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1.
Copper-induced changes in the growth, oxidative metabolism, and saponin production in suspension culture roots of Panax ginseng in bioreactors 总被引:3,自引:0,他引:3
Roots of Panax ginseng exposed to various concentrations of Cu (0.0, 5, 10.0, 25.0, and 50.0 μM) accumulated high amounts of Cu in a concentration-dependent and duration-dependent manner. Roots treated with 50 μM Cu resulted in 52% and 89% growth inhibition after 20 and 40 days, respectively. Saponin synthesis was stimulated at a Cu concentration between 5 and 25 μM but decreased at 50 μM Cu. Malondialdehyde content (MDA), lipoxygenase activity (LOX), superoxide ion (O2
•−) accumulation, and H2O2 content at 5 and 10 μM Cu-treated roots were not increased but strongly increased at 50 μM Cu resulting in the oxidation of ascorbate (ASC) and glutathione (GSH) to dehydroascorbate (DHA) and glutathione disulfide (GSSG), respectively indicating a clear oxidative stress. Seven well-resolved bands of superoxide dismutase (SOD) were detected in the gel and an increase in SOD activity seemed to be mainly due to the induction of Fe-SOD 3. Five to 10 μM Cu slightly induced activity of ascorbate peroxidase (APX) and dehydroascorbate reductase (DHAR), guaiacol peroxidase (G-POD) but inhibited monodehydroascorbate reductase (MDHAR) and glutathione reductase (GR) enzyme activities. No changes in catalase (CAT) activity and in activity gel were found up to 25 μM Cu, but both G-POD and CAT activities were inhibited at 50 μM Cu. Glutathione metabolism enzymes such as γ-glutamylcysteine synthetase (γ-GCS), glutathione-S-transferase (GST), and glutathione peroxidase activities (GPx) were activated at 5 and 10 μM Cu but were strongly inhibited at 50 μM Cu due to the Cu accumulation in root tissues. The strong depletion of GSH at 50 μM Cu was associated to the strong induction of γ-glutamyltranspeptidase (γ-GGT) activity. These results indicate that plant could grow under Cu stress (5–25 μM) by modulating the antioxidant defense mechanism for combating Cu induced oxidative stress. 相似文献
2.
Involvement of nitric oxide-induced NADPH oxidase in adventitious root growth and antioxidant defense in Panax ginseng 总被引:1,自引:0,他引:1
Rajesh Kumar Tewari Soohyun Kim Eun-Joo Hahn Kee-Yoeup Paek 《Plant biotechnology reports》2008,2(2):113-122
Nitric oxide (NO) affects the growth and development of plants and also affects plant responses to various stresses. Because
NO induces root differentiation, we examined whether or not it is involved in increased ROS generation. Treatments with sodium
nitroprusside (SNP), an NO donor, 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO), a specific NO scavenger,
and Nω-nitro-l-arginine methyl ester hydrochloride (l-NAME), an NO synthase (NOS) inhibitor, revealed that NO is involved in the adventitious root growth of mountain ginseng.
Supply of an NO donor, SNP, activates NADPH oxidase activity, resulting in increased generation of O2
·−, which subsequently induces growth of adventitious roots. Moreover, treatment with diphenyliodonium chloride (DPI), an NADPH
oxidase inhibitor, individually or with SNP, inhibited root growth, NADPH oxidase activity, and O2
·− anion generation. Supply of the NO donor, SNP, did not induce any notable isoforms of enzymes; it did, however, increase
the activity of pre-existing bands of NADPH oxidase, superoxide dismutase, catalase, peroxidase, ascorbate peroxidase, and
glutathione reductase. Enhanced activity of antioxidant enzymes induced by SNP supply seems to be responsible for a low level
of H2O2 in the adventitious roots of mountain ginseng. It was therefore concluded that NO-induced generation of O2
·− by NADPH oxidase seems to have a role in adventitious root growth of mountain ginseng. The possible mechanism of NO involvement
in O2
·− generation through NADPH oxidase and subsequent root growth is discussed. 相似文献
3.
C. S. Jeong H. N. Murthy E. J. Hahn H. L. Lee K. Y. Paek 《Acta Physiologiae Plantarum》2009,31(1):219-222
The effect of the root-inoculum size and axuin concentration on growth of adventitious roots and accumulation of ginsenosides
were studied during suspension cultures of ginseng (Panax ginseng C.A. Meyer). Of the various concentrations of indole-3-butyric acid (IBA) and γ-naphthaleneacetic acid (NAA) used as supplementary
growth regulators along with Murashige and Skoog medium, 25 μM IBA was found suitable for lateral root induction and growth,
as well as accumulation of ginsenosides. Inoculum size of 5 g L−1 was found suitable for optimal biomass (10.5 g L−1 dry biomass) and ginsenosides (5.4 mg g−1 DW) accumulation. Of the various length of root inocula tested (chopped to 1–3, 4–6, 7–10 mm and un-chopped), root inocula
of 7–10 mm was found suitable for biomass and ginsenoside accumulation. 相似文献
4.
Summary
Panax ginseng hairy root cultures were established by infecting petiole segments with Agrobacterium rhizogenes strain 15834. Hairy root segments including root tips placed onto phytohormone-free 1/2 Murashige and Skoog solid medium and stored at 4 °C in the dark for 4 months, resumed elongation when the temperature was raised to 25 °C in the dark. For cryopreservation, a vitrification method was applied. Root tips precultured with 0.1 mg/l 2,4-D for 3 days and dehydrated with PVS2 solution for 8 minutes prior to immersion into liquid nitrogen had a survival rate of 60 % and could regenerate. The hairy roots regenerated from cryopreserved root tips grew well and showed the same ginsenoside productivity and patterns as those of the control hairy roots cultured continuously at 25 °C. The conservation of T-DNAs in the regenerated hairy roots was proved by PCR analysis.Abbreviations 1/2 MS
a half strength Murashige and Skoog (1962)
- B5
Gamborg B5 (Gamborg et al. 1968)
- WP
woody plant (Lloyd and McCown 1980)
- RC
root culture (Thomas and Davey 1982)
- RCI
root culture medium containing 100 mg/l myoinositol
- HF
phytohormone-free
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- TIBA
2,3,5-triiodobenzoic acid
- PCR
polymerase chain reaction
- PVS2
plant vitrification solution 2 (Sakai et al., 1990)
- FDA
fluorecein diacetate 相似文献
5.
The involvement of NO in O2
·− generation, rootlet development and antioxidant defence were investigated in the adventitious root cultures of mountain ginseng.
Treatments of NO producers (SNP, sodium nitroprusside; SNAP, S-nitroso-N-acetylpenicillamine; and sodium nitrite with ascorbic acid), and NO scavenger (PTIO, 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl3-oxide)
revealed that NO is involved in the induction of new rootlets. Severe decline in number of new rootlets compared to the control
under PTIO treatment indicates that NO acts downstream of auxin action in the process. NO producers (SNP, SNAP and sodium
nitrite with ascorbic acid) activated NADPH oxidase activity, resulting in greater O2
·− generation and higher number of new rootlets in the adventitious root explants. Moreover, treatment of diphenyliodonium chloride,
a NADPH oxidase inhibitor, individually or along with SNP, inhibited root growth, NADPH oxidase activity and O2
·− anion generation. NO supply also enhanced the activities of antioxidant enzymes that are likely to be responsible for reducing
H2O2 levels and lipid peroxidation as well as modulation of ascorbate and non-protein thiol concentrations in the adventitious
roots. Our results suggest that NO-induced generation of O2
·− by activating NADPH oxidase activity is related to adventitious root formation in mountain ginseng. 相似文献
6.
Choi Sung Mee Ho Son Sung Rho Yun Seung Woung Kwon Oh Hoon Seon Jeong Yoeup Paek Kee 《Plant Cell, Tissue and Organ Culture》2000,62(3):187-193
A pilot-scale culture of multiple adventitious roots of ginseng was established using a balloon-type bubble bioreactor. Adventitious
roots (2 cm) induced from callus were cultured in plastic Petri dishes having 20 ml of solid Schenk and Hildebrandt (1972)
medium containing 3% sucrose, 0.15% gelrite, and 24.6 μM indole-3-butric acid. An average of 29 secondary multiple adventitious
roots were produced after 4 weeks of culture. These secondary roots were elongated on the same medium, reaching a length of
5 cm after 6 weeks of culture. A time course study revealed that maximum yields in 5-l and 20-l bioreactors were approximately
500 g and 2.2 kg at day 42 with 60 g and 240 g inoculations, respectively. Cutting twice during the culture increased the
total amount of biomass produced. The root biomass in a 20-l balloon-type bubble bioreactor was 2.8 kg at harvest with 240
g of inoculum after 8 weeks of culture. The total saponin content obtained from small-scale and pilot-scale balloon type bubble
bioreactors was around 1% based on dry weight. Inoculation of 500 g fresh weight of multiple adventitious roots into a 500
l balloon-type bubble bioreactor with cutting at 4 and 6 weeks after inoculation produced approximately 74.8 kg of multiple
roots. The ginsengnoside profiles of these multiple adventitious roots were similar to profiles of field-grown ginseng roots
when analyzed by HPLC.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
7.
Since jasmonates have been shown to mimic some of the plant'sresponses to stress, the effect of methyl jasmonate on antioxidant enzymes andcompounds was investigated in roots and shoots of light- and dark-grown canola(Brassica napus cv. Westar). The pattern of superoxidedismutase isoforms activity was also investigated. When enzyme activities werecalculated on a per gram of fresh weight basis, nearly all enzymes examinedshowed enhanced activity. However, when these activities were calculated basedon the amount of protein, methyl jasmonate induced an increase only insuperoxide dismutase activity in the roots of both light- and dark-grownseedlings. The ascorbate level was found to be higher in treated shoots,whereasthe glutathione level was found to be higher in treated roots. We conclude thatthe plant's antioxidant response to methyl jasmonate may be mainlydetermined by the type of tissue rather than by the light conditions. However,this last factor appeared to be involved in some antioxidant componentresponse,e.g. catalase activity and glutathione content. 相似文献
8.
Hairy root cultures have demonstrated great promise in terms of their biosynthetic capability toward the production of secondary
metabolites, but continue to constitute a major challenge with regard to large-scale cultures. In order to assess the possibility
of conducting mass production of biomass, and the extraction of useful metabolites fromPanax ginseng. P. ginseng hairy roots, transformed byRhizobium rhizogenes KCTC 2744, were used in bioreactors of different types and sizes. The most effective mass production of hairy roots was achieved
in several differently sized air bubble bioreactors compared to all other bioreactor types. Hairy root growth was enhanced
by aeration, and the production increased with increasing aeration rate in a 1 L bioreactor culture. It was determined that
the hairy root growth rate could be substantially enhanced by increases in the aeration rate upto 0.5 wm, but at aeration
rates above 0.5 wm, only slight promotions in growth rates were observed. In 20 L air bubble bioreactors, with a variety of
inoculum sizes, the hairy roots exhibited the most robust growth rates with an inoculum size of 0.1% (w/v), within the range
0.1 to 0.7% (w/v). The specific growth rates of the hairy roots decreased with increases in the inoculum size. 相似文献
9.
Bonfill Mercè Cusidó Rosa M. Palazón Javier Canut Esther Piñol M. Teresa Morales Carmen 《Plant Cell, Tissue and Organ Culture》2003,73(1):37-41
The changes of redox status during culture and their effect on organogenesis were investigated in ginseng callus cultures. Three kinds of Panax ginseng calluses (non-organogenic calluses, root-forming and bud-forming calli) 21 were obtained from each of the three examined auxins (2,4-D, IBA and NAA) at 2 mg l–1 (9.05 M 2,4-D; 9.84 M IBA; 10.74 M NAA) concentration and 0.1 mg l–1 (0.46 M) kinetin over 5 weeks. Peroxidase in the soluble fraction, which indicates oxidative status and shows a correlation with differentiation, was higher in bud-forming calluses cultured in the three types of auxins. On the other hand, reducing capacity (overall reducing capacity of glutathione, ascorbic acid, tocoferol, carotenoids and phenol) which indicates reductive status, was higher when these calluses grew on media containing 2,4-D but specially so in the case of non-organogenic calluses. With the three types of auxin used, the reducing capacity was always higher in non-organogenic calluses and lower in calluses with more organogenic capacity. It seems that organogenesis of calluses is conditioned by a high peroxidase activity in soluble fraction (equal to or more than 0.8 A430/mg protein per min) and therefore a low reducing capacity (less than 0.3 A760/g DW). On the other hand 2,4-D promotes proliferation whereas NAA promotes morphogenesis. 相似文献
10.
Rajesh Kumar Tewari Soo Yeon Lee Eun Joo Hahn Kee Yoeup Paek 《Plant biotechnology reports》2007,1(4):227-235
Nitric oxide (NO) is a diffusible, gaseous signaling molecule. In plants, NO influences growth and development, and it can
also affect plant responses to various stresses. Because NO induces root differentiation and interacts with reactive oxygen
species, we examined the temporal effect of NO elicitation on root growth, saponin accumulation and antioxidant defense responses
in the adventitious roots of mountain ginseng (Panax ginseng). The observations revealed that NO is involved in root growth and saponin production. Elicitation with sodium nitroprusside
(SNP) activated O2
−-generating NADPH oxidase (NOX) activity, which most probably subsequently enhanced growth of adventitious roots of mountain
ginseng. A severe inhibition of NOX activity and decline in dry weight of SNP elicited adventitious roots in the presence
of NOX inhibitor (diphenyl iodonium, DPI), which further supports involvement of NOX in root growth. Enhanced activities of
antioxidant enzymes by SNP appear to be responsible for low H2O2, less lipid peroxidation, and modulation of ascorbate and non-protein thiol statuses in the adventitious roots of mountain
ginseng. Dry mass, saponin content and NOX activity was related with NO content present in adventitious roots of mountain
ginseng. 相似文献
11.
Somatic embryogenesis of Panax ginseng in liquid cultures: a role for polyamines and their metabolic pathways 总被引:1,自引:0,他引:1
Claire Kevers Nathalie Le Gal Marta Monteiro Jacques Dommes Thomas Gaspar 《Plant Growth Regulation》2000,30(3):209-214
A callus with embryogenic capacity was generated fromroot sections of Panax ginseng and used as aninoculum source for embryogenic liquid cultures in athree-step process: – a suspension culture of cellaggregates in the presence of an auxin/cytokininmixture, – an induction medium containing auxin only(for 5 to 30 days), – a regeneration medium containingcytokinin only (for one month). Up to 25 embryos wererecovered per 2.5 g of aggregates in these conditions.Incorporation of polyamines or their precursorsarginine and ornithine into either the induction orregeneration media increased the number of embryosproduced by up to 4 times. Inhibitors of bothbiosynthesis and biodegradation of polyamines reducedthe number of embryos. These results support earlierfindings of the role of polyamines in the process ofsomatic embryogenesis. The success of these liquidcultures opens up the possibility of producing somaticembryos of Panax ginseng in bioreactors. 相似文献
12.
Ethephon at 50 μM enhanced both root growth and ginsenoside accumulation in ginseng (Panax ginseng C.A. Meyer) adventitious root cultures, but at 100 μM it inhibited only ginsenoside accumulation. Ginsenoside productivity with 50 μM ethephon was the highest at 1.7 mg l−1 d−1 after 8 days of elicitation. However, elicitation with 50 μM ethephon and 100 μM methyl jasmonate (MJ) improved productivity (6.3 mg l−1 d−1) whereas elicitation with 100 μM MJ alone gave only 2.9 mg l−1 d−1. 相似文献
13.
14.
Gorpenchenko TY Kiselev KV Bulgakov VP Tchernoded GK Bragina EA Khodakovskaya MV Koren OG Batygina TB Zhuravlev YN 《Planta》2006,223(3):457-467
Expression of the Agrobacterium rhizogenes rolC gene in Panax ginseng callus cells results in formation of tumors that are capable to form roots. The selection of non-root forming tumor clusters
yielded the embryogenic 2c3 callus line, which formed somatic embryos and shoots independently of external growth factors.
Although the 2c3 somatic embryos developed through a typical embryogenesis process, they terminated prematurely and repeatedly
formed adventitious shoot meristems and embryo-like structures. A part of the shoots and somatic embryos formed enlarged and
fasciated meristems. This is the first indication of the rolC gene embryogenic effect and, to our knowledge, the first indication that a single gene of non-plant origin can induce somatic
embryogenesis in plants. 相似文献
15.
The effects of methyl jasmonate (MJ) or salicylic acid (SA) on the sesquiterpene lactone content and biomass accumulation
were investigated in a hairy root culture of Cichorium intybus. The guaianolides crepidiaside B (1), 8-deoxylactucin (2), and the germacranolide sonchuside A (3) were quantified by RP-HPLC. Neither MJ nor SA affected the growth of examined hairy root culture. Jasmonate up-regulated
biosynthesis of the analysed sesqiterpene lactones in the culture (maximum after 72 h). SA caused a transient increase in
sonchuside A accumulation in the roots (up to twofold increase compared with the control) and decrease of guaianolide content. 相似文献
16.
This study was initiated to determine whether antibiotic pulse treatments (APT) could effectively eliminate internal infections of ginseng (Panax ginseng) root explants containing vascular tissue, and subsequently have post-treatment effects on changing explant behaviors in callus induction and organogenesis or somatic embryogenesis. For contamination control, a treatment of 40 min with an antibiotic solution consisting of 1000 mg/1 of penicillin-G and 1000 mg/1 of streptomycin immediately following Na-hypochlorite sterilisation significantly decreased contamination rate. Extending treatment time to 2–3 h further lowered the contamination rate to 30–40%. On the other hand, explants treated with antibiotics for 20 min or less were all contaminated. APT also had post-treatment effects; it delayed callus induction for 1–12 months depending on pulse duration and stimulated the generation of more hardand darker looking than fragile- and lighter looking callus. The induced callus proliferated at a depressed rate, increasing subculture intervals from 1 to several weeks, and not until after five subcultures did it fully recover. The regeneration ability of the recovered callus was also affected by APT: the regeneration of adventitious roots was promoted, whereas somatic embryos were not observed.Abbreviations
APT
Antibiotic pulse treatments
-
2,4-D
2,4-dichlorophenoxyacetic acid 相似文献
17.
Effect of polyploidy induction on biomass and ginsenoside accumulations in adventitious roots of ginseng 总被引:1,自引:0,他引:1
Adventitious roots ofPanax ginseng C.A. Meyer (a natural tetraploid) were treated with 50 or 100 mg L-1 colchicine for 12, 24,36, 48, or 60 h to induce polyploid (octoploid) roots. The largest number of octoploid roots was obtained
with a 100 mg L-1 colchicine treatment over 60 h. To verify that ginsenoside was being accumulated in the developing tissues, the tetraploid
(control) and octoploid roots were cultured for 40 d in Murashige and Skoog media that lacked NH4NO3 but was supplemented with 2 mg L-1 naphthaleneacetic acid and 50 g L-1 sucrose. Levels of fresh and dry biomass were greater in the octoploid roots. Although total ginsenoside and Rb-group ginsenoside
contents were less in the octoploid roots than in the tetraploids, the former had a higher amount of Rg-group ginsenosides
(especially Rg1). These results demonstrate the benefit that polyploid adventitious roots provide in enhancing the production
of secondary metabolites in ginseng. 相似文献
18.
ukasz Ku
ma Elbieta Bruchajzer Halina Wysokiska 《Enzyme and microbial technology》2009,44(6-7):406-410
Hairy root cultures of Salvia sclarea were grown in shake flasks and 10 L nutrient sprinkle bioreactor, running for 30 days and the effects of methyl jasmonate (MJ) on their growth and capacity to accumulate diterpenoids were measured. We found that MJ concentration and exposure time to the elicitor were factors that strongly affected the diterpenoid production. The highest diterpenoid accumulation (67.5 ± 7.1 mg g−1 dry weight, calculated as a sum of ferruginol, salvipisone, aethiopinone and 1-oxoaethiopinone) without reduction of biomass, was achieved when the 23-day-old hairy roots in bioreactor culture were exposed to 125 μM MJ for 7 days. The roots produced 9 and 3.8 times as much aethiopinone (40 ± 5.9 mg g−1 dry weight) and salvipisone (12.6 ± 0.4 mg g−1 dry weight), respectively, as roots cultured in shake flasks. Our results imply that cultivation of S. sclarea hairy roots in sprinkle bioreactor after elicitation with MJ may be valuable to enhance production of the bioactive diterpenoids. 相似文献
19.
Two oligosaccharides, a heptasaccharide (HS) and an octasaccharide (OS), isolated from Paris polyphylla var. yunnanensis, stimulated the growth and saponin accumulation of Panax ginseng hairy roots at 5–30 mg l−1. HS and OS at 30 mg l−1, fed separately to hairy root cultures at 10 days post-inoculation, increased the root biomass dry weight by more than 70%
to ∼20 g l−1 from 13 g l−1 and the total saponin content of roots by more than 1-fold to ∼3.5% from 1.6% (w/w). The results suggest that the two oligosaccharides
may have plant growth-regulatory activity in plant tissue cultures. 相似文献
20.
Francisco Palma Carmen Lluch Carmen Iribarne Jose M. García-Garrido Noel A. Tejera García 《Plant Growth Regulation》2009,58(3):307-316
It has been shown that salicylic acid (SA) acts as an endogenous signal molecule responsible for inducing abiotic stress tolerance
in plants. The effect of SA and sodium chloride (NaCl) on growth, metabolite accumulation, oxidative stress and enzymatic
and non-enzymatic antioxidant responses on common bean plants (Phaseolus vulgaris, cv. F-15) was studied. Results revealed that either SA or NaCl decrease, shoot, root and total plant dry weights. SA treatments
decreased the contents of proline, and reduced forms of ascorbate and glutathione, however, the content of soluble sugars
(TSS), thiobarbituric acid-reactive substances (TBARs) and oxidized ascorbate remained unaffected. On the other hand, salinity
significantly reduced the levels of endogenous SA but increased the content of proline, soluble sugars, TBARs, ascorbate and
glutathione, as well as all increasing the levels of antioxidant enzyme activities assayed, except CAT. The application of
SA improved the response of common bean plants to salinity by increasing plant dry weight and decreasing the content of organic
solutes (proline and TSS) and damage to the membrane (TBARs). Moreover, SA application under saline conditions decreased the
levels of antioxidant enzyme activities POX, APX and MDHAR which could indicate successful acclimatization of these plants
to saline conditions. 相似文献