Callus formation and plant regeneration from Hypericum perforatum leaves
01 Aug 2000-Plant Cell Tissue and Organ Culture (Kluwer Academic Publishers)-Vol. 62, Iss: 2, pp 107-113
TL;DR: The procedure reported here allows the micropropagation of H. perforatum in five months of culture and the proliferation of cell masses which could be used for studies on organic compounds of pharmaceutical interest.
Abstract: Use of Hypericum perforatum L. has increased in the past few years due to the antidepressant and antiviral activities found in extracts of this plant. As a result of its potential as a pharmaceutical, a new system was developed for in vitro culture of this species. Leaf explants were inoculated onto MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D, 0.45 or 4.5 μM) and 6-benzyladenine (BA, 0.44 or 4.4 μM) or kinetin (0.46 or 4.6 μM). Explants were cultivated under dark or light conditions to induce callus formation. Callus initiation was observed in all media evaluated and the highest cell proliferation was obtained from explants cultivated in the presence of 4.4 μM BA and 4.5 μM 2,4-D in the dark. Shoot induction was obtained from callus induced on 4.6 μM kinetin and 0.45 μM 2,4-D 6 weeks after transferring the callus to a MS medium supplemented with 4.4 μM BA. Roots were induced from shoots on full and half-strength MS media with or without indolebutyric acid (IBA, 4.9 μM) and the highest rooting frequencies were obtained on half-strength MS medium, regardless of the presence of IBA. Regenerated plants were easily acclimated in greenhouse conditions. The procedure reported here allows the micropropagation of H. perforatum in five months of culture and the proliferation of cell masses which could be used for studies on organic compounds of pharmaceutical interest.
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TL;DR: A dramatic increase in cell growth and hypericin production was observed after exposure to jasmonic acid (JA), however, other elicitors such as salicylic acid (SA) and fungal cell wall elicitors failed to show any stimulatory effect on either cell growth or hyperic in production.
153 citations
Cites background from "Callus formation and plant regenera..."
...It has previously been reported that both light conditions and growth regulator treatment influence the growth of cell cultures of H. perforatum in static media ( Pretto and Santarem, 2000 )....
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TL;DR: The results show that St. John's wort can be efficiently regenerated from root explants in a variety of bioreactors for aseptic production of bioactive compounds.
86 citations
TL;DR: Results indicated that the production of hypericin and pseudohypericin could be increased by carefully adapted in vitro cultures.
78 citations
Cites background from "Callus formation and plant regenera..."
...Accordingly, micropropagation of Hypericum has been reported through shoot regeneration from leaves [28], hypocotyls [23], shoot cuttings [5], stamens [17] and roots [38]....
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TL;DR: A system that could lower the cost of in vitro propagation by using liquid medium, as well as to evaluate the secondary metabolism in the systems tested, was established.
Abstract: Hypericum perforatum L. is a medicinal plant that has been extensively studied because of its bioactive properties. The objective of this study was to establish a system that could lower the cost of in vitro propagation by using liquid medium, as well as to evaluate the secondary metabolism in the systems tested. Nodal segments of H. perforatum were obtained from in vitro shoots and grown in three liquid culture systems: total immersion (TI), partial immersion (PI), and paper bridge support (PB). Semi-solid medium (3 g L−1 Phytagel™) was used as control (SS). The organogenic responses were evaluated, and phenolic compounds, hypericin, and the activity of polyphenol oxidases (PPO) and peroxidases (POX) were quantified. After 80 days of culture, induction and proliferation of adventitious shoots were similar in the PI and SS systems (65.3 and 71.3 shoots, respectively), whereas PB resulted in the fewest shoots per explant (29.5 shoots). Longer shoots were obtained under the PI conditions. Hyperhydricity was observed in the shoots from the TI system. Browning was visible in shoots from the TI and PB systems. The highest concentrations of phenolic compounds and hypericin were observed in shoots derived from PI and PB, at 80 days of culture. POX activity was higher in shoots cultured in PI at 40 days, whereas PPO was significantly more active at 80 days of culture. Likely, POX was more related to shoot growth, whereas PPO played a later role in response to the culture environment and medium stress.
71 citations
Cites background from "Callus formation and plant regenera..."
...Keywords Hypericin Liquid cultures Micropropagation Phenolic compounds Polyphenol oxidases Peroxidases...
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TL;DR: An efficient and reproducible protocol for high-frequency callus regeneration from immature leaf explants of T. indica was developed, and Histological studies of the regenerative callus revealed that shoot buds were originated from the outermost regions.
Abstract: Tylophora indica (Burm. f.) Merrill is a threatened medicinal climber distributed in the forests of northern and peninsular India. An efficient and reproducible protocol for high-frequency callus regeneration from immature leaf explants of T. indica was developed. Organogenic callus formation from immature leaf pieces was obtained by using Murashige and Skoog (MS) medium supplemented with 7 μM 2,4-dichlorophenoxyacetic acid and 1.5 μM 6-benzyladenine. On this medium 92% explants produced callus. The optimal hormone combination for plantlet regeneration was 8 μM thidiazuron, at which shoot regeneration was obtained from 100% of the cultures, with an average of 66.7 shoots per culture. Histological studies of the regenerative callus revealed that shoot buds were originated from the outermost regions. For root formation, half-strength MS medium supplemented with 3 μM indole-3-butyric acid was used. Plants were transferred to soil, where 92% survived after 3 mo. of acclimatization.
63 citations
Cites background from "Callus formation and plant regenera..."
...The promoting effect of auxin and cytokinin combinations on organogenic differentiation has been well documented (Lisowska and Wysokinska, 2000; Pereira et al., 2000; Pretto and Santarém, 2000; Koroch et al., 2002)....
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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
63,098 citations
"Callus formation and plant regenera..." refers methods in this paper
...The callus initiation medium contained MS salts and vitamins (Murashige and Skoog, 1962), 30 g l−1 sucrose and 3 g l−1 PhytagelTM....
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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.
9,342 citations
"Callus formation and plant regenera..." refers methods in this paper
...liense, the combination of cytokinins and auxins did not support callus growth and calluses were only obtained in the presence of 2,4D or NAA using either MS or B5 medium (Gamborg et al., 1968) under a 16-h photoperiod (Cardoso and Oliveira, 1996)....
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...D or NAA using either MS or B5 medium (Gamborg et al., 1968) under a 16-h photoperiod (Cardoso and Oliveira, 1996)....
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Book•
01 Aug 1991
TL;DR: This chapter discusses measurement and Sampling concepts, measurement practice, and the Basis of Statistical Testing as well as measuring Variability, Probability, and Correlations.
Abstract: Measurement and Sampling Concepts. Processing Data. Presenting Data. Measuring the Average. Measuring Variability. Probability. Probability Distributions as Models of Dispersion. The Normal Distribution. Data Transformation. How Good are Our Estimates? The Basis of Statistical Testing. Analysing Frequencies. Measuring Correlations. Regression Analysis. Comparing Averages. Analysis of Variance - ANOVA. Multivariate Analysis. Appendices. Bibliography and Further Reading. Index.
1,621 citations
01 Jan 1982
333 citations
"Callus formation and plant regenera..." refers background in this paper
...It has been reported that tissues cultured in the presence of light have increased activity of IAA-oxidase altering the endogenous balance between auxin/cytokinin and consequently decreasing the callus growth (Gaspar et al., 1982)....
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TL;DR: The data support the inhibition of arachidonic acid metabolism as one of the mechanisms by which flavonoids exert their anti-inflammatory effects.
Abstract: A group of flavonoids isolated from medicinal plants and which are selective inhibitors of lipoxygenase activityin vitro: sideritoflavone, cirsiliol, hypolaetin-8-O-β-d-glucoside, hypolaetin, oroxindin, quercetagetin-7-O-β-d-glucoside, gossypin, hibifolin and gossypetin, besides leucocyanidol, have been studied for their effects on acute responses induced by carrageenin in mice. The oral administration of flavonoids to mice inhibited dose-dependently the development of paw oedema at 1, 3 and 5 h after carrageenin injection. A similar administration of flavonoids induced a dose-dependent inhibition of leukocyte accumulation in inflammatory exudates following intraperitoneal injection of carrageenin into mice. Some of the flavonoids exhibited a potency against leukocyte infiltration similar to that seen for inhibition of carrageenin oedema at 3 h of induction. In agreement with data reported in rats, indomethacin was much more effective on inhibition of prostaglandin E2 (PGE2) formation than on leukocyte infiltration in mice. The selectivity of flavonoids towards lipoxygenase is not retainedin vivo since they behave as dual inhibitors of PGE2 and leukotriene B4 (LTB4) formation in peritoneal exudates. Our data support the inhibition of arachidonic acid metabolism as one of the mechanisms by which flavonoids exert their anti-inflammatory effects.
312 citations
"Callus formation and plant regenera..." refers background in this paper
...Species of the genus Hypericum have been used as traditional medicinal plants due to their wound healing (Yazaki and Okuda, 1990; Maisenbacher and Kovar, 1992), bactericide (Ishiguro et al., 1998), anti-inflammatory ( Fernandez and Alcaraz, 1991; Dias et al., 1998), diuretic and sedative properties (Holzl and Ostrowiski, 1987)....
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