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1.
Psoralea corylifolia is an endangered plant producing various compounds of medical importance. Adventitious roots and hairy roots were induced
in cultures prepared from hypocotyl explants. Psoralen content was evaluated in both root types grown either in suspension
cultures or on agar solidified medium. Psoralen content was ~3 mg g−1 DW in suspension grown hairy roots being higher than in solid grown hairy roots and in solid and suspension-grown adventitious
roots. 相似文献
2.
Kee-Won Yu Wen-Yuan Gao Sung-Ho Son Kee-Yoeup Paek 《In vitro cellular & developmental biology. Plant》2000,36(5):424-428
Summary Hairy root cultures of Panax ginseng, established after the infection of root sections with Agrobacterium rhizogenes KCTC 2703, were cultured in phytohormone-free Murashige and Skoog (MS) liquid medium containing different concentrations
of jasmonic acid and some other elicitors, in order to promote ginsenoside accumulation. Jasmonic acid in the range 1.0−5.0
mg l−1 (4.8–23.8 μM) strongly improved total ginsenoside production in ginseng hairy roots. Peptone (300 mg l−1) also showed some effect on ginsenoside improvement; however its effect was much weaker than that of jasmonic acid. Ginsenoside
content and productivity were 58.65 and 504.39 mg g−1, respectively. The Rb group of ginsenoside content was increased remarkably by jasmonic acid, while Rg group ginsenoside
content changed only slightly compared to controls. However, jasmonic acid also strongly inhibited ginseng hairy root growth. 相似文献
3.
Sucrose concentration in the culture medium affected chlorophyll content, trichome development and amount of solasodine in
regenerated plantlets of Solanum trilobatum. High chlorophyll content and glandular trichomes were observed in the plants grown on Murashige and Skoog basal medium supplemented
with 131.85 mM sucrose. The solasodine was quantified using reverse phase high performance liquid chromatography. The plantlets
cultivated on this medium yielded 35.97 mg g−1 (d.m.) solasodine whereas the field plants used as control yielded only 2.32 mg g−1 (d.m.) of solasodine. 相似文献
4.
Diana M. Cheng Gad G. Yousef Mary H. Grace Randy B. Rogers J. Gorelick-Feldman I. Raskin Mary Ann Lila 《Plant Cell, Tissue and Organ Culture》2008,94(1):73-83
In order to develop a sustainable source of metabolism-enhancing phytoecdysteroids, cell suspension and hairy root cultures
were established from shoot cultures of wild-harvested Ajuga turkestanica, a medicinal plant indigenous to Uzbekistan. Precursors of phytoecdysteroids (acetate, mevalonic acid cholesterol) or methyl
jasmonate (an elicitor) were added to subculture media to increase phytoecdysteroid accumulation. In cell suspension cultures,
20-hydroxyecdysone (20E) content increased 3- or 2-fold with the addition of 125 or 250 μM methyl jasmonate, respectively,
compared to unelicited cultures. Precursor addition, however, did not provoke phytoecdysteroid accumulation. In hairy root
cultures, addition of sodium acetate, mevalonic acid, and methyl jasmonate, but not cholesterol, increased phytoecdysteroid
content compared to unelicited cultures. Hairy root cultures treated with 150 mg l−1 sodium acetate, or 15 or 150 mg l−1 mevalonic acid, increased 20E content approximately 2-fold to 19.9, 20.4 or 21.7 μg mg−1, respectively, compared to control (10.5 μg mg−1). Older hairy root cultures, extracted after the seventh subculture cycle, also showed increases in 20E content (24.8 μg mg−1), turkesterone (0.9 μg mg−1) and cyasterone (8.1 μg mg−1) compared to control cultures maintained for a shorter duration of four subculture cycles. Doses of 10 or 20 μg ml−1 hairy root extract increased protein synthesis by 25.7% or 31.1%, respectively, in a C2C12 mouse skeletal cell line. These
results suggest that sustainable production of metabolically active phytoecdysteroid can be achieved through hairy root culture
systems.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
5.
Natasha Maurmann Carina M. B. De Carvalho Andréia Loviane Silva Arthur G. Fett-Neto Gilsane L. von Poser Sandra B. Rech 《In vitro cellular & developmental biology. Plant》2006,42(1):50-53
Summary
Valeriana glechomifolia is an endemic species of southern Brazil, capable of accumulating, in all of its organs, the terpene derivatives known as
valepotriates, the presumed sedative components of the roots of pharmaceutically used species of Valeriana. In vitro cultures of the plant were established and the accumulation of acevaltrate, didrovaltrate, and valtrate in callus, cell suspension,
and untransformed root cultures was studied. Leaves of in natura plants and roots of micropropagated plantlets were used as the explants for callus induction and root culture establishment,
respectively, on Gamborg B5 basal medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) alone or with kinetin (KIN).
Culture growth and secondary metabolite yields were enhanced with 2,4-D (4.52μM) and KIN (0.93μM). Maximum valepotriate contents, quantified by HPLC, of acevaltrate (ACE) 2.6mg g−1 DW, valtrate (VAL) 10.2mgg−1 DW, and didrovaltrate (DID) 2.9mg g−1 DW were observed in root cultures after 7–8wk of culture. 相似文献
6.
Summary Shoot regeneration in hairy root cultures of Solanum khasianum Clarke influences root growth, solasodine production. and permeabilization of solasodine into the medium. These parameters
are dependent on exogenously supplied auxin and cytokinin: the effect being both concentration-and clone-dependent. Hairy
root cultures with no shoot regeneration showed high permeabilization of solasodine into the medium by the sixth week of incubation,
suggesting the medium acts as a sink for the solasodine synthesized by the roots. Solasodine in the culture medium was toxic
to the transformed roots and caused browning of root tips. In a separate set of experiments, the hairy root cultures showed
regeneration of approximately 50–70 mm long shoots after treatment with indole-3-acetic acid and kinetin. These hairy root
cultures had inereased levels of solasodine production, compared to cultures without shoot regeneration. The plantlets formed
in the hairy root cultures accumulated some of the solasodine, thereby reducing its permcabilization into the medium. Transport
of solasodine from root to shoot reduced the toxic effect of solasodine in the root zone and extended the exponential growth
phase by 8-10d. 相似文献
7.
Establishment of in vitro plants, cell and tissue cultures from Oldenlandia affinis for the production of cyclic peptides 总被引:1,自引:1,他引:0
In vitro cultured plants from Oldenlandia affinis were established from seeds and grown on a hormone-free medium. In vitro plants produced the cyclic peptide kalata B1 in concentrations of 0.67 mg g−1 dry weight after growth of 30 days. This was approximately 50% of the concentration analysed in green house plants (shoot tips), where different concentrations have been determined in leaves (1.82 mg g−1), shoot tips (1.36 mg g−1), stems (0.36 mg g−1), and in flowers (0.16 mg g−1). Callus and cell suspension cultures could be initiated from aseptic root, stem and leaf explants of O. affinis seedlings and plants. Different light intensities were shown to affect culture growth as well as chlorophyll synthesis. The friable callus was then used for the establishment of a cell suspension culture. Fresh and dry weight measurements showed that growth was optimal on MS medium supplemented with 0.4 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-d). Leaf suspensions cultured on this medium showed a 4-fold increase of biomass by the first week of incubation. No quantifiable amounts of kalata B1 were produced under these conditions. Morphological differentiation seems to be essential for cyclic peptide production. Therefore, several undifferentiated as well as organised cell lines of O. affinis have been developed. These cell lines will constitute a worthwhile starting point for the optimisation of kalata B1 synthesis in liquid media to the objective of producing cyclic peptides under controlled and defined conditions in bioreactors. 相似文献
8.
Hairy roots cultures from different Solanaceous species have varying capacities to produce E. coli B-subunit heat-labile toxin antigen 总被引:1,自引:0,他引:1
The gene encoding enterotoxigenic Escherichia coli B-subunit heat-labile toxin (LTB) antigen was co-transformed into hairy root cultures of Nicotiana tabacum (tobacco), Solanum lycopersicum (tomato) and Petunia parodii (petunia) under the CaMV35S promoter. Tobacco and petunia roots contained ~65–70 μg LTB g−1 tissue whilst hairy roots of tomato contained ~10 μg LTB g−1. Antigen at ~600 ng ml−1 was detected in growth medium of tobacco and petunia. Tobacco roots with higher LTB levels showed growth retardation of ~80%
whereas petunia hairy roots with similar levels of LTB showed only ~35% growth retardation, relative to vector controls. Regeneration
of plants from LTB-containing tobacco hairy roots was readily achieved and re-initiated hairy roots from greenhouse-grown
plants showed similar growth and LTB production characteristics as the original hairy root cultures. 相似文献
9.
Scaled-up hairy root culture of Artemisia annua L. was established in three-liter Erlenmeyer flask. Both artemisinin and stigmasterol that derive from the common precursors
of isopentenyl diphosphate and farnesyl pyrophosphate were isolated from hairy roots. The production rate of artemisinin isolated
by column chromatography from hairy root cultures was 0.54% (mg.gDW−1). Stigmasterol was identified by mass spectrometry and nuclear magnetic resonance analysis. The production of stigmasterol
isolated by column chromatography from hairy root cultures was 108.3% (mg.gDW−1). In hairy root cultures, the production rate of stigmasterol was estimated to be 201 times greater than that of artemisinin.
Our results suggest that investigation of secondary metabolites may provide a new insight to study artemisinin production
in hairy root cultures.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
10.
Initiation and establishment of hairy root cultures from leaf or seedling hypocotyl explants of Solanum mauritianum Scop., using six strains of Agrobacterium rhizogenes was attempted. Success was only achieved following hypocotyl inoculation with strain LBA 9402. Transformation frequency was very low, with only one instance out of a possible 90 being recorded. Resultant hairy root cultures grew rapidly and could be maintained using a Murashige and Skoog (1962) medium supplemented with 0.1 g L–1
myo-inositol and 3% sucrose, either as a solid or liquid culture. Under these conditions, the roots had a solasodine content of 126 g g–1 DW. Lower levels of solasodine and decreased root growth rates were recorded when the medium strength was reduced by half or 3% glucose substituted for the 3% sucrose.Abbreviations MS
Murashige and Skoog's (1962) medium 相似文献
11.
In this paper, we would like to show unexpected morphogenic potential of cell suspensions derived from seedling explants of
Gentiana kurroo (Royle). Suspension cultures were established with the use of embryogenic callus derived from seedling explants (root, hypocotyl
and cotyledons). Proembryogenic mass proliferated in liquid MS medium supplemented with 0.5 mg l−1 2,4-D and 1.0 mg l−1 Kin. The highest growth coefficient was achieved for root derived cell suspensions. The microscopic analysis showed differences
in aggregate structure depending on their size. To assess the embryogenic capability of the particular culture, 100 mg of
cell aggregates was implanted on MS agar medium supplemented with Kin (0.0–2.0 mg l−1), GA3 (0.0–2.0 mg l−1) and AS (80.0 mg l−1). The highest number of somatic embryos was obtained for cotyledon-derived cell suspension on GA3-free medium, but the best morphological quality of embryos was observed in the presence of 0.5–1.0 mg l−1 Kin, 0.5 mg l−1 GA3 and 80.0 mg l−1 AS. The morphogenic competence of cultures also depended on the size of the aggregate fraction and was lower when size of
aggregates decreased. Flow cytometry analysis reveled luck of uniformity of regenerants derived from hypocotyl suspension
and 100% of uniformity for cotyledon suspension. 相似文献
12.
M. E. Estrada-Zúñiga F. Cruz-Sosa M. Rodríguez-Monroy J. R. Verde-Calvo E. J. Vernon-Carter 《Plant Cell, Tissue and Organ Culture》2009,97(1):39-47
Plant tissue cultures represent a potential source for producing secondary metabolites. In this work, Buddleja cordata tissue cultures were established in order to produce phenylpropanoids (verbascoside, linarin and hydroxycinnamic acids),
as these metabolites are credited with therapeutic properties. Highest callus induction (76.4–84.3%) was obtained in five
treatments containing 2,4-Dichlorophenoxyacetic acid (2,4-d: 0.45–9.05 μM) with Kinetin (KIN: 2.32, 4.65 μM), whereas highest root induction (79.6%) corresponded to the α-Naphthaleneacetic
acid (9.05 μM) with KIN (2.32 μM) treatment. Verbascoside was the major phenylpropanoid produced in in vitro cultures (root,
white and green callus) [66.24–86.26 mg g−1 dry weight (DW)], while linarin and hydroxycinnamic acid production was low (0.95–3.01 mg g−1 DW). Verbascoside and linarin production were improved in cell suspension culture (116 mg g−1 DW and 8.12 mg g−1 DW, respectively). 相似文献
13.
Salvia miltiorrhiza Bunge (Lamiaceae) hairy root cultures were inoculated (at 0.02 and 0.2% v/v) and co-cultured with Bacillus cereus bacteria. The root biomass growth was inhibited significantly by the bacteria inoculated to the root culture on the first
day (day 0) but not by the bacteria inoculated on days 14 or 21 (in a 28-day overall period). On the other hand, the growth
of the bacteria in the hairy root culture was also strongly inhibited by the hairy roots, partially because of the antibacterial
activity of the secondary compounds produced by the roots. Most interestingly, the tanshinone production was promoted by the
inoculation of bacteria at any of these days but more significantly by an earlier bacteria inoculation. With 0.2% bacteria
inoculated on day 0, for example, the total tanshinone content of roots was increased by more than 12-fold (from 0.20 to 2.67 mg
g−1 dry weight), and the volumetric tanshinone yield increased by more than sixfold (from 1.40 to 10.4 mg l−1). The tanshinone production was also stimulated by bacterial water extract and bacterial culture supernatant but less significantly
than by the inoculation of live bacteria. The results suggest that the stimulation of tanshinone production by live bacteria
in the root cultures may be attributed to the elicitor compounds originating from the bacteria, and the hairy root–bacteria
coculture may be an effective strategy for improving secondary metabolite production in plant tissue cultures. 相似文献
14.
Smita Srivastava Ashok Kumar Srivastava 《In vitro cellular & developmental biology. Plant》2012,48(1):73-84
Azadirachtin, a well-known biopesticide, is a secondary metabolite extracted from the seeds of Azadirachta indica. In the present study, azadirachtin was produced in hairy roots of A. indica, generated by Agrobacterium rhizogenes-mediated transformation of leaf explants. Liquid cultures of A. indica hairy roots were developed with a liquid-to-flask volume ratio of 0.15. The kinetics of growth and azadirachtin production
were established in a basal plant growth medium containing MS medium major and minor salts, Gamborg’s medium vitamins, and
30 g l−1 sucrose. The highest azadirachtin accumulation in the hairy roots (up to 3.3 mg g−1) and azadirachtin production (∼44 mg l−1) was obtained on Day 25 of the growth cycle, with a biomass production of 13.3 g l−1 dry weight. To enhance the production of azadirachtin, a Plackett–Burman experimental design protocol was used to identify
key medium nutrients and concentrations to support high root biomass production and azadirachtin accumulation in hairy roots.
The optimal nutrients and concentrations were as follows: 40 g l−1 sucrose, 0.19 g l−1 potassium dihydrogen phosphate, 3.1 g l−1 potassium nitrate, and 0.41 g l−1 magnesium sulfate. Concentrations were determined by a central composite design protocol and verified in shake-flask cultivation.
The optimized medium composition yielded a root biomass production of 14.2 g l−1 and azadirachtin accumulation of 5.2 mg g−1, which was equivalent to an overall azadirachtin production of 73.84 mg l−1, 68% more than that obtained under non-optimized conditions. 相似文献
15.
Hairy roots ofCatharanthus roseus obtained by co-cultivation of hypocotyl segments withAgrobacterium rhizogenes, and cultured in SH (Schenk and Hildebrandt) basal medium, formed two types of calli when subcultured in SH medium with 1 mg/1 -naphthaleneacetic acid and 0.1 mg/l kinetin. One of them, a compact callus, when re-subcultured in SH basal medium gave rise to hairy roots again. A rhizogenic cell suspension culture was established from this type of callus. When cultured in SH medium with growth regulators, the rhizogenic callus produced catharanthine at a level of 41% of the level in the initial hairy roots. Upon transfer to SH basal medium, regenerated hairy roots produced this alkaloid at the original level of 1.5 mg/g dry wt. Using this cell/hairy root interchange system a new management system for hairy root culture in bioreactors has been devised and examined involving production of biomass in the form of a cell suspension in medium supplemented with growth regulators, and catharanthine production by hairy roots regenerated from these cells in medium without growth regulators.Abbreviations NAA
-naphthaleneacetic acid
- SH
Schenk and Hildebrandt
- SHNK
SH medium + 1 mg 1–1 NAA + 0.1 mg 1–1 kinetin 相似文献
16.
A comparison between hairy root cultures and wild plants of Saussurea involucrata in phenylpropanoids production 总被引:2,自引:0,他引:2
Saussurea involucrata is an important medicinal plant that produces a few bioactive secondary metabolites, such as hispidulin, rutin, and syringin. Previously, we established a hairy root culture system for this species through Agrobacterium-mediated transformation. The present study addressed the issue as how hairy root cultures perform in phenylpronoid accumulation. From the ethanolic extract of a hairy root culture established for Saussurea involucrata, syringin, rutin and hispidulin, were isolated and their chemical structures were confirmed by HPLC-ESI-MS. A quantitative study of the compounds showed great levels of syringin and hispidulin (being 43.5±1.13 and 0.34±0.023 mg g−1 dry weight, respectively), about 40 and 3 times, respectively, higher than those from wild plants. But, the levels of rutin from hairy roots were much lower (0.71±0.043 vs. 6.59±0.56 mg g−1 dry weight). Compared with untransformed root cultures, syringin and hispidulin levels were also higher. An experiment on culture media showed that MS was superior to others for phenylpropanoids accumulation in hairy roots, a 28-day culture produced 405 mg l−1 syringin. 相似文献
17.
Arabidopsis halleri is increasingly employed as a model plant for studying heavy metal hyperaccumulation. With the aim of providing valuable
tools for studies on cellular physiology and molecular biology of metal tolerance and transport, this study reports the development
of successful and highly efficient methods for the in vitro regeneration of A. halleri plants and production of stable cell suspension lines. Plants were regenerated from leaf explants of A. halleri via a three-step procedure: callus induction, somatic embryogenesis and shoot development. Efficiency of callus proliferation
and regeneration depended on the initial callus induction media and was optimal in the presence of 1 mg L−1 2,4-dichlorophenoxyacetic acid, and 0.05 mg L−1 benzylaminopurine. Subsequent shoot and root regeneration from callus initiated under these conditions reached levels of
100% efficiency. High friability of the callus supported the development of cell suspension cultures with minimal cellular
aggregates. Characterization of regenerated plants and cell cultures determined that they maintained not only the zinc tolerance
and requirement of the whole plant but also the ability to accumulate zinc; with plants accumulating up to 50.0 μmoles zinc
g−1 FW, and cell suspension cultures 30.9 μmoles zinc g−1 DW. Together this work will provide the experimental basis for furthering our knowledge of A. halleri as a model heavy metal hyperaccumulating plant. 相似文献
18.
Latiporn Udomsuk Thaweesak Juengwattanatrakul Kanokwan Jarukamjorn Waraporn Putalun 《Acta Physiologiae Plantarum》2012,34(3):1093-1100
Miroestrol and deoxymiroestrol are highly active phytoestrogens derived from the tuberous roots of Pueraria candollei var. mirifica. To date, there have been no reports regarding the production of miroestrol and deoxymiroestrol in in vitro cell culture.
In this study, callus and cell suspension cultures were established for the purpose of investigating miroestrol and deoxymiroestrol
content in P. candollei var. mirifica cells. Stem-derived callus cultured on Murashige and Skoog (MS) medium supplemented with 0.1 mg l−1 thidiazuron (TDZ), 0.5 mg l−1 naphthaleneacetic acid (NAA), and 1.0 mg l−1 benzyladenine (BA) provided optimal conditions for the accumulation of deoxymiroestrol and total isoflavonoids. The calli
produced 184.83 ± 20.09 μg g−1 dry weight of total chromene and 20.72 ± 2.38 mg g−1 dry weight of total isoflavonoid. This is the first report to suggest that callus culture is a suitable alternative method
for producing miroestrol and deoxymiroestrol. Carbon sources were evaluated for the cell suspension cultures of P. candollei var. mirifica. Sucrose provided optimal conditions for biomass production, whereas fructose was the most suitable carbon source for deoxymiroestrol
and isoflavonoid production. The information from our study can be employed for enhancing the production of miroestrol, deoxymiroestrol,
and total isoflavonoids using in vitro cell culture of P. candollei var. mirifica. 相似文献
19.
Jualang Azlan G. Marziah M. Radzali M. Johari R. 《Plant Cell, Tissue and Organ Culture》2002,69(3):271-278
This report describes the technique used to induce the hairy roots in Physalis minima (Linn.). Different types of explants obtained from in vitro germinated seedlings were aseptically co-cultivated with A. rhizogenesstrain LBA9402 in different media. Root growth and production of physalins were investigated in various basal media grown under dark and light conditions, and compared to that of normal root cultures. Transformed hairy root cultures grew rapidly and reach stationary phase after 15 days on a B5 medium. HPLC analysis of extracts of hairy root cultures showed that the maximum content of physalin B and F was 1.82 and 4.15 mg g–1 DW, respectively, when grown under dark conditions. Normal root cultures produced higher physalin B (1.60–1.62 mg g–1 DW) and F (3.30–3.75 mg g–1 DW) under the same culture conditions. Physalin F synthesis in light-grown root cultures was reduced significantly. 相似文献
20.
S. Pattnaik C. Pradhan S. K. Naik P. K. Chand 《In vitro cellular & developmental biology. Plant》2000,36(5):407-411
Summary A complete and efficient protocol is presented for plant regeneration from cell-suspension cultures of Dalbergia sissoo Roxb., an economically important leguminous tree. Factors influencing callus initiation, establishment of cell-suspension
culture, callus formation from embredded microcolonies, and shoot organogenesis from suspension-derived callus were identified.
Of the two different auxins tested, callus induction was better on a medium containing naphthalene acetic acid (NAA). The
percentage of callus induction increased considerably when NAA at 2.0 mg l−1 (10.8 μM) was added in conjunction with 0.5 mg l−1 (2.2 μM) N6-benzyladenine (BA). Of the three different explants evaluated for callus induction, hypocotyl segments were most responsive.
Friable hypocotyl-derived callus from the second subculture passage was used to initiate the cell-suspension culture. Optimum
growth of the cell suspension was observed in MS medium supplemented with the same growth regulators as described above for
callus induction, with an initial inoculum cell density of 1%. The plating efficiency of the microcolonies was greatly influenced
by harvesting time and the gelling agent used for plating. Efficiency was highest (93%) with cells harvested at their exponential
growth phase and plated in 1.2 g l−1 Phytagel. Shoot organogenesis from callus cultures was higher on a medium supplemented with a combination of BA and NAA than
on BA alone. Seventy-one per cent of cultures exhibited shoot-bud differentiation on a medium containing 3.0 mg l−1 (13.3 μM) BA and 0.5 mg l−1 (2.7 μM) NAA. Regenerated shoots were rooted on half-strength MS medium containing 1 mg l−1 each of indole-3-acetic acid (5.7 μM), indole-3-butyric acid (4.9 μM) and indole-3-propionic acid (5.3 μM). Plantlets were acclimated and established in soil. 相似文献