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1.
In this study, morphological alterations, biomass growth, and secondary metabolite production of genetically transformed hairy
roots ofPanax ginseng C. A. Meyer, were evaluated after administration of plant growth regulators. The addition of benzylamino purine and kinetin
to the culture media increased biomass formation and phenolic compound biosynthesis in the hairy roots. α-Naphthaleneacetic
acid and indole-3-butyric acid inhibited hairy root growth, however, low concentrations of indole-3-acetic acid slightly increased
hairy root growth. Low concentrations of 2,4-Dichlorophenoxyacetic acid profoundly inhibited growth of hairy roots. The addition
of plant growth regulators, such as auxin, did not increase total phenolic compounds in hairy roots that did not contain gibberellic
acid and cytokinins. Callus formation was induced in cultures suspended in liquid medium amended with benzylamino purine and
kinetin. Hairy roots regenerated from these calluses exhibited an active growth pattern with extensive lateral branching in
non-amended medium, similar to the growth pattern of the original hairy roots. 相似文献
2.
Fredy R. Romero Kathleen Delate George A. Kraus Avery K. Solco Patricia A. Murphy David J. Hannapel 《In vitro cellular & developmental biology. Plant》2009,45(5):599-609
Hairy root cultures of Echinacea, one of the most important medicinal plants in the US, represent a valuable alternative to field cultivation for the production
of bioactive secondary metabolites. In this study, the three most economically important species of Echinacea (Echinacea purpurea, Echinacea pallida, and Echinacea angustifolia) were readily transformed with two strains of Agrobacterium that produce the hairy root phenotype. Transformed roots of all three species exhibited consistent accelerated growth and
increased levels of alkamide production. Optimization of the culture of Echinacea hairy roots was implemented to enhance both growth and alkamide production concomitantly. The use of half-strength Gamborg’s
B5 medium supplemented with 3.0% sucrose was twice as effective in maintaining hairy root production than any other media
tested. The addition of indolebutyric acid increased the growth rate of roots by as much as 14-fold. Alkamide production increased
severalfold in response to the addition of the elicitor, jasmonic acid, but did not respond to the addition of indolebutyric
acid. Induced accumulation of the important bioactive compounds, alkamides 2 and 8, was observed both in transformed roots
and in response to jasmonic acid treatments. The results of this study demonstrate the efficacy of hairy root cultures of
Echinacea for the in vitro production of alkamides and establish guidelines for optimum yield. 相似文献
3.
Hosakatte Niranjana Murthy Yun-Soo Kim So-Young Park Kee-Yoeup Paek 《Applied microbiology and biotechnology》2014,98(18):7707-7717
Caffeic acid derivatives (CADs) are a group of bioactive compounds which are produced in Echinacea species especially Echinacea purpurea, Echinacea angustifolia, and Echinacea pallida. Echinacea is a popular herbal medicine used in the treatment of common cold and it is also a prominent dietary supplement used throughout the world. Caffeic acid, chlorogenic acid (5-O-caffeoylquinic acid), caftaric acid (2-O-caffeoyltartaric acid), cichoric acid (2, 3-O-dicaffeoyltartaric acid), cynarin, and echinacoside are some of the important CADs which have varied pharmacological activities. The concentrations of these bioactive compounds are species specific and also they vary considerably with the cultivated Echinacea species due to geographical location, stage of development, time of harvest, and growth conditions. Due to these reasons, plant cell and organ cultures have become attractive alternative for the production of biomass and caffeic acid derivatives. Adventitious and hairy roots have been induced in E. pupurea and E. angustifolia, and suspension cultures have been established from flask to bioreactor scale for the production of biomass and CADs. Tremendous progress has been made in this area; various bioprocess methods and strategies have been developed for constant high-quality productivity of biomass and secondary products. This review is aimed to discuss biotechnological methods and approaches employed for the sustainable production of CADs. 相似文献
4.
Coleus forskohlii hairy root cultures were found to produce forskolin and rosmarinic acid (RA) as the main metabolites. The growth and RA production by C. forskohlii hairy root cultures in various liquid media were examined. The hairy root cultures showed good growth in hormone-free Murashige and Skoog medium containing 3% (w/v) sucrose (MS medium), and Gamborg B5 medium containing 2% (w/v) sucrose (B5 medium). RA yield reached 4.0 mg (MS medium) and 4.4 mg (B5 medium) after 5 weeks of culture in a 100 ml flask containing 20 ml of each medium. Hairy root growth and RA were also investigated after treatment with various concentrations of yeast extract (YE), salicylic acid (SA) and methyl jasmonic acid (MJA). RA production in a 100 ml flask containing 20 ml B5 medium reached 5.4 mg (1.9 times more than control) with treatment of 0.01 or 1% (w/v) YE, 5.5 mg (2.0 times more than control) with treatment of 0.1 mM SA, and the maximum RA content with 9.5 mg per flask (3.4 times more than control) was obtained in the hairy roots treated with 0.1 mM MJA. These results suggest that MJA is an effective elicitor for production of RA in C. forskohlii hairy root cultures. 相似文献
5.
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
6.
Transformation of <Emphasis Type="Italic">Saussurea medusa</Emphasis> for hairy roots and jaceosidin production 总被引:2,自引:0,他引:2
Axenically grown Saussurea medusa plantlets were inoculated with four Agrobacterium rhizogenes strains, and hairy root lines were established with A. rhizogenes strain R1601 in N6 medium. PCR and Southern hybridization confirmed integration of the T-DNA fragment of the Ri plasmid from A. rhizogenes into the genome of S. medusa hairy roots. In N6 medium, maximum biomass of the hairy root cultures was achieved [8 g (dry weight) per liter; growth ratio 35-fold] after 21 days of culture. The amount of jaceosidin extracted from the hairy root cultures was 46 mg/l (production ratio of 37-fold) after 27 days of culture. The maximum jaceosidin content obtained using N6 medium was higher than that obtained with Modified White, MS or B5 medium. In N6 medium, the tip segments were more efficient for hairy root growth and jaceosidin production than the middle and basal regions of the root.Abbreviations AS Acetosyringone - BA Benzyladenine - cef Cefotaxime sodium - DW Dry weight - FW Fresh weight - HPLC High-performance liquid chromatography - IAA Indole-3-acetic acid - km Kanamycin - NAA -Naphthaleneacetic acid - SDS Sodium dodecyl sulfate 相似文献
7.
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
8.
Harsh Pal Bais B. Suresh K. S. M. S. Rachavarao G. A. Ravishankar 《In vitro cellular & developmental biology. Plant》2002,38(6):573-580
Summary A transformed root culture of Cichorium intybus L. cv. Lucknow Local grown in different configurations of bioreactors was examined. The roots grown in an acoustic mist bioreactor
showed the best performance in terms of increased specific growth rate (0.072d−1) and esculin content (18.5gl−1), the latter of which was comparable to that of shake flask data. C. intybus hairy root cultures grown in an acoustic mist bioreactor produced nearly twice as much esculin as compared to roots grown
in bubble column and nutrient sprinkle bioreactors. Studies relating to on-line estimation of conductivity and osmolarity
to predict the growth of hairy root cultures are also discussed. The results demonstrate the efficacy and the advantages of
an acoustic mist bioreactor for the cultivation of hairy root cultures, especially with reference to C. intybus hairy roots. 相似文献
9.
The goals of this study were to investigate thidiazuron (TDZ)-induced morphogenesis of Echinacea purpurea L. and to assess the possibility of developing a liquid-based protocol for rapid micropropagation. Callus development and root organogenesis were observed on leaf explants cultured on media containing 2,4-dicholorophenoxyacetic acid or dicamba, but no plantlets were regenerated. Addition of TDZ to the culture medium as the sole growth regulator resulted in the production of regenerable callus cultures. The highest rate of regeneration was observed for explants cultured on medium with TDZ at 2.5 μM or higher. Tissue derived from 1.0 μM TDZ treatments was used to initiate liquid cultures. All liquid treatments produced a similar number of regenerants but significantly more healthy plants were obtained from cultures grown in the presence of 0.1 and 1.0 μM TDZ. This TDZ-based micropropagation system is the first liquid, large-scale propagation protocol developed for the mass production of E. purpurea plants. 相似文献
10.
Ignacio E. Maldonado-Mendoza Victor M. Loyola-Vargas 《Plant Cell, Tissue and Organ Culture》1995,40(3):197-208
Twelve different lines of Datura stramonium (normal and hairy) root cultures were subjected to conditions which induce photoautotrophy. Two of the hairy root lines responded to induction, showing clearly a diminished growth rate when compared to heterotrophic cultures, an increase in chlorophyll, a net O2 evolution, CO2 fixation, and de novo synthesis of the ribulose 1,5 biphosphate carboxylase enzyme. A time course of growth and tropane alkaloid levels in the tissue and medium, revealed a correlation between the development of the photosynthetic apparatus and the increase in scopolamine. Although normal cultures did not grow photosynthetically, they showed some greening response under the first step of the induction. The correlation between development of photosynthesis and increase in scopolamine synthesis were corroborated with normal root cultures. This experimental model is used for the basic study of the regulatory enzymes involved in the biosynthesis of tropane alkaloids, as well as for the study of their mechanisms of transport. 相似文献
11.
Induction of hairy root cultures from Gossypium hirsutum and Gossypium barbadense to produce gossypol and related compounds 总被引:2,自引:0,他引:2
Barbara A. Triplett Stephanie C. Moss John M. Bland Michael K. Dowd 《In vitro cellular & developmental biology. Plant》2008,44(6):508-517
Hairy root cultures were induced by inoculating cotyledonary leaves and hypocotyl segments from two cotton species, Gossypium hirsutum and Gossypium barbadense, with Rhizobium rhizogenes 15834. For both species, more hairy roots formed on inoculation sites on cotyledonary leaves than on hypocotyls. The addition
of sucrose to basal Murashige–Skoog media increased the frequency of hairy root formation, whereas the addition of naphthalene
acetic acid (0.54 μM) did not. After transfer to a liquid culture, hairy root growth was very rapid. After 3 wk in liquid
culture, both cotton species produced gossypol, a di-sesquiterpene secondary metabolite with known anticancer activity, and
two related methylated derivatives. Most (60–95%) gossypol produced by cultures was retained within the hairy root tissues,
but some was found in the media. The average gossypol level observed among 96 different cultures was 15 mg/g of dry culture
mass; however, some cultures produced >40 mg/g of dry culture mass. Variation in gossypol levels was greater for cultures
arising from different transformation events than for multiple subclones of a single transformant. The high level of gossypol
production attained by most of these cultures suggests that they will be valuable for studying the biochemical and molecular
aspects of gossypol biosynthesis, capable of producing large amounts of gossypol and related compounds, and useful for generating
modified forms of gossypol (e.g., radio-labeled gossypol) for understanding bioactivity mechanisms.
Mention of trade names or commercial products or vendors in this publication is solely for the purpose of providing specific
information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. 相似文献
12.
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
13.
Alkaline hydrolysis of cell wall material of tomato hairy roots yielded ferulic acid as the major phenolic compound. Other phenolics were 4-hydroxybenzoic acid, vanillic acid, 4-hydroxybenzaldehyde, vanillin and 4-coumaric acid. The content of phenolics was much higher at the early stage of hairy root growth. The ferulic acid content decreased up to 30 days and then sharply increased to 360 microg/g at 60 days of growth. Elicitation of hairy root cultures with Fusarium mat extract (FME) increased ferulic acid content 4-fold after 24 h. As the pathogen-derived elicitors have specific receptors in plants, FME may thus be used for inducing resistance against Fusarium oxysporum f. sp. lycopersici. 相似文献
14.
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
15.
Fed-batch cultures ofL. erythrorhizon hairy root were carried out by controlling sucrose concentration and media conductivity in a shake flask and a modified stirred
tank reactor. For the efficient product recovery from the culture,in situ adsorption by XAD-2 was also conducted. When sucrose was used as a carbon source, the highest shikonin production and hairy
root growth were obtained. When glucose or fructose was used instead, the growth was severely inhibited. In addition, it was
found that alternating feeding of sucrose could be used as an effective strategy for enhancing the productivity of shikonin
derivatives., As the XAD-2 amount was increased up to 1.5 g/L, shikonin production was enhanced by removing shikonin produced
and other products which might be inhibitory to cell growth. Most amount of shikonin produced was successfully recovered in
XAD-2 (Over 99%). Using hairy root culture in a modified stirred tank reactor, the shikonin productivity and hairy root growth
rate on the average were 9.34 mg/L day and 0.49 g DCW/L · day, respectively. 相似文献
16.
17.
Mondher Jaziri Kayo Yoshimatsu Jacques Homès Koichiro Shimomura 《Plant Cell, Tissue and Organ Culture》1994,38(2-3):257-262
Hairy root cultures of Atropa belladonna L. were established by infection either with Agrobacterium rhizogenes ATCC 15834 or MAFF 03-01724, and transgenic plants were obtained from both hairy root cultures. Doubly transformed roots were induced by re-infection of the leaf segments of transgenic Atropa belladonna plants (A. rhizogenes 15834) with MAFF 03-01724. Shoots and viviparous leaves were regenerated from the doubly transformed roots. The genetic transformation was determined by the opine assay (agropine, mannopine and/or mikimopine) and polymerase chain reaction. Physiological changes and tropane alkaloid biosynthesis in the hairy roots (singly and doubly transformed) were investigated. The alkaloid content in the doubly transformed root strain was intermediate as compared to the root strains which were singly transformed. On the other hand endogenous IAA levels in doubly transformed roots were significantly decreased compared to both singly transformed roots.Abbreviations BA
benzyladenine
- IAA
indoleacetic acid
- NAA
naphthaleneacetic acid
- PCR
polymerase chain reaction
-
t-ZR
trans-zeatin 相似文献
18.
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
19.
Transformed root cultures of Coluria geoides Ledeb. were established with the use of Agrobacterium rhizogenes LBA 9402. Both normal and transformed root cultures were investigated for their growth and yield of eugenol. Normal roots were grown in B5 medium-supplemented with 0.2 mg l-1 of kinetin and 0.2 mg l-1 of 1-naphthaleneacetic acid (NAA). Hairy roots grew well in hormone-free B5 medium. Both hairy roots and normal roots produced glycosidic bound eugenol. as with the roots of intact plants, eugenol was the main component of the total essential oils obtained from hairy root and normal root cultures. The yield of eugenol from normal roots was 0.1–0.25% of the dry wt. and depended on the development stage of the culture. Yield of eugenol from hairy roots was 0.08–0.1% of the dry wt. NAA modified the hairy root morphology and influenced the yield of eugenol.Abbreviations NAA
1-naphthaleneacetic acid 相似文献
20.
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