Effects of vitamin D-induced supernatant of placental explants from preeclamptic women on oxidative stress and nitric oxide bioavailability in human umbilical vein endothelial cells

The study evaluated the effect of the supernatant of placental explants from preeclamptic (PE) and normotensive (NT) pregnant women after tissue treatment with or without vitamin D (VD) on oxidative stress and nitric oxide (NO) bioavailability in human umbilical vein endothelial cells (HUVEC). Placental explants were prepared from eight NT and eight PE women, and supernatants were obtained after incubation with or without hydrogen peroxide (H2O2) and/or VD. HUVEC were cultured for 24 h with supernatants, and the following parameters were analyzed in HUVEC cultures: NO, nitrate (NO3-), and nitrite (NO2-) levels, lipid peroxidation, and intracellular reactive oxygen species (ROS). Results showed that the production of NO3-, NO2-, malondialdehyde (MDA), and ROS were significantly higher in HUVEC treated with explant supernatant from PE compared to NT pregnant women, while the supernatant of PE explants treated with VD led to a decrease in these parameters. A significantly high production of NO was detected in HUVEC cultured with control supernatant of NT group, and in cultures treated with supernatant of PE explants treated with VD. Taken together, these results demonstrated that cultures of placental explants from PE women with VD treatment generated a supernatant that decreased oxidative stress and increased the bioavailability of NO in endothelial cells.

Improving micropropagation of some grape cultivars via boron, calcium and phosphate

BACKGROUND: An efficient regeneration protocol is a priority for the successful application of plant biotechnology. Grape nodal explants were used to develop a micropropagation protocol for Thompson Seedless and Taify cvs. Explants were cultured on MS medium supplemented with Kinetin or benzylaminopurine (BA) and indolebutyric acid (IBA). RESULTS: For both cultivars, axillary buds were grown, only, on a medium enriched with kinetin, moreover, shoot tip necrosis and callus formation were observed on Thompson Seedless cv. cultures grown on a medium with BA. Supplementing the growth medium with 100 mM (boron) B and 2.5 mM (calcium) Ca successfully help overcome these phenomena. The highest regenerated shoot numbers (14 and 6.2 explant 1 ) for Taify and Thompson Seedless cvs., respectively, were on media supplemented with 13.2 mM BA + 4.9 mM IBA and BA 13.2 mM + 5.8 mM IBA, respectively. Moreover, these media supported the developing shoots to have the heaviest dry weights (1.46 and 0.72 mg explant 1 ) for Taify and Thompson Seedless cvs., respectively. Thompson Seedless cv. regenerated shoot numbers and their dry weights were significantly increased by increasing the MS medium PO4 concentration. However, these two parameters were significantly decreased for Taify cv. Developing shoots were elongated and rooted on MS medium enriched with 4.9 mM, IBA 100 mM B and 2.5 mM Ca. Plantlets were acclimatized and successfully transferred to the greenhouse conditions. CONCLUSIONS: A novel promising protocol for Thomson Seedless and Taify cvs. micropropagation using single nodes has been developed.

Effect of Rhustox on Micropropagation of Scoparia dulcis L. through Leaf Explants Culture

Aim: To investigate the effect of Rhus toxicodendron (30CH) along with different compositions of phytohormones (Auxin and Cytokinin) on the basis of growth and multiplication of explants under optimum temperature under in-vitro conditions. Study Design: To establish and design the standard protocol for the in-vitro propagation through leaf explant of Scoparia dulcis under stress of phytohormones and homeopathic medicine Rhus toxicodendron (30CH). Place and Duration of Study: The plant materials were procured from the Herbal Botanical Garden Patna Science College, Department of Botany, Patna University, Patna, Bihar. The experimental part was carried out in Plant Tissue Culture Laboratory, between December 2017 to August 2018 in Department of Botany P.U. Patna. Methodlogy: The sterilized leaf explants were inoculated into MS media fortified with different phytohormones (Auxin and Cytokinin) and Rhus tox(30CH) under aseptic environmental conditions for the growth and development of callus, embryoids etc. Result: The explants in MS medium supplemented with auxins phytohormones and Rhus tox(30CH) exhibited that IAA (0.10 to 2.0 mg/l) and BAP (0.10 to 2.5 mg/l) induces green and compact calli. Whereas at 0.30mg/l of IAA and 0.50 mg/l BAP induced brown friable calli. 2,4-D (1.5 mg/l) and Kinetin (1.5-6.5mg/l) concentrations induced brown and friable calli. Rhus tox(30CH) (100 µl/100 ml) enhances proliferation with 2,4-D and Kinetin (1.5/1.5 mg/l.). Conclusion: After 42 days of culture initiation and establishment the callus was 520.0±1.12 mg in the mixture of 2,4-D and Kinetin (1.5 mg/l) in Rhus tox free medium. Whereas weight of callus were found to be 1092±0.74 mg after 42 days in the same medium of 2, 4-D and Kinetin (1.5/5.5 mg/l) supplemented with Rhus tox (100 µl/100 ml). Hence, the investigation proponded that the Rhus tox (CH30) has increased the rate of callus development and plantlet regeneration.

A comparison of callus induction in 4 Garcinia species

Background: This research is intended to determine suitable types and concentrations of plant growth regulators (PGRs) to induce callus on stem and leaf sections of 4 species of the genus Garcinia, namely, Garcinia mangostana, Garcinia schomburgkiana, Garcinia cowa, and Garcinia celebica. The base medium was MS medium containing 30 g l -1 sucrose, 0.5 g l-1 polyvinylpyrrolidone (PVP), and 7 g l-1 agar, and for the different treatments, PGRs were added to the medium as follows: thidiazuron (TDZ) at concentrations of 0, 0.1, 0.5, 1, and 2 mg l-1; 6-(3- hydroxybenzylamino) purine (meta-topolin) at concentrations of 0, 0.5, 2.5, and 5 mg l-1; 4-amino-3,5,6- trichloro-2-pyridinecarboxylic acid (picloram) at concentrations of 0, 0.5, 2.5, and 5 mg l-1; and 2,4- dichlorophenoxyacetic acid (2,4-D) at concentrations of 0, 0.5, 1, 2, and 4 mg l-1. The occurrence of callus was observed after 4 weeks. Results: A maximum of 100% and 93% of G. mangostana leaf explants formed callus in the 0.5 mg l-1 and 1 mg l-1 TDZ treatments, respectively, while 100% of G. schomburgkiana stem explants formed callus in the 1 mg l-1 TDZ treatment and 89% of G. schomburgkiana leaf explants formed callus in the 0.5 mg l-1 picloram treatment. The highest callus induction rate for G. cowa was 62% in the 1 mg l-1 TDZ treatment and for G. celebica was 56% in the 0.5 mg l-1•mT-1 treatment. Conclusions: For all 4 species, the greatest amount of large nodular callus was observed in the TDZ treatments. White, friable callus was observed on most of the 2,4-D and picloram treatment groups. Most meta-topolin treatments resulted in minimal callus formation.

In vitro multiplication of cassava varieties / Multiplicação in vitro de variedades de mandioca / Multiplicación in vitro de variedades de mandioca

The implantation of a model of sustainable development for agriculture can happen with the contribution of the mandiocultura. But for this, culture needs to be strengthened. In vitro propagation is an instrument for this purpose. Micropropagation can provide growers with large quantities of vigorous and healthy cassava seedlings in a short time. The objective of this study was to evaluate the in vitro establishment of four varieties of cassava cultivated in the municipality of Colorado do Oeste, State of Rondônia, popularly known as Arara, Caturra, Cacau Vermelha and Roxinha. For that, an experiment was carried out in the Laboratory of In Vitro Cultivation at the Pole of Technological Innovation of the University of Cruz Alta (UNICRUZ), with a completely randomized design in a 4 x 2 factorial scheme, with 6 replications. The treatments consisted of explants grown in Murashige and Skoog (MS) medium without the presence of growth regulator and MS medium supplemented with of 6-benzylaminopurine (BAP). The results indicate that the mean contamination percentage of the explants was 47.19%, differing among the varieties. The best growth response in culture media, in the multiple comparison of means (Scott-Knott's test, 5%), was obtained with MS medium without BAP addition, with significant difference between varieties. Under the conditions of this experiment, it was evidenced that micropropagation is a viable tool for obtaining varieties of interest, with desired phytosanitary qualities, with varietal and large-scale authenticity.(AU)

A implantação de um modelo de desenvolvimento sustentável para a agricultura pode acontecer com a contribuição da mandiocultura. Mas para isso, a cultura precisa ser fortalecida. A propagação in vitro é um instrumento para este fim. A micropropagação pode proporcionar aos produtores grande quantidade de mudas de mandioca vigorosas e sadias em um curto espaço de tempo. O objetivo deste trabalho foi avaliar o estabelecimento in vitro de quatro variedades de mandioca cultivadas no município de Colorado do Oeste, Rondônia, popularmente conhecidas como Arara, Caturra, Cacau Vermelha e Roxinha. Para isso, foi realizado um experimento no Laboratório de Cultivo In Vitro no Polo de Inovação Tecnológica da Universidade de Cruz Alta (UNICRUZ), com delineamento inteiramente casualizado em esquema fatorial 4 x 2, com 6 repetições. Os tratamentos consistiram em explantes cultivados em meio Murashige e Skoog (MS) sem a presença de regulador de crescimento e meio MS suplementado com 6-benzilaminopurina (BAP). Os resultados indicam que a porcentagem média de contaminação dos explantes foi de 47,19%, diferindo entre as variedades. A melhor resposta de crescimento em meios de cultura, na comparação múltipla de médias (teste de Scott-Knott, 5%), foi obtida com meio MS sem adição de BAP, com diferença significativa entre as variedades. Nas condições deste experimento, ficou evidenciado que a micropropagação é uma ferramenta viável para obtenção de variedades de interesse, com qualidades fitossanitárias desejadas, com autenticidade varietal e em larga escala.(AU)

La implantación de un modelo de desarrollo sostenible para la agricultura puede suceder con el aporte de la mandiocultura. Pero, para eso, la cultura necesita ser fortalecida. La propagación in vitro es un instrumento para ese fin. La micropropagación puede proporcionar a los cultivadores grandes cantidades de plántulas de mandioca vigorosas y saludables en poco tiempo. El objetivo de esta investigación ha sido evaluar el establecimiento in vitro de cuatro variedades de mandiocas cultivadas en el municipio de Colorado del Oeste, Rondônia, popularmente conocidas como Arara, Caturra, Cacau Vermelha y Roxinha. Para eso, se realizó un experimento en el Laboratorio de cultivo in vitro en el Polo de Innovación Tecnológica de la Universidad de Cruz Alta (UNICRUZ), con delineamiento completamente casualizado en esquema factorial 4 x 2, con 6 repeticiones. Los tratamientos consistieron en explantes cultivados en medio Murashige y Skoog (MS) sin la presencia de regulador de crecimiento y medio MS suplementado con 6-bencilaminopurina (BAP). Los resultados indican que el porcentaje de contaminación promedio de los explantes fue de 47.19%, diferenciándose entre las variedades. La mejor respuesta de crecimiento en medios de cultivo, en la comparación múltiple de medias (prueba de Scott-Knott, 5%), se obtuvo con medio MS sin adición de BAP, con una diferencia significativa entre las variedades. Bajo las condiciones de este experimento, se evidenció que la micropropagación es una herramienta viable para obtener variedades de interés, con cualidades fitosanitarias deseadas, con autenticidad varietal y de gran escala.(AU)

Histopathological and ultrastructural findings induced by heat-inactivated Lactobacillus plantarum and the culture supernatant on the intestinal mucosa of piglets: an ex vivo approach / Achados histopatológicos e ultraestruturais induzidos por Lactobacillus plantarum inativado pelo calor na mucosa intestinal de leitões: estudo ex-vivo

In the present study, histological, morphometrical and ultrastructural analysis were performed to investigate intestinal mucosa changes in piglets jejunal explants exposed to two concentration of heat-inactivated Lactobacillus plantarum and their respective culture supernatants. Jejunal explants were incubated for 4 hours in DMEM culture medium with a) only culture medium (control group), b) heat-inactivated Lactobacillus plantarum strain1 - LP1 (1.1 x 108CFU/ml), c) heat-inactivated Lactobacillus plantarum strain2 - LP2 (2.0 x 109CFU/ml), d) heat-inactivated Lactobacillus plantarum strain1 culture supernatant (CS1), and e) heat-inactivated Lactobacillus plantarum strain2 culture supernatant (CS2). Explants exposed to heat-inactivated L. plantarum strain 1 and 2 showed multifocal to difuse villi atrophy, villi apical necrosis and enterocyte flattening. Morphological assessment revealed similar results with bacterial adhesion to mucus and intestinal epithelial cells and, morphometric analysis showed a decreased villi height compared to the control group. Alterations in explants treated with the culture supernatant of both strains include mild villi atrophy and mild enterocyte apical necrosis. Morphological assesment reveled numerous well delineated villi and, morphometric analysis showed a significant increase in villi height compared to the control group. In general, exposure to the culture supernatants improved the intestinal morphology.(AU)

No presente estudo, foram realizadas análises histológica, morfométrica e ultraestrutural para investigar as alterações da mucosa intestinal em explantes jejunais de leitões expostos a duas cepas e concentrações de Lactobacillus plantarum inativado pelo calor e seus sobrenadantes de cultura. Os explantes jejunais foram incubados durante quatro horas, em meio de cultura DMEM com: a) meio de cultura (grupo controle); b) Lactobacillus plantarum, cepa 1 - LP1 (1,1 x 108CFU/mL); c) Lactobacillus plantarum, LP2 (2,0 x 109CFU/mL); d) sobrenadante da cultura do Lactobacillus plantarum, cepa 1 (SC1); e e) sobrenadante da cultura do Lactobacillus plantarum, cepa 2 (SC2). Os explantes expostos às cepas 1 e 2 do L. plantarum inativado pelo calor mostraram atrofia difusa de vilosidades, necrose apical das vilosidades e achatamento de enterócitos. A avaliação morfológica revelou resultados semelhantes, com adesão bacteriana ao muco e às células epiteliais intestinais, e a análise morfométrica mostrou uma diminuição da altura das vilosidades em relação ao grupo controle. Alterações nos explantes tratados com o sobrenadante da cultura de ambas as cepas caracterizaram-se por atrofia leve das vilosidades e necrose apical leve dos enterócitos. A avaliação morfológica revelou vilosidades bem delineadas, e a análise morfométrica mostrou um aumento significativo na altura das vilosidades em comparação ao grupo controle. Em geral, a exposição aos sobrenadantes da cultura melhora a morfologia intestinal.(AU)

Kinetics and modeling of cell growth for potential anthocyanin induction in cultures of Taraxacum officinale G.H. Weber ex Wiggers (Dandelion) in vitro

Background: Taraxacum officinale G.H. Weber ex Wiggers is a wild plant used in folk medicine to treat several diseases owing to bioactive secondary metabolites present in its tissue. The accumulation of such molecules in plant cells can occur as a response against abiotic stress, but these metabolites are often deposited in low concentrations. For this reason, the use of a biotechnological approach to improve the yields of technologically interesting bioactive compounds such as anthocyanins is a compelling option. This work focuses on investigating the potential of in vitro T. officinale cultures as an anthocyanin source. Results: To demonstrate the suitability of anthocyanin induction and accumulation in calluses under specific conditions, anthocyanin was induced in the T. officinale callus. A specific medium of 5.5% sucrose supplemented with 6-benzylaminopurine /1-naphthaleneacetic acid in a 10:1 ratio was used to produce an anthocyanin yield of 1.23 mg g-1 fw. An in vitro dandelion callus line was established from this experiment. Five mathematical models were then used to objectively and predictably explain the growth of anthocyanin-induced calluses from T. officinale. Of these models, the Richards model offered the most suitable representation of anthocyanin callus growth in a solid medium and permitted the calculation of the corresponding kinetic parameters. Conclusions: The findings demonstrate the potential of an in vitro anthocyanin-induced callus line from T. officinale as an industrial anthocyanin source.

Cloning of the Coffea canephora SERK1 promoter and its molecular analysis during the cell-to-embryo transition

Background: Somatic embryogenesis receptor-like kinase 1 (SERK1) is a cell membrane receptor active in different plant tissues and involved in cell differentiation activities including somatic embryogenesis. The identification of promoter elements responsible for SERK1 expression during the onset of somatic embryogenesis can be useful to understand the molecular regulation of the cell-to embryo transition, and these promoter elements represent biotechnological tools in plant organ tissue culture. Results: A −1,620 bp DNA sequence located upstream of the Coffea canephora SERK1 gene homologue (CcSERK1) was isolated, and then, different segments containing key response elements (REs) for somatic embryogenesis onset and development were fused to the uidA (encoding a ß-glucuronidase, GUS) reporter gene to evaluate its expression in transgenic leaf explants. DNA segments of −1,620 and −1048 bp in length directed uidA expression with patterns in leaf explants similar to those occurring during somatic embryogenesis. When a −792-bp fragment was used, uidA expression disappeared only in leaf explants and pro-embryogenic mass but persisted in developing embryos. No uidA expression was detected in any embryogenic stage when a −618-bp fragment was used. Conclusion: DNA deletions showed that a −1048-bp sequence located upstream of the CcSERK1 gene is sufficient to direct gene expression during the onset and the development of C. canephora somatic embryogenesis. The DNA segment located between −1048 and −792 bp (containing BBM and WUS REs) is needed for gene expression before embryogenesis onset but not during embryo development. The promoter segment between −792 and −618 bp (including GATA, ARR1AT, and ANT REs) regulates gene expression in developing embryos.

Somatic embryogenesis in Carica papaya as affected by auxins and explants, and morphoanatomical-related aspects

ABSTRACT The aim of this study was to evaluate somatic embryogenesis in juvenile explants of the THB papaya cultivar. Apical shoots and cotyledonary leaves were inoculated in an induction medium composed of different concentrations of 2,4-D (6, 9, 12, 15 and 18 µM) or 4-CPA (19, 22, 25, 28 and 31 µM). The embryogenic calluses were transferred to a maturation medium for 30 days. Histological analysis were done during the induction and scanning electron microscopy after maturing. For both types of auxin, embryogenesis was achieved at higher frequencies with cotyledonary leaves incubated in induction medium than with apical shoots; except for callogenesis. The early-stage embryos (e.g., globular or heart-shape) predominated. Among the auxins, best results were observed in cotyledonary leaves induced with 4-CPA (25 µM). Histological analyses of the cotyledonary leaf-derived calluses confirmed that the somatic embryos (SEs) formed from parenchyma cells, predominantly differentiated via indirect and multicellular origin and infrequently via synchronized embryogenesis. The secondary embryogenesis was observed during induction and maturation phases in papaya THB cultivar. The combination of ABA (0.5 µM) and AC (15 g L-1) in maturation medium resulted in the highest somatic embryogenesis induction frequency (70 SEs callus-1) and the lowest percentage of early germination (4%).

Longitudinal changes in intraocular pressure and iris vessels in rats with bulb external scleral explants / 眼科新进展

Objective To observe longitudinal changes in the intraocular pressure (IOP),the cornea and iris blood vessels in normal rats with bulb external scleral explants.Methods Totally 18 normal rats were enrolled in the experiment.Conjunctival incisions of the right eyes of these rats were performed,followed by the placement of a balloon catheter under the lateral rectus muscle and inflation at 4-5 ATM.The IOP was measured and iris images were taken longitudinally before and after pressuring and pressure withdrawing respectively.Results IOP was elevated to (40.66 ± 10.55) mm-Hg (1 kPa =7.5 mmHg) immediately after pressing,which was over the baseline (8.45 ± 1.23)mmHg (P=0.000),and then decreased gradually at 15 min,30 min,45min,60 min after pressing,but the IOPs were still greater than the baseline.There were significant differences in IOPs at 15 min,30 min,45 min after pressing and the baseline (all P ＜ 0.05),while no difference in this variable at 60 min after pressing and the baseline (P =0.929).However,the IOP was quickly reduced to (6.09 ± 0.49) mmHg at once after pressure withdrawing,which was below the baseline (P =0.000),and then gradually returned to the baseline at 15 min,30 min after pressure withdrawing.The corneal edema and dilation of iris vessels were observed after pressuring,and following corneal winkle and dilated iris vessels after pressure withdrawing.Conclusion Bulb external scleral explant can result in elevated IOP immediately,then decreased gradually.With the increase of compressing,the IOP of the contralateral eye tends to increase.Bulb external scleral explant may affect uveoscleral outflow pathway of aqueous humor in rats.

Primary culture of hPDSCs with modified enzymatic digestion-explants method / 口腔疾病防治

Objective@#To explore methods using modified tissue enzymatic separations for culturing primary hDPSCs in vitro and further identify the cells produced. @*Methods @#Primary hDPSCs were cultured using the modified tissue enzymatic separation method, and cells were identified by morphology, cell surface markers, and differentiation potential and evaluated using flow cytometry and growth curves.@*Results @#The hDPSCs were successfully isolated using the modified tissue enzymatic separation method. The morphology of these cells was similar to that of fibroblasts and mesenchymal stem cells, and the growth curve was "S" -type. The results of cell phenotype analysis indicated that the cells were positive for surface markers of mesenchymal stem cells, including CD29, CD44, and CD90, and negative for markers of hematopoietic stem cells, including CD34, CD45, and CD106. The cells were capable of differentiating into multiple cell types.@*Conclusion @# The modified tissue enzymatic separation method can successful be used to culture primary hDPSCs in vitro.

In vitro propagation of Vitis vinifera L. cv. 'Monastrell'

Background: A protocol for the micropropagation of the grape (Vitis vinifera L.) cultivar 'Monastrell' was developed. Initial plant material was obtained from the sanitary selection of grapevine plants performed by real-time RT-PCR to confirm the absence of Grapevine fanleaf virus, Arabis mosaic virus, Grapevine leafroll-associated virus 1, Grapevine leafroll-associated virus 3, and Grapevine fleck virus. Results: The effects of the salt composition (comparing Lloyd and McCown woody plant medium and Murashige and Skoog medium 1/2 macronutrients) and the growth regulator benzylaminopurine (BAP), at 0 and 8.9 µM, on plant propagation were evaluated using nodes as explants. The most efficient procedure consisted of bud induction in the medium with Lloyd and McCown woody plant salts and 8.9 µM BAP for 30 d along with elongation in cytokinin-free medium for 60 d, which gave 22 nodes/explant (174 plants/initial plant). A second cycle of propagation in a medium without BAP for another 60 d could give approximately 10,000 nodes, which can be obtained after an additional 2 months of culture. All plants acclimatized after the second cycle of multiplication were successfully transferred to soil. Conclusion: We developed an optimal protocol for V. vinifera cv. 'Monastrell' micropropagation, the first described for this cultivar.

A minimal cost micropropagation protocol for Dianthus caryophyllus L.— a commercially significant venture.

In tissue culture, high production cost of the products restricts their reach. Though tissue culture is a major strength in floriculture it is marred by pricing issues. Hence, we developed a complete regeneration low cost micropropagation protocol for an economically important floriculture crop, carnation (Dianthus caryophyllus L.). Successful regeneration of carnation from nodal explants on cost-efficient medium indicates that psyllium husk, sugar and RO water can effectively replace the conventional medium comprising agar, sucrose and distilled water. The protocol can contribute to increased carnation production at comparatively reduced cost, and there by encourage wide scale adoption by the common growers.

Research progress on browning phenomenon in tissue culture of medicinal plants / 中草药

Browning of explants may be easily induced by various factors during tissue culture of medicinal plants, which not only affects the vitality of explants, but also inhibits the callus induction and regeneration, blocking quick accumulation seriously of active components and survival rate of tissue culture. Therefore, it would be important to develop a fast, effective, safe, and economic browning restraining method. This paper reviewed the status, mechanism, and impact factors of the medicinal plants browning and effective anti-browning measures, expecting to provide the scientific references for development of anti-beowning measures. Meanwhile, we prospect anti-browning technologies in the tissue culture of medicinal plants.

Investigating the In vitro Regeneration Potentiality of Three High Yielding Indica Rice (Oryza sativa L.) Varieties.

Aims: An efficient and reproducible In vitro regeneration protocol is vital for varietal improvement research. The current research was conducted to optimize the callus induction, shoot and root regeneration of three indica rice varieties. Place, Duration and Design of Study: The experiment was conducted in the tissue culture and biotechnology laboratory of the department of Genetics and Plant Breeding of Bangladesh Agricultural University using completely randomized experimental design. Methodology: Dehusked mature seeds of three indica rice varieties namely, BRRI dhan28, BRRI dhan29 and BINA dhan6 were cultured In vitro in MS medium supplemented with different concentrations and combinations of phytohormones. Results: The callus induction ranged from 14 - 84% which showed a general increasing trend with the increase in the concentration of 2,4-Dichlorophenoxyacetic acid (2,4-D) starting from 1.0 mg L-1 till 2.5 mg L-1. A further increase in the concentration of 2,4-D to 2.5 mg L-1, however, decreased the percentage of callus induction in all three varieties. MS medium supplemented with 2.5 mg L-1 2,4-D and 0.5 mg L-1 6-Benzylaminopurine (BAP) was better than any other composition for callus induction. For size of callus and nature of callus, however, MS medium supplemented with 2.0 mg L-1 2,4-D and 0.5 mg L-1 BAP was found to perfume best. The highest percentage of callus induction was observed in the variety BRRI dhan29 (84%) followed by BRRI dhan28 (74%). Almost all the varieties produced yellowish and compact calli except BINA dhan6 which produced creamy and friable calli. The desiccation treatment has shown to increase size but decrease the compactness of the callus. The differences are, however, not statistically significant. MS medium supplemented with 0.6 mg L-1 1-Naphthaleneacetic acid (NAA) and 6 mg L-1 Kinetin (Kn) showed highest shoot regeneration in BRRI dhan29 (85%) followed by BRRI dhan28 (60%). Higher frequency of root formation was observed in all three varieties using Indole-3-butyric acid (IBA). The survival rate of the plantlet in acclimatization chamber (96%) and in field condition (93.33%) was higher for BRRI dhan29. BINA dhan6 has shown the least regeneration potentiality for all the aforementioned traits. Conclusion: Of the three varieties, BRRI dhan29 and BRRI dhan28 has shown higher regeneration potentiality. This optimized protocol will thus be useful in genetic improvement of these varieties using biotechnological manipulations.

Progress in the isolation and purification methods of adipose-derived stem cells / 中国比较医学杂志

Adipose-derived stem cells ( ASCs ) as potential seeded cells have been widely used in tissue engineering.Thus to obtain enough, high activity, high purity adipose-derived stem cells is the particular important premise of the application in tissue engineering.In this paper, the isolation and purification methods of ASCs were reviewed and the merit and demerit of different methods were compared in order to provide theoretical basis for safe and high-effective isolation and purification of ASCs.

Morphogenesis in vitro of chrysanthemum (Dendranthema grandiflora tzelev) from stem and pedicel explants / Morfogênese in vitro de crisântemo (Dendranthema grandiflora tzelev) a partir de explantes de caule e pedicelo

Visando a adequação de um protocolo de morfogênese in vitro para plantas de crisântemo (Dendranthema grandiflora Tzelev) analisou-se três fatores que afetam a frequência da regeneração. Foram avaliadas três concentrações de ácido indolil-acético (AIA) e 6- Benzilaminopurina (BAP), a fonte de explante (caule e pedicelo) e a orientação de inoculação do explante no meio de cultivo. Ao final de 15-30 dias de cultivo, observou-se que a fonte de explante, o tipo de explante e a interação explante e meio de cultivo afetaram significativamente a frequência de regeneração (FR) e o número de ramos por explantes (NRE) em crisântemo. Os explantes de pedicelo apresentaram maiores frequências de regeneração (15-20 dias) em meio de cultivo suplementado com 0,5 mg L-1 de BAP e 2,0 mg L-1 de AIA e os explantes caulinares (30 dias) em meio com 2,0 mg L-1 de BAP e 0,5 mg L-1 de AIA. Respostas diferenciais em relação à morfogênese foram observadas quando as duas posições do pedicelo foram inoculadas no meio de cultivo, a região seccionada em contato com o meio de cultivo suplementado com 0,5 mg L-1 de BAP e 2,0 mg L-1 de AIA proporcionou maior frequência de brotos. Foram obtidas plantas com flores 45-60 dias após a transferência para a casa de vegetação. As flores apresentavam os mesmos padrões e características das flores das plantas doadoras de explantes. Podese concluir que a o tipo de explante, sua orientação em relação ao meio de cultura bem como as combinações de reguladores de crescimento são fatores-chave envolvidos no sucesso da regeneração de plantas de crisântemo in vitro.

Aiming the optimization of a regeneration protocol for chrysanthemum plants (Dendranthema grandiflora), experiments of regeneration in vitro were accomplished, stem explants and pedicel inoculated in several shoots induction mediuns. The stem explants and of pedicels they were sectioned longitudinally and inoculated with the face opposed to the section in the medium of cultivation with the basic salts MS, with different concentrations and combinations of growth regulators BAP and AIA. At the end of 15-30 days of cultivation, significant differences were observed in the regeneration frequency, number of shoots for explante and in the length of the shoots among the appraised explantes. Explants of pedicel presented potential the best to the explants stem. The pedicel explants presented larger regeneration frequencies in medium of cultivation contends basic salts of MS, with 0,5 mg L-1 of BAP and 2,0 mg L-1 of AIA and the explantes of stem in medium with basic salts of MS + 2,0 mg L-1 of BAP and 0,5 mg L-1 of AIA. They were obtained plants with flowers 45-60 days after the transfer to the vegetation house. In function of the obtained results, it can be concluded that the choice of the explant type and the medium of cultivation are factor-key involved in the success of the regeneration of plants for posterior use in experiments of genetic transformation.

Adventitious Bud Formation and Plantlet Regeneration of Achyrocline satureoides-A Multipurpose Medicinal Plant.

Adventitious bud regeneration was achieved from hypocotyls, cotyledons and leaf explants of Achyrocline satureioides. Organogenesis was induced from every explant cultured on Murashige and Skoog semisolid medium (plus sucrose 30g·L-1) containing different combinations of 6-benzyladenine (BA) and -naphtalenacetic acid (NAA) under 116mol·m2·s-1 photosynthetic photon flux density (PPFD), photoperiod 14 h and at 272ºC. The regeneration was similar for every tested explant and varied between 64 and 83%. The number of buds formed per regenerative explants was similar in every treatment (5-8 shoots/explant). In order to stimulate In vitro rooting, regenerative leaves were sub cultured from the best induction medium in MS lacking plant growth regulators for the same periods. Every plantlet raised In vitro was phenotypically normal and successfully hardened to ex vitro conditions. An experimental field plot with 60-day-old in vitro regenerated plants was established.

Progress in micropropagation of Passiflora spp. to produce medicinal plants: a mini-review

Micropropagation of Passiflora species and its hybrids may play an important role in the production of healthy and disease-free plants which can be a source of medicinal herbal products, nutritional fruits and ornamental flowers. The rapid multiplication of elite plants to obtain pharmacognostic material, containing valuable flavonoid C-glycosides, is possible by usingcontrolled in vitro conditions, constituents of the medium and the interactions of plant growth regulators (1-naphtaleneacetic acid, benzyladenine, gibberellin GA3,kinetin, indole-3-acetyl-L-aspartic acid, indole-3-butyric acid, thidiazuron) and influencing various chemical additives (silver nitrate, coconut water, activated charcoal). Investigations of specific requirements during stages of micropropagation, such as the establishment of primary cultures (including type of explants, age of donor plant), shoot multiplication (by direct and indirect organogenesis and embryogenesis), rooting and acclimatization of regenerated plants are summarized in this review. The following species were recently studied for micropropagation: P. alata, P. caerulea, P. cincinnata, P. edulis, P. foetida, P. setacea, P. suberosa. It seems that for awide range of applications of in vitro clones of Passiflora, interdisciplinary studies including genetic and phytochemical aspects are needed.

Fitorreguladores e posição de explantes foliares na indução à calogênese em cerejeira-do-mato / Phytoregulators and leaf explants position on callus induction in cerejeira-do-mato

O presente trabalho objetivou avaliar o efeito de diferentes formas de inoculação, no meio nutritivo, de explantes foliares de Eugenia involucrata DC., uma espécie florestal nativa com diversas potencialidades econômicas. Foram avaliadas as posições abaxial, adaxial, com e sem cortes no limbo foliar. Foi utilizado o meio de cultura MS acrescido de 10 µM de ANA isolado ou das combinações, em µM, de 2,4-D e BAP: 5-5 e 5-10. A posição dos explantes afeta a calogênese e a organogênese em segmentos foliares de E. involucrata, sendo mais adequada a abaxial sem cortes na região do limbo. A associação dos reguladores de crescimento 2,4-D + BAP na concentração de 5-10 µM foi mais promissora para a obtenção de calos, especialmente os nodulares, putativos à embriogênese somática.

The aim of this research was to evaluate the effect of different forms of leaf explants inoculation in nutritive medium of Eugenia involucrata DC., an native forest species with diverse economic potentials. Inoculations were evaluated at abaxial and adaxial positions, with and without cuts on the leaf surface in the MS nutritive medium supplemented with 10 µM NAA or combination, in µM, of 2.4-D and BAP: 5-5 and 5-10. The explants position inoculation affects the callus induction and organogenesis in E. involucrata leaf segments, being the abaxial the most suitable position, without cuts in the leaf surface. The combination of growth regulators 2.4-D and BAP, at concentrations of 5-10( µM) was most promising of callus obtaining, especially nodular callus, putative at somatic embryogenesis.