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1.
Three lines of hairy root culture of Ambrosia maritima induced by Agrobacterium rhizogenes ATCC15834 were established. Thiarubrine A, thiarubrine A epoxide, thiarubrine A diol and their precursor pentayneene were produced by the hairy roots after elicitation with methyl jasmonate, the common signal molecule in the plant defense and development. Thiarubrine A diol was the main form detected in the medium. Maximum yield was achieved when the 13-day-old hairy root cultures were exposed to 40 M methyl jasmonate for 72 h. Callus and cell suspension cultures were established and maintained on Murashige and Skoog medium supplied with -naphthylacetic acid (NAA) and kinetin. When the cell suspension cultures were elicited with methyl jasmonate, pentayneene was the only polyacetylene produced. The yield of pentayneene in hairy root cultures was much higher (9.6 times) than that of cell suspension cultures. 相似文献
2.
Hairy root cultures of Gentiana macrophylla were established by infecting the different explants four Agrobacterium rhizogenes strains namely A4GUS, R1000, LBA 9402 and ATCC11325, and hairy root lines were established with A. rhizogenes strain R1000 in 1/2 MS + B5 medium. Initially, 42 independent hairy root clones were maintained and seven clones belongs to different category were evaluated
for growth, morphology, integration and expression of Ri T-DNA genes, and alkaloid contents in dry root samples. On the basis
of total root elongation, lateral root density and biomass accumulation on solid media, hairy root clones were separated into
three categories. PCR and Southern hybridization analysis revealed both left and right T-DNA integration in the root clones
and RT-PCR analysis confirmed the expression of hairy root inducible gene. GUS assay was also performed to confirm the integration
of left T-DNA. The accumulation of considerable amounts of the root-specific secoiridoid glucosides gentiopicroside was observed
in GM1 ( and ) and the GM2 ( and DNA) type clones in considerably higher amount whether as two but callus-type clones (GM3) accumulated much less or only very negligible amounts of gentiopicroside. Out of four media composition
the 1/2 MS + B5 vitamin media was found most suitable. We found that initial establishment of root cultures largely depends on root:media ratio. Maximum growth rate was recorded in 1:50 root:media ratio. The maximum biomass in terms of fresh weight (33-fold) was achieved in 1/2 MS + B5 media composition after 35 days in comparison to sixfold increase in control. The biomass increase was most abundant maximum
from 15 to 30 days. Influence of A. rhizogenes strains and Ri plasmid of hairy root induction, the possible role of the TL-DNA and TR-DNA genes on growth pattern of hairy root, initial root inoculum:media ratio and effect of media composition is discussed. 相似文献
3.
Cultures of Solanum aviculare hairy roots were established after transformation with Agrobacterium rhizogenes A4. High levels of steroidal alkaloids measured as solasodine equivalents were produced in shake-flasks and bioreactor, even though relatively low concentrations are found in roots in vivo. In shake flasks the maximum alkaloid yield was 32 mg g-1 dry weight; in a 3-1 air-driven bioreactor the yield was 29 mg g-1. These yields represent a 5-fold increase over previous reports for in vitro production, and are comparable with levels found in the aerial parts of intact S. aviculare plants. Production of steroidal alkaloids was growth-associated. High sugar levels at stationary phase and insensitivity to increased levels of medium components suggest that root cultures were limited by oxygen mass-transfer. In Petri-dish culture with and without exogenous gibberellic acid, root length and number of root tips increased exponentially; growth proceeded with a constant length per root tip of about 35 mm. Addition of gibberellic acid enhanced growth but reduced the specific steroidal-alkaloid level. Taking into account both growth and alkaloid yield, accumulation of steroidal alkaloids was improved by about 40% at gibberellic-acid concentrations of 10 and 100 g l-1. 相似文献
4.
Hairy root cultures of Gynostemma pentaphyllum were established by infecting leaf discs with Agrobacterium rhizogenes. The dry biomass of hairy roots grown in MS medium for 49 days was 7.3 g l−1 with a gypenoside content of 38 mg g−1 dry wt. 相似文献
5.
Enhanced production of antimicrobial sesquiterpenes and lipoxygenase metabolites in elicitor-treated hairy root cultures of Solanum tuberosum 总被引:2,自引:0,他引:2
Komaraiah P Reddy GV Reddy PS Raghavendra AS Ramakrishna SV Reddanna P 《Biotechnology letters》2003,25(8):593-597
Potato (Solanum tuberosum) hairy root cultures, established by infecting potato tuber discs with Agrobacterium rhizogenes, were used as a model system for the production of antimicrobial sesquiterpenes and lipoxygenase (LOX) metabolites. Of the four sesquiterpene phytoalexins (rishitin, lubimin, phytuberin and phytuberol) detected in elicitor-treated hairy root cultures, rishitin (213 g g–1 dry wt) was the most predominant followed by lubimin (171 g g–1 dry wt). The elicitors also induced LOX activity (25-fold increase) and LOX metabolites, mainly 9-hydroxyoctadecadienoic acid and 9-hydroxyoctadecatrienoic acid, in potato hairy root cultures. The combination of fungal elicitor plus cyclodextrin was the most effective elicitor treatment, followed by methyl jasmonate plus cyclodextrin in inducing sesquiterpenes and LOX metabolites. 相似文献
6.
Pedro A.G. Santos A. Cristina Figueiredo Pedro M.L. Lourenço José G. Barroso Luis G. Pedro M. Margarida Oliveira Jan Schripsema Stanley G. Deans Johannes J.C. Scheffer 《Biotechnology letters》2002,24(12):1031-1036
Transformed root cultures of Anethum graveolens were induced by inoculation of aseptically grown seedlings with Agrobacterium rhizogenes carrying plasmid pRi 1855. The main component of the essential oils from the fruits and from the roots of the parent plant was carvone, whereas -phellandrene and apiole were dominant in the oil from, respectively, the aerial parts and the hairy roots. The essential oils from the fruits, aerial parts and roots of the parent plant were at 2%, 0.3% and 0.06% (v/w), respectively, but only 0.02% (v/w) in the hairy root cultures. Growth of the hairy root cultures reached 600 mg dry wt/50 ml medium after 50 days. The essential oil composition did not change significantly during their growth. 相似文献
7.
S. Banerjee-Chattopadhyay A. M. Schwemmin D. J. Schwemmin 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1985,71(2):258-262
Summary Organ culture, plant regeneration from callus culture, and hairy root disease caused by Agrobacterium rhizogenes were utilized as methods of rapid in vitro propagation in Lycopersicon peruvianum Mill. A detailed and comparative karyotype analysis of the resulting material under such in vitro conditions revealed karyotypic stability under organ culture method, ploidy change in callus derived plants, and minor structural alterations of chromosomes in roots transformed by A. rhizogenes.Abbreviations BAP
N6-benzylaminopurine
- NAA
naphthaleneacetic acid
- MS
Murashige and Skoog medium
- RG
regeneration medium
- SDS
sodium dodecyl sulfate 相似文献
8.
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
9.
The hairy root culture of Lippia dulcis Trev., Verbenaceae, was established by transformation with Agrobacterium rhizogenes A4. The transformed roots grew well in Murashige and Skoog medium containing 2% sucrose. The roots turned light green when they were cultured under 16 h/day light. The green hairy roots produced the sweet sesquiterpene hernandulcin (ca. 0.25 mg/g dry wt) together with 20 other mono- and sesquiterpenes, while no terpenes were detected in the nontransformed root cultures. The growth and hernandulcin production in the hairy root cultures were influenced by the addition of auxins to the medium. The addition of a low concentration of chitosan (0.2 – 10.0 mg / l) enhanced the production of hernandulcin 5-fold.Abbreviations Cht
chitosan
- IAA
indole-3-acetic acid
- NAA
1-naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog(1962) 相似文献
10.
Hypocotyl explants of Catharanthus roseus produced hairy roots when cultured on Murashige and Skoog (MS) basal medium after infection by Agrobacterium rhizogenes. Explants gave rise to adventitious shoots at a frequency of up to 80% when cultured on MS medium supplemented with 31.1 M 6-benzyladenine and 5.4 M -naphthaleneacetic acid. There was a significant difference in the frequency of adventitious shoot formation for each hairy-root line derived from a different cultivar. Plants derived from hairy roots exhibited prolific rooting and had shortened internodes. Approximately half of the plants had wrinkled leaves and an abundant root mass with extensive lateral branching, but otherwise appeared morphologically normal. Plants with hairy roots that were derived from the cultivar Cooler Apricot developed flowers with petals that were white in the proximal region, whereas the wild-type flower petals are red. PCR and Southern blot analyses revealed that plants derived from hairy roots retained the Ri TL-DNA.Abbreviations BA 6-Benzyladenine - MS Murashige and Skoog medium - NAA -Naphthaleneacetic acid - SH Schenk and Hildebrandt mediumCommunicated by I.S. Chung 相似文献
11.
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
12.
Summary Hairy root cultures of Hyoscyamus muticus were established using Agrobacterium rhizogenes ATCC 15834. In one out of 8 clones established, an unusual root tip formation was observed after transfer of cultures from half-strength Murashige and Skoog (1962) to White's medium (1939). This phenomenon was associated with the production of a fine brownish cell suspension culture. Hairy root development resumed after transfer of the root tips from White to half-strength Murashige and Skoog medium. After plating the isolated brownish cells on hormone-free half-strength Murashige and Skoog or White solid medium, callus proliferation was observed, and then redifferentiation of hairy roots occurred. The polymerase chain reaction analysis of the H. muticus hairy root (clone Z2) revealed that only the tl region of the T-DNA was integrated. The growth and the production of five tropane alkaloids by this clone were examined.Abbreviations PCR
Polymerase Chain Reaction
- MS medium
Murashige and Skoog Medium
- 1/2 MS medium
half-strength MS medium
- WP medium
Woody Plant medium
- RC medium
Root Culture medium
- WH medium
White medium
- HPLC
High Performance Liquid Chromatography
- wt.
weight 相似文献
13.
Sook Young Lee Hui Xu Yong Kyung Kim Sang Un Park 《World journal of microbiology & biotechnology》2008,24(7):969-972
Rosmarinic acid, an important phenolic active compound, is one of the main active constituents of Agastache rugosa Kuntze and has astringent properties, antioxidant capacity, anti-inflammatory activity, antimutagenic ability, antimicrobial
capacity, and antiviral properties. To investigate in vitro production of rosmarinic acid, we established a hairy root culture
of A. rugosa by infecting leaf and stem explants with Agrobacterium rhizogenes R1000, and tested the growth and rosmarinic acid production of these cultures. Hairy roots were cultured in Murashige and
Skoog liquid medium and maximum growth (14.1 g dry wt/l) was attained after 14 days of culture, at which time the content
of rosmarinic acid was 116 mg/g dry wt. The present results demonstrate that hairy root culture of A. rugosa is a valuable alternative approach for the production of rosmarinic acid. 相似文献
14.
Li Li Jing Wang Wei Wang Yang Lu Yuliang Wang Genyu Zhou Guoyin Kai 《Biotechnology and Bioprocess Engineering》2008,13(5):606-612
In this study, an efficient transformation system for the medicinal plant Anisodus acutangulus was successfully developed and optimized using Agrobacterium rhizogenes. Three bacterial strains, A4, R1601, and modified C58C1 and three explant types, leaf blade, petiole, and stem, were examined.
The highest transformation efficiency of 94.44% was achieved using strain C58C1 with stem explants. Over 20 independent hairy
root lines were successfully established with strain C58C1 using stem explants, all of which contained the ro/B and ro/C genes as confirmed by polymerase chain reaction (PCR). Out of four media compositions, the liquid 1/2 MS medium was found
the most suitable for hairy root growth. The maximum biomass of one hairy root line increased up to 80 times in liquid 1/2
MS medium after a 30 day culture period. Different hairy root lines displayed a varied capacity for tropane alkaloid production
and the best hairy root line (T4) from the C58C1-stem combination produced up to 10.21 mg/g (dw) of hyoscyamine, which was
about 1.5-fold higher than in the wild type plants. To our knowledge, this is the first report to demonstrate the production
of tropane alkaloids in hairy roots of A. acutangulus. 相似文献
15.
Crypopreservation methods were firstly developed for root-tips from hairy root cultures of Beta vulgaris, established after transformation by Agrobacterium rhizogenes. The effects of culture age, pre-growth, cryoprotection, freezing rate and post-freeze culture conditions were determined. The resulting freezing protocol was then used to cryopreserve transformed root cultures of Nicotiana rustica. Both species were viable after freezing (ca. 80%), according to fluorescein diacetate vital staining. However, on average the regeneration of proliferating roots from surviving root-tips was low (<20%). Growth rates, secondary metabolite production and T-DNA structure of a number of hairy root lines were examined and found to be unchanged after cryopreservation.Abbreviations DMSO
dimethyl sulphoxide
- 2,4-D
dichlorophenoxyacetic acid
- FDA
fluoroscein diacetate
- IAA
indole-3-acetic acid
- NAA
-napthalene acetic acid 相似文献
16.
Hairy root cultures ofTrigonella foenum-graecum L. were established withAgrobacterium rhizogenes strain A4. The hairy roots produce diosgenin, an important spirostanol for the semi-synthesis of steroid hormones. Fourteen different liquid media were investigated. The fastest growth was obtained in McCown's woody plant (WP) medium supplemented with 3% sucrose; the highest diosgenin content was observed in half-strength WP medium with 1% sucrose (0.040% dry weight), which represents almost twice the amount detected in the 8-month-old non-transformed roots (0.024%). A time-course study in WP liquid media supplemented with 3% sucrose was undertaken. In these conditions, 17 g diosgenin/g fresh weight were produced. The influence of cholesterol, medium pH and chitosan on diosgenin production was tested. The addition of 40 mg/l chitosan elevated the diosgenin content to three times that found in non-elicited hairy roots.Abbreviations
MS
Murashige and Skoog (1962) medium
-
WP
McCown's woody plant medium 相似文献
17.
Chae Young-Am Hee-Suk Yu Ji-Sook Song Hee-Kyung Chun Sang-Un Park 《Biotechnology letters》2000,22(19):1527-1530
The transformed root culture of Polygonum tinctorium Lour. was established by infecting leaf explants with Agrobacterium rhizogenes A4. These cultures were examined for their growth and indigo content under various culture conditions. Among the four different culture media tested, SH medium showed the highest yield for root growth (28 mg dry wt/30 ml) and indigo production (152 g/dry wt). In SH medium, 30 g sucrose l–1, 2500 mg KNO3 l–1, 300 mg NH4H2PO4 l–1 were the best conditions for indigo production at pH 5.7. The production of indigo in hairy roots slightly increased with the addition of 200 mg chitosan l–1 (186 g/dry wt) and 20 U pectinase l–1 (181 g/dry wt). 相似文献
18.
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
19.
Summary
In vitro culture of hairy roots of Phyllanthus amarus induced by Agrobacterium rhizogenes was established. Their growth and ability for in vitro inactivation of hepatitis B virus surface antigen was studied and compared with adventitious roots grown in vitro. The selected hairy root clone HR-1 was capable of growing at a very fast rate, and an approximately 900-fold increase in
weight of root biomass was achieved after 4 wk of culture in hormone-free quarter-strength liquid Murashige and Skoog medium
with continuous agitation. Non-transformed roots cultured in the presence of 1.0 mg l−1 (5.71 μM) indole-3-acetic acid increased by 330-fold. The immuno-inactive property of roots was maximal in the crude extract. The
hairy roots were shown to possess 85% inhibition (in contrast to 15% in the control) in binding of hepatitis B surface antigen
(HBsAg) to its antibody (anti-HBs) after 24 h of incubation with HbsAg-positive sera in vitro at 37°C. Out of three fractions selected on the basis of molecular weight components of the extract, the Fraction III containing
comparatively lower molecular weight substances (≤3500) yielded the highest activity. The extract from non-transformed roots
was found to possess similar efficiency (87% inhibition). The levels of activity in both types of in vitro-raised roots were higher than those of naturally occurring roots and leafy shoots. The ability of P. amarus hairy root cultures to yield high biomass with the anti-viral property at high levels may provide an alternative source of
raw material for more detailed study in the field of pharmaceutical research. 相似文献
20.
Toshio Aoki Hideki Matsumoto Youichi Asako Yuji Matsunaga Koichiro Shimomura 《Plant cell reports》1997,16(5):282-286
Hairy root cultures ofAtropabelladonna were established by transformation withAgrobacterium rhizogenes 15834. Five clones of them were employed to study the production of hyoscyamine, the main constituent of the plant, together with other tropane alkaloids. The growth and alkaloid production of each clone were differently affected by basal liquid culture media tested. The transgenic plants regenerated from each clone of the hairy roots had different phenotypes and diverse alkaloid productivity both in the cultured condition and in productivitiy both in the cultured condition and in hydroponics.Abbreviations ANOVA
analysis of variance
- B5 medium
Gamborg B5 medium
- BA
N6-benzyladenine
- B.S.
Balanced Solution
- dw
dry weight
- EC
electric conductivity
- fw
fresh weight
- GC/MS
gas chromatography-mass spectrometry
- HPLC
high performance liquid chromatography
- MS medium
Murashige and Skoog medium
- NAA
naphthalene-l-acetic acid
- PCR
polymerase chain reaction
- SDS
sodium dodecyl sulfate
- TMS
trimethylsilyl
- WP medium
Woody Plant medium 相似文献