Apoptosis‐inducing effects of Morinda citrifolia L. and doxorubicin on the Ehrlich ascites tumor in Balb‐c mice

Morinda citrifolia L. (Noni) is a herbal remedy with promising anti‐cancer properties. However, its effects on various cancers are to be investigated to make a firm conclusion before implementing it into the clinical practice. Therefore, we investigated the cytotoxic potential of noni on Ehrlich ascites tumor grown in female Balb‐c mice and also combined it with a potent anti‐cancer agent, doxorubicin. One group received noni only (n = 8), another one doxorubicin (n = 8), and the other one noni + doxorubicin (n = 8) for 14 days after the inoculation of cells. The control group (n = 7) received 0.9% NaCl only. We found that short and long diameters of the tumor tissues were about 40–50% smaller, compared to those in control group. This anti‐growth effect resulted from the induction of apoptosis, which was confirmed by the positive results from the Terminal Deoxynucleotidyl Transferase dUTP Nick End Labeling (TUNEL) analysis and the active caspase‐3 cells in tissues. Apoptosis also confirmed by caspase‐cleaved cytokeratin 18 elevation in serum of the treated groups. Further, the proliferation was decreased, which was immunohistochemically shown by the PCNA staining. We conclude that noni may be useful in the treatment of breast cancer either on its own or in combination with doxorubicin. Further studies are warranted to assess the dosage and safety of using noni fruit juice in conjuction with anti‐cancer drugs against breast cancer. Copyright © 2009 John Wiley & Sons, Ltd.     

Citrus tissue culture: stimulation of fruit explant cultures with orange juice

In vitro growth of explant (juice vesicle or albedo tissues) cultures from citron (Citrus medica), lemon (C. limon), grapefruit (C. paradisi), sweet orange (C. sinensis), and mandarin (C. reticulata) fruits was stimulated by addition of orange juice (10% v/v optimum) to a basal medium containing Murashige and Skoog salts, 50 grams per liter sucrose, 100 milligrams per liter myo-inositol, 5 milligrams per liter thiamine·HCl, 2 milligrams per liter 2,4-dichlorophenoxyacetic acid and 0.5 milligrams per liter kinetin. In analyzing this effect of orange juice on citron explant cultures, we failed to obtain increased yields by addition of appropriate concentrations of citric acid to the basal medium but obtained growth stimulation when the medium was supplemented with juice from an “acidless” orange variety (cv. Lima). These facts suggest that some component(s) other than citric acid is involved. Addition of the inorganic ash corresponding to 10% (v/v) orange juice to the basal medium had no effect on yields. Similarly, the stimulatory effect of orange juice could not be explained based on its content of sucrose or of organic growth factors already present in the basal medium.     

Determination and comparative analysis of major iridoids in different parts and cultivation sources of Morinda citrifolia

Introduction – Noni is a medicinal plant with a long history of use as a folk remedy in many tropical areas, and is attracting more attention worldwide. A comprehensive study on the major phytochemicals in different plant parts (fruit, leaf, seed, root and flower) and sources is of great value for fully understanding their diverse medicinal benefits. Objective – To quantitatively determine the major iridoid components in different parts of noni plants, and compare iridoids in noni fruits collected from different tropical areas worldwide. Methodology – The optimal chromatographic conditions were achieved on a C₁₈ column with gradient elution using 0.1% formic acid aqueous formic acid and acetonitrile at 235 nm. The selective HPLC method was validated for precision, linearity, limit of detection, limit of quantitation and accuracy. Results – Deacetylasperulosidic acid (DAA) was found to be the major iridoid in noni fruit. In order of predominance, DAA concentrations in different parts of the noni plant were dried noni fruit > fruit juice > seed > flower > leaf > root. The order of predominance for asperulosidic acid (AA) concentration was dried noni fruit > leaf > flower > root > fruit juice > seed. DAA and AA contents of methanolic extracts of noni fruits collected from different tropical regions were 13.8–42.9 and 0.7–8.9 mg/g, respectively, with French Polynesia containing the highest total iridoids and the Dominican Republic containing the lowest. Conclusion – Iridoids DAA and AA are found to be present in leaf, root, seed and flower of noni plants, and were identified as the major components in noni fruit. Given the great variation of iridoid contents in noni fruit grown in different tropical areas worldwide, geographical factors appear to have significant effects on fruit composition. The iridoids in noni fruit were stable at the temperatures used during pasteurisation and, therefore, may be useful marker compounds for identity and quality testing of commercial noni products. Copyright © 2010 John Wiley & Sons, Ltd.     

Preliminary results on in vitro rearing of the endoparasitoid Cardiochiles nigriceps from egg to second instar

The composition of an artificial medium and technical procedures used for in vitro rearing of the endophagous larval parasitoid Cardiochiles nigriceps Viereck (Hymenoptera, Braconidae), from post-germ band egg to the 2nd instar larva, are described. Amino acids, carbohydrates, salts, and vitamins were supplied in defined amounts as an aqueous solution which, when supplemented with 20 mg/ml of bovine albumin, 5 mg/ml of lactalbumin (enzymatic hydrolysate), 20% (v/v) fetal bovine serum, 20% (v/v) milk and 10% (v/v) chicken egg yolk, allowed for parasitoid growth and molting to the 2nd instar. Molting to the final instar was never observed.     

Immobilization of tomato (Lycopersicon esculentum) pectinmethylesterase in calcium alginate beads and its application in fruit juice clarification

Clarity of fruit juices is desirable to maintain an aesthetically pleasing quality and international standards. The most commonly used enzymes in juice industries are pectinases. A partially-purified pectinmethylesterase from tomato was entrapped in calcium alginate beads and used for juice clarification. The activity yield was maximum at 1 % (w/v) CaCl₂ and 2.5 % (w/v) alginate. The immobilized enzyme retained ~55 % of its initial activity (5.7 × 10^?2 units) after more than ten successive batch reactions. The K_m, pH and temperature optima were increased after immobilization. The most effective clarification of fruit juice (%T₆₂₀ ~60 %) by the immobilized enzyme was at 4 °C with a holding time of 20 min. The viscosity dropped by 56 % and the filterability increased by 260 %. The juice remains clear after 2 months of storage at 4 °C.     

Some Factors Influencing the in vitro Infectivity and Replication of Encephalitozoon cuniculi*

SYNOPSIS. Rabbit Encephalitozoon cuniculi were propagated in vitro using rabbit choroid plexus (RCP) cells. The organisms reached maximum titer and numbers by 15 days. The source and in vitro passage level of RCP cells moderately influenced the sensitivity of the cells to infection. Cells less than 1 week old were significantly less sensitive than older cells. A moderate increase in infectivity for RCP cells was demonstrated with increasing organism passage level in vitro. Rabbit E. cuniculi were not affected by penicillin-streptomycin or gentamicin in the culture medium. The organism survived more than 9 days in buffer at 37 C and least 24 days at 4 and 20 C. Storage at -70 C or in liquid nitrogen was successful for at least 6 months. Encephalitozoon cuniculi survived 60 but not 120 min at 56 C. They were killed after 10 min of autoclaving and by 2% (v/v) Lysol, 10% (v/v) formalin and 70% (v/v) ethyl alcohol. The organisms survived at least 24 h at pH 9 or pH 4 and were not affected by sonication, freezing and thawing, or distilled water but lost significant infectivity after 24 h in CsCl or 40% (w/v) sucrose.     

Stability ofAlternaria toxins in fruit juices and wine

Alternaria alternata is a common fungal parasite on fruits and other plants and produces a number of mycotoxins, including alternariol (3,7,9-trihydroxy-1-methyl-6H-dibenzo [b,d]pyran-6-one), alternariol monomethyl ether (3,7-dihydroxy-9-methoxy-1-methyl-6H-dibenzo[b,d]pyran-6-one), and the mutagen altertoxin I {[1S-(1α,12aβ,12bα)] 1,2,11,12,12a, 12b-hexahydro-1,4,9,12a-tetrahydroxy-3,10-perylenedione}. Alternariol and alternariol monomethyl ether have previously been detected in some samples of fruit beverages. Stability studies of these toxins as well as altertoxin I added to fruit juices and wine (10–100 ng/mL) were carried out. To include altertoxin I in the analysis, cleanup with a polymer-based Varian Abselut solid phase extraction column was used, as recoveries from C-18 columns were low. The stabilities of alternariol and alternariol monomethyl ether in a low acid apple juice containing no declared vitamin C were compared with those in the same juice containing added vitamin C (60 mg/175 ml); there were no apparent losses at room temperature over 20 days or at 80°C after 20 min. in either juice. Altertoxin I was moderately stable in pH 3 buffer (75% remaining after a two week period). Furthermore, altertoxin I was stable or moderately stable in three brands of apple juice tested over 1–27 day periods and in a sample of red grape juice over 7 days. It is concluded that altertoxin I is sufficiently stable to be found in fruit juices and should be included in methods for alternariol and alternariol monomethyl ether.     

Momordica charantia fruit juice stimulates glucose and amino acid uptakes in L6 myotubes

The fruit of Momordica charantia (family: Cucurbitacea) is used widely as a hypoglycaemic agent to treat diabetes mellitus (DM). The mechanism of the hypoglycaemic action of M. charantia in vitro is not fully understood. This study investigated the effect of M. charantia juice on either ³H-2-deoxyglucose or N-methyl-amino-a-isobutyric acid (¹⁴C-Me-AIB) uptake in L6 rat muscle cells cultured to the myotube stage. The fresh juice was centrifuged at 5000 rpm and the supernatant lyophilised. L6 myotubes were incubated with either insulin (100 nM), different concentrations (1–10 g ml^–1) of the juice or its chloroform extract or wortmannin (100 nM) over a period of 1–6 h. The results were expressed as pmol min^–1 (mg cell protein)^–1, n= 6–8 for each value. Basal ³H-deoxyglucose and ¹⁴C-Me-AIB uptakes by L6 myotubes after 1 h of incubation were (means ± S.E.M.) 32.14 ± 1.34 and 13.48 ± 1.86 pmol min^–1 (mg cell protein)^–1, respectively. Incubation of L6 myotubes with 100 nM insulin for 1 h resulted in significant (ANOVA, p < 0.05) increases in ³H-deoxyglucose and ¹⁴C-Me-AIB uptakes. Typically, ³H-deoxyglucose and ¹⁴C-Me-AIB uptakes in the presence of insulin were 58.57 ± 4.49 and 29.52 ± 3.41 pmol min^–1 (mg cell protein^–1), respectively. Incubation of L6 myotubes with three different concentrations (1, 5 and 10 g ml^–1) of either the lyophilised juice or its chloroform extract resulted in time-dependent increases in ³H-deoxy-D-glucose and ¹⁴C-Me-AIB uptakes, with maximal uptakes occurring at a concentration of 5 g ml^–1. Incubation of either insulin or the juice in the presence of wortmannin (a phosphatidylinositol 3-kinase inhibitor) resulted in a marked inhibition of ³H-deoxyglucose by L6 myotubes compared to the uptake obtained with either insulin or the juice alone. The results indicate that M. charantia fruit juice acts like insulin to exert its hypoglycaemic effect and moreover, it can stimulate amino acid uptake into skeletal muscle cells just like insulin. (Mol Cell Biochem 261: 99–104, 2004)     

In vitro culture of vanhoutte's spirea explants from ‘secondary cultures’ and dormant stems forced in solutions containing plant growth regulators

Explants from new growth of forced dormant stems and secondary cultures of Vanhoutte's spirea were cultured on Linsmaier and Skoog (L.S.) medium containing benzyladenine (BA), indoleacetic acid (IAA), thidiazuron (TDZ), and zeatin. The dormant stems were forced by immersing their basal portions in forcing solutions containing 626 µM 8-hydroxyquinoline citrate (8-HQC) and 2% sucrose. BA and gibberellic acid (GA₃) were also added into the forcing solutions to determine if explants obtained from the new growth will benefit from this treatment when culturedin vitro.L.S. medium supplemented with 5 µM BA alone, 5 µM BA plus 1 or 5 µM IAA, and 0.5 or 0.75 µM TDZ alone produced the best shoot proliferation for both sources of explants. BA and GA₃ appeared to be taken up from the forcing solution by the new softwood growth. BA in the forcing solution stimulatedin vitro shoot proliferation in different degrees depending on the period of treatment, while GA₃ caused lessin vitro shoot proliferation. It is proposed that forcing solutions containing plant growth regulators (P.G.R.) are a useful approach for manipulating responses of plant tissues culturedin vitro.     

Relative importance of maltose and sucrose supplied during a 2-step potato microtuberization process

A 2-stage in vitro tuberization process comprising first micropropagation via nodal explants and then tuber induction in the resultant in vitro plantlets was studied using 2 cultivars of potato, Iwa and Daeji. In particular, the effects on both plantlet growth and subsequent in vitro tuberization of Murashige and Skoog (1962) basal medium containing either sucrose or maltose, each at 3 % (w/v), used for micropropagation were investigated. Sucrose and maltose were found to be equally effective in supporting development of vigorous plantlets from the nodal explants of both potato cultivars. Upon transfer to a medium with an optimised level of sucrose (i.e. 8 %, w/v) for in vitro tuberization, only the plantlets previously grown in the sucrose-containing medium were capable of forming more microtubers of the larger size category (greater than 0.5 g). The relative importance of sucrose supply at the mircropropagation stage was further confirmed when the resultant plantlets grown in the 3 % sucrose-containing medium were transferred to an in vitro tuberization medium containing either sucrose or maltose, each at 8 % (w/v). In this experiment, maltose and sucrose had indistingushable effects on in vitro tuberization.     

A simple,economical, and high efficient protocol to produce in vitro miniature rose

Due to its high commercial value, many studies on rose (Rosa hybrida L.) micropropagation have been published. However, there are a limited number of studies on rose in vitro flowering. These studies only focused on the roles of plant growth regulators in the formation and morphogenesis of flowers. In this protocol, cytokinin was confirmed to positively function in the induction of in vitro rose flowers. In fact, more than 40% of in vitro shoots were induced to flower when they were grown on a medium supplemented with benzylaminopurine (BA) (2 mg L⁻¹) and IAA (0.1 mg L⁻¹). In addition, this study showed that the growth medium supplemented with only coconut water (15 or 20% v/v) was very efficient to induce flowering of in vitro miniature rose plants (> 70%) after 60 d of subculture. In addition, the in vitro flowers were normal and almost similar to ex vitro flowers in terms of flower shape and color. Based on these results, a detailed procedure for in vitro miniature rose flower production is provided.

     

Seed cryopreservation and micropropagation of the federally threatened species,Price’s potato-bean (Apios priceana B.L. Robins.)

Apios priceana, commonly known as Price’s potato-bean, is a perennial species native to the Southeastern US. Habitat destruction has caused A. priceana to become rare, and it is federally listed as threatened. Protocols developed for in vitro germination, shoot micropropagation, in vitro rooting, shoot establishment in soil, and seed cryopreservation will assist in the safeguarding and conservation of dwindling natural populations. Seeds were germinated in vitro on plant growth regulator (PGR)-free Murashige and Skoog (MS) medium after seed sterilization in H₂O₂ and seed nicking. Greatest shoot multiplication occurred on MS medium with BAP/IBA/GA₃ at 2.22/0.49/1.44 μM with 2.0 g/l Phytagel and pH adjusted to 5.7. Shoots rooted in vitro in MS medium with 3.2 μM IBA. Shoots rooted in vitro rapidly established in greenhouse potting mix, usually showing new growth within 2 wk, with tuber formation by the end of the growing season. Plants transferred to a forest setting in late winter survived, grew throughout the summer, and became dormant in the fall. In small post cryopreservation tests with A. americana and A. priceana seeds, air or desiccant-dried to water contents below 10% but above 2.5%, germination reached 87–90%.

     

Single cell-produced and in vitro-assembled anti-FcRH5/CD3 T-cell dependent bispecific antibodies have similar in vitro and in vivo properties

Bispecific antibody production using single host cells has been a new advancement in the antibody engineering field. We previously showed comparable in vitro biological activity and in vivo mouse pharmacokinetics (PK) for two novel single cell variants (v10 and v11) and one traditional dual cell in vitro-assembled anti-human epidermal growth factor receptor 2/CD3 T-cell dependent bispecific (TDB) antibodies. Here, we extended our previous work to assess single cell-produced bispecific variants of a novel TDB against FcRH5, a B-cell lineage marker expressed on multiple myeloma (MM) tumor cells. An in vitro-assembled anti- FcRH5/CD3 TDB antibody was previously developed as a potential treatment option for MM. Two bispecific antibody variants (designs v10 and v11) for manufacturing anti-FcRH5/CD3 TDB in single cells were compared to in vitro-assembled TDB in a dual-cell process to understand whether differences in antibody design and production led to any major differences in their in vitro biological activity, in vivo mouse PK, and PK/pharmacodynamics (PD) or immunogenicity in cynomolgus monkeys (cynos). The binding, in vitro potencies, in vitro pharmacological activities and in vivo PK in mice and cynos of these single cell TDBs were comparable to those of the in vitro-assembled TDB. In addition, the single cell and in vitro-assembled TDBs exhibited robust PD activity and comparable immunogenicity in cynos. Overall, these studies demonstrate that single cell-produced and in vitro-assembled anti-FcRH5/CD3 T-cell dependent bispecific antibodies have similar in vitro and in vivo properties, and support further development of single-cell production method for anti-FcRH5/CD3 TDBs and other single-cell bispecifics.     

基于ISSR技术研究诺丽种质资源的亲缘关系

为探讨我国诺丽(Morinda citrifolia)种质资源的亲缘关系,采用ISSR技术对诺丽种质资源的遗传关系进行分析。结果表明,10条ISSR引物对13份诺丽种质资源共扩增出183条带,其中多态性条带有159条,占86.9%。13份诺丽种质的遗传相似系数为0.464~0.784。聚类分析将13份诺丽种质资源聚为两类,其中诺丽小黑种单独聚为一类,与其他12份诺丽种质资源的亲缘关系较远。虽然按照外部形态特征不能将所有诺丽种质完全区分,但具有相同特征的多数种质还是聚在同一类或亚类中。     

The path of photosynthate translocation into citrus fruit

Abstract The path of [¹⁴C]photosynthate translocation into citrus fruit was examined to determine which anatomical and physiological features were involved in this process. Experiments were conducted during the final pre-harvest months of 2 years grapefruit crops (Citrus paradisi Macf. cv. ‘Marsh’). A source leaf nearest the fruit was exposed to ¹⁴CO₂ for 1 h + 5 h ambient air, followed by dissection of vascular and phloem-free tissues in the fruit quarter directly aligned with the source. Radioactivity in each tissue was quantified after separation and extraction in boiling 80% ethanol. Peel (flavedo+albedo) contained an average 35% of the label in the quarter fruit, but an additional 20% was localized entirely in dorsal vascular bundles along exterior walls of juice segments. Less [¹⁴C]photosynthate was recovered from other vascular tissues and was nearly absent from adjacent mature seeds. Radioactivity in the single layer of segment epidermis, however, averaged 17% of that in the quarter fruit. Juice tissues interior to this accumulated only 17% of the total. No phloem tissue was evident in either the segment epidermis or juice tissues, but over 70% of the [¹⁴C]assimilates in the latter were localized in thread-like stalks which attach juice vesicles to dorsal vascular bundles. In addition, labelled hexose/sucrose ratios in these structures increased with distance from the vascular bundle. The majority of photosynthates, therefore, entered citrus fruit via dorsal vascular bundles and were partially hydrolysed during slow transfer through non-vascular segment epidermis and juice stalks.     

Use of alfalfa residual juice as a substrate for propagation of the red yeast Phaffia rhodozyma

Summary Alfalfa residual juice (ARJ) supported good growth of the yeast Phaffia rhodozyma but formation of astaxanthin was inhibited. Supplementary nutrients did not reverse the inhibition, indicating that the the juice probably contained some inhibitor of astaxanthin biosynthesis. Six strains of P. rhodozyma were tested and found to be susceptible to the inhibitory effects of the juice. Concentrations of ARJ above 1.25% (v/v) were inhibitory to pigmentation of the yeast. Above approximately 3.7%, total inhibition of astaxanthin formation was observed but some chromogenic components of the juice were adsorbed on Phaffia cells and appeared as artefacts in astaxanthin analyses. Phaffia biomass produced in ARJ showed greater susceptibility to autolysis than that produced in a peptone-glucose-salts medium. Supplementation of ARJ with glucose enhanced yield of cell mass and minimised the autolytic phenomenon, and is potentially useful for producing Phaffia biomass for use as a source of single cell protein.Unsupplemented brewer's malt wort and molasses, separately and in a suitable combination, were compared with ARJ and were found suitable for growth and pigmentation of P. rhodozyma.     

In situ and in vitro suppressive effect of agricultural composts and their water extracts on some phytopathogenic fungi

In situ and in vitro experiments were carried out to determine the effect of various composts (leafy fruit compost (LFC), garden compost (GC), and crops compost (CC)) and their water extract on Pythium debaryanum, Fusarium oxysporum f.sp. lycopersici, Sclerotium bataticola. Compost water extract (CWE) of LFC, GC, and CC were found to contain Bacillus spp., Micrococcus spp., Staphylococcus spp. and Corynebacterium spp., and the fungi Aspergillus spp., Rhizopus spp., and Drechslera spp., and various Actinomycetes. In situ results indicated considerable decrease in fungal growth around the unautoclaved compost especially in the case of S. bataticola and F. oxysporum f.sp. lycopersici, compared to the autoclaved compost. In vitro tests showed that concentration of CWE at 5, 10 and 15% (v/v) suppressed the hyphal growth of S. bataticola by 83% using 5% CC and by 94.4% using 5% LFC or 10% GC, and F. oxysporum f.sp. lycopersici by 94.4% using either composts. CWE of GC decreased fungal dry weight of F. oxysporum f.sp. lycopersici by 97.7%, P. debaryanum by 92.8%, and S. bataticola by 84.4%; CC decreased F. oxysporum f.sp. lycopersici by 94%, P. debaryanum by 86.2%, and S. bataticola by 63.3%, while CWE of LFC was the least effective against the tested fungi. CWE produced clear inhibition zones against all the tested fungi. Microflora found in CWE have an important role in suppressing the growth of tested fungi. CWE contained neither antibiotics nor siderophores. The presence of protease, chitinase, lipase and -1,3 glucanase (lysogenic enzymes) in CWE indicates a possible role in fungal degradation.     

Alkylresorcinols isolated from rye bran by supercritical fluid of carbon dioxide and suspended in a food-grade emulsion show activity against Penicillium expansum on apples

Apple cultivars are attacked by many fungal diseases, both on the tree and during storage. One of the most serious is blue mould, caused by Penicillium expansum. In this study, 5-(n)-alkylresorcinols (AR) were isolated from rye bran by the supercritical fluid of carbon dioxide and were used for the preparation of bioactive emulsions. These emulsions were applied to harvested fruit of five apple varieties to determine the levels of antifungal activity. A significant inhibition of disease symptoms was obtained after spraying some of the prepared AR emulsions on fruits that had been experimentally infected with Penicillium expansum. The most effective emulsions consisted of 0.025% (m/v) ARs, 0.1% (m/v) xanthan gum, 0.5% (m/v) Synperonic 91/6 or PDMs-copolymer, 0.2% (m/v) Tween 20, 1% (m/v) Trioleate, 2% (m/v) oleylalcohol, 2% (m/v) PEG 400, 5% (m/v) CaCl₂ or NaCl suspended in water.     

Effect of sucrose supplementation on aflatoxin production by Aspergillus parasiticus 2999 cultures in soybeans and cashew fruit juice

The aflatoxigenic potential ofAspergillus parasiticus 2999 on soybeans (raw and cooked) and cashew fruit juice (ripe, unripe, raw and cooked) variously supplemented with sucrose (0–20g) has been evaluated. Aflatoxin production showed a dual increase with increasing sucrose (0–20g) and soybeans (0.1–2.0g) concentrations which probably indicates the limited availability of suitable carbon sources in soybeans. This may be partly responsible for its resistance to aflatoxin synthesis. Two to five per cent (w/v) sucrose supplementation was optimal for maximal toxin production in cashew juice. Above this range aflatoxin production dropped steeply. Cooked soybeans supported higher yields of toxin than raw, in direct contrast with cashew fruit juice. Ripe cashew juice produced a greater quantity of toxin than the unripe. More fluorescent metabolites were synthesized in cashew fruit media than in soybeans. These results have been discussed in relation to the limiting role of the carbon source and the resistance to aflatoxin production on natural substrates.     

In vitro propagation of Notocactus magnificus

Most commercially grown cacti can be easily propagated by seed and/or cuttings. A group of rare and endangered species does not fit into this category and is therefore a good candidate for in vitro propagation productions as a tool to overcome habitat and plant-destruction. The number of rare and endangered species of Cacti goes into about 100. Many show a low production and germination of seeds and plantlets are prone to damping-off, making the in vitro propagation a feasible alternative for the multiplication and conservation of their germplasm. The aim of the present investigation is to establish a protocol for the in vitro culture and plant regeneration of Notocactus magnificus, the blue cactus, a highly ornamental species, native to Brazil. The surface sterilization of the explants was achieved with immersion for 10 min in sodium hypochlorite solution for either seeds (0.25% v/v) or ribs segments (1% v/v). Callus formation was observed when explants were cultured on MS medium supplemented with sucrose at 2% (w/v), 2,4-dichlorophenoxyacetic acid 0.5 μM, benzylaminopurine 4.4 μM, thiamine HCl 0.4 mg l⁻¹ and i-inositol 100 mg l⁻¹. The regeneration of shoots was carried out on MS medium supplemented with either different concentrations of benzylaminopurine and 1-naphthaleneacetic acid, or kinetin and indole-3-acetic acid. The highest number of shoots occurred when MS medium was supplemented with benzylaminopurine 22.2 μM, sucrose 3% (w/v) and agar 0,6% (w/v). In vitro spontaneous rooting of shoots was observed after eight months under culture on MS medium. Only in vitro rooted shoots developed into normal plants under glasshouse culture conditions. This in vitro protocol should be useful for the conservation as well as mass propagation of Notocactus magnificus.