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.

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.

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.

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.

<i>p</i>-Coumaric acid induces jasmonic acid-mediated phenolic accumulation and resistance to black rot disease in <i>Brassica napus</i>

Islam, Md Tabibul,Lee, Bok-Rye,La, Van Hien,Lee, Hyo,Jung, Woo-Jin,Bae, Dong-Won,Kim, Tae-Hwan Elsevier 2019 Physiological and molecular plant pathology Vol.106 No.-

<P><B>Abstract</B></P> <P>To investigate <I>p</I>-coumaric acid (<I>p</I>CA)-induced plant resistance, the regulation of jasmonic acid (JA)-mediated phenylpropanoid biosynthesis pathway was assessed in response to pre-treatment with <I>p</I>CA or an inhibitor of 4-coumarate–CoA ligase (4CL), 3,4-(methylenedioxy) cinnamic acid (MDCA), in <I>Brassica napus</I> cultivar (cv. Mosa) following <I>Xanthomonas campestris</I> pv. <I>campestris</I> (<I>Xcc</I>) inoculation. At seven days post inoculation with <I>Xcc</I>, necrotic lesions decreased in response to <I>p</I>CA pre-treatment. JA content and expression of signaling genes (<I>COI1</I> and <I>PDF1.2</I>) were enhanced in <I>p</I>CA-pre-treated plants, along with an increase in the MYB transcriptional factor, production of anthocyanin pigment 1 (<I>PAP1</I>), which regulates phenylpropanoid biosynthesis in <I>Xcc</I>-inoculated plants. Following <I>Xcc</I> inoculation, the accumulation of epigallocatechin and epigallocatechin gallate was increased in soluble forms; however, induced accumulation of <I>p</I>CA, ferulic acid, and sinapic acid was observed in both soluble and cell wall-bound forms in response to <I>p</I>CA pre-treatment, whereas this decreased in control or MDCA pre-treated plants. The expression of phenylpropanoid biosynthesis-related genes was upregulated in plants pre-treated with <I>p</I>CA, whereas it decreased or did not change in control and MDCA pre-treated plants, following <I>Xcc</I> inoculation. These results indicate that treatment with <I>p</I>CA primed the JA-signaling mediated induction of phenylpropanoid biosynthesis to provoke disease resistance in <I>B. napus</I> to <I>Xcc</I>.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Pre-treatment of <I>p</I>-coumaric acid (<I>p</I>CA) enhanced jasmonic acid (JA) signaling. </LI> <LI> <I>p</I>CA primed <I>PAP1</I>-induced upregulation of phenylpropanoid biosynthesis. </LI> <LI> <I>p</I>CA-induced alleviation of black rot disease relied on JA-mediated phenolic accumulation. </LI> </UL> </P>

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.

Effects of Biocontrol Agents on Suppression of Damping-Off in Cucumis sativus L. Caused by Rhizoctonia solani

Hoon Hwangbo,김길용,최현석 한국원예학회 2016 Horticulture, Environment, and Biotechnology Vol.57 No.2

Biocontrol agents (Pseudomonas aeruginosa HB-15 and Pantoea sp. HB-22, alone or in combination) were applied on cucumber (Cucumis sativus L.) plants to evaluate their effect on suppressing damping-off caused by the fungal pathogen Rhizoctonia solani. Plant biomass, percent root mortality, phosphorus uptake, and the levels of stress-indicators NH3-NH4+ and proline in the plants were measured following the treatments. Additionally, gene expression induced by the biocontrol agents was compared to the gene expression induced by jasmonic acid and salicylic acid treatments. The biocontrol treatments increased fresh weight and phosphorus uptake, and reduced root mortality of the cucumber plants inoculated with the pathogen. No differences in plant ecophysiological responses were detected among biocontrol agent treatments. Concentrations of NH3-NH4+ and proline in the leaves and roots were higher in the control than in the biocontrol treatments. Salicylic acid- and Pantoea sp. HB-22-treated cucumbers showed increased expression of the acidic peroxidase-encoding gene and the phenylalanine ammonia homologue, while the P. aeruginosa HB-15 treatment did not induce an increase in expression of any of the genes studied.