PROSPECÇÃO E AVALIAÇÃO DO POTENCIAL BIOTECNOLÓGICO DE LINHAGENS COMERCIAIS DE SHIITAKE

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.

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Phosphite-based Products in the In vitro Colletotrichum musae Control

Journal of Experimental Agriculture International ◽

10.9734/jeai/2019/v33i430148 ◽

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.

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Obtaining monokaryotic and dikaryotic mycelial cultures of two Amazonian strains of Geastrum (Geastraceae, Basidiomycota)

Acta Amazonica ◽

10.1590/1809-4392201901341 ◽

2020 ◽

Vol 50 (1) ◽

pp. 61-67

Author(s):

Marcos Diones Ferreira SANTANA ◽

Ruby VARGAS-ISLA ◽

Janaina da Costa NOGUEIRA ◽

Thiago ACCIOLY ◽

Bianca Denise Barbosa da SILVA ◽

...

Keyword(s):

Culture Medium ◽

Culture Media ◽

Brazilian Amazon ◽

Potato Dextrose Agar ◽

Morphological Description ◽

Agar Culture ◽

Biotechnological Potential ◽

Sexual Crosses ◽

High Diversity

ABSTRACT The high diversity of the genus Geastrum and the difficulty of obtaining mycelial cultures impairs the study of the ecophysiology and the exploration of the biotechnological potential of the taxon. In this study, different culture media were tested to obtain mycelial cultures for G. lloydianum and G. subiculosum collected in the Brazilian Amazon. Data on spore germination, and isolation of monokaryotic cultures and in vitro sexual reproduction are presented, as well as a brief morphological description of the cultures obtained. For both species, Potato Dextrose Agar (PDA) was the most promising of the tested culture media. The highest growth in agar culture ever recorded for this genus is reported (4.9 mm per week for G. lloydianum and 7.5 mm for G. subiculosum). In the PDA culture medium, spores germinated after 35-40 days of incubation and the isolation of monokaryotic cultures of the two species, as well as in vitro sexual crosses, were successfully performed.

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

Brazilian Archives of Biology and Technology ◽

10.1590/s1516-89132007000400004 ◽

2007 ◽

Vol 50 (4) ◽

pp. 587-595 ◽

Cited By ~ 7

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.

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

Brazilian Journal of Biology ◽

10.1590/1519-6984.253436 ◽

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.

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

Canadian Journal of Botany ◽

10.1139/b82-111 ◽

1982 ◽

Vol 60 (6) ◽

pp. 862-868 ◽

Cited By ~ 9

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.

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

Agrologia ◽

10.30598/a.v1i1.293 ◽

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.

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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

Polysaccharides ◽

10.3390/polysaccharides2020032 ◽

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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Biochemical markers for pregnancy in the spent culture medium of in vitro produced bovine embryos

Biology of Reproduction ◽

10.1093/biolre/ioab095 ◽

2021 ◽

Author(s):

Gabriela de Oliveira Fernandes ◽

Marcella Pecora Milazzotto ◽

Andrei Antonioni Guedes Fidelis ◽

Taynan Stonoga Kawamoto ◽

Ligiane de Oliveira Leme ◽

...

Keyword(s):

Mass Spectrometry ◽

Biochemical Markers ◽

Culture Medium ◽

Culture Media ◽

Ionization Mass ◽

Metabolic Profiles ◽

Bovine Embryos ◽

The Media

Abstract The present study aimed to identify biomarkers to assess the quality of in vitro produced (IVP) bovine embryos in the culture media. IVP embryos on Day (D) 5 of development were transferred to individual drops, where they were maintained for the last 48 h of culture. Thereafter, the medium was collected and the embryos were transferred to the recipients. After pregnancy diagnosis, the media were grouped into the pregnant and nonpregnant groups. The metabolic profiles of the media were analyzed via electrospray ionization mass spectrometry, and the concentrations of pyruvate, lactate, and glutamate were assessed using fluorimetry. The spectrometric profile revealed that the media from embryos from the pregnant group presented a higher signal intensity compared to that of the nonpregnant group; the ions 156.13 Da [M + H]+, 444.33 Da [M + H]+, and 305.97 Da [M + H]+ were identified as biomarkers. Spent culture medium from expanded blastocysts (Bx) that established pregnancy had a greater concentration of pyruvate (p = 0.0174) and lesser concentration of lactate (p = 0.042) than spent culture medium from Bx that did not establish pregnancy. Moreover, pyruvate in the culture media of Bx can predict pregnancy with 90.9% sensitivity and 75% specificity. In conclusion, we identified markers in the culture media that helped in assessing the most viable IVP embryos with a greater potential to establish pregnancy.

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MODIFICAÇÕES NO MEIO DE CULTURA, FOTOPERÍODO E TEMPO DE CULTIVO AFETAM O ALONGAMENTO E ENRAIZAMENTO IN VITRO DE BANANEIRA CV. PACOVAN

Nativa ◽

10.31413/nativa.v6i1.4731 ◽

2018 ◽

Vol 6 (1) ◽

pp. 27

Author(s):

Marcos Vinícius Marques Pinheiro ◽

Ana Cristina Portugal Pinto De Carvalho ◽

Fabrina Bolzan Martins

Keyword(s):

Plant Height ◽

Culture Medium ◽

Culture Media ◽

Dry Weight ◽

Salt Mixture ◽

Musa Spp ◽

Fresh Biomass ◽

Shoot Dry Weight ◽

Number Of Leaves

No intuito de elevar as taxas de sobrevivência durante a etapa de aclimatização e posterior plantio a campo, avaliou-se o enraizamento in vitro de bananeira cv. Pacovan, em diferentes concentrações de sais MS e de sacarose. Utilizou-se DIC, esquema fatorial (6x2x3), com seis meios de cultura [sendo três concentrações de nutrientes do meio MS (100%; 50% de macronutrientes; e 50% dos sais macro e micronutrientes), e duas concentrações de sacarose (1,5/3,0%)], dois fotoperíodos (12/16 h) e três tempos de cultivo (21, 28 ou 35 dias) e seis repetições/tratamento. Analisaram-se: altura da planta, número de folhas/planta, massas frescas e secas das partes aérea e radicular. Para altura da planta, massa fresca da parte aérea e radicular, o meio MS 50% dos sais + sacarose (1,5%) com fotoperíodo de 16 h e tempo de cultivo de 35 dias foi satisfatório. Para massa seca da parte aérea foi MS 50% de sais + sacarose (3%), e para massa seca da parte radicular, MS 100% + sacarose (3%) (em 12hs/28 dias e 16hs/21 dias). Para o alongamento/enraizamento in vitro da bananeira cv. Pacovan sugere-se MS 50% de sais (macro e micronutrientes), redução ou manutenção de sacarose (1,5 ou 3%) em 16h/35 dias de cultivo.Palavra-chave: Musa spp., propagação in vitro, sistema radicular. CHANGES IN CULTURE MEDIUM, PHOTOPERIOD AND TIME OF CULTIVATION AFFECT THE IN VITRO ELONGATION AND ROOTING OF BANANA CV. PACOVAN ABSTRACT:In order to achieve high rates of survival during the acclimatization and later planting in the field, was evaluated the in vitro of banana cv. Pacovan plants under different concentrations of sucrose and MS basal salt mixture. The experiment was assembled in a DIC, in 6x2x3, six different culture media [three different MS salt mixture concentrations (100%; 50% of macronutrients; and 50% of macro/micronutrients) and two sucrose concentrations (1.5/3%)], two photoperiods (12/16 hours) and three cultivation times (21, 28 or 35 days). Each treatment was composed by 6 replicates. Plant height, number of leaves/plant, fresh and dry weight of roots and shoots, were analyzed. Satisfactory results for plant height and shoot and root fresh biomass were observed in MS with macro/micronutrients (50%) + sucrose (3%), 16 hours/35 days. The highest values of shoot dry weight were observed in MS with macro/micronutrients (50%) + sucrose (3%); the highest root dry weight was achieved with MS 100% + sucrose (3%) (12hs/28 and 16hs/21 days). The suggested medium for the in vitro elongation and rooting stage of banana cv. Pacovan is the MS with 50% of salts (macro and micronutrients), reduction or maintenance of sucrose (1.5 or 3%) in 16h/35 days of cultivation.Keywords: Musa spp., in vitro propagation, root system. DOI:

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In vitro answer of Bulgarian pepper (Capsicum annuum L.)

Genetika ◽

10.2298/gensr0602129r ◽

2006 ◽

Vol 38 (2) ◽

pp. 129-136

Author(s):

Velichka Rodeva ◽

Stanislava Grozeva ◽

Velichka Todorova

Keyword(s):

Culture Medium ◽

Culture Media ◽

Casein Hydrolysate ◽

Basal Medium ◽

Regeneration Ability ◽

Stem Explants ◽

Media Composition ◽

Breeding Lines ◽

High Level

Callusogenesis and regeneration ability of cotyledon and hypocotyl explants from three Bulgarian pepper varieties in MS basal medium supplemented with l-3mg/l BAP. l.0mg/1 IAA and 0.5mg/l GA3 was studied. In the different variants of culture medium was registered high level of callusogenesis and organogenesis in both type of explants from the all varieties. The highest percentage of plant-regenerants is established in cotyledon explants (from 3.3 to 18.3) in variant 3 of the culture medium containing 3mg/l BA. In the process of micropropagation by stem explants of the same studied pepper varieties the addition of the vitamins C. B12. Casein hydrolysate and Ferulic acid had a stimulation effect on the plant growth in height and rooting. In result of anther cultivation from three pepper varieties and four breeding lines the highest percentage of embryo structure formation was registered in varieties Albena and Strjama (12.0 and 13.8 respectively). The Bulgarian peppers are recalcitrant and their in vitro answer is different depending from the explants type, genotype and the culture media composition.

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