Background Mammary gland(MG)infections(mastitis)are frequent diseases of dairy cows that affect milk quality,animal welfare and farming profitability.These infections are commonly associated with the bacteria Escheric...Background Mammary gland(MG)infections(mastitis)are frequent diseases of dairy cows that affect milk quality,animal welfare and farming profitability.These infections are commonly associated with the bacteria Escherichia coli and Staphylococcus aureus.Different in vitro models have been used to investigate the early response of the MG to bacteria,but the role of the teat in mastitis pathogenesis has received less attention.In this study,we used punch-excised teat tissue as an ex vivo model to study the immune mechanisms that arise early during infection when bacteria have entered the MG.Results Cytotoxicity and microscopic analyses showed that bovine teat sinus explants have their morphology and viability preserved after 24 h of culture and respond to ex vivo stimulation with TLR-agonists and bacteria.LPS and E.coli trigger stronger inflammatory response in teat when compared to LTA and S.aureus,leading to a higher produc-tion of IL-6 and IL-8,as well as to an up-regulation of proinflammatory genes.We also demonstrated that our ex vivo model can be applied to frozen-stored explants.Conclusions In compliance with the 3Rs principle(replacement,reduction and refinement)in animal experimenta-tion,ex vivo explant analyses proved to be a simple and affordable approach to study MG immune response to infec-tion.This model,which better reproduces organ complexity than epithelial cell cultures or tissue slices,lends itself particularly well to studying the early phases of the MG immune response to infection.展开更多
We developed a system for the regeneration of Lombardy poplar (Populus nigra L. var. italica) shoots from internodal stem explants. Using this system, shoots regenerated from 87% of the stem explants placed on Murashi...We developed a system for the regeneration of Lombardy poplar (Populus nigra L. var. italica) shoots from internodal stem explants. Using this system, shoots regenerated from 87% of the stem explants placed on Murashige and Skoog (MS) medium supplemented with 0.1 mg/L indole-3-acetic acid and 0.5 mg/L benzylaminopurine without undergoing callus formation. About 80% of the in vitro regenerated shoots developed roots on MS medium supplemented with 0.5 mg/L indole-3-butyric acid and 0.02 mg/L 1-naphthylacetic acid. Well-rooted seven-to eight-week-old regenerated plants could be transferred to soil for further growth and the survival rate of such plants after three weeks was 88%. The protocol presented here is simple and economical because it does not rely on pre-incubation in callus induction medium or repeated subculture in shoot induction medium containing trans-zeatin, an expensive substance. The in vitro regeneration system presented here could be used for evaluation of radiation sensitivity for Lombardy poplar tissues.展开更多
Adventitious shoots were successfully regenerated from hypocotyl explants of in vitro cultures of Euonymus japonicusCu zhi. Hypocotyl slices were cultured on Murashige and Skoog (MS) and B5 basal medium supplemented w...Adventitious shoots were successfully regenerated from hypocotyl explants of in vitro cultures of Euonymus japonicusCu zhi. Hypocotyl slices were cultured on Murashige and Skoog (MS) and B5 basal medium supplemented with variedconcentration of different plant growth-regulators, e.g., α-naphthaleneacetic acid (NAA) and indole-3-butyric acid (IBA)in combination with 6-benzylaminopurine (6-BA) and kinetin. The study showed that shoots could be directly regeneratedfrom hypocotyl explants without the intervening callus phase; MS medium was more suitable for adventitious shootsregeneration. The ability of hypocotyls segments to produce shoots varied depending upon their position on the seedlings.The highest regeneration rate was obtained with hypocotyl segments near to the cotyledon cultured on MS basal mediumsupplemented with 1.5 mg L-1 6-BA and 0.05 mg L-1 NAA (63.64%). The regenerated shoots were readily elongated on thesame medium as used for multiplication and rooted on half-strength MS basal medium supplemented with 1.0 mg L-1 IBAand 100 mg L-1 activated carbon. After being transferred to greenhouse conditions, 96% of the plantlets were successfullyacclimatized. This regeneration system is applied for genetic transformation now.展开更多
We have previously shown that capacitively coupled electrical stimulation of either normal bovine articular chondrocytes or osteoarthritic human articular cartilage explants resulted in up-regulation of cartilage matr...We have previously shown that capacitively coupled electrical stimulation of either normal bovine articular chondrocytes or osteoarthritic human articular cartilage explants resulted in up-regulation of cartilage matrix gene expression and down-regulation of metalloproteinase gene expression. In addition, collagen and proteoglycan protein levels were also elevated. To determine visually the effect of specific electric fields on modifying cartilage structure, freshly harvested human full-thickness osteoarthritic cartilage explants were stimulated in the absence or presence of interleukin-1β, an inflammatory cytokine, and were examined photographically and spectrophotometrically. Hexosamine and hydroxyproline contents were also determined. Spectrophotometric analysis was used to quantify any changes in the depth of defects in the cartilage ranging from surface level (red-colored) to the deepest affected layer (blue-colored). Interleukin-1β treatment alone caused significant additional cartilage erosion. Electrical stimulation alone resulted in significant decreases in the cartilage defects. Electrical stimulation in the presence of interleukin-1β resulted in a small, but significant, surface improvement. Meta-analysis also confirmed a significant increase in the hexosamine and hydroxyproline contents (indicating matrix deposition). It was concluded that an appropriate electric field could modify osteoarthritic lesions in full-thickness cartilage plugs by increasing matrix production and/or by decreasing matrix destruction. Furthermore, it appears that spectrophotometric analysis is a relatively easy method for quantifying the “filling in” or healing of articular cartilage defects.展开更多
In the present investigation, an attempt has been made to study the influence of ethylene inhibitor silver nitrate on direct shoot regeneration in Sphaeranthus indicus, an important antijaundice medicinal plant, by us...In the present investigation, an attempt has been made to study the influence of ethylene inhibitor silver nitrate on direct shoot regeneration in Sphaeranthus indicus, an important antijaundice medicinal plant, by using in vitro raised shoot tip explants. The effect of various concentrations of kinetin, BAP (0.5 - 3.0 mg/l), and NAA (0.1 - 0.5 mg/l) along with AgNO<sub>3</sub> (0.1 - 1.0 mg/l) was studied. Among the combinations tested MS medium augmented with kinetin (1.0 mg/l), NAA (0.1 mg/l) and AgNO<sub>3</sub> (0.4 mg/l) was found to be optimum for production of multiple shoots (34.3 ± 0.36). Addition of AgNO<sub>3</sub> to the media not only increases shoot number in all the concentrations tested but also shoot length. AgNO<sub>3</sub> at the concentration of 0.4 mg/l produced 35% more number of multiple shoots when compared to multiple shoots (10.8 ± 0.12) produced in control. In the present study by the addition of ethylene inhibitor silver nitrate and growth regulators, more number of multiple shoots (three folds) and shoot length was observed compared to control. These in vitro raised shoots were transferred to the rooting medium containing different concentrations of auxins such as NAA and IAA along with AgNO<sub>3</sub> (0.1 - 0.6 mg/l). Better rooting response (21.6) was observed on NAA (2.0 mg/l) and AgNO<sub>3</sub> (0.4 mg/l) containing media. The healthy rooted plantlets were transferred to polybags containing soil and vermiculate in 1:1 ratio for hardening. Finally the hardened plants were transferred to field environment for utmost survivability.展开更多
Objective:To develop the reproducible in vitro propagation protocols for the medicinally important plants viz.,Achyranthes aspera(A.aspera)L.and Achyranthes bidentata(A.bidentata)Blume using nodal segments as explants...Objective:To develop the reproducible in vitro propagation protocols for the medicinally important plants viz.,Achyranthes aspera(A.aspera)L.and Achyranthes bidentata(A.bidentata)Blume using nodal segments as explants.Methods:Young shoots of A.aspera and A.bidentata were harvested and washed with running tap water and treated with 0.1%bavistin and rinsed twice with distilled water.Then the explants were surface sterilized with 0.1%(w/v)HgCl_2 solutions for I min.After rinsing with sterile distilled water for 3-4 times,nodal segments were cut into smaller segments(1 cm)and used as the explants.The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog(MS)medium supplemented with 3%sucrose,0.6%(w/v)agar(HiMedia,Mumbai)and different concentration and combination of 6-benzyl amino purine(BAP),kinetin(Kin),naphthalene acetic acid(NAA)and indole acetic acid(IAA)for direct regeneration.Results:Adventitious proliferation was obtained from A.aspera and A.bidentata nodal segments inoculated on MS basal medium with 3%sucrose and augmented with BAP and Kin with varied frequency.MS medium augmented with 3.0 mg/L of BAP showed the highest percentage(93.60±0.71)of shootlets formation for A.aspera and(94.70±0.53)percentages for A.bidentata.Maximum number of shoots/explants(10.60±0.36)for A.aspera and(9.50±0.56)for A.bidentata was observed in MS medium fortified with 5.0 mg/L of BAP.For A.aspera,maximum mean length(5.50±0.34)of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A.bidentata(5.40±0.61)was observed in the very same concentration.The highest percentage,maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of 1BA.Seventy percentages of plants were successfully established in polycups.Sixty eight percentages of plants were well established in the green house condition.Sixty five percentages of plants were established in the field.Conclusions:The results have shown that use of nodal buds is an alternative reproducible and dependable method for clonal propagation of A.aspera and A.bidentata.The high rate of direct shoot-root multiplication and their high rate of post-hardening survival indicate that this protocol can he easily adopted for commercial large scale cultivation.展开更多
Retinitis pigmentosa is a hereditary retinal disease that affects rod and cone photoreceptors,leading to progressive photoreceptor loss.Previous research supports the beneficial effect of electrical stimulation on pho...Retinitis pigmentosa is a hereditary retinal disease that affects rod and cone photoreceptors,leading to progressive photoreceptor loss.Previous research supports the beneficial effect of electrical stimulation on photoreceptor survival.This study aims to identify the most effective electrical stimulation parameters and functional advantages of transcorneal electrical stimulation(tcES)in mice affected by inherited retinal degeneration.Additionally,the study seeked to analyze the electric field that reaches the retina in both eyes in mice and post-mortem humans.In this study,we recorded waveforms and voltages directed to the retina during transcorneal electrical stimulation in C57BL/6J mice using an intraocular needle probe with rectangular,sine,and ramp waveforms.To investigate the functional effects of electrical stimulation on photoreceptors,we used human retinal explant cultures and rhodopsin knockout(Rho^(-/-))mice,demonstrating progressive photoreceptor degeneration with age.Human retinal explants isolated from the donors’eyes were then subjected to electrical stimulation and cultured for 48 hours to simulate the neurodegenerative environment in vitro.Photoreceptor density was evaluated by rhodopsin immunolabeling.In vivo Rho^(-/-)mice were subjected to two 5-day series of daily transcorneal electrical stimulation using rectangular and ramp waveforms.Retinal function and visual perception of mice were evaluated by electroretinography and optomotor response(OMR),respectively.Immunolabeling was used to assess the morphological and biochemical changes of the photoreceptor and bipolar cells in mouse retinas.Oscilloscope recordings indicated effective delivery of rectangular,sine,and ramp waveforms to the retina by transcorneal electrical stimulation,of which the ramp waveform required the lowest voltage.Evaluation of the total conductive resistance of the post-mortem human compared to the mouse eyes indicated higher cornea-to-retina resistance in human eyes.The temperature recordings during and after electrical stimulation indicated no significant temperature change in vivo and only a subtle temperature increase in vitro(~0.5-1.5°C).Electrical stimulation increased photoreceptor survival in human retinal explant cultures,particularly at the ramp waveform.Transcorneal electrical stimulation(rectangular+ramp)waveforms significantly improved the survival and function of S and M-cones and enhanced visual acuity based on the optomotor response results.Histology and immunolabeling demonstrated increased photoreceptor survival,improved outer nuclear layer thickness,and increased bipolar cell sprouting in Rho^(-/-)mice.These results indicate that transcorneal electrical stimulation effectively delivers the electrical field to the retina,improves photoreceptor survival in both human and mouse retinas,and increases visual function in Rho^(-/-)mice.Combined rectangular and ramp waveform stimulation can promote photoreceptor survival in a minimally invasive fashion.展开更多
Fucoidan,a sulfate polysaccharide obtained from brown seaweed,has various bioactive properties,including anti-inflammatory,anti-cancer,anti-viral,anti-oxidant,anti-coagulant,anti-thrombotic,anti-angiogenic,and anti-He...Fucoidan,a sulfate polysaccharide obtained from brown seaweed,has various bioactive properties,including anti-inflammatory,anti-cancer,anti-viral,anti-oxidant,anti-coagulant,anti-thrombotic,anti-angiogenic,and anti-Helicobacter pylori properties.However,the effects of low-molecular-weight fucoidan(LMW-F)on melanoma cell lines and three dimensional(3D)cell culture models are not well understood.This study aimed to investigate the effects of LMW-F on A375 human melanoma cells and cryopreserved biospecimens derived from patients with advanced melanoma.Ultrasonic wave was used to fragment fucoidan derived from Fucus vesiculosus into smaller LMW-F.MTT and live/dead assays showed that LMW-F inhibited cell proliferation in both A375 cells and patientderived melanoma explants in a 3D-printed collagen scaffold.The PTEN/AKT pathway was found to be involved in the anti-melanoma effects of fucoidan.Western blot analysis revealed that LMW-F reduced the phosphorylation of Bcl-2 at Thr 56,which was associated with the prevention of anti-apoptotic activity of cancer cells.Our findings suggested that LMW-F could enhance anti-melanoma chemotherapy and improve the outcomes of patients with melanoma resistance.展开更多
Explant browning is one of the major problems in the tissue culture process, and polyphenol oxidase(PPO), is the major proteases involved in plant tissue browning. We investigated the effects of polyphenol oxidase on ...Explant browning is one of the major problems in the tissue culture process, and polyphenol oxidase(PPO), is the major proteases involved in plant tissue browning. We investigated the effects of polyphenol oxidase on the early stage of browning in explants of the orchid Phalaenopsis. Our results show that PPO activity was significantly higher in explants cultured for 3 d than in the 0 h control. The levels of PPO transcripts and PPO protein were significantly higher in explants cultured for 6 h compared to the 0 h control; these high expression levels were maintained over increasing cultivation time. Cycloheximide(CHX) treatment reduced PPO transcript levels, PPO protein levels, and PPO enzyme activity. High levels of PPO m RNA and PPO protein were detected in the cytoplasm and vascular bundles of Phalaenopsis explants cultured for 6 h compared to explants cultured for 0 h, 24 h, and 3 d. CHX treatment did not significantly affect the distribution of PPO m RNA and PPO protein in explant tissues, but their levels were significantly lower than those of the untreated control.展开更多
[Objectives]This study was conducted to increase the reproduction coefficient of Japanese honeysuckle(Lonicera japonica)to keep the character of optimal benign.[Methods]The young leaves of medicinal Japanese honeysuck...[Objectives]This study was conducted to increase the reproduction coefficient of Japanese honeysuckle(Lonicera japonica)to keep the character of optimal benign.[Methods]The young leaves of medicinal Japanese honeysuckle were selected as explants,and MS was used as the basic culture medium.Suitable culture concentrations and conditions were screened through different concentration gradients of growth regulators and cytokinin.[Results]As the concentration of 6-BA in the culture medium increased,the browning rate increased,and the browning phenomenon occurred earlier.On the contrary,a lower concentration of 6-BA was suitable for the differentiation and growth of young leaves,and the browning response was slow.However,if the cultivation time was too long and the materials were not transferred in a timely manner,browning would also occur.The optimal combination of levels was obtained through a 3×3 orthogonal experiment(three parallel groups for each of 6-BA and NAA).The culture conditions included a constant temperature of 26℃and light intensity of 1200 lx.The optimal medium for inducing callus proliferation was MS+6-BA 0.5 mg/L+NAA 0.5 mg/L;and the optimal medium for inducing bud differentiation was MS+6-BA 1.0 mg/L+NAA 0.1 mg/L.[Conclusions]This study provides a theoretical basis for accelerating the development of the honeysuckle industry.展开更多
The skin is a formidable physical and biological barrier which communicates continuously with the outside of the body. And the stratum corneum, the outermost layer of human epidermis, plays a central role in the inter...The skin is a formidable physical and biological barrier which communicates continuously with the outside of the body. And the stratum corneum, the outermost layer of human epidermis, plays a central role in the interaction between the cutaneous tissue and the external environment. The horny layer, and more generally the whole skin layers, avoid the penetration of harmful exogenous agents, produce molecules named anti-microbial peptides which impact the composition of the cutaneous microbiota, regulate the internal corporal temperature, avoid the water loss from the inside of the body and constitute an incredible efficient anti-oxidant network. Nevertheless, nowadays, the skin is more and more solicited by the different elements of the cutaneous exposome, including atmospheric pollution and solar radiations, which can cause a dramatic acceleration of the skin ageing process. As a consequence, due to the multifunctional protective role of the skin, during the recent decade the cosmetic industry invested massively in the development of new raw materials and end-products (dermo-cosmetics) able to preserve an optimal state of the skin regarding the external environment. Based on their physical-chemical properties thermal spring waters, which are extremely rich in inorganics ions, are interesting and powerful candidates to be part, as integral component, of new efficient dermo-cosmetic formulations dedicated to protect the skin from the external stimuli. The aim of the present work was to investigate and characterize the activity of Jonzac thermal spring water on the skin. Using different models, we proved for the first time that Jonzac thermal spring water reinforces the barrier function of the skin by modulating the expression of key markers including filaggrin and human beta defensin 2 on ex vivo human skin. The ex vivo and in vivo hydration activity, by Raman spectroscopy and corneometry respectively, has been also demonstrated. We have also shown that Jonzac thermal spring water ameliorates significantly the cutaneous microrelief in vivo. To conclude, we characterize the soothing effect of Jonzac thermal spring water by the analysis of histamine release in Substance P treated skin explants and by measuring the redness of the skin following UV exposure of the skin in vivo. We observed that both parameters decreased following a preventive treatment of the skin with Jonzac thermal spring water. Taken together our results indicate that Jonzac thermal spring water is a promising and powerful dermo-cosmetic which can be used to preserve an optimal state of the cutaneous tissue.展开更多
In this study,we investigated the preferable explant types and sterilization method for tissue culture in Chinese chestnut,in order to provide a technical support for the asexual reproduction of Chinese chestnut.The b...In this study,we investigated the preferable explant types and sterilization method for tissue culture in Chinese chestnut,in order to provide a technical support for the asexual reproduction of Chinese chestnut.The base,middle and apex parts of annual shoots with buds in Chinese chestnut were sampled and cut to 2-3 cm stem segments each with one bud,then sterilized orderly with different duration in 2%NaClO plus 0.1%HgCl_(2)solutions.The results indicated that the duration of 2%NaClO for 20 min+0.1%HgCl_(2)for 15 min exerted an effective disinfection property on the middle parts of annual shoots with buds,and achieved a contamination rate under 5%and a survival rate over 90%.展开更多
This article presents data on the introduction of in vitro culture and microclonal propagation of plants identified in the group of hyperhalophytes belonging to the species Suaeda arcuata Bunge. This study was carried...This article presents data on the introduction of in vitro culture and microclonal propagation of plants identified in the group of hyperhalophytes belonging to the species Suaeda arcuata Bunge. This study was carried out to optimize the composition of nutrient media for the main stages of reproduction in vitro, as well as studies on the rooting and adaptation of regenerants for species of the genus Suaeda obtained from axillary or apical buds, but more often from stem segments with a node. In this work, hormones of the cytokinin and auxin series, or a combination of them, were added to the nutrient environment for growth activation. The analyzing of microplants for the content of soluble in water B vitamins was carried out. As a result of the research, it was found that intact Suaeda arcuata plants in their natural habitats produce a large amount of soluble in water vitamins: riboflavin—0.062% and thiamine up to 0.006%. And in regenerated plants obtained on media without hormones, the content of vitamins was: B2 0.053%, B1 0%, respectively, and with a combination of 1/2 MS + 1 mg/l 6-BAP + 0.3 mg/l IAA + 2, 4-D, the content of vitamins varied as follows: riboflavin—0.059%, folic acid—0.030%, and thiamine was not detected. The cultivation of regenerates on the environment 1/2 MS + 1 mg/l 6-BAP + 0.3 mg/l IAA + 2,4-D showed the best effect on the growth of regenerants, created the possibility of obtaining the maximum amount of biomass and accumulation of B vitamins.展开更多
Objective:To determine the effects of different cytokinins at various concentrations onin vitro shoot multiplication of an important medicinal plant.Methods:Nodal explants(1.5-2.0 cm)of Sophora tonkinensiswere used.Mu...Objective:To determine the effects of different cytokinins at various concentrations onin vitro shoot multiplication of an important medicinal plant.Methods:Nodal explants(1.5-2.0 cm)of Sophora tonkinensiswere used.Multiple shoots were induced from nodal explants cultured onthe Murashige and Skoog(MS)medium supplemented with 0.0,0.5,1.0,2.0,4.0,8.0,or 16.0μmol2-isopentyladenine(2iP),N6 benzyladenine,kinetin or thiadiazuron.Results:Among the fourinvestigated cytokinins,2iP showed the best response for shoot multiplication.Maximum shootinduction(75%)was achieved on the MS medium supplemented with 2.0μmol 2iP,with a meannumber of 5.0 shoots per explant.In comparison to other cytokinins tried,2iP showed the highestshoot elongation with a mean shoot length of 4.8 cm.Root initiation was observed within 15 dwithin the transfer of shoots onto the MS basal medium,and the rooting percentage was 100%with a mean number of 5.4 roots per shoot and root length of 6.2 cm over a period of 4 weeks.Thehealthy plants,hardened and transferred to a greenhouse for proper acclimatization,exhibited100%survival.Conclusions:It can be summarized that 2iP is the optimal plant growth regulatorforSophoramultiplication.展开更多
Salvadora oleoides Decne. is a pharmaceutically important plant. Owing to poor seed formation, viability and, germination, and to anthropogenic disturbances, this species is on the verge of extinction. A reproducible ...Salvadora oleoides Decne. is a pharmaceutically important plant. Owing to poor seed formation, viability and, germination, and to anthropogenic disturbances, this species is on the verge of extinction. A reproducible micropropagation protocol to increase the population through tissue culture has been standardized and the results are reported here. Callus tissues were initiated from young leaves and stem explants. Leaf calluses proliferated with 1.5 mg/L BAP and 0.9 mg/L 2, 4-D with additives and continuous slow proliferation up to 15 weeks on 0.5 mg/L BAP and additives with 200 mg/L activated charcoal.Direct shoot initiation took place from stem node explants after 12 days; 4–5 shoots per node were produced in 30 days. Shoot clumps elongated and grew further on MS media supplemented with 2 mg/L BAP, 0.2 mg/L NAA and additives, which generated 20–23 shoots. The elongated shoots induced tap roots with 4 mg/L NAA and200 mg/L activated charcoal in 12 days. In vitro raised plants produced secondary roots when transferred to pots containing vermiculite maintained at 28–35 ℃. The plantlets successfully acclimatised in pots containing soil in natural conditions.展开更多
Retinal ganglion cell(RGC) axons provide the only link between the light sensitive and photon transducing neural retina and visual centers of the brain.RGC axon degeneration occurs in a number of blinding diseases and...Retinal ganglion cell(RGC) axons provide the only link between the light sensitive and photon transducing neural retina and visual centers of the brain.RGC axon degeneration occurs in a number of blinding diseases and the ability to stimulate axon regeneration from surviving ganglion cells could provide the anatomic substrate for restoration of vision.OTX2 is a homeoprotein transcription factor expressed in the retina and previous studies showed that,in response to stress,exogenous OTX2 increases the in vitro and in vivo survival of RGCs.Here we examined and quantified the effects of OTX2 on adult RGC axon regeneration in vitro and in vivo.The results show that exogenous OTX2 stimulates the regrowth of axons from RGCs in cultures of dissociated adult retinal cells and from explants of adult retinal tissue and that RGCs respond directly to OTX2 as regrowth is observed in cultures of purified adult rat RGCs.Importantly,after nerve crush in vivo,we observed a positive effect of OTX2 on the number of regenerating axons up to the optic chiasm within 14 days post crush and a very modest level of acuity absent in control mice.The effect of OTX2 on RGC survival and regeneration is of potential interest for degenerative diseases affecting this cell type.All animal procedures were approved by the local "Comié d'éιthique en expérimentation animale n°59" and authorization n° 00702.01 delivered March 28,2014 by the French "Ministére de l'enseignement supérieur et de la recherche".展开更多
Shoot organogenesis is critical for the shortening of long breeding cycles and circumvent the barrier of cloning mature Eucalyptus cloeziana trees.It enables large-scale production of plants from transformed tissues.T...Shoot organogenesis is critical for the shortening of long breeding cycles and circumvent the barrier of cloning mature Eucalyptus cloeziana trees.It enables large-scale production of plants from transformed tissues.This study evaluates the effect ofα-naphthaleneacetic acid(NAA),thidiazuron(TDZ)and benzylaminopurine(BAP)on the organogenesis of E.cloeziana from hypocotyls and cotyledonary leaves.In the induction stage,hypocotyls and cotyledonary leaves were established in a Murashige and Skoog(MS)culture medium supplemented with NAA or TDZ.Callus tissues were cultivated in a MS culture medium containing only BAP or different concentrations of BAP/NAA in the differentiation stage.Adventitious buds were multiplied in vitro and elongated in a WPM culture medium supplemented with 0.89μM BAP and 0.05μM NAA.Cotyledonary leaves exhibited the best in vitro regeneration.The induction of adventitious buds occurred only in calluses induced from TDZ.In the differentiation stage,4.4μM BAP treatment promoted an increase of adventitious bud regeneration.Micro-cuttings from regenerated shoots were acclimatized and rooted ex vitro in mini-incubators.The results confirm the establishment of an efficient protocol for the in vitro regeneration of E.cloeziana by indirect organogenesis,providing new insights regarding cloning of this species.展开更多
[Objectives] This study was conducted to develop a rapid propagation method for red sandalwood ( Pterocarpus santalinus ) by tissue culture.[Methods] The well-grown red sandalwood seed embryos were inoculated into thr...[Objectives] This study was conducted to develop a rapid propagation method for red sandalwood ( Pterocarpus santalinus ) by tissue culture.[Methods] The well-grown red sandalwood seed embryos were inoculated into three kinds of culture media after aseptic treatment,and the aseptic explants were obtained and inoculated into six kinds of media for light culture.[Results] In the best disinfection schemes of red sandalwood,disinfecting with HgCl 2 for 8 min achieved the highest germination and survival rates;when the medium for inducing red sandalwood explants was MS+0.2 mg/L IBA,the induction rate reached a maximum value;and when the culture medium for inducing stem segments of aseptic red sandalwood plantlets was MS+3.0 mg/L 6-BA+0.3 mg/L IBA,the growth of the stem segments achieved the best effect.The optimal medium for inducing red sandalwood explants was MS+IBA 0.2 mg/L,and the optimal medium for inducing stem segments of red sandalwood was MS+3.0 mg/L 6-BA+0.3 mg/L IBA.[Conclusions] The results of this study have a large reproductive coefficient,simple process and low cost,which have outstanding value for promoting the breeding and promotion of red sandalwood seedlings.展开更多
The present study aimed to develop a protocol for somatic embryogenesis and encapsulation of coffee embryos(Coffea arabica L.),for the conservation of genotypes with characteristics of commercial interest.Somatic embr...The present study aimed to develop a protocol for somatic embryogenesis and encapsulation of coffee embryos(Coffea arabica L.),for the conservation of genotypes with characteristics of commercial interest.Somatic embryos were induced from leaf explants in Murashige and Skoog medium(MS)supplemented with 1 mg·L^(−1) of 2,4-dichlorophenoxiacetic acid(2,4-D)combined with 2 mg·L^(−1) of benzyladenine(BA).Somatic embryos(SE)at the globular stage were encapsulated in a sodium alginate matrix;two treatments were tested:MS+5 mg·L^(−1) BA+1 mg·L^(−1) NAA+3%(w/v)alginate,and MS+7 mg·L^(−1) BA+5.7 mg·L^(−1) indoleacetic acid(IAA)+3%(w/v)alginate.Alginate was complexed with 100 mM calcium chloride(CaCl_(2)).Viability of the encapsulated SE was determined by staining with 0.01%fluorescein diacetate(FDA)after 0,15,30,and 45 days of storage at 4℃.Embryo viability was 100%in both treatments.展开更多
There is no efficient tracking system available for the therapeutic molecules delivered to cartilage.The dense matrix covering the cartilage surface is the main biological barrier that the therapeutic molecules must o...There is no efficient tracking system available for the therapeutic molecules delivered to cartilage.The dense matrix covering the cartilage surface is the main biological barrier that the therapeutic molecules must overcome.In this study,we aimed to establish a system that can dynamically and effectively track the therapeutic molecules delivered to cartilage.To this aim,we adopted bovine and human cartilage explants as ex vivo models for chondrocyte-targeted exosome dispersion.The efficiency of drug delivery was evaluated using frozen sections.The results of this study showed that the penetration and distribution of chondrocyte-targeted exosomes in cartilage explants can be tracked dynamically.Thus,ex vivo cartilage explants provide an effective and economic system to evaluate therapeutic drugs encapsulated in chondrocyte-targeted exosomes in preclinical studies.展开更多
基金FEDER/Region Centre Val de Loire ANIMALT grant (FEDER convention number EX007516, Region Centre convention number 2019–00134936, research program number AE-2019–1850)a post-doc fellowship from INRAE–Département Santé Animale
文摘Background Mammary gland(MG)infections(mastitis)are frequent diseases of dairy cows that affect milk quality,animal welfare and farming profitability.These infections are commonly associated with the bacteria Escherichia coli and Staphylococcus aureus.Different in vitro models have been used to investigate the early response of the MG to bacteria,but the role of the teat in mastitis pathogenesis has received less attention.In this study,we used punch-excised teat tissue as an ex vivo model to study the immune mechanisms that arise early during infection when bacteria have entered the MG.Results Cytotoxicity and microscopic analyses showed that bovine teat sinus explants have their morphology and viability preserved after 24 h of culture and respond to ex vivo stimulation with TLR-agonists and bacteria.LPS and E.coli trigger stronger inflammatory response in teat when compared to LTA and S.aureus,leading to a higher produc-tion of IL-6 and IL-8,as well as to an up-regulation of proinflammatory genes.We also demonstrated that our ex vivo model can be applied to frozen-stored explants.Conclusions In compliance with the 3Rs principle(replacement,reduction and refinement)in animal experimenta-tion,ex vivo explant analyses proved to be a simple and affordable approach to study MG immune response to infec-tion.This model,which better reproduces organ complexity than epithelial cell cultures or tissue slices,lends itself particularly well to studying the early phases of the MG immune response to infection.
文摘We developed a system for the regeneration of Lombardy poplar (Populus nigra L. var. italica) shoots from internodal stem explants. Using this system, shoots regenerated from 87% of the stem explants placed on Murashige and Skoog (MS) medium supplemented with 0.1 mg/L indole-3-acetic acid and 0.5 mg/L benzylaminopurine without undergoing callus formation. About 80% of the in vitro regenerated shoots developed roots on MS medium supplemented with 0.5 mg/L indole-3-butyric acid and 0.02 mg/L 1-naphthylacetic acid. Well-rooted seven-to eight-week-old regenerated plants could be transferred to soil for further growth and the survival rate of such plants after three weeks was 88%. The protocol presented here is simple and economical because it does not rely on pre-incubation in callus induction medium or repeated subculture in shoot induction medium containing trans-zeatin, an expensive substance. The in vitro regeneration system presented here could be used for evaluation of radiation sensitivity for Lombardy poplar tissues.
文摘Adventitious shoots were successfully regenerated from hypocotyl explants of in vitro cultures of Euonymus japonicusCu zhi. Hypocotyl slices were cultured on Murashige and Skoog (MS) and B5 basal medium supplemented with variedconcentration of different plant growth-regulators, e.g., α-naphthaleneacetic acid (NAA) and indole-3-butyric acid (IBA)in combination with 6-benzylaminopurine (6-BA) and kinetin. The study showed that shoots could be directly regeneratedfrom hypocotyl explants without the intervening callus phase; MS medium was more suitable for adventitious shootsregeneration. The ability of hypocotyls segments to produce shoots varied depending upon their position on the seedlings.The highest regeneration rate was obtained with hypocotyl segments near to the cotyledon cultured on MS basal mediumsupplemented with 1.5 mg L-1 6-BA and 0.05 mg L-1 NAA (63.64%). The regenerated shoots were readily elongated on thesame medium as used for multiplication and rooted on half-strength MS basal medium supplemented with 1.0 mg L-1 IBAand 100 mg L-1 activated carbon. After being transferred to greenhouse conditions, 96% of the plantlets were successfullyacclimatized. This regeneration system is applied for genetic transformation now.
文摘We have previously shown that capacitively coupled electrical stimulation of either normal bovine articular chondrocytes or osteoarthritic human articular cartilage explants resulted in up-regulation of cartilage matrix gene expression and down-regulation of metalloproteinase gene expression. In addition, collagen and proteoglycan protein levels were also elevated. To determine visually the effect of specific electric fields on modifying cartilage structure, freshly harvested human full-thickness osteoarthritic cartilage explants were stimulated in the absence or presence of interleukin-1β, an inflammatory cytokine, and were examined photographically and spectrophotometrically. Hexosamine and hydroxyproline contents were also determined. Spectrophotometric analysis was used to quantify any changes in the depth of defects in the cartilage ranging from surface level (red-colored) to the deepest affected layer (blue-colored). Interleukin-1β treatment alone caused significant additional cartilage erosion. Electrical stimulation alone resulted in significant decreases in the cartilage defects. Electrical stimulation in the presence of interleukin-1β resulted in a small, but significant, surface improvement. Meta-analysis also confirmed a significant increase in the hexosamine and hydroxyproline contents (indicating matrix deposition). It was concluded that an appropriate electric field could modify osteoarthritic lesions in full-thickness cartilage plugs by increasing matrix production and/or by decreasing matrix destruction. Furthermore, it appears that spectrophotometric analysis is a relatively easy method for quantifying the “filling in” or healing of articular cartilage defects.
文摘In the present investigation, an attempt has been made to study the influence of ethylene inhibitor silver nitrate on direct shoot regeneration in Sphaeranthus indicus, an important antijaundice medicinal plant, by using in vitro raised shoot tip explants. The effect of various concentrations of kinetin, BAP (0.5 - 3.0 mg/l), and NAA (0.1 - 0.5 mg/l) along with AgNO<sub>3</sub> (0.1 - 1.0 mg/l) was studied. Among the combinations tested MS medium augmented with kinetin (1.0 mg/l), NAA (0.1 mg/l) and AgNO<sub>3</sub> (0.4 mg/l) was found to be optimum for production of multiple shoots (34.3 ± 0.36). Addition of AgNO<sub>3</sub> to the media not only increases shoot number in all the concentrations tested but also shoot length. AgNO<sub>3</sub> at the concentration of 0.4 mg/l produced 35% more number of multiple shoots when compared to multiple shoots (10.8 ± 0.12) produced in control. In the present study by the addition of ethylene inhibitor silver nitrate and growth regulators, more number of multiple shoots (three folds) and shoot length was observed compared to control. These in vitro raised shoots were transferred to the rooting medium containing different concentrations of auxins such as NAA and IAA along with AgNO<sub>3</sub> (0.1 - 0.6 mg/l). Better rooting response (21.6) was observed on NAA (2.0 mg/l) and AgNO<sub>3</sub> (0.4 mg/l) containing media. The healthy rooted plantlets were transferred to polybags containing soil and vermiculate in 1:1 ratio for hardening. Finally the hardened plants were transferred to field environment for utmost survivability.
文摘Objective:To develop the reproducible in vitro propagation protocols for the medicinally important plants viz.,Achyranthes aspera(A.aspera)L.and Achyranthes bidentata(A.bidentata)Blume using nodal segments as explants.Methods:Young shoots of A.aspera and A.bidentata were harvested and washed with running tap water and treated with 0.1%bavistin and rinsed twice with distilled water.Then the explants were surface sterilized with 0.1%(w/v)HgCl_2 solutions for I min.After rinsing with sterile distilled water for 3-4 times,nodal segments were cut into smaller segments(1 cm)and used as the explants.The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog(MS)medium supplemented with 3%sucrose,0.6%(w/v)agar(HiMedia,Mumbai)and different concentration and combination of 6-benzyl amino purine(BAP),kinetin(Kin),naphthalene acetic acid(NAA)and indole acetic acid(IAA)for direct regeneration.Results:Adventitious proliferation was obtained from A.aspera and A.bidentata nodal segments inoculated on MS basal medium with 3%sucrose and augmented with BAP and Kin with varied frequency.MS medium augmented with 3.0 mg/L of BAP showed the highest percentage(93.60±0.71)of shootlets formation for A.aspera and(94.70±0.53)percentages for A.bidentata.Maximum number of shoots/explants(10.60±0.36)for A.aspera and(9.50±0.56)for A.bidentata was observed in MS medium fortified with 5.0 mg/L of BAP.For A.aspera,maximum mean length(5.50±0.34)of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A.bidentata(5.40±0.61)was observed in the very same concentration.The highest percentage,maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of 1BA.Seventy percentages of plants were successfully established in polycups.Sixty eight percentages of plants were well established in the green house condition.Sixty five percentages of plants were established in the field.Conclusions:The results have shown that use of nodal buds is an alternative reproducible and dependable method for clonal propagation of A.aspera and A.bidentata.The high rate of direct shoot-root multiplication and their high rate of post-hardening survival indicate that this protocol can he easily adopted for commercial large scale cultivation.
基金supported by The Norwegian Research CouncilDepartment of Ophthalmology,Oslo University Hospital,Oslo,Norway(to TPU)+10 种基金Department of Medical Biochemistry,Oslo University Hospital,Oslo,Norway(to TPU)The Norwegian Association for the Blind and Partially Sighted(to TPU)The Ministry of Science and Technology of Taiwan,China MOST 105-2917-I-002-031,MOST 109-2917-I-564-032(to KC)The Scientific and Technological Research Council of Turkiye-TUBITAK(to KG)BrightFocus Foundation(to KSC)the Massachusetts Lions Foundation(to KSC)National Eye Institute Grant EY031696(to DFC)Harvard NeuroDiscovery Center Grant(to DFC)Department of Defense(USA)HT9425-23-1-1045(to DFC and AL)Core Grant for Vision Research from NIH/NEI to the Schepens Eye Research Institute(P30EY003790)South-Eastern Norway Regional Health Authority and the Norwegian Society of the Blind(to TPU).
文摘Retinitis pigmentosa is a hereditary retinal disease that affects rod and cone photoreceptors,leading to progressive photoreceptor loss.Previous research supports the beneficial effect of electrical stimulation on photoreceptor survival.This study aims to identify the most effective electrical stimulation parameters and functional advantages of transcorneal electrical stimulation(tcES)in mice affected by inherited retinal degeneration.Additionally,the study seeked to analyze the electric field that reaches the retina in both eyes in mice and post-mortem humans.In this study,we recorded waveforms and voltages directed to the retina during transcorneal electrical stimulation in C57BL/6J mice using an intraocular needle probe with rectangular,sine,and ramp waveforms.To investigate the functional effects of electrical stimulation on photoreceptors,we used human retinal explant cultures and rhodopsin knockout(Rho^(-/-))mice,demonstrating progressive photoreceptor degeneration with age.Human retinal explants isolated from the donors’eyes were then subjected to electrical stimulation and cultured for 48 hours to simulate the neurodegenerative environment in vitro.Photoreceptor density was evaluated by rhodopsin immunolabeling.In vivo Rho^(-/-)mice were subjected to two 5-day series of daily transcorneal electrical stimulation using rectangular and ramp waveforms.Retinal function and visual perception of mice were evaluated by electroretinography and optomotor response(OMR),respectively.Immunolabeling was used to assess the morphological and biochemical changes of the photoreceptor and bipolar cells in mouse retinas.Oscilloscope recordings indicated effective delivery of rectangular,sine,and ramp waveforms to the retina by transcorneal electrical stimulation,of which the ramp waveform required the lowest voltage.Evaluation of the total conductive resistance of the post-mortem human compared to the mouse eyes indicated higher cornea-to-retina resistance in human eyes.The temperature recordings during and after electrical stimulation indicated no significant temperature change in vivo and only a subtle temperature increase in vitro(~0.5-1.5°C).Electrical stimulation increased photoreceptor survival in human retinal explant cultures,particularly at the ramp waveform.Transcorneal electrical stimulation(rectangular+ramp)waveforms significantly improved the survival and function of S and M-cones and enhanced visual acuity based on the optomotor response results.Histology and immunolabeling demonstrated increased photoreceptor survival,improved outer nuclear layer thickness,and increased bipolar cell sprouting in Rho^(-/-)mice.These results indicate that transcorneal electrical stimulation effectively delivers the electrical field to the retina,improves photoreceptor survival in both human and mouse retinas,and increases visual function in Rho^(-/-)mice.Combined rectangular and ramp waveform stimulation can promote photoreceptor survival in a minimally invasive fashion.
基金supported by the Priority Research Centers Program through the National Research Foundation of Korea(NRF)funded by the Ministry of Education,Science and Technology(Grant 2017R1A6A03015562 and RS-2023-00237386).
文摘Fucoidan,a sulfate polysaccharide obtained from brown seaweed,has various bioactive properties,including anti-inflammatory,anti-cancer,anti-viral,anti-oxidant,anti-coagulant,anti-thrombotic,anti-angiogenic,and anti-Helicobacter pylori properties.However,the effects of low-molecular-weight fucoidan(LMW-F)on melanoma cell lines and three dimensional(3D)cell culture models are not well understood.This study aimed to investigate the effects of LMW-F on A375 human melanoma cells and cryopreserved biospecimens derived from patients with advanced melanoma.Ultrasonic wave was used to fragment fucoidan derived from Fucus vesiculosus into smaller LMW-F.MTT and live/dead assays showed that LMW-F inhibited cell proliferation in both A375 cells and patientderived melanoma explants in a 3D-printed collagen scaffold.The PTEN/AKT pathway was found to be involved in the anti-melanoma effects of fucoidan.Western blot analysis revealed that LMW-F reduced the phosphorylation of Bcl-2 at Thr 56,which was associated with the prevention of anti-apoptotic activity of cancer cells.Our findings suggested that LMW-F could enhance anti-melanoma chemotherapy and improve the outcomes of patients with melanoma resistance.
基金supported by Natural Science Foundation in China(31070618)
文摘Explant browning is one of the major problems in the tissue culture process, and polyphenol oxidase(PPO), is the major proteases involved in plant tissue browning. We investigated the effects of polyphenol oxidase on the early stage of browning in explants of the orchid Phalaenopsis. Our results show that PPO activity was significantly higher in explants cultured for 3 d than in the 0 h control. The levels of PPO transcripts and PPO protein were significantly higher in explants cultured for 6 h compared to the 0 h control; these high expression levels were maintained over increasing cultivation time. Cycloheximide(CHX) treatment reduced PPO transcript levels, PPO protein levels, and PPO enzyme activity. High levels of PPO m RNA and PPO protein were detected in the cytoplasm and vascular bundles of Phalaenopsis explants cultured for 6 h compared to explants cultured for 0 h, 24 h, and 3 d. CHX treatment did not significantly affect the distribution of PPO m RNA and PPO protein in explant tissues, but their levels were significantly lower than those of the untreated control.
基金Supported by Bureau of Science and Technology of Huizhou Municipality(2022CQ0100252021SC040202004).
文摘[Objectives]This study was conducted to increase the reproduction coefficient of Japanese honeysuckle(Lonicera japonica)to keep the character of optimal benign.[Methods]The young leaves of medicinal Japanese honeysuckle were selected as explants,and MS was used as the basic culture medium.Suitable culture concentrations and conditions were screened through different concentration gradients of growth regulators and cytokinin.[Results]As the concentration of 6-BA in the culture medium increased,the browning rate increased,and the browning phenomenon occurred earlier.On the contrary,a lower concentration of 6-BA was suitable for the differentiation and growth of young leaves,and the browning response was slow.However,if the cultivation time was too long and the materials were not transferred in a timely manner,browning would also occur.The optimal combination of levels was obtained through a 3×3 orthogonal experiment(three parallel groups for each of 6-BA and NAA).The culture conditions included a constant temperature of 26℃and light intensity of 1200 lx.The optimal medium for inducing callus proliferation was MS+6-BA 0.5 mg/L+NAA 0.5 mg/L;and the optimal medium for inducing bud differentiation was MS+6-BA 1.0 mg/L+NAA 0.1 mg/L.[Conclusions]This study provides a theoretical basis for accelerating the development of the honeysuckle industry.
文摘The skin is a formidable physical and biological barrier which communicates continuously with the outside of the body. And the stratum corneum, the outermost layer of human epidermis, plays a central role in the interaction between the cutaneous tissue and the external environment. The horny layer, and more generally the whole skin layers, avoid the penetration of harmful exogenous agents, produce molecules named anti-microbial peptides which impact the composition of the cutaneous microbiota, regulate the internal corporal temperature, avoid the water loss from the inside of the body and constitute an incredible efficient anti-oxidant network. Nevertheless, nowadays, the skin is more and more solicited by the different elements of the cutaneous exposome, including atmospheric pollution and solar radiations, which can cause a dramatic acceleration of the skin ageing process. As a consequence, due to the multifunctional protective role of the skin, during the recent decade the cosmetic industry invested massively in the development of new raw materials and end-products (dermo-cosmetics) able to preserve an optimal state of the skin regarding the external environment. Based on their physical-chemical properties thermal spring waters, which are extremely rich in inorganics ions, are interesting and powerful candidates to be part, as integral component, of new efficient dermo-cosmetic formulations dedicated to protect the skin from the external stimuli. The aim of the present work was to investigate and characterize the activity of Jonzac thermal spring water on the skin. Using different models, we proved for the first time that Jonzac thermal spring water reinforces the barrier function of the skin by modulating the expression of key markers including filaggrin and human beta defensin 2 on ex vivo human skin. The ex vivo and in vivo hydration activity, by Raman spectroscopy and corneometry respectively, has been also demonstrated. We have also shown that Jonzac thermal spring water ameliorates significantly the cutaneous microrelief in vivo. To conclude, we characterize the soothing effect of Jonzac thermal spring water by the analysis of histamine release in Substance P treated skin explants and by measuring the redness of the skin following UV exposure of the skin in vivo. We observed that both parameters decreased following a preventive treatment of the skin with Jonzac thermal spring water. Taken together our results indicate that Jonzac thermal spring water is a promising and powerful dermo-cosmetic which can be used to preserve an optimal state of the cutaneous tissue.
基金Supported by The Sub-project of Hebei Collaborative Innovation Center of Chestnut Industry(202202)The Sub-project of Research and Development Center of Horticultural Crop Breeding Application Technology of Hebei University(YF201404)+1 种基金Chestnut Science and Technology Backyard of Hebei QinglongChina Rural Special Technology Association。
文摘In this study,we investigated the preferable explant types and sterilization method for tissue culture in Chinese chestnut,in order to provide a technical support for the asexual reproduction of Chinese chestnut.The base,middle and apex parts of annual shoots with buds in Chinese chestnut were sampled and cut to 2-3 cm stem segments each with one bud,then sterilized orderly with different duration in 2%NaClO plus 0.1%HgCl_(2)solutions.The results indicated that the duration of 2%NaClO for 20 min+0.1%HgCl_(2)for 15 min exerted an effective disinfection property on the middle parts of annual shoots with buds,and achieved a contamination rate under 5%and a survival rate over 90%.
文摘This article presents data on the introduction of in vitro culture and microclonal propagation of plants identified in the group of hyperhalophytes belonging to the species Suaeda arcuata Bunge. This study was carried out to optimize the composition of nutrient media for the main stages of reproduction in vitro, as well as studies on the rooting and adaptation of regenerants for species of the genus Suaeda obtained from axillary or apical buds, but more often from stem segments with a node. In this work, hormones of the cytokinin and auxin series, or a combination of them, were added to the nutrient environment for growth activation. The analyzing of microplants for the content of soluble in water B vitamins was carried out. As a result of the research, it was found that intact Suaeda arcuata plants in their natural habitats produce a large amount of soluble in water vitamins: riboflavin—0.062% and thiamine up to 0.006%. And in regenerated plants obtained on media without hormones, the content of vitamins was: B2 0.053%, B1 0%, respectively, and with a combination of 1/2 MS + 1 mg/l 6-BAP + 0.3 mg/l IAA + 2, 4-D, the content of vitamins varied as follows: riboflavin—0.059%, folic acid—0.030%, and thiamine was not detected. The cultivation of regenerates on the environment 1/2 MS + 1 mg/l 6-BAP + 0.3 mg/l IAA + 2,4-D showed the best effect on the growth of regenerants, created the possibility of obtaining the maximum amount of biomass and accumulation of B vitamins.
基金This study was carried out with the support of"On-Site Cooperative Agriculture Research Project(Grant No.006330)",RDA,Republic of Korea
文摘Objective:To determine the effects of different cytokinins at various concentrations onin vitro shoot multiplication of an important medicinal plant.Methods:Nodal explants(1.5-2.0 cm)of Sophora tonkinensiswere used.Multiple shoots were induced from nodal explants cultured onthe Murashige and Skoog(MS)medium supplemented with 0.0,0.5,1.0,2.0,4.0,8.0,or 16.0μmol2-isopentyladenine(2iP),N6 benzyladenine,kinetin or thiadiazuron.Results:Among the fourinvestigated cytokinins,2iP showed the best response for shoot multiplication.Maximum shootinduction(75%)was achieved on the MS medium supplemented with 2.0μmol 2iP,with a meannumber of 5.0 shoots per explant.In comparison to other cytokinins tried,2iP showed the highestshoot elongation with a mean shoot length of 4.8 cm.Root initiation was observed within 15 dwithin the transfer of shoots onto the MS basal medium,and the rooting percentage was 100%with a mean number of 5.4 roots per shoot and root length of 6.2 cm over a period of 4 weeks.Thehealthy plants,hardened and transferred to a greenhouse for proper acclimatization,exhibited100%survival.Conclusions:It can be summarized that 2iP is the optimal plant growth regulatorforSophoramultiplication.
基金financially supported from Central University of Punjab,Bathinda,India
文摘Salvadora oleoides Decne. is a pharmaceutically important plant. Owing to poor seed formation, viability and, germination, and to anthropogenic disturbances, this species is on the verge of extinction. A reproducible micropropagation protocol to increase the population through tissue culture has been standardized and the results are reported here. Callus tissues were initiated from young leaves and stem explants. Leaf calluses proliferated with 1.5 mg/L BAP and 0.9 mg/L 2, 4-D with additives and continuous slow proliferation up to 15 weeks on 0.5 mg/L BAP and additives with 200 mg/L activated charcoal.Direct shoot initiation took place from stem node explants after 12 days; 4–5 shoots per node were produced in 30 days. Shoot clumps elongated and grew further on MS media supplemented with 2 mg/L BAP, 0.2 mg/L NAA and additives, which generated 20–23 shoots. The elongated shoots induced tap roots with 4 mg/L NAA and200 mg/L activated charcoal in 12 days. In vitro raised plants produced secondary roots when transferred to pots containing vermiculite maintained at 28–35 ℃. The plantlets successfully acclimatised in pots containing soil in natural conditions.
基金supported by Fovea-Pharmaceuticals,and Global Research Laboratory Program Grant 2009-00424 from the Korean Ministry of Education,Science,and Technology,HOMEOSIGN:ERC-2013-AdG n°339379 and Neuropr Otx:ANR-16-CE16-0003-02。
文摘Retinal ganglion cell(RGC) axons provide the only link between the light sensitive and photon transducing neural retina and visual centers of the brain.RGC axon degeneration occurs in a number of blinding diseases and the ability to stimulate axon regeneration from surviving ganglion cells could provide the anatomic substrate for restoration of vision.OTX2 is a homeoprotein transcription factor expressed in the retina and previous studies showed that,in response to stress,exogenous OTX2 increases the in vitro and in vivo survival of RGCs.Here we examined and quantified the effects of OTX2 on adult RGC axon regeneration in vitro and in vivo.The results show that exogenous OTX2 stimulates the regrowth of axons from RGCs in cultures of dissociated adult retinal cells and from explants of adult retinal tissue and that RGCs respond directly to OTX2 as regrowth is observed in cultures of purified adult rat RGCs.Importantly,after nerve crush in vivo,we observed a positive effect of OTX2 on the number of regenerating axons up to the optic chiasm within 14 days post crush and a very modest level of acuity absent in control mice.The effect of OTX2 on RGC survival and regeneration is of potential interest for degenerative diseases affecting this cell type.All animal procedures were approved by the local "Comié d'éιthique en expérimentation animale n°59" and authorization n° 00702.01 delivered March 28,2014 by the French "Ministére de l'enseignement supérieur et de la recherche".
基金CAPES (Coordination for the Improvement of Higher Personnel, Brazil) for their scholarshipCNPq (National Counsel of Technological and Scientific Development, Brazil)IPEF (Forestry Science and Research Institute, Brazil) for technical support
文摘Shoot organogenesis is critical for the shortening of long breeding cycles and circumvent the barrier of cloning mature Eucalyptus cloeziana trees.It enables large-scale production of plants from transformed tissues.This study evaluates the effect ofα-naphthaleneacetic acid(NAA),thidiazuron(TDZ)and benzylaminopurine(BAP)on the organogenesis of E.cloeziana from hypocotyls and cotyledonary leaves.In the induction stage,hypocotyls and cotyledonary leaves were established in a Murashige and Skoog(MS)culture medium supplemented with NAA or TDZ.Callus tissues were cultivated in a MS culture medium containing only BAP or different concentrations of BAP/NAA in the differentiation stage.Adventitious buds were multiplied in vitro and elongated in a WPM culture medium supplemented with 0.89μM BAP and 0.05μM NAA.Cotyledonary leaves exhibited the best in vitro regeneration.The induction of adventitious buds occurred only in calluses induced from TDZ.In the differentiation stage,4.4μM BAP treatment promoted an increase of adventitious bud regeneration.Micro-cuttings from regenerated shoots were acclimatized and rooted ex vitro in mini-incubators.The results confirm the establishment of an efficient protocol for the in vitro regeneration of E.cloeziana by indirect organogenesis,providing new insights regarding cloning of this species.
基金Supported by National Natural Science Foundation of China(31270674)Guangdong College Students’Science and Technology Innovation and Cultivation Project(pdjhb0541)
文摘[Objectives] This study was conducted to develop a rapid propagation method for red sandalwood ( Pterocarpus santalinus ) by tissue culture.[Methods] The well-grown red sandalwood seed embryos were inoculated into three kinds of culture media after aseptic treatment,and the aseptic explants were obtained and inoculated into six kinds of media for light culture.[Results] In the best disinfection schemes of red sandalwood,disinfecting with HgCl 2 for 8 min achieved the highest germination and survival rates;when the medium for inducing red sandalwood explants was MS+0.2 mg/L IBA,the induction rate reached a maximum value;and when the culture medium for inducing stem segments of aseptic red sandalwood plantlets was MS+3.0 mg/L 6-BA+0.3 mg/L IBA,the growth of the stem segments achieved the best effect.The optimal medium for inducing red sandalwood explants was MS+IBA 0.2 mg/L,and the optimal medium for inducing stem segments of red sandalwood was MS+3.0 mg/L 6-BA+0.3 mg/L IBA.[Conclusions] The results of this study have a large reproductive coefficient,simple process and low cost,which have outstanding value for promoting the breeding and promotion of red sandalwood seedlings.
基金Project financed by the Tecnologico Nacional de Mexico in the call Support for Scientific and Applied Research Projects,Technological Development and Innovation 2015(CI-02-2015).
文摘The present study aimed to develop a protocol for somatic embryogenesis and encapsulation of coffee embryos(Coffea arabica L.),for the conservation of genotypes with characteristics of commercial interest.Somatic embryos were induced from leaf explants in Murashige and Skoog medium(MS)supplemented with 1 mg·L^(−1) of 2,4-dichlorophenoxiacetic acid(2,4-D)combined with 2 mg·L^(−1) of benzyladenine(BA).Somatic embryos(SE)at the globular stage were encapsulated in a sodium alginate matrix;two treatments were tested:MS+5 mg·L^(−1) BA+1 mg·L^(−1) NAA+3%(w/v)alginate,and MS+7 mg·L^(−1) BA+5.7 mg·L^(−1) indoleacetic acid(IAA)+3%(w/v)alginate.Alginate was complexed with 100 mM calcium chloride(CaCl_(2)).Viability of the encapsulated SE was determined by staining with 0.01%fluorescein diacetate(FDA)after 0,15,30,and 45 days of storage at 4℃.Embryo viability was 100%in both treatments.
基金National Natural Science Foundation of China(Nos.81972116,81972085,81772394,31900046)Key Program of Natural Science Foundation of Guangdong Province(No.2018B0303110003)+3 种基金Guangdong International Cooperation Project(No.2021A0505030011)Shenzhen Science and Technology Projects(Nos.GJHZ20200731095606019,JCYJ20170817172023838,JCYJ20170306092215436,JCYJ20170413161649437)China Postdoctoral Science Foundation(No.2020M682907)Special Funds for the Construction of High Level Hospitals in Guangdong Province.
文摘There is no efficient tracking system available for the therapeutic molecules delivered to cartilage.The dense matrix covering the cartilage surface is the main biological barrier that the therapeutic molecules must overcome.In this study,we aimed to establish a system that can dynamically and effectively track the therapeutic molecules delivered to cartilage.To this aim,we adopted bovine and human cartilage explants as ex vivo models for chondrocyte-targeted exosome dispersion.The efficiency of drug delivery was evaluated using frozen sections.The results of this study showed that the penetration and distribution of chondrocyte-targeted exosomes in cartilage explants can be tracked dynamically.Thus,ex vivo cartilage explants provide an effective and economic system to evaluate therapeutic drugs encapsulated in chondrocyte-targeted exosomes in preclinical studies.
