scholarly journals Providing inoculum for Didymella bryoniae studies: the effect of light spectrum and storing at low temperature

2024 ◽  
Vol 84 ◽  
Author(s):  
M. C. S. Virtuoso ◽  
E. H. C. Silva ◽  
E. M. Silva ◽  
T. S. Valente ◽  
P. F. Vargas ◽  
...  
Keyword(s):  
Mycelial Growth ◽  
Culture Medium ◽  
Culture Media ◽  
Medium Composition ◽  
Didymella Bryoniae ◽  
Light Spectrum ◽  
Storage Duration ◽  
Light Spectra ◽  
Fungal Structure

Abstract The in vitro sporulation of Didymella bryoniae is of great importance for studies that require pure inoculum and in large quantities. Thus, the objectives of this study were to identify the best condition for D. bryoniae sporulation combining different light spectra (UV-A or UV-B light, white light, and continuous dark), with distinct culture media (PDA, V8, ML, and PDAB) and, to evaluate fungus’ survivability stored at -20°C over time. The fungus samples were only able to sporulate when subjected to the UV-B light treatment, regardless of the culture medium. The highest appearance of spores conidium type was observed in the PDAB medium, and the lowest production occurred in the ML medium. Reproductive structures, such as perithecia and pycnidia, were observed in all culture media. However, there was considerable variation in the amount of each structure between the different culture media. The ML and V8 media showed a greater number of perithecia and the PDA and PDAB media presented a greater proportion of pycnidia compared to perithecia. The storage duration at -20°C did not affect mycelial growth or mycelial growth rate. In conclusion, the UV-B light is essential for D. bryoniae in vitro sporulation. Moreover, the culture medium composition influences the type of fungal structure produced, as well as spores’ size and quantity. Freezing at -20°C is an efficient technique that can be used to store D. bryoniae for at least five months without loss of viability.

Pertumbuhan Dan Perkembangan Anggrek Dendrobium anosmum Pada Media Kultur In Vitro Dengan Beberapa Konsentrasi Air Kelapa

Agrologia ◽  
2018 ◽  
Vol 1 (1) ◽  
Author(s):  
S. Tuhuteru ◽  
Meity L Hehanussa ◽  
Simon H.T Raharjo
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Water Concentration ◽  
Medium Composition ◽  
Coconut Water ◽  
Orchid Species ◽  
Randomized Design ◽  
Wet Weight ◽  
Completely Randomized Design

Dendrobium anosmum is one of natural orchids in Indonesia. Optimization of medium composition for orchid propagation through in vitro culture is necessary to enhance propagule multiplication capabilities and quality. This study was aimed to study the influence of concentration of coconut water in culture medium on in vitro growth and development of D. anosmum orchid species and to determine the optimal coconut water concentration in culture media.  The experiment were arranged in a Completely Randomized Design with four treatments and eight replications. The treatments consisted of the addition of coconut water with concentrations: 0 ml•l -1 (control), 50 ml•l-1, 100 ml•l-1 and 150 ml•l-1. The results showed that addition of coconut water in culture medium gave different effect on shoot growth and multiplication of D. anosmum orchids.  Coconut water concentration of 100 ml•l-1 was the best concentration for growth and multiplication of D. anosmum orchids, based on both shoots and roots growth, plantlet height and wet weight.

scholarly journals Influence of Light Conditions and Medium Composition on Morphophysiological Characteristics of Stevia rebaudiana Bertoni In Vitro and In Vivo

Horticulturae ◽  
2021 ◽  
Vol 7 (7) ◽  
pp. 195
Author(s):  
Alla A. Shulgina ◽  
Elena A. Kalashnikova ◽  
Ivan G. Tarakanov ◽  
Rima N. Kirakosyan ◽  
Mikhail Yu. Cherednichenko ◽  
...  
Keyword(s):  
Culture Media ◽  
Stevia Rebaudiana ◽  
Adventitious Shoot ◽  
Optimal Growth ◽  
Medium Composition ◽  
Light Spectra ◽  
Stevia Rebaudiana Bertoni ◽  
Multiplication Coefficient

We investigated the influence of different conditions (light composition and plant growth regulators (PGRs) in culture media) on the morphophysiological parameters of Stevia rebaudiana Bertoni in vitro and in vivo. Both PGRs and the light spectra applied were found to significantly affect plant morphogenesis. During the micropropagation stage of S. rebaudiana, optimal growth, with a multiplication coefficient of 15, was obtained in an MS culture medium containing 2,4-epibrassinolide (Epin) and indole-3-acetic acid (IAA) at concentrations of 0.1 and 0.5 mg L−1, respectively. During the rooting stage, we found that the addition of 0.5 mg L−1 hydroxycinnamic acid (Zircon) to the MS medium led to an optimal root formation frequency of 85% and resulted in the formation of strong plants with well-developed leaf blades. Cultivation on media containing 0.1 mg L−1 Epin and 0.5 mg L−1 IAA and receiving coherent light irradiation on a weekly basis resulted in a 100% increase in the multiplication coefficient, better adventitious shoot growth, and a 33% increase in the number of leaves. S. rebaudiana microshoots, cultured on MS media containing 1.0 mg L−1 6-benzylaminopurine (BAP) and 0.5 mg L−1 IAA with red monochrome light treatments, increased the multiplication coefficient by 30% compared with controls (white light, media without PGRs).

Phosphite-based Products in the In vitro Colletotrichum musae Control

2019 ◽  
pp. 1-9
Author(s):  
Maria Luísa Mendes Rodrigues ◽  
Edson Hiydu Mizobutsi ◽  
Paola Junayra Lima Prates ◽  
Paula Virgínia Leite Duarte ◽  
Regina Cássia Ferreira Ribeiro ◽  
...  
Keyword(s):  
Mycelial Growth ◽  
Culture Medium ◽  
Culture Media ◽  
Petri Dish ◽  
State University ◽  
Randomized Design ◽  
Colletotrichum Musae ◽  
Study Laboratory ◽  
Vitro Effect

Aims: The aim of this study was to evaluate the in vitro effect of different phosphite formulations and concentrations on the development of Colletotrichum musae. Sample: to evaluate the inhibition of germination, mycelial growth and sporulation of Colletotrichum musae. Study Design:  Treatments were conducted in a completely randomized design, with 4 replicates, each replicate consisting of 1 Petri dish. Place and Duration of Study:  Laboratory of Post-Harvest Pathology, State University of Montes Claros, between March and October 2017. Methodology: Three different phosphite formulations were used: FCu1 (4% Cu + 20% P2O5), FCu2 (4% Cu + 22% P2O5) at concentrations of 0.5;1.0; 1.5 and 2.0 mL L-1 and FK (42% P2O5 + 27.7% K2O) at concentrations of 0.5; 1.0; 1.5 and 2.0 mg.L-1. Products were incorporated into the respective culture media. Culture medium alone and culture medium + imazalil were used as controls. Petri dishes were housed in BOD chamber at 25°C under a 12 hours photoperiod. Results: Results were submitted to analysis of variance and regression, and means were compared by the Tukey test (P <0.05). Control was compared to the other treatments by the Dunnet's test (P <0.05). Among the tested phosphite formulations, copper and potassium phosphites were found to reduce the mycelial growth of Colletotrichum musae. FCu2 presents a fungicide-like effect from the concentration of 0.5 m.L-1 in the control of conidia production. As for the FCu1, a fungicide-like effect was observed in the control of germination from the concentration of 1.5 mL.L-1. Conclusion: A significant fungistatic effect was observed between the concentrations of the products in the mycelial growth, sporulation and germination obtaining control of up to 100% of the development of C. musae. Copper phosphites were as effective as fungicide in inhibiting fungal development.

scholarly journals PROSPECÇÃO E AVALIAÇÃO DO POTENCIAL BIOTECNOLÓGICO DE LINHAGENS COMERCIAIS DE SHIITAKE

2021 ◽  
Vol 35 (4) ◽  
pp. 640-649
Author(s):  
Dalvan Pereira Abilio ◽  
Olívia Gomes Martins ◽  
Giovana Silva ◽  
Meire Andrade
Keyword(s):  
Wheat Bran ◽  
Radial Growth ◽  
Mycelial Growth ◽  
Culture Medium ◽  
Lentinula Edodes ◽  
Culture Media ◽  
Biotechnological Potential ◽  
E Mail ◽  
Eucalyptus Sawdust

PROSPECÇÃO E AVALIAÇÃO DO POTENCIAL BIOTECNOLÓGICO DE LINHAGENS COMERCIAIS DE SHIITAKE   DALVAN PEREIRA ABÍLIO1, OLÍVIA GOMES MARTINS2, GIOVANA CRISTINA PINTO ALVES DA SILVA3, MEIRE CRISTINA NOGUEIRA DE ANDRADE4   1 Graduando em Ciências Biológicas, Centro Universitário Sagrado Coração – Unisagrado, R. Irmã Arminda, 10-50 - Jardim Brasil, 17011-160 Bauru, SP, Brasil. E-mail: [email protected] 2 Doutoranda em Agronomia – Energia na Agricultura, Faculdade de Ciências Agronômicas, Universidade Estadual Paulista "Julio de Mesquita Filho" (UNESP), Av. Universitária, 3780 - Altos do Paraíso, 18610-034, Botucatu, SP, Brasil. E-mail: [email protected] 3 Mestre em Agronomia – Energia na Agricultura, Faculdade de Ciências Agronômicas, Universidade Estadual Paulista "Julio de Mesquita Filho" (UNESP), Av. Universitária, 3780 - Altos do Paraíso, 18610-034, Botucatu, SP, Brasil. E-mail: [email protected] 4 Docente da Faculdade Gran Tietê, Av. 15 de Novembro, 125 - Centro, 17340-000, Barra Bonita, SP, Brasil. E-mail: [email protected]   RESUMO: O objetivo deste trabalho foi avaliar e comparar o crescimento micelial in vitro de quatro linhagens comerciais de Lentinula edodes (LE-241, LE-242, LE-243 e LE-244) em meio de cultura à base de serragem de eucalipto, suplementado com bagaço de malte ou farelo de trigo. O delineamento experimental foi inteiramente casualizado, em esquema fatorial 2x4 (substratos x linhgens), totalizando oito tratamentos, cada um com cinco repetições. Os substratos foram preparados com 20% de suplementação, acrescidos de calcário (tamponante), umidificados e esterilizados. Os meios de cultura foram preparados a partir da infusão dos substratos, filtração, adição de ágar e esterilização. As placas foram inoculadas com as linhagens e o crescimento radial do micélio na superfície do meio de cultura foi mensurado com paquímetro. O menor desenvolvimento micelial ocorreu com a linhagem LE-241 e com a linhagem LE-243 no substrato suplementado com farelo de trigo. Os maiores desenvolvimentos foram obtidos com as linhagens LE-242 e LE-244 no substrado suplementado com bagaço de malte. O desenvolvimento de todas as linhagens foi maior no substrato suplementado com bagaço de malte. Sendo assim, a utilização do bagaço de malte para o crescimento micelial é uma alternativa viável à suplementação tradicional com farelo de trigo.   Palavras-chave: fungos, cogumelo, Lentinula edodes, micélio, bagaço de malte.   PROSPECTING AND EVALUATING THE BIOTECHNOLOGICAL POTENTIAL OF SHIITAKE COMMERCIAL STRAINS   ABSTRACT: This study aimed to evaluate and compare the in vitro mycelial growth of four commercial Lentinula edodes strains (LE-241, LE-242, LE-243 and LE-244) in a culture medium based on eucalyptus sawdust, supplemented with malt bagasse or wheat bran. The experimental design was completely randomized, in a 2x4 factorial scheme (substrates x strains), totaling eight treatments, each with five repetitions. The substrates were prepared with 20% supplementation, added with lime (buffer), humidified and sterilized. The culture media were prepared from the infusion of substrates, filtration, addition of agar and sterilization. The plates were inoculated with the strains and the radial growth of the mycelium on the surface of the culture medium was measured using a caliper. The smallest mycelial growth occurred with the LE-241 strain and with the LE-243 strain on the substrate supplemented with wheat bran. The greatest growths were obtained with the LE-242 and LE-244 strains in the substrate supplemented with malt bagasse. The development of all strains was greater in the substrate supplemented with malt bagasse. Therefore, the use of malt bagasse for mycelial growth is a viable alternative to traditional wheat bran supplementation.   Keywords: fungi, mushroom, Lentinula edodes, mycelium, malt bagasse.

scholarly journals In vitro organogenesis in some citrus species

2011 ◽  
Vol 33 (2) ◽  
pp. 526-531 ◽  
Author(s):  
Evandro Henrique Schinor ◽  
Fernando Alves de Azevedo ◽  
Francisco de Assis Alves Mourão Filho ◽  
Beatriz Madalena Januzzi Mendes
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Adventitious Shoots ◽  
Medium Composition ◽  
Naphthaleneacetic Acid ◽  
In Vitro Organogenesis ◽  
Basal Culture Medium ◽  
Medium Supplementation ◽  
Media Supplementation

In vitro organogenesis of Citrus was studied for the genotypes Citrus sinensis cv. 'Natal', C. limonia, C. volkameriana, and C. aurantium, with the use of epicotyl segments-derived explants, cultured in MT salts and vitamins medium supplemented with different concentrations of 6-benzylaminopurine (BAP - 0.0; 0.5; 1.0; 1.5 or 2.0 mg L-1). For the recalcitrant genotypes C. limonia and C. aurantium the in vitro organogenesis was also studied with internodal segments-derived explants, cultured in MT salts and vitamins medium supplemented with 0; 0.5; 1.0; 2.0, or 4.0 mg L-1 of BAP. The efficiency of culture medium supplementation with the combination of BAP (0.0; 1.0, or 2.0 mg L-1) and NAA (1-naphthaleneacetic acid - 0.0; 0.3, or 0.5 mg L-1) in the development of adventitious shoots was evaluated for C. aurantium. Culture medium supplementation with BAP is not essential for the adventitious shoots development in the four genotypes studied when epicotyl segments-derived explants are used. In general, culture media supplementation with BAP decreased the percentage of responsive explants excepted for C. sinensis cv. 'Natal' and C. limonia when the concentrations of 1.5 and 2.0 mg/L were used. The presence of cytokinin, in concentrations up to 2 mg/L, stimulated the in vitro organogenesis when internodal segments-derived explants were used for C. limonia and C. aurantium. For C. aurantium no adventitious shoots developed in explants (internodal segments) cultured in basal culture medium, without BAP supplementation. Although no statistic differences could be detected, culture media supplementation with the combination of BAP and NAA favored the development of adventitious shoots in C. aurantium. The best concentration of NAA varied according to BAP concentration. The results presented herein, show that Citrus in vitro organogenesis depends on the interaction of culture medium composition, explant differentiation level, and genotype.

scholarly journals Isolation and mycelial growth of Diehliomyces microsporus: effect of culture medium and incubation temperature

2007 ◽  
Vol 50 (4) ◽  
pp. 587-595 ◽  
Author(s):  
José Soares do Nascimento ◽  
Augusto Ferreira da Eira
Keyword(s):  
Fruiting Body ◽  
Mycelial Growth ◽  
Culture Medium ◽  
Incubation Temperature ◽  
Culture Media ◽  
Growth Speed ◽  
Three Stages ◽  
False Truffle ◽  
Incubation Temperatures

The false truffle is one of the main problems in the production of the Agaricus brasiliensis in Brazil and the control of this fungal competitor has been rather difficult due to difficulties in the isolation and cultivation of this pathogen. This experiment was conducted in three stages, the first consisting of the isolation of Diehliomyces microsporus starting from portions of the fruiting body and through the ascospores suspension; second, D. microsporus cultivated in vitro at 15, 20, 25, 30 and 35ºC in six different culture media (CSDA, OCDA, PCDA, ODA, PDA, CDA); third, D. microsporus was inoculated on sterilized compost for formation of the fruiting body. The colony formation from tissue of D. microsporus starting from portions of fruiting body was more efficient than germination of the ascospores. Compost medium (CDA) allowed a larger diameter of the D. microsporus colony, followed by the medium made up of compost and potato mixture, favoring a denser composition. The largest mycelial growth speed of D. microsporus occurred when the culture was incubated at 28 and 30ºC. Incubation temperatures lower than 15ºC or above 35ºC inhibited the mycelial growth of D. microsporus completely. The fruiting bodies were obtained easily in sterilized compost and later inoculated along with mycelial competitor.

scholarly journals Adjustment of culture media to optimize in vitro rhizogenesis of domestic plums of various origins

2021 ◽  
Vol 34 ◽  
pp. 03001
Author(s):  
Natalia Kovalenko ◽  
Nadezhda Polivara
Keyword(s):  
Nutrient Medium ◽  
Culture Medium ◽  
Root Formation ◽  
Culture Media ◽  
Medium Composition ◽  
Ms Medium ◽  
Growth Substances ◽  
Nutrient Media ◽  
Plum Varieties

This paper presents the results of the adjustment of 18 modifications of culture medium based on MS medium composition – Murashige and Skoog (1962) for in vitro rhizogenesis of 10 varieties of domestic plum. There were used 4 nutrient media with a full amount of macro- and microelements – MS1 with a different combination of phytohormones, 1 medium without hormones – MSc, 13 media – MS2 with half the amount of macronutrients, differing in hormonal composition. It was found that the maximum number of rooted microshoots from 21.2 to 52.2 % was on MS2–8 medium, from 11.2 to 43.1 % on MS2–5. The analysis showed that into the medium increases the percentage of microplants by 12-17 % comparing with the medium MS2–5 and MS2–7 without FA. The distinctiveness of nutrient media by the type of auxin (IAA, NAA, IBA) made it possible to clarify that IBA is the most optimal of the auxins, and the concentration of 1.7 mg/l is borderline for the growth reactions of plum varieties. It was revealed that in vitro root formation depends not only on the compositions of growth substances in the nutrient medium, but also on the genotype of the variety.

Growth and sporulation in vitro of Cercospora apii, Cercospora arachidicola, Cercospora kikuchii, and other species of Cercospora

1982 ◽  
Vol 60 (6) ◽  
pp. 862-868 ◽  
Author(s):  
N. E. El-Gholl ◽  
S. A. Alfieri Jr. ◽  
W. H. Ridings ◽  
C. L. Schoulties
Keyword(s):  
Mycelial Growth ◽  
Culture Medium ◽  
Culture Media ◽  
Single Point ◽  
Potato Dextrose Agar ◽  
Multiple Point ◽  
Cercospora Arachidicola ◽  
Cercospora Kikuchii ◽  
Conidial Production

Cercospora apii Fres., Cercospora arachidicola Hori, and Cercospora kikuchii (Matsumoto & Tomoyasu) Gardn. were single spored, and the resultant colonies were characterized. All isolates of C. arachidicola were morphologically identical, whereas those of C. apii and C. kikuchii were either smooth or had radial folds. Variability in sporulation was also evident. Multiple-point inoculation of culture media in plates with conidia helped to overcome variability in the amount of conidial production. Maximum sporulation was obtained at 4 days with C. apii and C. kikuchii, and from 7 to 10 days with C. arachidicola. Single-point inoculation of a V-8 culture medium in plates from peripheral dark mycelial growth in potato dextrose agar (PDA) culture resulted in increased sporulation by C. kikuchii. Sporulation of C. kikuchii was maintained throughout a 15-month study by selective subculturing; however, conidial production decreased over time. A total of 15 other species of Cercospora sporulated within a week following a V-8 inoculation of culture medium in plates with mycelial agar blocks.

Differentiation of mesenchymal stem cells from humans and animals into insulin-producing cells: An overview in vitro induction forms

2020 ◽  
Vol 16 ◽  
Author(s):  
Bruna O. S. Câmara ◽  
Bruno M. Bertassoli ◽  
Natália M. Ocarino ◽  
Rogéria Serakides
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Culture Medium ◽  
Adipogenic Differentiation ◽  
Medium Composition ◽  
Cell Therapies ◽  
Insulin Producing Cells ◽  
Msc Differentiation

The use of stem cells in cell therapies has shown promising results in the treatment of several diseases, including diabetes mellitus, in both humans and animals. Mesenchymal stem cells (MSCs) can be isolated from various locations, including bone marrow, adipose tissues, synovia, muscles, dental pulp, umbilical cords, and the placenta. In vitro, by manipulating the composition of the culture medium or transfection, MSCs can differentiate into several cell lineages, including insulin-producing cells (IPCs). Unlike osteogenic, chondrogenic, and adipogenic differentiation, for which the culture medium and time are similar between studies, studies involving the induction of MSC differentiation in IPCs differ greatly. This divergence is usually evident in relation to the differentiation technique used, the composition of the culture medium, the cultivation time, which can vary from a few hours to several months, and the number of steps to complete differentiation. However, although there is no “gold standard” differentiation medium composition, most prominent studies mention the use of nicotinamide, exedin-4, ß-mercaptoethanol, fibroblast growth factor b (FGFb), and glucose in the culture medium to promote the differentiation of MSCs into IPCs. Therefore, the purpose of this review is to investigate the stages of MSC differentiation into IPCs both in vivo and in vitro, as well as address differentiation techniques and molecular actions and mechanisms by which some substances, such as nicotinamide, exedin-4, ßmercaptoethanol, FGFb, and glucose, participate in the differentiation process.

scholarly journals Rheological and Microstructural Features of Plant Culture Media Doped with Biopolymers: Influence on the Growth and Physiological Responses of In Vitro-Grown Shoots of Thymus lotocephalus

2021 ◽  
Vol 2 (2) ◽  
pp. 538-553
Author(s):  
Natacha Coelho ◽  
Alexandra Filipe ◽  
Bruno Medronho ◽  
Solange Magalhães ◽  
Carla Vitorino ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Culture Medium ◽  
Culture Media ◽  
Average Pore Size ◽  
Physiological Responses ◽  
Gum Arabic ◽  
Shoot Elongation ◽  
Hydroxyethyl Cellulose ◽  
Plant Culture

In vitro culture is an important biotechnological tool in plant research and an appropriate culture media is a key for a successful plant development under in vitro conditions. The use of natural compounds to improve culture media has been growing and biopolymers are interesting alternatives to synthetic compounds due to their low toxicity, biodegradability, renewability, and availability. In the present study, different culture media containing one biopolymer (chitosan, gum arabic) or a biopolymer derivative [hydroxyethyl cellulose (HEC), carboxymethyl cellulose (CMC)], at 100 or 1000 mg L−1, were tested regarding their influence on the growth and physiological responses of Thymus lotocephalus in vitro culture. Cellulose-based biopolymers (HEC and CMC) and gum arabic were used for the first time in plant culture media. The results showed that CMC at 100 mg L−1 significantly improved shoot elongation while chitosan, at the highest concentration, was detrimental to T. lotocephalus. Concerning only the evaluated physiological parameters, all tested biopolymers and biopolymer derivatives are safe to plants as there was no evidence of stress-induced changes on T. lotocephalus. The rheological and microstructural features of the culture media were assessed to understand how the biopolymers and biopolymer derivatives added to the culture medium could influence shoot growth. As expected, all media presented a gel-like behaviour with minor differences in the complex viscosity at the beginning of the culture period. Most media showed increased viscosity overtime. The surface area increased with the addition of biopolymers and biopolymer derivatives to the culture media and the average pore size was considerably lower for CMC at 100 mg L−1. The smaller pores of this medium might be related to a more efficient nutrients and water uptake by T. lotocephalus shoots, leading to a significant improvement in shoot elongation. In short, this study demonstrated that the different types of biopolymers and biopolymer derivatives added to culture medium can modify their microstructure and at the right concentrations, are harmless to T. lotocephalus shoots growing in vitro, and that CMC improves shoot length.

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