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Background: Micro-propagation is the primary technique for the mass propagation of greenhouse orchids. However, various factors, including culture media and cultivation systems, influence the scaling-up and efficient commercialization of in vitro techniques. The utilization of liquid cultivation systems and bioreactors are relatively cost-effective and has attracted significant attention for mass production. In this study, we evaluated the effects of eight culture media, in both semi-solid and liquid forms, on the growth of in vitro mini-plantlets of Phalaenopsis orchids. We subsequently assessed the performance of four selected media: half-strength modified Murashige and Skoog (1/2 MMS), modified FAST (MFAST), and two simplified media (SM1 and SM2), across four types of cultivation systems, including semi-solid media in glass jars, liquid media in a permanent immersion system using Erlenmeyer flasks on a shaker (PIS), and a temporary immersion system (TIS) in two forms: FA-Bio bioreactor (TIS-FA-Bio) and RITA® bioreactor (TIS-RITA®). Results: The results indicated that the optimal culture medium for orchid growth depends on the cultivation system used. Among the eight evaluated semi-solid and liquid media, the highest plant growth was observed in the SM2 medium, attributed to the presence of additives (banana powder and activated charcoal), two types of sugar (with high carbon content), and small amounts of nickel and ascorbic acid. The high carbon content in the SM2 liquid medium, combined with the temporary immersion explants in a large volume of medium in the TIS-RITA®, resulted in the highest total carbohydrate content and enhanced plant growth. The maximum quantum efficiency of photosystem II (Fv/Fm) and the number of plantlets per liter in SM2 semi-solid medium were higher than those in the 1/2 MMS semi-solid medium (control), leading to a 61.6% reduction in production costs. Furthermore, the number of plantlets per liter in the TIS-FA-Bio containing SM2 (10.8 USD) was higher than in TIS-RITA® with 1/2 MMS (39.3 USD), resulting in a 72.5% decrease in production costs due to a lower volume of medium and the use of inexpensive equipment. Conclusions: We recommend the use of SM2 in TIS-FA-Bio and TIS-RITA® for economical and efficient mass propagation of Phalaenopsis orchid. [ABSTRACT FROM AUTHOR]

Abstract Background Micro-propagation is the primary technique for the mass propagation of greenhouse orchids. However, various factors, including culture media and cultivation systems, influence the scaling-up and efficient commercialization of in vitro techniques. The utilization of liquid cultivation systems and bioreactors are relatively cost-effective and has attracted significant attention for mass production. In this study, we evaluated the effects of eight culture media, in both semi-solid and liquid forms, on the growth of in vitro mini-plantlets of Phalaenopsis orchids. We subsequently assessed the performance of four selected media: half-strength modified Murashige and Skoog (1/2 MMS), modified FAST (MFAST), and two simplified media (SM1 and SM2), across four types of cultivation systems, including semi-solid media in glass jars, liquid media in a permanent immersion system using Erlenmeyer flasks on a shaker (PIS), and a temporary immersion system (TIS) in two forms: FA-Bio bioreactor (TIS-FA-Bio) and RITA® bioreactor (TIS-RITA®). Results The results indicated that the optimal culture medium for orchid growth depends on the cultivation system used. Among the eight evaluated semi-solid and liquid media, the highest plant growth was observed in the SM2 medium, attributed to the presence of additives (banana powder and activated charcoal), two types of sugar (with high carbon content), and small amounts of nickel and ascorbic acid. The high carbon content in the SM2 liquid medium, combined with the temporary immersion explants in a large volume of medium in the TIS-RITA®, resulted in the highest total carbohydrate content and enhanced plant growth. The maximum quantum efficiency of photosystem II (Fv/Fm) and the number of plantlets per liter in SM2 semi-solid medium were higher than those in the 1/2 MMS semi-solid medium (control), leading to a 61.6% reduction in production costs. Furthermore, the number of plantlets per liter in the TIS-FA-Bio containing SM2 (10.8 USD) was higher than in TIS-RITA® with 1/2 MMS (39.3 USD), resulting in a 72.5% decrease in production costs due to a lower volume of medium and the use of inexpensive equipment. Conclusions We recommend the use of SM2 in TIS-FA-Bio and TIS-RITA® for economical and efficient mass propagation of Phalaenopsis orchid.

The palm Euterpe precatoria holds great social, cultural, and environmental importance. The heart of palm and the fruit are the main products used for industrialization due to their energetic properties. Thus, the aim of this study was to establish a suspension cultivation protocol for the species using different explant sources. For this, eight lineages of E. precatoria embryogenic calluses were tested, with five in liquid medium Murashige and Skoog (MS) with 5 μM Picloram and three for comparison in semisolid medium MS with 20 μM Picloram and 5 μM 2iP. The growth curve was obtained by weighing the calli from 60 to 180 days of cultivation. The Gompertz model was applied, and growth kinetics were evaluated. At 100 days, the contents of total soluble sugars (TSSs) and total soluble proteins (TSPs) were determined. Principal components (PCA) were measured. According to the analysis of the data, the cultivation of E. precatoria lineages in liquid medium was successfully carried out, and the establishment was achieved. The model can be considered adequate since the R2 values found describe more than 90% of the growth kinetics of the lineages. In the liquid system, lineages L1 (from leaf explants and multiplied in semisolid medium—SM), L2 (from leaf explants and multiplied in SM), and L6 (from zygotic embryo explants and multiplied in liquid medium—LM) showed the shortest time to double the biomass accumulation. Multivariate analysis reveals a significant increase in masses in liquid cultures, represented by lineages L6 and L2. There was statistical difference in the amount of TSSs extracted; the highest TSS levels were observed in lineages cultivated in LM. The protein content found was very low, showing statistical differences among the lineages. In this work, the establishment and multiplication of embryogenic calli of E. precatoria are described for the first time, and they emerge as viable alternatives for the vegetative propagation of the species. [ABSTRACT FROM AUTHOR]

Reynoutria japonica Houtt. (Japanese knotweed) is an invasive plant belonging to the Polygonaceae family. However, being native to East Asia, it has been used in natural medicine for ages because of its broad range of biological activity. Although R. japonica is known as a rich source of phenolic compounds, plant biomass collected from the field may be contaminated with toxic elements like heavy metals, and the level of metabolite accumulation depends on environmental conditions. Therefore, the aim of this study was to derive Japanese knotweed tissue cultures and investigate biomass production and phenolic compound synthesis in in vitro conditions. Plants were cultivated in a traditional agar-solidified medium, in a liquid medium with rotary shaking (agitated culture), and in a temporary immersion bioreactors Plantform™, as well as in soil (ex vitro conditions). Analyses of the growth index and dry weight accumulation were performed on the collected material. In the extracts obtained from examined plants, qualitative and quantitative analysis of phenolic derivatives using DAD-HPLC was conducted to determine the sum of phenolic compounds, as well as the quantity of selected phenolic acids, catechins, and other flavonoids. Results have shown that agitated cultures and temporary immersion bioreactors increased biomass accumulation compared to solid medium cultures. Tissue cultures of R. japonica had increased synthesis of phenolic compounds compared to plants from ex vitro conditions. Shoots and roots from agitated cultures were 2.8- and 3.3-fold richer in catechins, respectively, compared to plants cultivated in soil. Based on the obtained results it can be concluded that agitated and bioreactor cultures are the best source of Japanese knotweed biomass rich in valuable secondary metabolites. Key message: Reynoutria japonica Houtt. plants in agitated culture and temporary immersion bioreactor are a rich source of flavonoids, including catechin derivatives. [ABSTRACT FROM AUTHOR]

Blueberries are high-value fruits. The traditional method of propagation by cuttings cannot supply the modern market with large quantities of seedlings. The method of micropropagation of plants in vitro makes it possible to bring the production of blueberry seedlings to the highest level. Blueberries have not been sufficiently studied in in vitro culture, so the search for the simplest and most cost-effective methods of micropropagation remains relevant. The problem of accelerated micropropagation of blueberries can be solved using rocker-type bioreactors, which differ from other models in terms of simplicity of design and low cost. A study was carried out to evaluate the effectiveness of micropropagation of Vaccinium corymbosum 'Bluecrop' in rocker bioreactors. Two types of bioreactors were compared: the bioreactor of the Platform system and the TIS rocker bioreactor modified by the author. As a control, blueberries were grown on a semi-solid medium. The effectiveness of blueberry micropropagation was evaluated by the following indicators: multiplication coefficient, shoot length, and proportion of vitrified shoots. Experiments were conducted on WPM medium, with zeatin supplementation at a concentration of 1.0 mg/l, resulting in optimal results. It is shown that the rocker bioreactor is slightly inferior to the plantform bioreactor in micropropagation but outperforms the method of micropropagation on semisolid media. The rocker bioreactor can be fully utilized for production purposes. In order to reduce costs and increase technical reliability, the working principle of the mechanical drive of the author's model of a rocker-type bioreactor was changed. [ABSTRACT FROM AUTHOR]

Plant growth regulators play a crucial role in the proliferation of protocorm-like bodies through somatic embryogenesis in Phalaenopsis orchids. Additionally, media components and cultural systems can impact their proliferation and reduce costs. This research aimed to enhance proliferation rates of orchid protocorm–like bodies and decrease costs of both media providing and culture systems. To achieve this, protocorm explants were cultivated in four different systems: solid, liquid permanent immersion (Erlenmeyer flasks on a shaker), and temporary immersion (RITA® and FA-Bio bioreactor, a new laboratory mini-bioreactor). The media used included half-strength Murashige and Skoog (1/2 MS), modified FAST medium (MFAST), and low-cost medium (LC). The solid medium contained thidiazuron at concentrations of 0, 1.5, or 3.0 mg L−1. Solid 1/2 MS without thidiazuron was the control. Protocorm-like bodies in solid LC and MFAST exhibited high chlorophyll and anthocyanin content. Their proliferation rates increased by 32.6% and 29.6%, respectively, while culture costs decreased by 45.2% and 43.1%, respectively, compared to solid 1/2 MS with 3.0 mg L−1 thidiazuron. The production of protocorm-like bodies and their carbohydrate content were significantly higher in MFAST and LC media, compared to 1/2 MS media across all culture systems. The RITA® bioreactor with liquid LC showed higher proliferation rates than the other treatments. Liquid LC reduced costs by 57.6% compared to solid LC, making it a cost-effective option for mass propagation. The FA-Bio bioreactor reduced costs by 70.5% compared to RITA® for laboratory research evaluations in liquid culture. MFAST media in FA-Bio bioreactors can be used commercially for cost-effective propagation of Phalaenopsis orchids. [ABSTRACT FROM AUTHOR]

Plant in vitro cultures can be an effective tool in obtaining desired specialized metabolites. The purpose of this study was to evaluate the effect of light-emitting diodes (LEDs) on phenolic compounds in Rhaponticum carthamoides shoots cultured in vitro. R. carthamoides is an endemic and medicinal plant at risk of extinction due to the massive harvesting of its roots and rhizomes from the natural environment. The shoots were cultured on an agar-solidified and liquid-agitated Murashige and Skoog's medium supplemented with 0.1 mg/L of indole-3-acetic acid (IAA) and 0.5 mg/L of 6-benzyladenine (BA). The effect of the medium and different treatments of LED lights (blue (BL), red (RL), white (WL), and a combination of red and blue (R:BL; 7:3)) on R. carthamoides shoot growth and its biosynthetic potential was observed. Medium type and the duration of LED light exposure did not affect the proliferation rate of shoots, but they altered the shoot morphology and specialized metabolite accumulation. The liquid medium and BL light were the most beneficial for the caffeoylquinic acid derivatives (CQAs) production, shoot growth, and biomass increment. The liquid medium and BL light enhanced the content of the sum of all identified CQAs (6 mg/g DW) about three-fold compared to WL light and control, fluorescent lamps. HPLC-UV analysis confirmed that chlorogenic acid (5-CQA) was the primary compound in shoot extracts regardless of the type of culture and the light conditions (1.19–3.25 mg/g DW), with the highest level under R:BL light. BL and RL lights were equally effective. The abundant component was also 3,5-di-O-caffeoylquinic acid, accompanied by 4,5-di-O-caffeoylquinic acid, a tentatively identified dicaffeoylquinic acid derivative, and a tricaffeoylquinic acid derivative 2, the contents of which depended on the LED light conditions. [ABSTRACT FROM AUTHOR]

A procedure for plant regeneration from cell suspension cultures through somatic embryogenesis is described for Sapindus trifoliatus, a commercially and medicinally important tree. Callus was induced from leaf disc on agar-solidified MS medium with 5.0 mg l−1 2, 4-D and 0.01 mg l−1 Kin. Embryogenic cell suspension cultures were established by placing leaf-derived friable calli in PGR-free full-strength MS liquid medium with 3% sucrose. The growth of cell suspension culture was significantly affected by the strength of the MS mineral solution and L-glutamine. Plating of the suspension on semisolid MS medium resulted in the formation of globular structures. These embryogenic globular structures differentiated into secondary globular structures or somatic embryos on a semisolid MS medium. The differentiation of globular structures and different stages of somatic embryos (from globular to cotyledonary) was enhanced by the addition of 200 mg l−1 L-glutamine in the medium. Sucrose at relatively high concentrations (5%) or ABA (0.01 mg l−1) promoted somatic embryo maturation. The highest percentage (about 90%) of germination of somatic embryo and plantlet conversion was achieved on a half-strength MS medium containing 2% sucrose. The plants were hardened and established in soil with a 90% survival rateKey message: The present study demonstrates the establishment of suspension culture from leaf-derived callus in Sapindus trifoliatus and plant regeneration via somatic embryogenesis [ABSTRACT FROM AUTHOR]

Hydrothermal carbonization (HTC) of wetland plant could achieve the recovery of phosphorus (P) via the production of P-enriched hydrochar to alleviate the crisis of phosphate resources, while the migration and transformation of P should be determined. In this study, Canna indica was derived into hydrochar through HTC at different temperatures (200°C-260°C) and liquid mediums (H2O, CaCl2, and NaOH). The P forms were systematically characterized using P K-edge X-ray absorption near edge structure (XANES), 31P liquid nuclear magnetic resonance (NMR), and sequential extraction. The total P content in hydrochar was up to 23 mg g−1 with mainly inorganic P (> 97.8%), and the recovery rate was almost 100% during NaOH-mediated HTC. The P species, monoester-P and soluble orthophosphate (ortho-P), in biomass were transformed to more stable ortho-P in hydrochars, which was highly dependent on temperature and liquid medium. With increasing temperature, Al/Mg-P was gradually replaced by Ca/Fe-P. The CaCl2 solution facilitated the transformation of Ca(H2PO4)2 into CaHPO4 by elevating the Ca/P ratio. While for the NaOH-mediated HTC, the CaHPO4 and Ca(H2PO4)2 were transformed to hydroxyapatite (74.3%-81.5%), and the proportion of MgHPO4 elevated with increasing temperature. The diffusive gradients in thin films (DGT) results implied that the addition of hydrochar greatly elevated the soil available P content, which was further promoted by high temperature and NaOH medium. These results indicate that the species and availability of P in hydrochar could be adjusted through varying liquid medium and reaction temperature, which provide guidance for the target design of P-enriched hydrochar and P reclamation. Highlight: • P-enriched hydrochar was prepared from the wetland plant by the modified HTC. • Liquid medium and reaction temperature affected the speciation dynamics of P. • P was stabilized with high temperature, Ca2+ addition, and alkaline liquid medium. • Hydrochar addition greatly elevated the soil available P content via DGT analysis. [ABSTRACT FROM AUTHOR]

Hibiscus moscheutos L. 'Berry Awesome' is a complex hybrid of the new Proven Winners Summerific series of varieties with highly ornamental characteristics. Micropropagation of highly ornamental varieties is important for mass production of planting material for commercial purposes. The traditional methods for propagating Hibiscus varieties, such as cuttings or seed propagation, however, do not guarantee high rates of production of high-quality seedlings. To solve this problem, an attempt was made to develop protocols for micropropagation of Hibiscus moscheutos L. 'Berry Awesome' in vitro on agar and liquid medium using a bioreactor system, followed by ex vitro adaptation of the regenerants. The optimal method for sterilization of nodal explants as well as the optimal composition of the initiation medium for shoot proliferation and rooting were determined. For micropropagation on a liquid medium, a rocker-type bioreactor was used, and its advantages over micropropagation on an agar medium were demonstrated. The results showed that the best sterilization method for nodal segment explants was as follows: pretreatment by rinsing with running tap water, sterile water, and distilled water for 70 min and soaking for 5 min in a mixture of solutions of ethyl alcohol (96%), hydrogen peroxide (38%), and water in a ratio of 1:1:2. In this case, live and sterile explants accounted for 62.6%. The optimal initiation medium for axillary buds in nodal segments was the Murashige and Skoog (MS) medium supplemented with 0.1 mg L−1 N-(2-chloro-4-pyridinyl)-N'-phenylurea (CPPU), which resulted in 73.3% of axillary buds being induced. The optimal solid proliferation medium was MS medium supplemented with 0.1 mg L−1 CPPU with a proliferation coefficient of 5.8. In a liquid medium, the optimal concentration of CPPU was 0.05 mg L−1 with a proliferation coefficient of 9.2. The best medium for rooting/shoots with agar and in bioreactors was MS medium with the addition of 0.1 mg L−1 indole-3-butyric acid (IBA). The highest rooting rate was 99.0% in both types of media, and the survival rate of plantlets was 88.7% in solid media and 98.7% in the bioreactor. [ABSTRACT FROM AUTHOR]

Boesenbergia rotunda is a well-known Southeast Asian herbal that has been used as a condiment, traditional medicine, and more recently in certain modern pharmaceutical goods. At Forest Research Institute Malaysia (FRIM), mass production of B. rotunda was obtained using tissue culture technique known as temporary immersion bioreactor system (TIBs), SETIS™, an efficient plant production system. However, it was observed that the young leaves were unable to open or unfurl due to lack of space in the vessel. This created unhealthy plant development and caused high mortality during acclimatisation period, as the unfurled leaves might have insufficient cuticular layer on their surface. A technique to manipulate the plant height was needed to facilitate leaf development and enhance high survival rates. Therefore, this study aimed to investigate the effect of paclobutrazol (PBZ) in the manipulation of medium culture formulations towards B. rotunda plant height. Young shoots of B. rotunda were treated in MS medium supplemented with 2 mg/L BAP and different concentrations of PBZ using SETIS™. Results indicated a reduction in plant height, where samples treated with 0.1 mg/L and 1.0 mg/L gave a plant height of 16.93 ± 2.00 cm and 20.17 ± 3.20 cm, respectively, compared to 34.27 ± 2.73 cm in control. The photosynthetic rates showed an increment in samples treated with 0.1 mg/L and 1.0 mg/L PBZ (0.56 ± 0.09 and 0.56 ± 0.07 µmol/m²/s), respectively, compared to the control (0.18 ± 0.04 µmol/m²/s). The PBZ treatments successfully inhibited B. rotunda plant heights to facilitate the development in vitro and increase survival rates during acclimatisation process. [ABSTRACT FROM AUTHOR]

A simple in vitro shoot regeneration method was developed for Solanum tuberosum L. 'Yansi' using a simple continuous immersion airlift bioreactor culture system with Murashige and Skoog (MS) medium. Growth characteristics such as fresh weight, dry weight, length of shoots, and the number of nodes per shoot, length, and width of leaves were compared between cultures raised in bioreactor cultures and solid cultures. Shoots developed in bioreactor cultures showed higher values for these parameters than shoots grown in solid cultures. The leaf thickness and width of the upper epidermis, lower epidermis, palisade, and spongy tissues were also higher with shoots cultured in bioreactor cultures. Microscopic observation of the lower epidermal surface indicated that the stomata of leaves grown on liquid medium were normal and elliptical with narrow pores, whereas leaves grown on solid medium showed round, abnormal, and closed stomata. Studies on the optimization of the culture medium of bioreactor cultures were carried out. Of the varied strengths of MS media (0.5, 1, 1.5, and 2.0 ×) tested, 2.0 × MS medium was found suitable for shoot cultures. Similarly, varied levels of ammonium (NH4+) and nitrate (NO3−) (20/100, 40/80, 60/60, 80/40, and 100/20 mM) were tested for the growth of shoots in bioreactor cultures; 20/100 mM ratio of NH4+:NO3− was found suitable. The results indicate that bioreactor cultures with modified MS medium could be used for mass propagation of Solanum tuberosum L. 'Yansi' shoots. [ABSTRACT FROM AUTHOR]

Plant in vitro cultures can be an effective tool in obtaining desired specialized metabolites. The purpose of this study was to evaluate the effect of light-emitting diodes (LEDs) on phenolic compounds in Rhaponticum carthamoides shoots cultured in vitro. R. carthamoides is an endemic and medicinal plant at risk of extinction due to the massive harvesting of its roots and rhizomes from the natural environment. The shoots were cultured on an agar-solidified and liquid-agitated Murashige and Skoog’s medium supplemented with 0.1 mg/L of indole-3-acetic acid (IAA) and 0.5 mg/L of 6-benzyladenine (BA). The effect of the medium and different treatments of LED lights (blue (BL), red (RL), white (WL), and a combination of red and blue (R:BL; 7:3)) on R. carthamoides shoot growth and its biosynthetic potential was observed. Medium type and the duration of LED light exposure did not affect the proliferation rate of shoots, but they altered the shoot morphology and specialized metabolite accumulation. The liquid medium and BL light were the most beneficial for the caffeoylquinic acid derivatives (CQAs) production, shoot growth, and biomass increment. The liquid medium and BL light enhanced the content of the sum of all identified CQAs (6 mg/g DW) about three-fold compared to WL light and control, fluorescent lamps. HPLC-UV analysis confirmed that chlorogenic acid (5-CQA) was the primary compound in shoot extracts regardless of the type of culture and the light conditions (1.19–3.25 mg/g DW), with the highest level under R:BL light. BL and RL lights were equally effective. The abundant component was also 3,5-di-O-caffeoylquinic acid, accompanied by 4,5-di-O-caffeoylquinic acid, a tentatively identified dicaffeoylquinic acid derivative, and a tricaffeoylquinic acid derivative 2, the contents of which depended on the LED light conditions.

Objective: To assess a temporary immersion system for the in vitro propagation of Agave potatorum Zucc., compared with the traditional micropropagation technique that uses a solid medium. Design/Methodology/Approach: The effect of treatments in a solid medium with low and high doses of the BA (Benzylaminopurine) growth regulator (0.5 mg L-1 and 2 mg L-1) on the number of sprouts per explant was assessed in a first phase. Since the best treatment was 2 mg L-1 of BA, three forms of propagation were considered: solid medium, liquid medium in a paper bridge, and liquid medium in a temporary immersion system. Results: From the initial test, an average of 6.6 shoots per explant were obtained with 2 mg L-1 of BA. Regarding the different systems, the solid medium, the paper bridge, and the temporary immersion system recorded 6.4, 7.2, and 14.4 shoots per explant, respectively. Findings/Conclusions: Mass sprout production is higher in the temporary immersion system, as a consequence of the use of a liquid medium that increases the absorption of nutrients and regulators, combined with the injection of air with oxygen that can accelerate cellular processes. [ABSTRACT FROM AUTHOR]

Context: The rationalization of acid rain formation steps is fundamental for mitigating its effects. It is believed the hydroxysulfinyl radical is an intermediate species for the production of atmospheric sulfuric acid. Two stable configurations HOSO and HSO2 have been reported for such a radical in the gas phase. This work aims at studying these isomers in the aqueous medium. The electrical and reactivity quantities — electronic chemical potential (μ ), chemical hardness (η ), and electrophilicity (ϵ ) — are here calculated and compared. Considering first solvation shells (15 H2O for HSO2 and 23 H2O for HOSO), an increase of 25% in the dipole moment of HSO2 was obtained, while the dipole moment of HOSO decreases in 11%. Both solvated isomers grow softer (η decreases) contrasted to the gas phase. Methods: HOSO and HSO2 are studied through a sequential Monte Carlo/quantum mechanics approach. Lennard–Jones plus the Coulomb potentials were used to represent intermolecular potential interaction in the frame of the DICE package. Molecular structure calculations were performed at CASPT2/aug - cc - pV(T + d)Z level of theory using the MOLPRO suite of programs. [ABSTRACT FROM AUTHOR]

Plasma cleaning with simultaneous polishing by a jet electric discharge is more efficient than electrochemical polishing. The paper discusses the processes arising during the interaction of an electric discharge with solutions of inorganic substances, and studies the propagation of an inorganic discharge in a liquid on the surface of various materials (NaCl, NH4NO3, and discharge in glycerol). A NH4NO3 solution is shown to be the most effective electrolyte. We report on current‒voltage characteristics obtained for an electric discharge between a jet electrolytic cathode and a solid anode, as well as for a multichannel discharge in a jet electrolytic cathode at different jet lengths and diameters. [ABSTRACT FROM AUTHOR]

The use of a liquid medium in near-dry electrical discharge machining (EDM) with a two-phase gas‒liquid medium endows it with desirable characteristics such as a stable discharge process and high material removal rate. However, the effect mechanism of the liquid medium in near-dry EDM is not well understood, which limits its development. In this present, a variant of the near-dry EDM technology, namely, mixed-gas atomization discharge ablation processing (MA-DAP) of titanium alloy, is used to reveal the effect mechanism of a liquid medium in the process of near-dry EDM. This technology uses a mixed gas containing oxygen and a liquid medium to form the dielectric medium, so as to obtain a higher effective material erosion ability than near-dry EDM. Through the combination of experimental and theoretical modeling and analysis, it can be concluded that liquid dielectric water has the functions of endothermic cooling, inducing an intermittent gas supply environment between electrodes, efficient etching product removal, improving oxygen utilization efficiency, controlling oxygen diffusion speed, and increasing the discharge gap between electrodes, which can significantly improve the inter-electrode discharge state and material removal rate. Compared with conventional EDM, the material removal rate associated with MA-DAP technology increases by 16 times, and the relative wear rate of the electrode is reduced by more than 93%. [ABSTRACT FROM AUTHOR]

The preparation and modulation of bare nano-diamonds (NDs) are of great significant for further investigation into their intrinsic structures, exploring more underlying properties, unlocking latent performances and exploiting more extensive applications. In this study, bare NDs have been successfully synthesized and modulated through 343 nm femtosecond laser ablation in liquids (LAL). When laser scanning point interval (LSPI) was reduced from 4 μm to 0.5 μm, the average size of NDs decreased from 4.1 nm to 3 nm. While it remained around 3.2 nm for NDs obtained at different energies. Notable, sp3-C within NDs accounted for above 71 % at laser energy of diamond damage threshold (118 mW), and then transformed into sp2-C as laser energy increased. NDs prepared in water and ethanol differed in terms of their size distribution, intrinsic structure, and surface state. NDs@ethanol sized 3.8 nm exhibited higher sp3/sp2 ratio and more complex surface states than NDs@water sized 3.3 nm. These findings not only can provide valuable guidance for precise regulation of NDs preparation, but also can offer diverse structural NDs for exploring their potential applications. [Display omitted] [ABSTRACT FROM AUTHOR]

Hibiscus moscheutos L. ‘Berry Awesome’ is a complex hybrid of the new Proven Winners Summerific series of varieties with highly ornamental characteristics. Micropropagation of highly ornamental varieties is important for mass production of planting material for commercial purposes. The traditional methods for propagating Hibiscus varieties, such as cuttings or seed propagation, however, do not guarantee high rates of production of high-quality seedlings. To solve this problem, an attempt was made to develop protocols for micropropagation of Hibiscus moscheutos L. ‘Berry Awesome’ in vitro on agar and liquid medium using a bioreactor system, followed by ex vitro adaptation of the regenerants. The optimal method for sterilization of nodal explants as well as the optimal composition of the initiation medium for shoot proliferation and rooting were determined. For micropropagation on a liquid medium, a rocker-type bioreactor was used, and its advantages over micropropagation on an agar medium were demonstrated. The results showed that the best sterilization method for nodal segment explants was as follows: pretreatment by rinsing with running tap water, sterile water, and distilled water for 70 min and soaking for 5 min in a mixture of solutions of ethyl alcohol (96%), hydrogen peroxide (38%), and water in a ratio of 1:1:2. In this case, live and sterile explants accounted for 62.6%. The optimal initiation medium for axillary buds in nodal segments was the Murashige and Skoog (MS) medium supplemented with 0.1 mg L−1 N-(2-chloro-4-pyridinyl)-N’-phenylurea (CPPU), which resulted in 73.3% of axillary buds being induced. The optimal solid proliferation medium was MS medium supplemented with 0.1 mg L−1 CPPU with a proliferation coefficient of 5.8. In a liquid medium, the optimal concentration of CPPU was 0.05 mg L−1 with a proliferation coefficient of 9.2. The best medium for rooting/shoots with agar and in bioreactors was MS medium with the addition of 0.1 mg L−1 indole-3-butyric acid (IBA). The highest rooting rate was 99.0% in both types of media, and the survival rate of plantlets was 88.7% in solid media and 98.7% in the bioreactor.

Agarwood (Aquilaria malaccensis Lamk.) is a forest product commodity with high economic value, but its availability in nature is decreasing due to deforestation and overexploitation. The availability of high-quality seedlings is necessary to meet the demand and conserve the sources of agarwood plants. Therefore, this study investigates the interaction effect between media types and 1-Naphthaleneacetic acid (NAA) concentrations on agarwood shoot development. It also aims to determine the best medium type and NAA concentration to stimulate agarwood shoot development. This research had two stages, namely the shoot induction and development. The shoot induction stage was conducted experimentally, using a completely randomized design (CRD) on a factorial treatment pattern of 2 factors i.e., the types of media and NAA concentrations. The shoots produced from the induction stage were cultured on Murashige-Skoog (MS) media with the same media types but without any NAA addition. Furthermore, the variables observed in both stages were the development of agarwood shoots, and the parameters measured included the number of shoots, number of leaves and shoot lengths. The results showed that the development of agarwood shoots was controlled by the type of medium used. Liquid MS medium supported with filter paper bridge produced the highest average number of shoots with 4.93 and 5.87 shoots explants-1 and length at 3.25 and 3.64 cm explant-1 in the induction and development stage, respectively. These findings will facilitate mass propagation of agarwood shoots, and in turn the availability of agarwood plantlets supports its conservation and production.

Orchid mycorrhizal fungi (OMF) from the rhizoctonia aggregate are generally considered to be soil saprotrophs, but their ability to utilize various nutrient sources has been studied in a limited number of isolates cultivated predominantly in liquid media, although rhizoctonia typically grow on the surface of solid substrates. Nine isolates representing the key OMF families (Ceratobasidiaceae, Tulasnellaceae and Serendipitaceae), sampled in Southern France and the Czech Republic, were tested for their ability to utilize carbon (C), nitrogen (N) and phosphorus (P) sources in vitro in both liquid and solid media. The isolates showed significant inter- and intra-familiar variability in nutrient utilization, most notably in N sources. Isolates produced generally larger amounts of dry biomass on solid medium than in liquid one, but some isolates showed no or limited biomass production on solid medium with particular nutrient sources. The largest amount of biomass was produced by isolates from the family Ceratobasidiaceae on most sources in both medium types. The biomass production of Tulasnellaceae isolates was affected by their phylogenetic relatedness on all sources and medium types. The ability of isolates to utilize particular nutrients in a liquid medium but not a solid one should be considered when optimizing solid media for symbiotic orchid seed germination and in understanding of OMF functional traits under in situ conditions. [ABSTRACT FROM AUTHOR]

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Salix viminalis L. is a species with high capacity for micropropagation and acclimation and could therefore be used to evaluate emergent techniques in the field of plant propagation. The aims of this study were to propagate willow in liquid medium with a continuous immersion system, to explore the application of photoautotrophic conditions and to investigate the adaptation of willow plantlets to different soils that could be used as alternatives to commercial peat. For proliferation, we used 3% sucrose or sugar-free medium, and as substrates, we used commercial peat, a soil from an oak forest with high organic matter content and a crop soil with low organic matter content. The effect of sugar supplementation during proliferation and the soil characteristics during acclimation and growth were evaluated on the basis of aerial and root growth and the hydrolytic and dehydrogenase enzymatic activities of the soils. The results indicate that under photoautotrophic conditions, the supplementation of sucrose during micropropagation did not affect the subsequent growth of the plantlets. All plants acclimated without loss, but the type of soil influenced the height and vigor. Plants produced the highest shoots in peat, whereas the most root development occurred in crop soil. Soil enzyme activities were more influenced by the type of soil than by the presence of plants. [ABSTRACT FROM AUTHOR]

The aim of this research was to investigate the inhibitory effect of Bacillus subtilis P5-6 against Staphylococcus aureus on different states of standard medium with the presence of sodium chloride at 5% (v/v), to develop an effective protective culture in food. Halophile Bacillus subtilis P5-6 isolated from Plara (Thai traditional salt fermented fish) contained both the genes encoding subtilin (spaS) and subtilosin (sboA). Only subtilosin gene expression was detected along with housekeeping gene BA-rpoB when co-cultured with Staphylococcus aureus. With the spot on lawn assay, cell suspension from the P5-6 culture showed inhibitory effect against Staphyloccus aureus, while no inhibition was observed when cell free supernatant was used. In liquid co-culture, the inhibitory effect of P5-6 on Staphylococcus aureus was observed when its inoculum size (population density of 8 log CFU/ mL) was double that of S.aureus. In solid medium, Bacillus subtilis P5-6 could exert higher antagonistic action against this target pathogen. Bacillus subtilis P5-6 displayed an inhibitory effect even when its population was 2 log CFU/ mL lower than that of Staphylococcus aureus. Solid state cultivation with the presence of sodium chloride could enhance production and/or activity of bacteriocin of P5-6. The observation reflects an importance of the self-inhibitory effect as found in liquid and solid medium cultivation system. The data obtained could be fundamental importance in bacteriocin production development and application of protective culture from P5-6 to protect food against Staphyloccus aureus-one of the most common cause of foodborne disease.

A unique method was developed to significantly improve the strength of Bi(1−x)Sbx single crystals, the most effective thermoelectric (TE) materials in the temperature range from 100 to 200 K due to their plastic deformation by extrusion. After plastic deformation at room temperature under all-round hydrostatic compression in a liquid medium, n-type Bi–Sb polycrystalline solid solutions show a significant increase in mechanical strength compared to Bi–Sb single crystals in the temperature range from 300 to 80 K. The significantly higher strength of extruded alloys in comparison with Bi–Sb single crystals is associated with the development of numerous grains with a high boundary surface as well as structural defects, such as dislocations, that accumulate at grain boundaries. Significant stability of the structure of extruded samples is achieved due to the uniformity of crystal plastic deformation under all-round hydrostatic compression and the formation of the polycrystalline structure consisting of grains with the orientation of the main crystallographic directions close to the original single crystal. The strengthening of Bi–Sb single crystals after plastic deformation allows for the first time to create workable TE devices that cannot be created on the basis of single crystals that have excellent TE properties, but low strength.

Somatic embryogenesis is an efficient micro-propagation technique, which can lead to mass multiplication of tea with a tap root. Hence, the development of a viable somatic embryogenesis and multiplication protocol through liquid culture conditions is needed to increase the efficiency of the protocol. Cotyledon-derived somatic embryos, leaf and stem callus of TRI 5001 cultivar, were tested with different liquid MS media. A significantly higher cotyledon-derived somatic embryo multiplication rate was observed in MS +3 mg/L Thidiazuron (TDZ) medium. Callus proliferation and compact and embryonic callus formation were comparatively higher in MS + 1.1 mg/L TDZ, 1.86 mg/L NAA medium, and subsequent somatic embryo formation was observed in MS + 0.1 mg/L NAA,1 mg/L BAP and 0.2 mg/L GA3 medium on callus produced on leaf explants. Higher nodal callus proliferation and compact callus formation were recorded in MS+0.11 mg/L TDZ 0.1 mg/L IBA and higher embryonic callus was observed in MS+0.1 mg/L TDZ, 1.86 mg/L NAA media. Plant regeneration was resulted in MS+3 mg/LBAP 0.1 mg/L NAA medium. Identified treatment combinations can be used as a foundation to establish somatic embryogenesis protocol through different explants of tea. [ABSTRACT FROM AUTHOR]

Objective. To improve stool sample culture and isolation of anaerobic flora, including Clostridioides difficile in the routine microbiological laboratory practice at the children’s oncology hospital. Materials and Methods. A total of 517 stool samples collected from patients in children’s oncology hospital from 2013 to 2015 were studied. All samples were analyzed by ELISA for C. difficile toxins and by culture according to dedicated 5 schemes for isolation of anaerobic bacteria, including C. difficile. Statistical significance of differences in isolation rates between the studied groups (culture schemes) was assessed by Pearson test. Results. Culture in liver broth and covering with technical agar followed by culture on anaerobic agar yielded 100% isolation rate of toxigenic C. difficile strains. This culture scheme is also suitable for isolating concomitant anaerobic flora: non-toxigenic C. difficile strains, Clostridium perfringens, other Clostridia spp. and Bacteroides spp. Conclusions. Use of the liquid accumulation medium and covering with technical agar make it possible to isolate anaerobic flora from stool samples and increase an isolation rate of toxigenic C. difficile strains to 100% of ELISA-positive samples.

The study aimed to introduce in vitro propagation techniques for Vanilla phaeantha , focusing on the effectiveness of the double-phase technique and temporary immersion bioreactor systems for clonal multiplication. Optimal conditions for plantlets acclimatization were also sought. Endophytic bacteria occurring in the culture were isolated, analyzed, and controlled. Explants obtained from greenhouse mother plants underwent a decontamination process using sodium ampicillin at different concentrations (0, 250, and 500 mg L−1) for up to five subcultures. Bacteria present during establishment phase were isolated and analyzed via 16 S rRNA sequencing. The study also encompassed an evaluation of clonal multiplication strategies, which included assessing the influence of different consistencies of culture media (liquid, double-phase, and semisolid), and the use of three temporary immersion bioreactor models (RITA®, RALM® and TIB®). The resulting plantlets underwent acclimatization in a mixture of commercial substrate and coconut fiber with varying composition and were kept in a growth chamber before being transferred to a greenhouse. It was verified that the use of ampicillin sodium led to a significant reduction in bacterial contamination over five subcultures. Sequencing using the 16 S rRNA gene enabled the identification of some contaminant bacteria, including species of the genera Rhizobium , Herbaspirillum , and Methylobacterium. Clonal multiplication strategies were evaluated, with the double-phase method proving superior in shoot production per explant (7.0 ± 0.4) compared to the semisolid medium (5.3 ± 0.3). Among temporary immersion bioreactor models tested, RALM® was most effective in shoot formation when compared to RITA® and TIB® (8.2 ± 0.3; 4.9 ± 0.2, and 1.2 ± 0.1, respectively). The use of substrates composed of commercial substrate and coconut fiber, in any combination tested, ensuring 100 % survival of vanilla plantlets in the greenhouse. The findings provide a micropropagation protocol for Vanilla phaeantha , offering a diversification avenue for vanilla cultivation. • This study describes innovative techniques for the in vitro propagation of vanilla species. • Endophytic bacteria were isolated, identified, and controlled through the continuous use of antibiotics. • The double-phase system is more efficient than tradicional multiplication in a conventional culture medium. • This study test a new temporary immersion bioreactor model, proving efficient for enhancing in vitro vanilla propagation. [ABSTRACT FROM AUTHOR]

The aim of this study was to propagate axillary shoots of Cannabis sativa L. using liquid medium in temporary immersion bioreactors. The effect of immersion frequency (3 or 6 immersions per day), explant type (apical or basal sections), explant number (8, 10, and 16 explants), mineral medium (Murashige and Skoog half-strength nitrates, β -A and β -H, all supplemented with 2-μM metatopoline), sucrose supplementation (2, 0.5, and 0% sucrose), culture duration (4 and 6 weeks), and bioreactor type (RITA® and Plantform™) were investigated. As a result, we propose a protocol for the proliferation of cannabis apical segments in RITA® or Plantform™ bioreactors. The explants (8 per RITA® and 24 per Plantform™) are immersed for 1 min, 3 times per day in β -A medium supplemented with 2-μM metatopoline and 0.5% of sucrose and subcultured every 4 weeks. This is the first study using temporary immersion systems in C. sativa production, and our results provide new opportunities for the mass propagation of this species. [ABSTRACT FROM AUTHOR]

The NeuMoDx HPV assay is a novel fully automated, real-time PCR-based assay for the qualitative detection of high-risk human papillomavirus (HPV) DNA in cervical specimens. The assay specifically identifies HPV16 and HPV18 and concurrently detects 13 other high-risk HPV types at clinically relevant infection levels. Following the international guidelines, the clinical performance of the NeuMoDx HPV assay for cervical intraepithelial neoplasia grade 2 or worse (CIN2+) against the reference standard Hybrid Capture 2, as well as intra- and inter-laboratory reproducibility were assessed on PreservCyt samples. The clinical accuracy of the assay was additionally evaluated against the clinically validated Alinity m HR HPV and COBAS 4800 HPV Test on PreservCyt samples, and against the clinically validated HPV-Risk assay on SurePath samples. The NeuMoDx HPV assay performance for CIN2+ was non-inferior to the reference methods on both sample types (all p < 0.05), and showed excellent intra- and inter-laboratory reproducibility (95.7%; 95% CI: 93.9–97.3; kappa value 0.90 (95% CI: 0.86–0.94); and 94.5%; 95% CI: 92.6–96.2; kappa value 0.87 (95% CI: 0.82–0.92), respectively). In conclusion, the NeuMoDx HPV assay meets international guideline criteria for cross-sectional accuracy and reproducibility, and performs equally well on cervical screening specimens collected in two widely used collection media. The NeuMoDx HPV assay fulfils the requirements to be used for primary cervical screening. [ABSTRACT FROM AUTHOR]

A classification of the heat exchangers of capacitance apparatuses with rotary mixers for use in mixing liquid media is described. A critical survey of the most characteristic heat exchangers now available is performed. [ABSTRACT FROM AUTHOR]

In the work, the study and determination of the acoustic parameters of the movement of a cavitation bubble in a liquid medium according to discrete and continuous models was carried out. The research is based on the hypothesis that the determination of the effective parameters of the work process of acoustic processing is implemented by applying a transitional physical model from a discrete to a continuous type of processing of the technological environment. The obtained analytical dependences allow to calculate the amplitude of oscillations and the frequency of natural oscillations. With the help of the specified formulas, it is possible to determine the zones of amplification or attenuation of the amplitude of oscillations for different frequencies of oscillations. The proposed formula for determining the frequency of natural oscillations, which takes into account changes in the properties of the medium from homogeneous at the initial stage to the appearance of cavitation bubbles at the specified frequency of natural oscillations. Numerical values of intensity, pressure, amplitude of oscillations, velocity, acceleration, viscosity and maximum bubble radius are given. The obtained numerical values can be used in practical calculations of acoustic processing parameters of different nature and properties of technological environments. [ABSTRACT FROM AUTHOR]

The in vitro shoot proliferation of endemic Begonia pavonina in three culture conditions i.e semisolid medium (SM), liquid culture medium (LM) and in temporary immersion medium; RITA System (TIM) was analyzed in this study. To minimize contamination rates, seeds were surface sterilized and cultured on MS basal media. The clean raised shoots were then used as explants for inoculation onto the tested culture conditions. For this experiment, the explants were maintained in MS medium supplemented with 0.1mgL-1 BAP for shoot multiplication. After 4 weeks of incubation, higher regeneration rates were observed in TIM as compared to other medium conditions. The propagation rate in TIM was nearly doubled as compared to liquid as well as in solid medium. Shoot hyperhydration was also lowest in a TIM system. Overall, TIM has shown to produced higher shoot multiplications combined with healthy morphological characteristics of plantlets. Shoot cultures from the all cultures successfully rooted in vitro and acclimatized well in the greenhouse.

The aim of this study was to propagate axillary shoots of Cannabis sativa L. using liquid medium in temporary immersion bioreactors. The effect of immersion frequency (3 or 6 immersions per day), explant type (apical or basal sections), explant number (8, 10, and 16 explants), mineral medium (Murashige and Skoog half-strength nitrates, β-A and β-H, all supplemented with 2-μM metatopoline), sucrose supplementation (2, 0.5, and 0% sucrose), culture duration (4 and 6 weeks), and bioreactor type (RITA® and Plantform™) were investigated. As a result, we propose a protocol for the proliferation of cannabis apical segments in RITA® or Plantform™ bioreactors. The explants (8 per RITA® and 24 per Plantform™) are immersed for 1 min, 3 times per day in β-A medium supplemented with 2-μM metatopoline and 0.5% of sucrose and subcultured every 4 weeks. This is the first study using temporary immersion systems in C. sativa production, and our results provide new opportunities for the mass propagation of this species.

In this study, we propagated two old Galician plum varieties in liquid medium using a temporary immersion system with RITA© bioreactors. Environmental variables including culture system, light intensity, CO2 enrichment, immersion frequency and sucrose supplementation were evaluated in relation to in vitro proliferation, physiological status and ex vitro performance. Bioreactors were superior to jars for culturing shoots in photomixotrophic conditions, producing up to 2 times more shoot numbers and up to 1.7 times more shoot length (depending on the genotype) using shoot clusters. The number and quality of shoots were positively influenced by the sucrose concentration in the medium, plus by the light and gaseous environment. For individual apical sections the best response occurred with 3% sucrose, 150 µmol m−2 s−1 photosynthetic photon flux density and 2000 ppm CO2, averaging 2.5 shoots per explant, 26 mm shoot length and 240 mm2 leaf area, while with 50 µmol m−2 s−1 light and ambient CO2 (400 ppm) values decreased to 1.2 shoots per explant, 14 mm of shoot length and 160 mm2 of leaf area. Shoots cultured photoautotrophically (without sucrose) were successfully rooted and acclimated despite of showing limited growth, low photosynthetic pigments, carbohydrate, phenolic and antioxidant contents during the multiplication phase. [ABSTRACT FROM AUTHOR]

Orchid mycorrhizal fungi (OMF) from the rhizoctonia aggregate are generally considered to be soil saprotrophs, but their ability to utilize various nutrient sources has been studied in a limited number of isolates cultivated predominantly in liquid media, although rhizoctonia typically grow on the surface of solid substrates. Nine isolates representing the key OMF families (Ceratobasidiaceae, Tulasnellaceae and Serendipitaceae), sampled in Southern France and the Czech Republic, were tested for their ability to utilize carbon (C), nitrogen (N) and phosphorus (P) sources in vitro in both liquid and solid media. The isolates showed significant inter- and intra-familiar variability in nutrient utilization, most notably in N sources. Isolates produced generally larger amounts of dry biomass on solid medium than in liquid one, but some isolates showed no or limited biomass production on solid medium with particular nutrient sources. The largest amount of biomass was produced by isolates from the family Ceratobasidiaceae on most sources in both medium types. The biomass production of Tulasnellaceae isolates was affected by their phylogenetic relatedness on all sources and medium types. The ability of isolates to utilize particular nutrients in a liquid medium but not a solid one should be considered when optimizing solid media for symbiotic orchid seed germination and in understanding of OMF functional traits under in situ conditions.

Salix viminalis L. is a species with high capacity for micropropagation and acclimation and could therefore be used to evaluate emergent techniques in the field of plant propagation. The aims of this study were to propagate willow in liquid medium with a continuous immersion system, to explore the application of photoautotrophic conditions and to investigate the adaptation of willow plantlets to different soils that could be used as alternatives to commercial peat. For proliferation, we used 3% sucrose or sugar-free medium, and as substrates, we used commercial peat, a soil from an oak forest with high organic matter content and a crop soil with low organic matter content. The effect of sugar supplementation during proliferation and the soil characteristics during acclimation and growth were evaluated on the basis of aerial and root growth and the hydrolytic and dehydrogenase enzymatic activities of the soils. The results indicate that under photoautotrophic conditions, the supplementation of sucrose during micropropagation did not affect the subsequent growth of the plantlets. All plants acclimated without loss, but the type of soil influenced the height and vigor. Plants produced the highest shoots in peat, whereas the most root development occurred in crop soil. Soil enzyme activities were more influenced by the type of soil than by the presence of plants.

The study reports a low-cost micropropagation protocol for Nardostachys jatamansi , a high value, critically endangered Himalayan herb. Shoot-bud bearing rhizome segments of the plant were made to sprout and elongate on 0.8% agar-gelled Murashige and Skoog's medium containing 3% sucrose. The sprouted shoots were then multiplied on agar-gelled as well as liquid MS medium containing various concentrations and combinations of kinetin (Kn) and 1-phenyl-3- (1,2,3-thiadiazol-5-yl) urea (thidiazuron or TDZ). Maximum number of shoots developed per explant after 30 days on 100 ml agar gelled (6.7 shoots) and 30 ml liquid (6 shoots) MS medium containing 0.5 mg l−1 Kn and 0.75 mg l−1 TDZ. Although rooting of in vitro shoots was attempted on full as well as half-strength MS medium containing different auxins, rooting was achieved in 100% shoots (3.07 roots/ shoot) after 30 days on half-strength MS medium supplemented with 1.0 mg l−1 indole-3- acetic acid (IAA). Different potting mixtures were used to acclimatize the micropropagated plantlets under polyhouse condition, but a maximum of 83% survival was recorded in sand and sand plus soil. The genetic fidelity of the in vitro plantlets was also assessed using Start Codon Targeted Polymorphism (SCoT) and Directed Amplification of Minisatellite DNA (DAMD) markers, and a total of 8.03% polymorphism was recorded. Eventually, the cost of micropropagation was reduced by 6.4 times during shoot multiplication and further by half during rooting. The present micropropagation method developed for N. jatamansi can have potential applications in medicinal plant-based industry. [Display omitted] • A low-cost micropropagation method for critically endangered Himalayan medicinal herb. • Cost of in vitro shoot multiplication reduced by 6.4 times in liquid shoot multiplication medium. • In vitro raised plants showed 8.03% polymorphism as compared to mother plant in genetic variability analysis. [ABSTRACT FROM AUTHOR]

Blueberry (Vaccinium spp. L.) plants exhibit high potential of regeneration via adventitious shoot formation on a semi-solid medium followed by shoot elongation in a liquid medium under bioreactor systems. To find out whether DNA methylation plays a role during shoot elongation, we compared DNA methylation level in the regenerants of two in vitro-grown half-high blueberry (V. corymbosum L. × V. angustifolium Ait.) cultivars Patriot and Chippewa on a semi-solid medium (SSM) in glass bottles and in a liquid medium in temporary immersion bioreactors (TIB), via methylation-sensitive amplification polymorphism (MSAP) technique. The SSM was separately fortified with various combinations of two plant growth regulators, zeatin and thidiazuron for shoot regeneration but elongation was carried out using the same medium under both SSM and TIB systems with only zeatin. Zeatin at 9.2 µM produced the maximum shoots in SSM and TIB for both cultivars, which varied from 30 to 33 in SSM and from 28 to 29 in TIB for Patriot and Chippewa, respectively. However, shoots proliferated in TIB were more vigorous. Noticeable changes in the methylation profiles were detected using MSAP in the regenerants grown in each type of culture system. The TIB system exhibited a significant increase in total methylation percentage (47–50%) in comparison to SSM (25–32%) in both cultivars. Similarly, the plants regenerated from TIB system were more polymorphic than those from SSM system. We describe here the effects of culture in vitro on DNA methylation that induced during the process of shoot elongation in two half-high blueberry cultivars. Key message: In vitro shoot culture was achieved in half-high blueberry cultivars using a semi-solid and a liquid medium; plantlets developed from liquid medium showed higher level of DNA methylation alterations. [ABSTRACT FROM AUTHOR]

The formation of food liquid medium structures containing at least 70 % of disperse particles with high dispersiveness has been considered. The possible formation mechanisms of food liquid medium structure when slow (hydrodynamic) and quick (acoustic) processes create favorable conditions for cavitation have been studied. The possibility to control these processes for initiation of mechanical and kinetic reactions that change the structure of the medium has been demonstrated. The invert syrup has been selected as the study object. The change in the invert syrup structure before and after such cavitation treatment has been recorded with the use of metallographic microscope Nikon Eclipse MA100. The decrease in disperse phase sizes from 2–3 µm to 0.1–0.4 µm along with establishing the high uniformity of component distribution as compared to the syrup without cavitation process treatment has been detected.

The preparation of porous ceramics by freezing casting has received increasing attention in recent years because of the advantages of green economy, controllable pore structure and excellent material properties. Based on the brief introduction to the principle of freezing casting process, the pore forming mechanism and conditions of freezing casting process were elaborated. The effect of liquid medium and freezing conditions on pore structure of porous ceramics was discussed in details. The material system of porous ceramics prepared by freezing casting and the typical technological conditions were summarized. Finally, it was pointed out that the research directions of freeze casting in the future are the effective control of pore structure and the preparation of new functionalized porous ceramics.

A new Diels-Alder type adduct named morushalunin (1), together with three similar compounds, guangsangon E (2), chalcomoracin (3), and kuwanon J (4) were isolated from the EtOAc extract of liquid medium of Morus alba var. shalun root cultures. The structures of the isolated compounds were determined based on spectroscopic data, including 1D, 2D NMR, and MS data. Cytotoxic properties of compounds 1–4 were evaluated against murine leukemia P-388 cells. Morushalunin (1), guangsangon E (2), and chalcomoracin (3) were found to have significant cytotoxicity with IC50 values 0.7, 2.5, and 1.7 μg/mL, respectively, while kuwanon J (4) showed weak cytotoxicity (IC50 = 5.9 μg/mL). [ABSTRACT FROM AUTHOR]

The aim of the present study was to assess the transcription of Listeria monocytogenes key virulence genes, namely sigB, prfA, hly, plcA, plcB, inlA, inlB, inlC and inlJ during subsistence on the surface of tomato, cucumber and carrot stored at 4, 10 and 30 °C for 0, 0.5, 6 and 24 h. Gene relative transcription was assessed through reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The results obtained, indicated that the relative transcription of plcA, plcB and inlB was more affected during subsistence on cucumber surface whereas the relative transcription of sigB, prfA, hly, inlA and inlC was more affected on tomato surface. Subsistence of the pathogen on carrot surface had only marginal effect on the relative transcription of the virulence genes assessed in the present study. In the majority of the cases, the aforementioned effects were dependent on the storage temperature employed. [ABSTRACT FROM AUTHOR]

Scaevola taccada (Gaertn.) Roxb. is a hemi-sclerophyllous littoral shrub of the family Goodeniaceae. It is a salt-tolerant plant and used in soil reclamation and coastal landscaping programs to control beach erosion. The present article reports a reproducible in vitro regeneration system using meta-topolin (mT) through liquid medium employing nodal segment cultures. The highest number of axillary shoots (5.8 ± 0.29) was differentiated on Murashige and Skoog's (MS) medium containing meta-topolin (1.5 mg L−1). The amplification of shoots (49.8 ± 0.37) and fresh growth was achieved by recurrent subcultures on liquid MS medium supplemented with 0.25 mg L−1mT and 0.15 mg L−1 indole-3-acetic acid (IAA) in the presence of additives (50 mg L−1 ascorbic acid + 25 mg L−1 citric acid, adenine sulphate and l-arginine). Rooting and acclimatization was simultaneously achieved using concurrent ex vitro rooting and acclimatization (CEVRA) technique by pulse treating the shoot base with 300 mg L−1 IBA for 5 min. The in vitro developed shoots represented a maximum percentage (100%) of rooting in autoclaved soilrite in a greenhouse. After 2 mo, the plantlets were established in the field with cent percentage survival success. The genetic fidelity analysis using inter simple sequence repeats (ISSR) DNA molecular markers developed maximum of 55 monomorphic bands and revealed the genetic homogeneity of in vitro regenerated plantlets with the mother plant. [ABSTRACT FROM AUTHOR]

Date palm micropropagation is commonly performed on gelled media. However, it’s typically a labour-intensive system and consequently plantlets production cost is very high. Therefore, it is necessary to develop cost effective alternatives without compromising the quality of produced plant material. New technologies based on liquid media in bioreactors have been developed to reduce the handling time, while increasing the multiplication rates and plant quality. The present research focuses on the comparison between Temporary Immersion System (TIS) and gelled media (GM) culture systems of two Moroccan date palm varieties ‘Mejhool’ and ‘Boufeggous’. Obtained results indicated that shoot and root lengths as well as shoot fresh and dry weights were significantly (P < 0.05) higher in TIS compared to GM. Moreover, the vigour of obtained shoots was better in TIS compared to GM. Therefore, TIS-derived plantlets have shown an acclimatization rate of 95% while this rate for GM-derived plantlets was 82%. Hence, bioreactors, as a growing system based on TIS, can be a valid alternative to conventional systems for in vitro culture, resulting in a reduction of cost, shelving area requirements, labour and time for the mass propagation of date palm cultivars.

Tissue culture have been used for plant propagation generally, and the medium has been important role in its growth. Vegetative propagation on Phalaenopsis sp orchids can be through the protocorm like bodies (PLB). Medium of affect on propagation of PLB was carried out on medium type, kind of basal medium and concentrations ratio of naphtaleneacetic acid (NAA) and benzylamino purine (BAP). The experiment used Completely Randomized Factorial Design with 3 replications and continued with the Duncan Multiple Range Test (DMRT) if there were significant differences. The results showed that the best callus formed in a combination of solid medium type and Murashige & Skoog (MS) basal medium was 100%. The most number of PLB produced from a combination of liquid medium types and a concentration ratio of NAA 1 mgL-1 and BAP 5 mgL-1, the most number of plantlet produced from a combination of MS basal medium and the concentrations ratio of NAA 0.1 mgL-1 and BAP 0.1 mgL-1, the number of PLB germination and PLB with leaves were influenced by each single factor.

The NeuMoDx HPV assay is a novel fully automated, real-time PCR-based assay for the qualitative detection of high-risk human papillomavirus (HPV) DNA in cervical specimens. The assay specifically identifies HPV16 and HPV18 and concurrently detects 13 other high-risk HPV types at clinically relevant infection levels. Following the international guidelines, the clinical performance of the NeuMoDx HPV assay for cervical intraepithelial neoplasia grade 2 or worse (CIN2+) against the reference standard Hybrid Capture 2, as well as intra- and inter-laboratory reproducibility were assessed on PreservCyt samples. The clinical accuracy of the assay was additionally evaluated against the clinically validated Alinity m HR HPV and COBAS 4800 HPV Test on PreservCyt samples, and against the clinically validated HPV-Risk assay on SurePath samples. The NeuMoDx HPV assay performance for CIN2+ was non-inferior to the reference methods on both sample types (all p < 0.05), and showed excellent intra- and inter-laboratory reproducibility (95.7%; 95% CI: 93.9–97.3; kappa value 0.90 (95% CI: 0.86–0.94); and 94.5%; 95% CI: 92.6–96.2; kappa value 0.87 (95% CI: 0.82–0.92), respectively). In conclusion, the NeuMoDx HPV assay meets international guideline criteria for cross-sectional accuracy and reproducibility, and performs equally well on cervical screening specimens collected in two widely used collection media. The NeuMoDx HPV assay fulfils the requirements to be used for primary cervical screening.

In this study, we propagated two old Galician plum varieties in liquid medium using a temporary immersion system with RITA© bioreactors. Environmental variables including culture system, light intensity, CO2 enrichment, immersion frequency and sucrose supplementation were evaluated in relation to in vitro proliferation, physiological status and ex vitro performance. Bioreactors were superior to jars for culturing shoots in photomixotrophic conditions, producing up to 2 times more shoot numbers and up to 1.7 times more shoot length (depending on the genotype) using shoot clusters. The number and quality of shoots were positively influenced by the sucrose concentration in the medium, plus by the light and gaseous environment. For individual apical sections the best response occurred with 3% sucrose, 150 µmol m−2 s−1 photosynthetic photon flux density and 2000 ppm CO2, averaging 2.5 shoots per explant, 26 mm shoot length and 240 mm2 leaf area, while with 50 µmol m−2 s−1 light and ambient CO2 (400 ppm) values decreased to 1.2 shoots per explant, 14 mm of shoot length and 160 mm2 of leaf area. Shoots cultured photoautotrophically (without sucrose) were successfully rooted and acclimated despite of showing limited growth, low photosynthetic pigments, carbohydrate, phenolic and antioxidant contents during the multiplication phase.