bean sprout was about 2.3 and 2.2 times higher than that of control, respectively. However, the increasing patterns of isoflavone in cotyledon and hypocotyl and root were not identical. The major increase among isoflavone fractions in cotyledon was led by the increase in malonylglycitin and malonyldaidzin level. Whereas, the increase in hypocotyl and root was led by malonyldaidzin. Methyl jasmonic acid did not show statistically significant increase in mung bean sprout. With this result, we were able to propose the non-transgenic method, which can control the isoflavone production in germinating mung bean

Jasmonic acid improves ginsenoside accumulation in adventitious root culture of Panax ginseng C.A. Meyer

Yua, Kee-Won,Gaob, Wenyuan,Hahn, Eun-Joo,Paeka, Kee-Yoeup 충북대학교 한국과학재단 지정 첨단원예기술개발 연구센터 2002 연구보고서 Vol.6 No.-

Jasmonic acid significantly increased ginsenoside content in ginseng adventitious roots in flask and bioreactor cultures. In flask culture, ginsenoside content increased by the addition of jasmonic acid, resulting in the highest ginsenoside content at 10 mgl-1 jasmonic acid. However, the root growth was strongly inhibited by increasing jasmonic acid concentration. Fresh weight, dry weight, and growth rate of the roots decreased as jasmonic acid concentration increased. The highest ginsenoside yield was obtained at 2.0mgl-1 jasmonic acid. In a two-stage culture ina balloon type airlift bioreactor, the content of total ginsenoside and protopanaxadiol ginsenosides increased by over 5 times after 7 days of jasmonic acid treatment but the contents of Rg group ginsenosides increased insignificantly.

Chitosan과 Jasmonic acid 처리에 의한 인삼 부정근의 Ginsenosides의 생산성 증대

이범수,인준교,송원섭,양덕춘 한국자원식물학회 2004 한국자원식물학회지 Vol.17 No.1

인삼 부정근의 생장과 ginsenosides 함량에 미치는 elicitor의 영향을 조사한 결과 chitosan처리구의 생장량은 10 mg/L에서 가장 양호하였으며, ginsenosides의 함량은 5 mg/L처리농도에서 14.48 mg/g $.$ DW로 대조구에 비해 약9%의 함량증대를 가져왔다. 세포내에서 이차대사계를 활성화시키는 중간 신호전달 물질(signal transducer)로 알려진 jasmonic acid를 인삼 부정근 배양에 처리한 결과 부정근의 생장량은 10 uM 처리구에서 ginsenosides함량이 가장 양호하였으나, 인삼 부정근의 생장은 다소 억제되었다. In order to investigate the effects of elicitors on the growth and ginsenosides biosynthesis of ginseng adventitious roots, chitosan and jasmonic acid were treated with various concentrations. The growth rate of adventitious roots was increased with the addition of chitosan at higher concentrations (10 mg/L), but the best accumulation of ginsenosides was observed at the lower concentration (5 mg/L). Jasmonic acid was an effective elicitor for ginsenosides biosynthesis in ginseng adventitious roots. The maximum accumulation of ginsenosides was observed at the treatment of 10 uM jasmonic acid. But the jasmonic acid was found to decrease the growth rate of adventitious roots.

Jamonic acid carboxyl methyltransferase: a key enzyme for jasmonate-regulated plant responses

Choi, Yang-Do 서울대학교 농업개발연구소 2000 농업생명과학연구 Vol.4 No.-

Methy1 jasmonate is a plant volatile that acts as an important cellular regulator mediating diverse developmental processes and defense responses. We have cloned the novel gene JMT encoding a S-adenosy1-L-methionine: jasmonic acid carboxyl methyltransferase from Arabidopsis thaliana. Recombinant JMT protein expressed in E. coli catalyzed the formation of methyl jasmonate from jasmonic acid with Km value of 38.5 mM. JMT was not detected in young seedlings but expressed in rosettes, cauline leaves and developing flowers. In addition, expression of the gene was induced both locally and systemically by wounding or methyl jasmonate treatment. This result suggests that JMT can perceive and respond to local and systemic signals generated by external stimuli, and that the signals may include methyl jasmonate itself. Transgenic Arabidopsis overexpressing the JMT contained 3-fold elevated level of endogenous methyl jasmonate without altering jasmonic acid content. The transgenic plants exhibited constitutive expression of jasmonate-responsive genes including VSP and PDF1.2 Furthemore, the transgenic plants showed enhanced level of resistance against the virulent fungus Botrytis cinerea. Thus, our data suggest that the jasmonic acid carboxyl methyltransferase is a key enzyme for the jasmonate-regulated plant responses. Activation of JMT expression leads to production of methyl jasmonate that could act as an intracellular regulator, a diffusible intercellular signal transducer, and an airborne signal mediating intra-and inter- plant communications.

Overexpression of jasmonic acid carboxyl methyltransferase increases tuber yield and size in transgenic potato

Sohn, Hwang-Bae,Lee, Han-Yong,Seo, Ju-Seok,Jung, Choon-Kyun,Jeon, Jae-Heung,Kim, Jeong-Han,Lee, Yin-Won,Lee, Jong-Seob,Cheong, Jong-Joo,Choi, Yang-Do The Korean Society of Plant Biotechnology 2011 Plant biotechnology reports Vol.5 No.1

Jasmonates control diverse plant developmental processes, such as seed germination, flower, fruit and seed development, senescence and tuberization in potato. To understand the role of methyl jasmonate (MeJA) in potato tuberization, the Arabidopsis JMT gene encoding jasmonic acid carboxyl methyltransferase was constitutively overexpressed in transgenic potato plants. Increases in tuber yield and size as well as in vitro tuberization frequency were observed in transgenic plants. These were correlated with JMT mRNA level-- the higher expression level, the higher the tuber yield and size. The levels of jasmonic acid (JA), MeJA and tuberonic acid (TA) were also higher than those in control plants. Transgenic plants also exhibited higher expression of jasmonate-responsive genes such as those for allene oxide cyclase (AOC) and proteinase inhibitor II (PINII). These results indicate that JMT overexpression induces jasmonate biosynthesis genes and thus JA and TA pools in transgenic potatoes. This results in enhanced tuber yield and size in transgenic potato plants.

Ginsenoside Production by Hairy Root Cultures of Panax Ginseng C.A. Meyer in Bioreactors

Hahn, Eun-Joo,Yu, Kee-Won,Paek, Kee-Yoeup 충북대학교 첨단원예기술개발연구센터 2000 연구보고서 Vol.5 No.-

Hairy roots infected with Agrobacterium rhizogenes were induced from roots, stems, and leaves of Korean ginseng (Panax ginseng C.A Meyer) on half strength MS medium supplemented with 300 mgㆍ1-1 Cefotaxin sodium. DNA extraction was carried out and PCR results showed that the induced hairy roots by A. rhizogenesis KCTC 2703 were confirmed to have rol C gene in T-DNA. Selected root lines were propagated sucrose in 5-liter cone type bubble bioreactors containing MS media supplemented with 20 mgㆍ1-1 NAA and 30 mgㆍ1-1 sucrose. To increase ginsenoside contents, jasmonic acid with various concentrations was added to the culture medium after 30 days of culture then the roots were cultured for 7 more days until the harvest. Total ginsenoside content increased with increasing jasmonic acid but high concentrations of jasmonic acid inhibited root growth. Ginsenosid productivity was greatest at 2.0 mgㆍ1-1 jasmonic acid. On the other hand, ginsenosides in Rb group mainly increased, while those in Rg group did not increase. Particularly, high concentrations (5 and 10 mgㆍ1-1) of jasmonic acid decreased Rg1 content almost half of that in control but significantly increased Rb1 content in Rb group. Among the ginsenosides in Rb group, Rb1 content increased more than Rb2, Rc, and Rd. There were gradual increases in ginsenosides in Rb group but those in Rg group fluctuated then slightly decreased at the end of the culture period. Further studies are required to raise the contents of ginsenosides in Rg group and to determine optimal type of elicitors as well as optimal time for the elicitor treatment for accelerating ginsenoside production.

Cloning and characterization of jasmonic acid carboxyl methyltransferase genes

Choi, Yang Do 서울대학교 농업개발연구소 2001 농업생명과학연구 Vol.5 No.-

Methyl jasmonate is an important cellular regulator that mediates diverse developmental processes and defense responses. We cloned the novel genes JMT and NTR1 encoding S-adenosyl-L-methionine: jasmonic acid carboxyl methyltransferase from Arabidopsis thaliana and Brassica campestris L. ssp. perkinensis, respectively. Recombinant JMT and NTR1 proteins expressed in E. coli catalyzed the formation of methly jasmonate from jasmonic acid. Kinetic properties, thermal characteristics, optimal pH, and ion-dependency of the NTR1 activity were almost identical to those of Arabidopsis JMT, indicating that these two proteins are orthologues of each other. Transgenic Arabidopsis overexpressing the JMT contained elevated level of endogenous methyl jasmonate, and exhibited constitutive expression of jasmonate-responsive genes and enhanced level of resistance against a virulent fungus. Expression of these genes initiated and limited in developing flower, but propagated systemically, through phloem in vascular bundles, down along with the floral and main stems to reach to the primary root. Moreover, the genes were induced both locally and systemically by wounding or methyl jasmonate treatment. Expression pattern and function of the genes are consistent with previous observations of jasmonate distributions among developing tissues and of jasmonate-responsive defense responses, indicating that the gene activation is a key control point for the jasmonate-regulated responses. Our data suggests that JMT can perceive and respond to local and systemic signals generated during flowering process or by external stimuli, and that the signals may include methyl jasmonate itself. Thus, the gene activation leads to production of methyl jasmonate that could act as a diffusible intra- and inter-cellular signal transducer.

오이(Cucumis sativus L.) 떡잎의 노쇠화와 관련된 유전자에 영향을 미치는 Jasmonic acid의 역할에 관한 연구

이정선 ( Jeong Seon Lee ),김대재 ( Dae Jae Kim ) 충북대학교 과학교육연구소 2011 과학교육연구논총 Vol.27 No.1

New plant leaf begins from leaf primordia to develop a leaf. Leaf senescence is one type of programed cell death(PCD) that is controlled by genetic program. Previously, studies identified many cucumber (Cucumis sativus L.) SAGs (senescence-associated genes). In this study, cucumber SAGs were selected to examine the gene expression patterns and to characterize of Jasmonic acid response which may affect in these genes. The SAGs were sen 4-1(pheophytinase encoding gene), sen 15-8(L-asparaginase encoding gene) and sen 053(ATPase subunitⅢ encoding gene). The most probable function of genes sequences was identified throughout NCBI-GenBank database search using tbalstx or blastx finding module. The sen 4-1 peptide sequence was matched completely with pheophytinase. The sen 15-8 peptide sequence matched in 89% with L-asparaginase. The sen 053 peptide sequence matched completely with ATPase subunitⅢ. Jasmonic acid was treated at the 9th day of after seeds sowing and then cucumber cotyledons were collected after 3 days passed of phytohormone treatment. Therefore, cotyledons were detached at the 14th, 21th, 28th day after seeds sowing and senescence stage Ⅰ(50% yellow), stage Ⅱ(70% yellow) until 45 days of cucumber development. We examined the activity of the genes by RT-PCR approach. As a result of the experiment, Sen 4-1 gene and Sen 053 genes were repressed by jasmonic acid. Sen 15-8 gene did not show any effect of jasmonic acid treatment.

오이(Cucumis sativus L.) 떡잎의 노쇠화 관련 유전자의 발현 특성과 그 유전자에 미치는 자스몬산의 영향에 관한 연구

한지수 ( Jee Soo Han ),김대재 ( Dae Jae Kim ) 충북대학교 과학교육연구소 2011 과학교육연구논총 Vol.27 No.1

Plant senescence is the last stage of development and leads to their death an organ and an organism. Senescence is a programmed cell death(PCD) that controlled by genetic program. Several genes are identified senescence - associated genes(SAGs) in cucumber(Cucumis sativus L.). The SAGs selected to study of the gene expression to jasmonic acid which affects in these genes. The genes were sen 8-6((1-4)-β-mannan endohydrolase encoding gene), sen 11-8(temperature-induced lipocalin encoding gene) and sen 218(hypothetical protein encoding gene). Function of the sen cDNA sequences was identified through NCBI-GenBank database investigation using blastx searching module. Natural senescence was examined from the days at 14th, 21th, 28th day after seeds sowing and senescence stage Ⅰ(50% yellow; SⅠ), stage Ⅱ(70% yellow; SⅡ) according to chlorophyll contents in cucumber cotyledons. Cotyledons treated jasmonic acid was 14th, 21th, 28th, seeds sowing after day and senescence stage Ⅰ(50% yellow; SⅠ), stage Ⅱ(70% yellow; SⅡ). The cotyledons were used for total RNA purification. Then cDNA from mRNA using reverse transcription (RT) and performed PCR using gene primer. In natural senescence, sen 8-6 wasn`t reaction and sen 11-8 was expressed in all stage. Sen 218 showed action in 14th, 21th, 28th SⅠ. In senescence with jasmonic acid, sen 8-6 was similar to that of the natural senescence result. The sen 11-8 and sen 218 showed stage Ⅱ inconsistent expression. Sen 11-8 in natural senescence was expressed consistently, but stage Ⅱ of senescence with jasmonic acid wasn`t active. Therefore sen 11-8 was affected by jasmonic acid.