Number Of Subcultures Research Articles

Enhanced micropropagation protocol of ex vitro rooting of a commercially important crop plant Simmondsia chinensis (Link) Schneider

A micropropagation protocol was developed by further improvement of prevailing methods using proven germplasm for ex vitro rooting and addressed the effect of the number of subcultures on the rooting ability of shoots. A comparative study was done between in vitro rooting method and ex vitro rooting method. Using the ex vitro rooting method a plantlet could be produced in 135 days, which was in a shorter time compared to the in vitro rooting method - 180 days. The best axillary shoot bud induction was observed on Murashige and Skoog (MS) medium supplemented with 4.6 μM thidiazuron (TDZ) with 5 shoot buds per explant. In the shoot cluster, which was subcultured on MS medium supplemented with 2.3 μM TDZ, the rate of shoot multiplication increased in the 3rd subculture. The maximum mean number of shoots per explant (20) was obtained at the 3rd subculture on the same medium. Shoots were harvested at the 1st, 3rd and 5th subculture and pulse treated for root induction. The highest rooting (95%) was achieved from the 3rd subculture onwards with pulse treated shoots for fifteen days. The rooted plants could be established in a greenhouse with 99% survival. Ex vitro rooting is a promising method to reduce the time for plant generation. The resultant plantlets well established in pots and fruiting was observed within a year.

Patronage and territoriality: Islamizing space in the Western city

Today’s cosmopolitan Western City contains large numbers of sub-cultures within its neighbourhoods, streets and buildings that are re-configuring the norms of citizenship and place. This paper examines the role of citizenship as a sense of cultural identity, territorial belonging and place attachment in the building of three mosques in Adelaide across a 100-year time span. Starting with the colonial period and the building of the first mosque in Australia up until the post-colonial period of large-scale Muslim immigration to Australia, the paper demonstrates the forms of social organization and patronage that were created as a strategy for collective action to make the building of these mosques possible. Set against the marginal political position of Muslims in Australia throughout this period, this paper shows the creativity of social and resource mobilization to stake a claim for the ownership of land in the Western City and to define a Muslim ethnic boundary.

MICROPROPAGATION OF CZECH APPLE CULTIVARS

The objective of this study was to investigate micropropagation protocols for use with three apple cultivars (‘James Grieve Compact’, ‘Jarka’ and ‘Mivibe’) bred in the Czech Republic. Apple shoot tips (5 to 10 mm in length) were established in vitro using 0.15% HgCl2 as sterilization solution. Contamination rate, survival and development of shoots from excised shoot tips were analyzed after sterilization. For the three cultivars, the effect of three growth regulators 1, 2 and 4 mg L-1 BAP (6-benzylaminopurine), 0.5 and 1 mg L-1 TDZ or 10 mg L-1 2iP in basal MS medium on proliferation, callus formation and shoot morphology is shown. Values of proliferation rate varied between 1.1 and 3.6. Generally, the highest rate was obtained for cultivar ‘Jarka’ that produced 3.6 new shoots on MS medium containing 4 mg L-1 BAP. The lowest number of newly formed shoots (1.1) was noted for cultivars ‘James Grieve Compact’ and ‘Mivibe’ on MS medium with low concentration of BAP (1 mg L-1). In vitro plants of ‘James Grieve Compact’ were of poor quality and developed symptoms of yellowing or chlorosis of leaves in later subcultures. It was possible to maintain actively growing in vitro cultures of ‘James Grieve Compact’ on all tested multiplication media only for a limited number of subcultures (usually, 8-9 subcultures; total culture period about 10 months), after which they either ceased to proliferate and multiply or eventually turned yellow-brown and subsequently died out. The effects of indole-3-butyric acid, indole-3-acetic acid or naphtalene acetic acid in concentration 1 mg L-1 on root induction were tested in MS medium for cultivars ‘Jarka’ and ‘Mivibe’. Rooting was induced for both genotypes tested. However, the root induction was relatively low from 0 to 44% of in vitro rooted plants.

Drivers of attitudes toward luxury brands

Purpose– Limited attention has been paid to the cultural influences on the formation of consumer attitudes toward luxury brands (LUX). The purpose of this paper is to investigate this relationship by developing a model that additionally employs the constructs of susceptibility to normative interpersonal influence (SNII) and brand consciousness (BCO).Design/methodology/approach– Sample data were gathered through surveys administered to 383 college students in the UK and Taiwan. The model of cultural influences on attitudes toward luxury brands was empirically tested using multi-group structural equation modeling to evaluate its applicability across the two countries.Findings– Results are presented in two parts: first, the exogenous construct part of the model establishing the reliability and validity of the cultural dimension constructs (horizontal individualism, vertical individualism, horizontal collectivism, and vertical collectivism) that are antecedent to consumer SNII and 2) the endogenous part of the model in which consumer SNII affects LUX through the mediating role of BCO.Research limitations/implications– The findings in the current study are limited to a sample of college students in the UK and Taiwan, which, through representing western and Asian countries, each housing different cultures, do not span the greater number of cultures found across these countries, much less across the world. Furthermore it is assumed that there are a number of subcultures in both the UK and Taiwan that are not accounted for in this study.Practical implications– An individual level of cultural orientation (e.g. horizontalism and verticalism) rather than traditionally adopted regionally defined or nationally based (Hofstede, 1980) cultural criteria should be investigated to identify more accurate market demand patterns in order to best target consumers in these markets (Sharma, 2010). In addition, appealing, vertical ad messages would be more effective in stimulating consumer motivations for consumption of luxury brands. Conversely, horizontal ad messages would be effective in demarketing approaches.Originality/value– The current study is the first of its kind to explore the effect of cultural-orientation on the formation of LUX cross-nationally. As such it provides future cross-cultural researchers with valid and reliable culturally based constructs that can be used to predict consumer SNII in developing LUX. In addition, establishing the mediating role of BCO in the relationship between SNII and LUX helps marketers better understand the equity of their luxury brands, particularly in Asian countries.

Organ and plantlet regeneration of Menyanthes trifoliata through tissue culture

The conditions for the regeneration of plants through organogenesis from callus tissues of <em>Menyanthes trifoliata</em> are described. The shoot multiplication rate was affected by basal culture media, the type and concentration of cytokinin and subculture number. The best response was obtained when caulogenic calli were cultured on the modified Schenk and Hildebrandt medium (SH-M) containing indole-3-acetic acid (IAA 0,5 mg/l) and 6-benzyladenine (BA 1 mg/l) or zeatin (2 mg/l). Under these conditions ca 7 shoots (mostly 1 cm or more in length) per culture in the 5th and 6th passages could be developed. In older cultures (after 11-12 passages) there was a trend for more numerous but shorter shoot formation. All regenerated shoots could be rooted on the SH-M medium supplemented with 0.5 mg/l IAA within 6 weeks; 80% of in vitro rooted plantlets survived their transfer to soil.

Estabilidade fenotípica de genótipos de morangueiro submetidos a número variável de subcultivos in vitro

Objetivando analisar o número de subcultivos que permita a multiplicação do morangueiro, sem que ocorram alterações nas características fenotípicas dos clones submetidos a esse processo, explantes dos genótipos 'Aromas', 'Camarosa' e 'Camino Real' foram submetidos, em uma primeira etapa, a 12 ciclos de subcultivos in vitro e, no ano seguinte, explantes dos mesmos genótipos, das mesmas plantas matrizes, foram subcultivados por três ciclos. Após a fase de aclimatização, as mudas foram transplantadas para canteiros, e o experimento conduzido segundo delineamento em blocos casualizados em esquema fatorial 3x2 (genótipos e níveis de subcultivos). As características avaliadas foram: altura da planta e comprimento da raiz (cm), massas fresca e seca da parte aérea e da raiz (g) e número de estolões por planta. Na avaliação a campo das características fenotípicas, não foram observadas diferenças significativas entre os dois níveis de subcultivos. As cultivares 'Camarosa' e 'Camino Real' apresentaram maior variação entre os dois níveis de subcultivos, sendo as maiores variações observadas em plantas submetidas a três subcultivos. Com 12 subcultivos in vitro de plantas de morangueiro, das cultivares 'Aromas', 'Camarosa' e 'Camino Real', é possível obter maior número de mudas micropropagadas sem alterações nas características fenotípicas. A cultivar 'Aromas' apresentou uma maior estabilidade nas características fenotípicas estudadas em relação aos diferentes ciclos de subcultivo in vitro.

Gauging the genetic changes occurring across globe artichoke micropropagation towards an appropriate variety registration and nursery production

Micropropagation of globe artichoke is an alternative method for production of large-scale vegetative material, particularly for spring cultivars. In spite of the widespread use of in vitro propagated material both for artichoke large-scale commercial production and for germplasm preservation, there is a lack of information about the consequences of in vitro propagation on genetic stability of globe artichoke. In the present work leaves from mother plants grown in the field and from in vitro multiplicated shoots were analyzed over two years to determine genetic stability and to evaluate the influence of the number of subcultures on the generation of somaclonal variants. Genetic stability was assessed using ISSR and SSR molecular markers on four ecotypes and two lines derived by crosses. The primers used exhibited a varying ability to detect genetic variation as shown by their polymorphic information content (PIC) values. It was found that ISSRs can be applied effectively in the evaluation of genetic stability/variability, including in vitro, detected by an automated sequencer. The technique was effective in detecting changes occurring during the micropropagation process. Statistically significant differences were revealed among clones, i.e. there were clones accumulating a greater amount of changes while others showed a lower degree of changes during subculture. Nevertheless, the general trend of changes across the subculture was similar, reaching a maximum rate between the 2nd and 4th subcultures. These results are important to both the theoretical and practical points of view, for breeders in order to proceed for variety registration, and their implications are discussed.

Micropropagation of an elite F1 watermelon (Citrullus lanatus) hybrid from the shoot tip of field grown plants

The aim of this work was to develop a protocol for rapid micropropagation of an elite F1 hybrid watermelon cultivar using shoot tip of field-grown plants. Maximum frequency (73%) of shoot tip showed growth response in MS medium supplemented with 5 mg l-1 benzyl adenine (BA) and 0.1 mg l-1 indole-3 acetic acid (IAA). Upon transfer to cytokinin-enriched medium, the cultures produced multiple shoots and 2.0 mg l-1 BA was optimum in this respect. Addition of gibberellic acid (GA3) in the multiplication medium resulted in better growth of shoots. Rooting rate was 100% when shoots were obtained from second subculture were cultured in medium with 1.0 mg l-1 indole-3 butyric acid (IBA). The shoots produced more roots with increasing number of subcultures. About 72% of the regenerated plantlets acclimatized successfully and survived in the soil condition.

Period, subculture number and culture color influence on the isolated Magnaporthe oryzae sporulation at the Tocantins State

El hongo Magnaporthe oryzae , causante de lo anublo del arroz (Bruzone), presenta una serie de limitaciones a los cultivos bajo condiciones controladas. Algunas caracteristicas pueden ser variables, incluso para un mismo aislado y requieren innovaciones por los investigadores para permitir lo desarrollo de las pruebas. Este estudio tuvo como objetivo verificar, en esporas aislados M. oryzae raza IA-1 recogidas en el Proyecto Rio Formoso, municipalidad de Formoso do Araguaia, la durabilidad de los cultivos almacenados en el laboratorio, la cantidad maxima de repicagens para mantener la produccion satisfactoria de las conidias por las colonias y la relacion de diferentes colores de las culturas en medio BDA con la produccion de conidias. Con este fin, se llevo a cabo tres experimentos en el diseno completamente al azar. En el estudio I, se utilizaron las culturas con 10, 14, 18, 22, 26 y 30 dias despues de la repicagen. En el estudio II, se utilizaron las culturas con 0, 1, 2, 3 y 4 repicagens de la cultura original. En el estudio III, los cultivos fueron clasificados como Negro, centro Gris con bordes anchos, Gris, centro Gris con bordes Blancos estrechos y Blanco. Mayores esporulaciones de M. oryzae se obtuvieron a los 14 dias despues del trasplante, en la segunda repicagen y de la coloracion negra.

TISSUE CULTURE PROPAGATION OF LEMON VERBENA (Aloysia triphylla)

The current research was conducted at the experimental station and tissue culture laboratory of Vegetable and Floriculture Department, Faculty of Agriculture, MansouraUniversity during the seasons of 2008 – 2010. A protocol is described for rapid and large-scale propagation of the woody aromatic and medicinal shrub lemon verbena (Aloysia triphylla) by in vitro culture of shoot tips or nodal segments from mature plants. Nodal segments were found to be more efficient than shoot tips for lemon verbena shoots regeneration on MS medium (Murashig and Skoog 1962) supplemented with BAP at 0.1 mg/L. Of the three different cytokinins ,6-benzyl aminopurine (BAP), kinetin (Kin) and thidiazrone (TDZ) evaluated as supplements to Murashige and Skoog medium (MS), BAP at concentration of 0.5 mg/L was found to be the most effective cytokinin in inducing multiple shoots. The frequency of shoot proliferation was markedly influenced by number of subcultures, since the highest percentage of shoot multiplication (100 %) as well as large the number of shoots/ explant (11.5 shoots) were obtained in the 4th subculture, after which there was a declined in shoot development. Rooting was best induced (92 %) in shoots cultured on MS medium augmented with indole-3-butyric acid (IBA) at 0.5 mg/L. Mixture of soil: sand: peatmoss (1: 1: 1) was the most suitable planting substrate for hardening and its use ensured high survival frequency (100 %) of regenerated plants prior to outdoor transfer.

Effects of successive subculturing on stability, virulence, conidial yield, germination and shelf-life of entomopathogenic fungi

To determine the stability and conidial yield of two strains of the entomopathogenic fungus Metarhizium anisopliae and one strain of M. brunneum, being developed for the control of insect pests. The conidial yields and the shelf-life of the conidia of two commercially viable strains of M. anisopliae V275 (=F52) and ARSEF 4556 and one strain of M. brunneum (ARSEF 3297) were determined after harvesting conidia from in vitro subcultures on Sabouraud dextrose agar (SDA) and broken basmati rice. The strains were stable and showed no decline in virulence against Tenebrio molitor, even when subcultured successively 12 times on SDA. Conidia-bound Pr1 protease activity decreased in conidia harvested from SDA and mycosed cadavers after the 1st subculture, but increased in conidia produced on rice. The C:N ratio of conidia from mycosed cadavers was lower than that of conidia from rice or SDA. Irrespective of the number of subcultures, strain ARSEF 4556 produced significantly higher conidial yields than ARSEF 3297 and V275. The 12th subculture of V275 and ARSEF 3297 produced the lowest conidial yield. Shelf-life studies showed that conidia of strain ARSEF 4556 had a higher conidial viability than V275 and ARSEF 3297 after 4 months, stored at 4°C. The current study shows that determining strain stability and conidial yield through successive subculturing is an essential component for selecting the best strain for commercial purposes. This is the first study to compare quality control parameters in the production of conidia on rice, and it shows that the level of Pr1 is comparatively high for inoculum produced on rice.

In vitro regeneration of Centaurium erythraea Rafn from shoot tips and other seedling explants

Various explants from 30-day-old seedlings of <em>Centaurium erythraea </em>Rafn were evaluated for their morphogenetic capacity under in vitro culture conditions. Shoot formation from shoot tip explants was achieved mainly through adventitious bud differentiation. The highest number of shoots (up to 43.3 ± 2.2 from a single shoot tip) was obtained on Murashige and Skoog medium (MS) supplemented with indole-3-acetic acid (IAA) (0.57 μM) and 6-benzylaminopurine (BAP) (4.4 μM). Adventitious shoot regeneration was also achieved through organogenesis from calluses obtained from hypocotyls, cotyledons, roots and leaves on MS medium containing IAA (2.85 μM) and BAP (0.88 μM). Significant differences were noted between explant types in their effects on shoot regeneration. In the primary culture, the best response was obtained either from calluses derived from roots or leaves (44.4 ± 4.5 and 40.2 ± 6.0 shoots per callus, respectively). The number of subcultures of inoculated calluses affected both the multiplication rate (the number of shoots/explant) and shoot morphology (the frequency of shoot hyperhydricity). Shoots rooted with the frequency of 94-100% after culture on MS medium without growth regulators. Plantlets were successfully acclimatized (97%) under high relative humidity and then moved to the greenhouse.

Public Space, Public Morality: The Media Construction of Sex in Public Places

Public sex has been the focus of a number of academic texts in recent years, but while some comment more generally on how the media have presented specific forms of public sexual activity, it remains unclear how the media have constructed these undertakings. The phenomenon consists of a number of subcultures aimed at providing sexual enjoyment in public settings in different ways. There has been evidence that those engaging in public sex undertake significant risk that, while motivating for those taking part, can lead to a number of tragic consequences. Some of these consequences include physical attacks by members of the public as well as susceptibility to robbery and even abduction. This article will explore one of the possible reasons behind an unfavourable view of public sex in the public conscience- the mass media, and in particular, newspapers in the United Kingdom. The way in which the media have constructed various forms of public sex will be explored, as well as the information provided in reports, and in particular, the detailing of locations of Public Sex Environments (PSEs).

Helping Ordnance Survey stand out from the crowd

Purpose – Ordnance Survey was established in 1791 as a paper map maker for Britain's Armed Forces. Two hundred and nineteen years on, it has evolved to a high‐tech geographic data specialist, and the impact of this on its culture, identity and vision has been massive. In 2008 Ordnance Survey found itself struggling to recruit and retain in technology and commercial fields. It also saw great challenges in motivating staff and bringing together a number of subcultures that had developed over time. This paper aims to investigate this issue.Design/methodology/approach – It was critical to understand the current employer brand strengths and weaknesses, how Ordnance Survey is perceived externally, what it is like in reality, and its vision. This was achieved through solid research conducted internally and externally, including focus groups with existing staff, depth interviews with senior management and consultation with recruitment consultants, short‐service leavers and recent joiners.Findings – The research h...

Content of Neutral Lipids and Fatty Acids in Callus Cultures and Leaves of Intact Plants of Ajuga Genevensis and Ajuga Chia

ABSTRACTThe comparative study of quantity and composition of neutral lipids and their fatty acids in the callus cultures and leaves of intact plants of Ajuga genevensis and Ajuga chia was carried out. The dependence of composition and content of lipids and fatty acids on the species belonging, the origin of calli and the culture age (number of subcultures) was revealed.

Stability of transgenes in long-term micropropagation of plants of transgenic birch (Betula platyphylla)

The stability of integration and expression level of transgenes in long-term micropropagation clones of transgenic birch (Betula platyphylla Suk.) was examined. Multiplexed PCR and reverse primer PCR demonstrated stable integration of transgenes into regenerated plants. Expression levels of the bgt and gus genes among shoot plantlets, subcultured 4, 7, 9 and 15 times, were significantly different. The transcriptional expression level of extraneous genes in regenerated plants decreased with increasing subculture number. Transcriptional gene silencing (TGS) occured in regenerated transgenic lines. The silencing rate of GUS in the 5th subculture plants was 22-65%. TGS in regenerated plants could be reactivated with 5-azacytidine (Azac) at 50-200 microM. GUS and BGT protein expression was reactivated in the micropropagated transgenic birch plants when treated with Azac. A decrease in expression level with increasing number of subcultures is thus associated with DNA methylation.

Factors influencing somatic embryogenesis and regeneration ability in somatic tissue culture of spring and winter rye

Rye is an important crop in Northern and Eastern Europe. However, the application of various biotechnologies in rye breeding has been limited duo to its recalcitrant in tissue culture. In order to improve somatic tissue effi ciency, key factors affecting somatic embryogenesis and reproducible green plant regeneration of rye (Secale cereale L.) were evaluated and optimised. In this study, a total 27 rye genotypes including 10 spring and 17 winter genotypes were involved in the investigation. Genotype, culture medium, sugar, gel agent and auxin infl uenced somatic embryogenesis of immature embryo signifi cantly. One-two weeks cold pretreatment of young embryo enhanced somatic embryogenesis and green plant regeneration. In culture of immature embryos, infl orescences and leaf segments of the seedlings, explants signifi cantly infl uenced the culture effi ciency. Highest embryogenic callus yield resulted from rye immature embryo as explant compared to young infl orescence and leaf segment of seedling. Developmental stage of embryo played an important role in somatic embryogenesis. Late spherical coleoptile stage (embryo size 0.5–1mm in length) was optimal developmental stage of immature embryo for culture. Morphogenetic potential of embryogenic callus decreased with an increasing number of subcultures, and this ability could be maintained in vitro for a maximum of 8 months of culturing.;

The Englishness of English Punk: Sex Pistols, Subcultures, and Nostalgia

This article considers the Englishness of English Punk, or, more specifically, the Englishness of the Sex Pistols and the cultural productions associated with them. It will consider whether the challenge that they posed to conventional, Establishment, consensus notions of Englishness has merely been recuperated as an entertaining diversion within a broader hegemonic nationalist history or whether this challenge has had a more fundamental impact. It will argue that the Pistols facilitated a reframing and a re‐imagining of English culture and left a legacy, which has been drawn upon by a number of subsequent art and music subcultures.

Anthraquinones from in vitro root culture of Morinda royoc L.

Morinda royoc L. (Rubiaceae) root cultures were established for the production of anthraquinones (AQs). Three independent experiments were performed to evaluate the effects of different levels of indole-3-acetic acid (0–22.8 μM), culture duration (15–75 days) and subculture number (0–4). The following indicators were recorded: root fresh weight per Erlenmeyer and intracellular and extracellular AQ production. The experiments performed in this study allowed an increase of intracellular AQ content up to a maximum of 4.5 mg g−1 of fresh mass, after 30 days of culture in a medium 5.7 μM of IAA. In addition, isolation and identification of seven AQs from M. royoc L. roots is described, one of them being reported for the first time for this species. The structures of isolated compounds were determined from 1H-NMR data. To the best of our knowledge, this is the first report on AQ production from root culture of this plant.

CONCENTRAÇÃO DE BENZILAMINOPURINA E AVALIAÇÃO DE PROTOCOLO PARA MULTIPLICAÇÃO IN VITRO DE GENÓTIPOS DE BANANEIRA

The purpose of this study was to evaluate a protocol for in vitro multiplication of four Musa spp. genotypes. Previous tests indicated the utilization of an ideal benzilaminopurin concentration of 5.0 mg L-1; 5.0 mg L-1; 4.5 mg L-1 e 4.0 mg L-1 in culture medium MS, during the multiplication of Caipira, Thap maeo, PV03-44 and FHIA-01 genotypes, respectively. After that, the multiplication rate, the size of shoots produced per explant, fungal and bacterial contamination rate, oxidation rate, and abnormality occurrence in the multiplication phases were evaluated. The multiplication rate and the size of shoots produced per explant varied among genotypes in the five subcultures studied. The highest contamination rate was caused by bacteria and was more frequent in the in vitro establishment, tending to be reduced with the number of subcultures. The most of the genotypes did not present problems of microorganism contamination, except for Caipira cultivar which presented high bacterial contamination rate during in vitro establishment. This problem is a limitation of the protocol to obtain micropropagated plantlets of this genotype. KEY-WORDS: Musa spp. ; micropropagation; tissue culture; benzilaminopurin.