Forskolin synthesis in in vitro cultures of Coleus forskohlii Briq transformed with Agrobacterium tumefaciens.
TL;DR: F Forskolin synthesis and accumulation in tumorous C. forskohlii cultures may permit the elucidation of diterpene metabolism in this species.
Abstract: Agrobacterium tumefaciens mediated tumor tissue and shooty teratomas of Coleus forskohlii were cultured in vitro. Forskolin was detected in tumorous callus (0.002%), rhizogenic callus (0.011%) and root cultures (0.014%), but not in shooty teratomas. Forskolin synthesis and accumulation in tumorous C. forskohlii cultures may permit the elucidation of diterpene metabolism in this species.
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TL;DR: It is suggested that the use of the root endophyte fungus P. indica in sustainable agriculture will enhance the medicinally important chemical production.
Abstract: This study was undertaken to investigate the influence of plant probiotic fungus Piriformospora indica on the medicinal plant C. forskohlii. Interaction of the C. forskohlii with the root endophyte P. indica under field conditions, results in an overall increase in aerial biomass, chlorophyll contents and phosphorus acquisition. The fungus also promoted inflorescence development, consequently the amount of p-cymene in the inflorescence increased. Growth of the root thickness was reduced in P. indica treated plants as they became fibrous, but developed more lateral roots. Because of the smaller root biomass, the content of forskolin was decreased. The symbiotic interaction of C. forskohlii with P. indica under field conditions promoted biomass production of the aerial parts of the plant including flower development. The plant aerial parts are important source of metabolites for medicinal application. Therefore we suggest that the use of the root endophyte fungus P. indica in sustainable agriculture will enhance the medicinally important chemical production.
120 citations
Cites methods from "Forskolin synthesis in in vitro cul..."
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TL;DR: Suspension cultures derived from gall calli which were obtained following infection with Agrobacterium tumefaciens were established in Coleus forskohlii and showed continuous and stable production of forskolin.
Abstract: Suspension cultures derived from gall calli which were obtained following infection with Agrobacterium tumefaciens (C58) were established in Coleus forskohlii. Cell line selection following single cell cloning or cell aggregate cloning was carried out to select cell lines capable of fast growth and for producing high level of forskolin. A fast growing cell line (GSO-5/7) thus selected was found to accumulate 0.021% forskolin in 42 days. The effect of cultural conditions on cell growth was studied to identify factors influencing biomass yield. Cell growth in suspension was found to be influenced significantly by carbon source, initial cell density and light or dark condition. Optimal cell growth (20 fold increase in biomass in a 42 day period) was obtained when the cells were grown in dark condition in B5O media containing 3% sucrose as sole carbon source with an initial cell density of 1.5 x 10(5) cells per ml. Forskolin accumulation was maximum (0.021%) in the stationary phase of cell growth. These suspension cultures showed continuous and stable production of forskolin.
43 citations
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TL;DR: Transformed organ cultures of Withania somnifera established following infection with wild type nopaline and octopine strains of Agrobacterium tumefaciens found shooty teratomas of altered phenotype spontaneously developed, finding the main difference was found in the nature of the galls formed and in their subsequent morphological competence.
Abstract: Transformed organ cultures of Withania somnifera were established following infection with wild type nopaline and octopine strains of Agrobacterium tumefaciens. The oncogenic strains had different levels of virulence on two genotypes studied, the main difference was found in the nature of the galls formed and in their subsequent morphological competence. Ten percent of the galls obtained following infection with nopaline strain N2/73 spontaneously developed shooty teratomas of altered phenotype. The shooty teratomas grew in unsupplemented basal medium and were able to synthesize both the major withanolides of the parent plants. Withanolide synthesis in shooty teratomas was much higher (0.07–0.1% withaferin A and 0.085–0.025% withanolide D) than in non-transformed shoot cultures.
37 citations
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TL;DR: Transformed organ cultures of Artemisia annua were established following infection with two wild type nopaline strains of Agrobacterium tumefaciens: Parameters like explant type, strain type, age of the plant source for explants, affected the tumorigenesis frequency significantly.
Abstract: Transformed organ cultures of Artemisia annua were established following infection with two wild type nopaline strains of Agrobacterium tumefaciens . Parameters like explant type, strain type, age of the plant source for explants, affected the tumorigenesis frequency significantly. Crown galls were formed both on in vivo and in vitro plants: 2–3% of the in vitro galls regenerated spontaneously to produce shooty teratoma of altered phenotype. Artemisinin contents were measured in all transformed as well as non-transformed clones. While shooty teratomas synthesized 0.063 ± 0.002 g/100 g DW artemisinin, non-transformed shoots were found to synthesize only 0.0179 ± 0.002 g/100 g DW of the compound.
29 citations
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TL;DR: The significance of the present micropropagation protocol of C. forskohlii is the formulation of growth regulators which effected very fast multiplication of the plant (time reduced to one-third of the hitherto known methods).
Abstract: A protocol has been developed that leads to the development of complete plantlets of Coleus forskohlii within 35–40 d by culturing stem tip explants in MS medium containing 0.57 μM indole-3-acetic acid and 0.46 μM kinetin through direct multiplication at the rate of 12.5 shoots per explant. About 100% shoots rooted and micropropagated plants were successfully established in soil after hardening with a high survival rate. The significance of the present micropropagation protocol of C. forskohlii is the formulation of growth regulators which effected very fast multiplication of the plant (time reduced to one-third of the hitherto known methods).
27 citations
References
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TL;DR: In vivo redox biosensing resolves the spatiotemporal dynamics of compartmental responses to local ROS generation and provide a basis for understanding how compartment-specific redox dynamics may operate in retrograde signaling and stress 67 acclimation in plants.
Abstract: In experiments with tobacco tissue cultured on White's modified medium (basal meditmi hi Tnhles 1 and 2) supplemenk'd with kiticthi and hidoleacctic acid, a slrikin^' fourlo (ive-told intTease iu yield was ohtaitu-d within a three to Tour week j^rowth period on addition of an aqtteotis exlrarl of tobacco leaves (Fi^'ures 1 and 2). Subse(iueutly it was found Ihiit this jnoniotiou oi' f^rowih was due mainly though nol entirely to inorj^auic rather than organic con.stitttenls in the extract. In the isolation of Rrowth factors from plant tissues and other sources inorj '̂anic salts are fre(|uently carried along with fhe organic fraclioits. When tissue cultures are used for bioassays, therefore, il is necessary lo lake into account increases in growth which may result from nutrient elements or other known constituents of the medium which may he present in the te.st materials. To minimize interference trom rontaminaitis of this type, an altempt has heen made to de\\eh)p a nieditmi with such adequate supplies of all re(iuired tnineral nutrients and cotntnott orgattic cottslitueitls that no apprecial»le change in growth rate or yield will result from the inlroduclion of additional amounts in the range ordinarily expected to be present in tnaterials to be assayed. As a point of referetice for this work some of the culture media in mc)st common current use will he cotisidered briefly. For ease of comparis4)n Iheir mineral compositions are listed in Tables 1 and 2. White's nutrient .solution, designed originally for excised root cultures, was based on Uspeuski and Uspetiskaia's medium for algae and Trelease and Trelease's micronutrieni solution. This medium also was employed successfully in the original cttltivation of callus from the tobacco Iiybrid Nicotiana gtauca x A', tanijadorffii, atitl as further modified by White in 194̂ ^ and by others it has been used for the
60,055 citations
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TL;DR: The nutrient requirements of suspension cultures from soybean root have been investigated, and a simple medium consisting of mineral salts, sucrose, vitamins and 2,4-dichlorophenoxyacetic acid (2, 4- d) has been designed.
Abstract: The nutrient requirements of suspension cultures from soybean root have been investigated, and a simple medium consisting of mineral salts, sucrose, vitamins and 2,4-dichlorophenoxyacetic acid (2,4- d ) has been designed. The cells required thiamine, 2,4- d and ammonium in addition to the usual mineral salts and sucrose. Optimum concentrations of nitrate and ammonium were 25 and 2 m M respectively. The highest yield of cells was achieved at an initial pH of 4.5–5.5. During the growth cycle the pH gradually increased to 6.0–6.2.
9,034 citations
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TL;DR: Comparison of T-DNA-encoded transcripts present in crown gall tumors showing teratoma-like growth (BT37) with those from an unorganized tumor line (W38C58) reveals that this difference in phenotype is accompanied by a difference in the expression of the T- DNA.
Abstract: Up to thirteen T-DNA-encoded, polyadenylated transcripts of different relative abundance were detected by Northern blot hybridization in the tobacco nopaline BT37 crown gall teratoma tissue Their sizes range from 900 to 2,700 bases The polarity of eight of the thirteen transcripts was assigned by hybridization of labeled RNA to single-stranded DNA fragments of the T-region obtained by cloning in an M13 vector Both strands of the T-DNA are transcribed Our data indicate that most, if not all, transcripts are generated via independent promoter and poly(A)-addition sites on the T-DNA Comparison of T-DNA-encoded transcripts present in crown gall tumors showing teratoma-like growth (BT37) with those from an unorganized tumor line (W38C58) reveals that this difference in phenotype is accompanied by a difference in the expression of the T-DNA T-DNA sequences common to both octopine and nopaline tumors encode at least five, and probably six, cross-hybridizing transcripts of the same size, location, polarity and function These transcripts are involved in the process of plant tumor formation and maintenance
154 citations
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TL;DR: Agrobacterium tumefaciens strains, known to induce tobacco crown galls that spontaneously develop shoots, were used to induce galls on cultured shoots of a tetraploid potato cultivar and most of the tumour-derived shoots formed roots, did not produce opines and were indistinguishable from the parental plants on the basis of morphology and chromosome numbers.
Abstract: Agrobacterium tumefaciens strains, known to induce tobacco crown galls that spontaneously develop shoots, were used to induce galls on cultured shoots of a tetraploid potato cultivar (Solanum tuberosum cv. ‘Maris Bard’). Shoots also appeared spontaneously from the induced potato galls, although only after 2–4 months. The shoots were excised and cultured separately. Some of these frequently developed side-shoots from their axillary buds. They did not form roots and they produced opines, a strong indication that they were transformed and carried T-DNA. Grafts of the transformed plants were still able to develop tubers. Most of the tumour-derived shoots, however, formed roots, did not produce opines and were indistinguishable from the parental plants on the basis of morphology and chromosome numbers (48 chromosomes per cell). The results are discussed in relation to the origin of previously described variation among protoplast-derived potato plants and with respect to genetic engineering of tetraploid potato cultivars.
54 citations
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TL;DR: Root cultures of COLEUS FORSKOHLII Briq.
Abstract: Root cultures of COLEUS FORSKOHLII Briq. were initiated from primary callus or IBA-treated suspension cultures and maintained on Gamborg's B5 medium containing 1 mg/l IBA. Transformed root cultures were established by infecting surface-sterilized leaves with AGROBACTERIUM RHIZOGENES strain 15834. Transformation was confirmed by mannopine detection. These cultures displayed the typical characteristics of hairy root cultures, with the sole exceptions of slow growth in hormone-free medium and accelerated growth on medium containing phytohormones. All root cultures examined formed forskolin and its derivatives in amounts ranging from 500 to 1300 mg/kg dry weight, corresponding to about 4 to 5 mg/l. During cultivation roots could be cut into small pieces without affecting growth and forskolin production. Scale-ups of the cultivation procedure were performed in 20-l glass jars with a working volume of 10 to 13l. Forskolin production in bioreactors was better than in shake flasks. Levels of almost 14 mg/l could be reached after 21 d of cultivation. As in the shake flask experiments cutting the roots did not affect growth or productivity in a negative way.
34 citations