The young juvenile nodal explants of Adhatoda vasica cultured on Murashige and Skoog’s (MS) medium supplemented with CW (15% v/v) + BAP (5 mg/l) started proliferating multiple shoot buds (14±2) in 3 to 4 weeks and the shoots became 3 cm long in 6 to 8 weeks. The regenerated plants produced well developed roots on transfer to MS + IBA 1 mg/l in 10 to 12 days. The rooted plantlets were acclimatized before transfer to soil and 80% of the transferred plants survived (Raageeva Bimal and M.d. Shahnawaz, 2011). Earlier not much work has been studied in A. beddomei plant. In this context, our work mainly deals establishment of a plant tissue culture technique which can provide numerous plant production within a stipulated time period that to disease free and at any season.
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A protocol has been developed for in vitro plant regeneration from a nodal explant of Knock out rose variety, an economically important variety for cut rose. The present study utilized 6-benzyl adenine (BA) and napthaleneacetic acid (NAA) for the induction of shoot organogenesis on nodal explants. The highest frequency of shoot regeneration (71.67± 2.9 %) and number of shoots per explant (8.00 ± 1.00) were obtained on medium supplemented with 4.44 µM BA and 0.14 µM NAA. Rooting was achieved on half strength MS solid medium supplemented with 2.46 µM indole-3-butyric acid (IBA) produced (6.0 ± 1.0) roots with an average height of 4.9 ± 0.26 cm after 30 days of culture. The rooted plantlets were transferred for hardening, 80 per cent of plantlets survived and resumed growth in the mixture of soil, vermiculite and farmyard manure (1:1:1). Antibacterial activity of ethanolic flower extract was measured by agar disc diffusion method. Different concentrations of ethanolic flower extract of Knock out rose was tested for the anti-bacterial activity. Ethanolic flower extract of Knock out rose showed the maximum zone of inhibition (17 mm) and (15 mm) against Pseudomonas aeruginosa and Escherichia coli respectively.
The inoculation was carried out in the vicinity of flame. The surface sterilized explants were aseptically transferred to the respective culture media in the Laminar Flow Chamber. The explants were taken out from beaker and at the same time the cotton plug of the culture tube was slightly opened in front of the spirit lamp flame, the explant was put it the medium and immediately covered with cotton plug. The explants with nodal regions were inserted in the medium vertically. Cultures were transferred to fresh media with the same hormone concentration at 4 week intervals.
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There was some reported work on direct shoot regeneration of med icina l plants. By e liminating unnecessary callus formation in Phalaenopsisorchids direct shoot regeneration from nodes was also reported . In vitro shoot induction and plant regeneration fro m nodal segments of Balanitesaegyptiaca on Murashige and Skoog (MS) mediu m fort ified with 6-benzyladenine (BA), thid ia zuron (TDZ) and kinetin (Kin) was also observed . A rapid and effic ient protocol for h igh frequency direct shoot proliferation was observed in Andrographispaniculata, a valuable medic inal plant by using nodal e xplant. Direct shoot regeneration fro m nodal e xp lant also reported in SidacordifoliaLinn. , PogostemoncablinBenth. .Germinat ion and development of 30 days old seedling, in itiation of nodal e xp lant to elongation of 2-3 c m long shoots were depicted in Fig. 1.
Abstract: Vernonia amygdalina is a vegetable and medicinal plant used to treat various ailments such as diabetes, malaria, gastrointestinal disorders, and parasitic infections. The present study investigated the effect of supplementing different concentrations of 6-benzylaminopurine (BAP) and 1-naphthaleneacetic acid (NAA) either alone or in combination to Murashige and Skoog (MS) medium on in vitro plantlet regeneration of V. amygdalina from nodal explants. Control treatment without plant growth regulators was ideal for in vitro plantlet regeneration of V. amygdalina. In vitro plantlets regenerated from nodal explants supplemented with BAP and/or NAA showed growth abnormalities including chlorosis, basal callus, and excessive adventitious rooting. Callus cultures were induced from leaf explants on MS medium supplemented with different concentrations of BAP, NAA, and 2,4-dichlorophenoxyacetic acid (2,4-D) either alone or in combination. Maximum callus induction frequency (100%) was recorded in leaf explants cultured on MS medium supplemented with 0.5 – 2.0 mg/L 2,4-D. Fresh weight of calli increased up to 11-fold when treated with 0.5 mg/L 2,4-D after 8 weeks of culture.
achieved in maximum number of 12 shoot on MS + BAP 1.0 mg/L+ Kin 0.5 mg/L+ IBA 1.5 mg/L followed by 10 shoots on MS + BAP 0.5 mg/L +Kin 0.5 mg/L+ IAA 2.0 mg/Lout of 25 explants inoculated. Lemos and Baker (1998) used internodal explants of A. muricata to induce shoot regeneration on Nitsch media. The highest percentage (7%) of explants produced shoots in sorbitol 10-30 g/Las carbon source supplemented with BAP 2.0 mg/L+ NAA 0.5 mg/L. Zobayed et al. (2001) investigated A. muricata, nodal explants and produced multiple shoots in MS + BA 1.0 mg/L+ NAA 0.1 mg/L. Done Dipre et al. (2012) produced multiple direct organogenesis in A. muricata from immature hypocotyl explants in MS + BAP 2.25 mm + IBA 2.5 mm concentration. Bejoy and Hariharan (1992) used half strength MS medium with 20 g/L sucrose and 7 g/L agar + BA (6 benzyladenine) 2.2-13.3 mM + NAA 0.27-2.7 mm concentration for shoot induction. The present report shows variation in shoot formation, probably due to the medium used was MS and carbon source was sucrose, since the role of Kin and IBA are vital along with BAP in shoot regeneration.
was found to be the most effective media for maximum (95%) regeneration of nodal explant at 25±1°C and 16/8 hours cycle of light (2000 lux fluorescent tubes) and dark. Nodal segment showed maximum (9) shoot induction per explant. Twenty-nine shoots per nodal segment explant were observed on MS+0.75 mg l -1 BAP+0.01 mg l -1 IAA within 3 weeks. The rooting percentage (66%) and average number
The morphogenetic responses of nodal segment explants to various cytokinins (BA/KN/2-iP) are summarized in Table 2. When Explants cultured on MS medium with- out cytokinins failed to produce shoots even after 4 weeks of inoculation, but the explants when cultured on MS medium supplemented with different cytokinins at different concentrations showed variation in the regen- eration percentage and number of shoots formed. Nodes cultured on half strength MS basal medium showed no visible signs of tissue differentiation. This was possibly due to a greater demand of nitrogen and potassium-con- taining compounds, which induce greater amount of new proteins . These components are lower in half strength MS basal medium compared to full strength MS basal medium. Initial induction of shoots was noted after 10 - 12 days of inoculation (Figure 1(a)). Data on different growth parameters from different treatments were re- corded after 4 weeks of culture initiation following one transfer to the new medium. Among the three cytokinins tested, the best response (91.10%) was obtained in the presence of 1.0 mg/l BA and was found to be signifi- cantly higher than shoots induced per nodal explant in other concentrations of cytokinins (KN/2-iP) in the pre- sent study (Figure 1(b)). Nodes cultured on MS medium with different concentrations of KN and 2-iP showed lower induction of axillary shoot bud proliferation. The maximum number of multiple shoots was obtained (5.17 ± 0.04) in the medium containing 1.0 mg/l of BA. The shoots developed in this medium also attained maximum
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The shoots obtained from nodal explant were excised and cultured on MS liquid medium containing ascorbic acid (50mgl -1 ), citric acid (25mgl -1 ) and cysteine (25mgl -1 ) additives, auxin NAA 0.25mgl -1 and different concentrations of cytokinin BAP 2.5-5.0 mg -1 and TDZ 0.25-0.5mgl -1 for 5 passages. Sub culturing was carried out at an interval of 2 weeks on fresh liquid medium. The average number of propagules derived at the end of each subculture cycle out of a single propagule (5-6 shoot clusters) was regarded as rate of multiplication or the multiplication fold.
Initiation of multiple shoots/ microshoots occurred first on the 3 rd day after inoculation. Maximum number of microshoots was formed after a week time. Of the five different concentrations of BAP tested, 2 mg/L proved better in shoot induction with 7.63 (±3.0) shoots per explant (Table 1; Fig. 2C). Increase in BAP amount could not enhance the shoot formation anymore. Second best response was found at 5 mg/L. Concentration at 2 mg/L was effective both in multiple shoots formation and growth of shoots. Besides these, the shoot length as well as the number of leaves produced were higher, 3.67(±1.2) cm and 12.29(±7.0), respectively. From the previous studies, it is learned that, BAP is the most preferred growth regulator for direct shoot induction in tissue culture experiments using nodal explant of A. aspera (Gnanaraj et al., 2012; Sen et al., 2013a), and the indirect shoot formation from A. aspera internode, leaf and root derived callus (Sen et al., 2014), and potato cv. Granula (Laboney et al., 2013).
The MS media recorded better results in 2009 and 2010 on the examined parameters, followed by the OM and WPM (Ruggini, 1984). These findings appeared to be in contradiction with those observed on cv. Moraiolo, where OM proved to be significantly superior than WPM and yielded the highest number of shoots per proliferated explant (0.84) (Ansar et al., 2009). Regarding the concentration of the growth regulator (BAP), the largest quantity used (4mg/l) had a negative impact on the multiplication rate that also declined with subculture. This finding is consistent with Ansar et al., (2004) who indicated that a high concentration of growth regulators limited significantly shoot multiplication on OM and WPM. The shoot height in Leccino cv. decreased with the increase in the cytokininconcentration levels (BAP, 2ip and Kinetin) (Ramzan Khan et al., 2002). Dimassi-Theriou (1994) found that BAP alone resulted in bettershoot proliferation than 2ip, and when added at a concentration of 5 to 7.5 ml/l, BAP produced 1.2-1.8 shoots/explant and did not impact the height of Kalamon olive (Rama and Pontikis, 1990). MS medium produced the longest explants, followed by OM and WPM, respectively. These results were consistent with those recorded on Moraiolo cv. where OM yielded the highest shoot length (2.25cm) in comparison with WPM (1.24cm). In Moroccan Picholine cv., media had a significant impact on the length of developed shoots after 2subcultures. OM produced the longest shoots (1.24cm), followed by MS/2 (0.89cm) then WPM (0.3cm) (Brhadda et al., 2003). The year had an important influence on the multiplication rate of Bshaaleh explants, potentially resulting from the use of foliar fertilizers during the 2009 season, which improved vegetative growth and produced well-developed twigs on the tree regardless of the seasons
The nodal density of the modeled domain was varied, that is, the number of nodes used to represent the area of analysis (discretization). The software's possibilities in terms of density are: thick, medium, fine, very fine and personalized. Nodal density represents the degree of refinement of the cloud of points on which the solution to the problem is obtained. In the modeled problem thick and medium density is adopted due to the limitations imposed by the trial version; the analysis is carried out for the two nodal density situations and results are obtained. We expect the results with the average mesh to be more adjusted; however, we consider the importance of observing the variations between the densities of the two meshes in order to conclude about the justification of possible increases in computational cost due to refinement.
assumption that there was no false-positive nodal staging in clinical practice. We regard this assumption as reason- able, because pathologists must have substantial expertise and follow pathological guidelines to discriminate a nodal- positive patient. Moreover, a group judgment, instead of single-minded decision, is needed to assert a nodal disease. Therefore, it is statistically and clinically reasonable to compute NSS using false-negative probability derived from beta-binomial model under such an assumption.
Inoculation of plants with endophytic bacterium Nodal segments (length: 0.5 cm) of Bacopa monnieri were disinfected by sonicating in water for 20 min and dipping in 70% ethanol for 1 min, followed by 25 min of sodium hypochlorite (25%) / Tween 80 (0.01%) solution and rinsed three times with distilled and sterile water (Alderete et al., 2006). The explants were cultured onto a hormone free Murashige– Skoog (MS) medium (Murashige and Skoog, 1962) supplemented with 20 gl _1 sucrose, 7 gl _1 agar, and the pH was adjusted to 5.7 with KOH with the addition of 200μl of endophytic B.cereus. Then, the explants were cultured under a photoperiod of 16 h of light and 8 h of dark under an irradiance of 52 mmol m _2 seg _1 . The explants without endophytic B.cereus were marked as control explants. This experimental setup was repeated for 10 times. All the data collected from these experiments were subjected to an analysis of variance (ANOVA) using Microsoft Excel 2000.
Once the optimum physico-chemical conditions for induction and growth of shoots from axillary bud explant were defined, the in vitro shoot cultures were excised as such in the form of a cluster and were cut into two or more bunches each consisted of 4-5 shoots and each bunch was called a propagule. In this way if one mother propagule produced ten shoots (Fig.8 ) after a period of 30-35 days incubation then the same propagule was cut into two daughter propagules the number of daughter propagules (4-5 shoots) achieved and from each mother propagule was presented as rate of multiplication to multiplication (M-M). At the time of sub-culture of mother propagule the elongated shoots (3-4cm) were excised individually and were transferred for rooting. The number of shoots harvested for rooting from each mother propagule was presented as rate of multiplication to rooting (M-R) in the results. The experiments for shoot multiplication were conducted in liquid as well as in MS media but chlorosis and browning of shoots were not observed in semi-solid media in multiplication experiments, therefore, only semisolid MS media were used for subsequent experimentations of multiplication.
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The buried pipelines of water supply network have the following characteristic: wide spatial distribution, huge number and indiscoverable failure position. When the network suffers from disasters, most of the nodes which are damaged slightly are hard to be found and recovered, however, only the nodes damaged very seriously and obviously can be found and recovered in time. Because of this, the water supply network is always in the working state of leakage and low pressure after disasters. In order to meet the water demand right after disaster, the minimum water pressure of nodes is one of the most important hydraulic conditions. If the actual nodal pressure is still higher than the required minimum pressure, usually the users can accept it. However, if the actual pressure is lower than the allowable value, the water demand would not be guaranteed. Due to the cascading failure of network, if some nodes or edges were damaged in the disaster, most of nodes in the network would be affected. In a sense, the APR can reflect the disaster reliability and importance of the node. Therefore, nodal average value of water pressure reducing is also selected from resource supply factors as an evaluation index of node importance.
There were also similarities in cell transduction results from the murine rectal in vivo transduction and human explant ex vivo transduction models. In both cases, VSV- G-pseudotyped LV exhibited the ability to transduce CD4-positive cell population. Notably, LV pseudotyped with the VSV-G envelope, which only requires binding to phospholipid constituents of most mammalian cells , achieved much more efficient infection of T cells than is possible with identical vectors pseudotyped with ampho- tropic retrovirus envelope . However, CD4 is not only expressed by helper T lymphocytes and T regulatory cells, but also by macrophages and dendritic cells at lower levels. All of these cell types generally reside in the LP, which is where the predominant staining was observed in humans. It is not clear why the LV was unable to trans- duce CD8 positive cells and this finding warrants addi- tional in vitro studies with purified T cell populations.
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Visual impairment is becoming recognized in PD pa- tients [11, 61], which has a correlation with gait impair- ment  and akinesia and rigidity . In the present study, disrupted occipital nodal degree centrality and ef- ficiency were observed in MPD patients in all three os- cillation frequencies. It has been described that PD patients show a higher dependence on visual informa- tion for motor control , which was confirmed by the evidence that visual cueing can improve walking in PD . Though the mechanism of visual-motor loop in PD has not been fully studied , the significant correla- tions between impaired occipital nodes and motor sever- ity dominated by akinesia/rigidity not tremor specifically within slow-3 oscillation frequency further supported the critical role of visual modulation in parkinsonian akinesia and rigidity. Thus, our result suggested that the oscillation-specific occipital nodes whose disruption ac- companied with motor exacerbation might be a critical target to modulate motor controls.
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The response of D. gangeticum nodal explants cultured on different shoot proliferation media over a period of six weeks is presented in table 1. Culture medium devoid of growth regulators (control) failed to stimulate the bud break response in the cultured explants even when the cultures were maintained beyond the normal observation period of four weeks. MS medium with growth regulator supplements produced better results in terms of percentage explant response, shoots/ explant, average shoot length and average number of nodes produced per shoot. In such media combinations bud break was noticed within 6-8 days of culture (Figure 1 A & B). Of the combination tested, MS + BA (1.0 mg/l) + IAA (0.25 mg/l) elicited optimal response in which an average of 5.5 ± 0.3 shootlets (Figure 1C) with a mean shoot length of 3.5 ± 0.3 cm per explant was recorded. The second best shoot multiplication (4.5 ± 0.2) was obtained in the medium MS + BA (1.0 mg/l) with a mean shoot length of 2.5 ± 0.3 cm. Higher concentration of BA (1.5 mg/l) in combination with IAA (0.5 mg/l) showed callusing explants with fewer shoots. In such cultures shoots were stunted with a mean shoot length of 1.4 ± 0.2 cm. The dependence of cultured explants on bud break response and shoot multiplication has already been established and extensively discussed (George and Sherrington 1984). This has also been recently reported in the case of micropropagation of many other medicinal plants like Hemidesmus indicus (Patnaik and Debata 1996), Gmelina arborea (Thirunavoukkarasu and Debata 1998) and Plumbago zeylanica (Sahoo and Debata 1998).
In the zebrafish genome, there are three nodal-related genes: cyclops, squint and southpaw. Of these, squint/nodal-related 1 tran- script expression is detected in pre-blastula stage zebrafish embryos, whereas cyclops/nodal-related 2 is expressed from late blastula stages in the blastoderm margin, in the axial mesendo- derm during gastrulation, and subsequently in the left diencephalon and left lateral plate mesoderm (LPM) [46–49]. Expression of southpaw/nodal-related 3 is observed in the left LPM and left diencephalon during late somitogenesis . Among other core nodal pathway components, transcripts encoding the Nodal receptors, co-receptor One-eyed pinhead (oep), the downstream effectors Smad2 and Smad3, and FoxH1 are expressed both maternally and zygotically, whereas the Nodal inhibitors lefty1 and lefty2 are detected after the mid-blastula transition (MBT) in the early embryo [25,51–58].