RESUMEN

A high Mw (5012 kDa) polysaccharide (PNPS) from the fruiting body of Pleurotus nebrodensis was isolated using water extraction followed by ethanol precipitation. The structural characteristics and in vitro fermentation behaviors of this polysaccharide was investigated. Chemical composition analysis showed the total sugar content of PNPS was up to 97.20 ± 1.80 wt%. Monosaccharide composition analysis showed PNPS contained mainly glucose (89.22 ± 5.70 mol%) while small percentage of mannose (5.60 ± 0.74 mol%) and galactose (5.18 ± 0.33 mol%) were also detected. According to the linkage pattern analysis (methylation analysis), PNPS comprised mainly 4-ß-D-Glcp (58.90 mol%), while other residues including α-D-Glcp, 6-α-D-Galp, 3,6-α-D-Manp, 3-ß-D-Glcp and 6-α-D-Glcp were detected with a comparable amount. Combined with results from 1D and 2D NMR spectrum, a proposed structure of PNPS was presented. In vitro fermentation of PNPS by gut microbiota showed total SCFA production of all treatment groups was higher than negative control group (NC) significantly (p < 0.05) after 48 h of fermentation. The formation of SCFAs was mainly acetic acid, followed by propionic acid and butyric acid, and the pH was decreased from 6.95 to 4.70. After 72 h, the total sugar content decreased from 5.813 ± 0.87 mg/L to 0.23 ± 0.065 mg/L, and the molecular weight of PNPS decreased.

Asunto(s)

Fenómenos Químicos , Fermentación , Pleurotus/química , Polisacáridos/química , Polisacáridos/aislamiento & purificación , Animales , Espectroscopía de Resonancia Magnética con Carbono-13 , Cromatografía Líquida de Alta Presión , Ácidos Grasos Volátiles/biosíntesis , Concentración de Iones de Hidrógeno , Metilación , Peso Molecular , Monosacáridos/análisis , Espectroscopía de Protones por Resonancia Magnética , Relación Estructura-Actividad , Azúcares/análisis , Porcinos

RESUMEN

The effects of a polysaccharide (PNPA) from the fruiting bodies of Pleurotus nebrodensis on hepatic ischemia/reperfusion (I/R) injury in rats were explored. Severe liver injury was shown in rats following I/R with high content of serum AST and ALT, as evidenced by histological examination, whereas less damage was noted after PNPA treatment compared to rats in the I/R group. Also, pre-treatment with PNPA (400mg/kg) could reduce the level of MDA in liver tissue and increase the activity of ROS scavengers (T-AOC, SOD, CAT, GSH-PX), Nitric oxide synthase (T-NOS and iNOS), and GSH. Moreover, PNPA could still reduce the production of I/R-induced inflammatory cytokines (IL-1ß, IL-6, TNF-α and NF-κB) in liver, along with MPO. I/R-induced high level of pro-apoptotic indicators (cytochrome c, Bax, and caspase 3) and low level of anti-apoptotic indicators Bcl-2 were reversed after PNPA pretreatment. Therefore, PNPA might be a promising candidate for preventing hepatic I/R injury.

Asunto(s)

Hígado/efectos de los fármacos , Hígado/lesiones , Pleurotus/química , Polisacáridos/farmacología , Daño por Reperfusión/tratamiento farmacológico , Animales , Apoptosis/efectos de los fármacos , Citocinas/metabolismo , Hígado/metabolismo , Hígado/patología , Masculino , Estrés Oxidativo/efectos de los fármacos , Polisacáridos/uso terapéutico , Ratas , Ratas Sprague-Dawley , Daño por Reperfusión/metabolismo , Daño por Reperfusión/patología

RESUMEN

Three laccase isoenzymes (Lac1, Lac2 and Lac3) have been purified to homogeneity from Pleurotus nebrodensis in our previous study. Lac2 was shown to be the dominant isoform, capable of oxidizing the majority of laccase substrates and manifesting good thermostability and pH stability. Hence, Lac2 was selected to decolourize structurally different dyes and the colour removal efficiencies of Lac2 and the crude extract of P. nebrodensis were compared. By monitoring the λmax of the reaction system during the course of biotransformation, clear hypsochromic shifts were observed for most of the dyes examined, illustrating that at least one peak disappeared as a result of laccase treatment. In general, Lac2 was more efficient within a short time (1 h) and the crude extract, in general, could achieve similar or even higher efficiency when the duration of treatment was extended to 24 h. Malachite green (MG) was chosen to study the detoxifying potential of Lac2, because of the relatively simple structure and high toxicity of the dye towards microorganisms. The toxicity of MG towards both bacteria (Bacillus subtilis, Bacillus licheniformis, Pseudomonas fluorescens and Escherichia coli) and fungi (Fusarium graminearum and Trichoderma harzianum) was dramatically decreased and the potential mechanism was estimated by GC-MS as to remove four methyl groups firstly and the two newly formed amine groups would be degraded or polymerized further. The present study facilitates an understanding of the application of P. nebrodensis laccases and furnishes evidence for the safety of their utilization in the treatment of wastewater emanating from textile industries.

Asunto(s)

Colorantes/química , Contaminantes Ambientales/química , Proteínas Fúngicas/química , Lacasa/química , Pleurotus/enzimología , Colorantes de Rosanilina/química , Bacillus licheniformis/efectos de los fármacos , Bacillus licheniformis/crecimiento & desarrollo , Bacillus subtilis/efectos de los fármacos , Bacillus subtilis/crecimiento & desarrollo , Biodegradación Ambiental , Colorantes/toxicidad , Mezclas Complejas/química , Contaminantes Ambientales/toxicidad , Estabilidad de Enzimas , Escherichia coli/efectos de los fármacos , Escherichia coli/crecimiento & desarrollo , Proteínas Fúngicas/aislamiento & purificación , Fusarium/efectos de los fármacos , Fusarium/crecimiento & desarrollo , Humanos , Concentración de Iones de Hidrógeno , Isoenzimas/química , Isoenzimas/aislamiento & purificación , Lacasa/aislamiento & purificación , Viabilidad Microbiana/efectos de los fármacos , Oxidación-Reducción , Pleurotus/química , Pseudomonas fluorescens/efectos de los fármacos , Pseudomonas fluorescens/crecimiento & desarrollo , Colorantes de Rosanilina/toxicidad , Trichoderma/efectos de los fármacos , Trichoderma/crecimiento & desarrollo

RESUMEN

The characterization of three laccase isoforms from Pleurotus nebrodensis is described. Isoenzymes Lac1, Lac2 and Lac3 were purified to homogeneity using ion exchange chromatography on DEAE-cellulose, CM-cellulose and Q-Sepharose and a gel filtration step on Superdex 75. The molecular weights of the purified laccases were estimated to be 68, 64 and 51 kDa, respectively. The isoenzymes demonstrated the same optimum pH at 3.0 but slightly different temperature optima: 50-60 °C for Lac1 and Lac3 and 60 °C for Lac2. Lac2 was always more stable than the other two isoforms and exposure to 50 °C for 120 min caused 30% loss in activity. Lac2 was relatively less stable than the other two isoforms when exposed to the pH range of 3.0-8.0 for 24 h, but inactivation only occurred initially, with around 70% residual activity being maintained during the whole process. Oxidative ability towards aromatic compounds varied substantially among the isoforms and each of them displayed preference toward some substrates. Kinetic constants (Km, Kcat) were determined by using a 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assay, with Lac3 showing the best affinity and Lac2 displaying the highest catalytic efficiency. Amino acid sequences from peptides derived from digestion of isoenzymes showed great consistency with laccases in the databases.

Asunto(s)

Lacasa/aislamiento & purificación , Lacasa/metabolismo , Pleurotus/enzimología , Secuencia de Aminoácidos , Activación Enzimática , Estabilidad de Enzimas , Proteínas Fúngicas/genética , Proteínas Fúngicas/aislamiento & purificación , Proteínas Fúngicas/metabolismo , Isoenzimas/aislamiento & purificación , Lacasa/química , Peso Molecular , Pleurotus/química

RESUMEN

In the present study, the effects of Pleurotus nebrodensis polysaccharide (PN-S) on the immune functions of immunosuppressed mice were determined. The immunosuppressed mouse model was established by treating the mice with cyclophosphamide (40 mg/kg/2d, CY) through intraperitoneal injection. The results showed that PN-S administration significantly reversed the CY-induced weight loss, increased the thymic and splenic indices, and promoted proliferation of T lymphocyte, B lymphocyte, and macrophages. PN-S also enhanced the activity of natural killer cells and increased the immunoglobulin M (IgM) and immunoglobulin G (IgG) levels in the serum. In addition, PN-S treatment significantly increased the phagocytic activity of mouse peritoneal macrophages. PN-S also increased the levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interferon-γ (INF-γ), and nitric oxide (NOS) in splenocytes. qRT-PCR results also indicated that PN-S increased the mRNA expression of IL-6, TNF-α, INF-γ, and nitric oxide synthase (iNOS) in the splenocytes. These results suggest that PN-S treatment enhances the immune function of immunosuppressed mice. This study may provide a basis for the application of this fungus in adjacent immunopotentiating therapy against cancer and in the treatment of chemotherapy-induced immunosuppression.

Asunto(s)

Productos Biológicos/farmacología , Inmunidad/efectos de los fármacos , Factores Inmunológicos/farmacología , Terapia de Inmunosupresión , Macrófagos/efectos de los fármacos , Pleurotus/química , Polisacáridos/farmacología , Animales , Antineoplásicos Alquilantes , Productos Biológicos/uso terapéutico , Línea Celular , Ciclofosfamida , Factores Inmunológicos/uso terapéutico , Interferón gamma/metabolismo , Interleucina-6/metabolismo , Macrófagos/metabolismo , Masculino , Ratones Endogámicos BALB C , Neoplasias/tratamiento farmacológico , Neoplasias/inmunología , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fagocitosis/efectos de los fármacos , Polisacáridos/uso terapéutico , Factor de Necrosis Tumoral alfa/metabolismo

RESUMEN

In this study, we isolated a polysaccharide (PNPA), with a molecular weight of 105kDa, from the fruiting bodies of Pleurotus nebrodensis. It had a backbone consisting of 1,3-linked-d-glucpyranosyl and 1,3,6-linked-d-galactopyranosyl residues, which was terminated with 1-linked-d-mannopyranosyl terminal at O-3 position of 1,3,6-linked-d-galactopyranosyl unit along the main chain in the ratio of 4:1:1. We further examined the effect of PNPA on myocardial ischemia-reperfusion (I/R) injury in rats and elucidated the underlying mechanism. Pretreatment with PNPA (100 and 400mg/kg) for 30 days significantly attenuated myocardial infarct size as compared to I/R model group. A decrease in superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) levels, as well as an increased malondialdehyde (MDA) content were observed in both myocardial serum and tissues of control I/R group, whereas pretreatment with PNPA markedly restored these change, and also relieved myocardial cell apoptosis. These results suggested that PNPA achieved protective effect on myocardial I/R injury in part through improving endogenous antioxidants and suppressing myocardial cell apoptosis.

Asunto(s)

Citoprotección/efectos de los fármacos , Daño por Reperfusión Miocárdica/prevención & control , Pleurotus/química , Polisacáridos/farmacología , Animales , Antioxidantes/metabolismo , Apoptosis/efectos de los fármacos , Glicosilación , Masculino , Peso Molecular , Infarto del Miocardio/complicaciones , Infarto del Miocardio/prevención & control , Daño por Reperfusión Miocárdica/complicaciones , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/patología , Polisacáridos/química , Polisacáridos/aislamiento & purificación , Ratas , Ratas Wistar

RESUMEN

A novel Pleurotus nebrodensis polysaccharide (PN-S) was purified and characterized, and its immune-stimulating activity was evaluated in RAW264.7 macrophages. PN-S induced the proliferation of RAW264.7 cells in a dose-dependent manner, as determined by the MTT assay. After exposure to PN-S, the phagocytosis of the macrophages was significantly improved, with remarkable changes in morphology being observed. Flow cytometric analysis demonstrated that PN-S promoted RAW264.7 cells to progress through S and G2/M phases. PN-S treatment enhanced the productions of interleukin-6 (IL-6), nitric oxide (NO), interferon gamma (INF-γ), and tumor necrosis factor-α (TNF-α) in the macrophages, with up-regulation of mRNA expressions of interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS), interferon gamma(INF-γ) and tumor necrosis factor-α (TNF-α) being observed in a dose-dependent manner, as measured by qRT-PCR. In conclusion, these results suggest that the purified PN-S can improve immunity by activating macrophages.

Asunto(s)

Polisacáridos Fúngicos/farmacología , Macrófagos/inmunología , Pleurotus , Animales , Ciclo Celular/inmunología , Línea Celular , Proliferación Celular/efectos de los fármacos , Inmunidad/efectos de los fármacos , Interferón gamma/biosíntesis , Interferón gamma/metabolismo , Interleucina-6/biosíntesis , Interleucina-6/metabolismo , Macrófagos/metabolismo , Ratones , Óxido Nítrico/biosíntesis , Óxido Nítrico Sintasa de Tipo II/metabolismo , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Necrosis Tumoral alfa/biosíntesis , Factor de Necrosis Tumoral alfa/metabolismo , Regulación hacia Arriba

RESUMEN

A novel Pleurotus nebrodensis polysaccharide (PN-S) was purified and characterized, and its immune-stimulating activity was evaluated in RAW264.7 macrophages. PN-S induced the proliferation of RAW264.7 cells in a dose-dependent manner, as determined by the MTT assay. After exposure to PN-S, the phagocytosis of the macrophages was significantly improved, with remarkable changes in morphology being observed. Flow cytometric analysis demonstrated that PN-S promoted RAW264.7 cells to progress through S and G2/M phases. PN-S treatment enhanced the productions of interleukin-6 (IL-6), nitric oxide (NO), interferon gamma (INF-γ), and tumor necrosis factor-α (TNF-α) in the macrophages, with up-regulation of mRNA expressions of interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS), interferon gamma(INF-γ) and tumor necrosis factor-α (TNF-α) being observed in a dose-dependent manner, as measured by qRT-PCR. In conclusion, these results suggest that the purified PN-S can improve immunity by activating macrophages.

Asunto(s)

Animales , Ratones , Ciclo Celular , Alergia e Inmunología , Línea Celular , Proliferación Celular , Polisacáridos Fúngicos , Farmacología , Inmunidad , Interferón gamma , Metabolismo , Interleucina-6 , Metabolismo , Macrófagos , Alergia e Inmunología , Metabolismo , Óxido Nítrico , Óxido Nítrico Sintasa de Tipo II , Metabolismo , Pleurotus , ARN Mensajero , Metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Necrosis Tumoral alfa , Metabolismo , Regulación hacia Arriba

RESUMEN

In the present study, the effects of Pleurotus nebrodensis polysaccharide (PN-S) on the immune functions of immunosuppressed mice were determined. The immunosuppressed mouse model was established by treating the mice with cyclophosphamide (40 mg/kg/2d, CY) through intraperitoneal injection. The results showed that PN-S administration significantly reversed the CY-induced weight loss, increased the thymic and splenic indices, and promoted proliferation of T lymphocyte, B lymphocyte, and macrophages. PN-S also enhanced the activity of natural killer cells and increased the immunoglobulin M (IgM) and immunoglobulin G (IgG) levels in the serum. In addition, PN-S treatment significantly increased the phagocytic activity of mouse peritoneal macrophages. PN-S also increased the levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interferon-γ (INF-γ), and nitric oxide (NOS) in splenocytes. qRT-PCR results also indicated that PN-S increased the mRNA expression of IL-6, TNF-α, INF-γ, and nitric oxide synthase (iNOS) in the splenocytes. These results suggest that PN-S treatment enhances the immune function of immunosuppressed mice. This study may provide a basis for the application of this fungus in adjacent immunopotentiating therapy against cancer and in the treatment of chemotherapy-induced immunosuppression.

Asunto(s)

Animales , Masculino , Antineoplásicos Alquilantes , Productos Biológicos , Farmacología , Usos Terapéuticos , Línea Celular , Ciclofosfamida , Inmunidad , Factores Inmunológicos , Farmacología , Usos Terapéuticos , Terapia de Inmunosupresión , Interferón gamma , Metabolismo , Interleucina-6 , Metabolismo , Macrófagos , Metabolismo , Ratones Endogámicos BALB C , Neoplasias , Quimioterapia , Alergia e Inmunología , Óxido Nítrico , Metabolismo , Óxido Nítrico Sintasa de Tipo II , Metabolismo , Fagocitosis , Pleurotus , Química , Polisacáridos , Farmacología , Usos Terapéuticos , Factor de Necrosis Tumoral alfa , Metabolismo

RESUMEN

Polysaccharides derived from edible fungi inhibit the proliferation of tumor cells. In this study, we investigated the effects of a polysaccharide (PN50G) from Pleurotus nebrodensis on A549 cell proliferation and apoptosis. MTT assay showed that PN50G induced apoptosis in the A549 cells in a dose-dependent. However, PN50G did not affect the proliferation viability of human fetal lung fibroblast cells MRC-5. Scanning electro microscopy (SEM) results indicate that PN50G induced a typical apoptotic morphological feature in A549. DNA accumulation and fragmentation were determined by acridine orange/ethidium bromide (AO/EB) staining. Flow cytometric analysis demonstrated that PN50G caused A549 cell apoptosis via cell arrest at the S phase. PN50G also extended the comet tail length in single-cell gel electrophoresis and disrupted the mitochondrial membrane potential as determined by Rdamine-123 staining. Further analysis by qRT-PCR showed that the expression of caspase-3 and caspase-9 mRNA increased. These findings suggest that PN50G can inhibit A549 cell proliferation and induce apoptosis mainly by activating the intrinsic mitochondrial pathway.

Asunto(s)

Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Polisacáridos Fúngicos/farmacología , Pleurotus/química , Caspasa 3/genética , Caspasa 3/metabolismo , Caspasa 9/genética , Caspasa 9/metabolismo , Línea Celular , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Humanos , Potencial de la Membrana Mitocondrial

RESUMEN

Pleurotus nebrodensis is an edible and commercially available mushroom in Korea. This study was conducted in order to evaluate the anticancer and immunopotentiating activities of crude polysaccharides, extracted in methanol, neutral saline, and hot water (hereafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of P. nebrodensis. ß-Glucan and protein contents in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. nebrodensis ranged from 23.79~36.63 g/100 g and 4.45~6.12 g/100 g, respectively. Crude polysaccharides were not cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at a range of 10~2,000 µg/mL. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 11.76~27.06% in mice previously inoculated with sarcoma 180. Treatment with Fr. NaCl resulted in an increase in the numbers of spleen cells by 1.49 fold at the concentration of 50 µg/mL, compared with control. Fr. HW improved the immuno-potentiating activity of B lymphocytes through an increase in alkaline phosphatase activity by 1.65 fold, compared with control at 200 µg/mL. Maximum production of nitric oxide (14.3 µM) was recorded in the Fr. NaCl fraction at 200 µg/mL. Production of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), and interleukin-6 (IL-6) was significantly higher, compared to control, and IL-6 production was highest, in contrast to TNF-α, IL-1ß, and positive control, concanavalin at the tested concentration of the various fractions. Results of the current study suggest that polysaccharides extracted from P. nebrodensis have a strong anticancer effect and may be useful as an ingredient of biopharmaceutical products for treatment of cancer.

RESUMEN

Pleurotus nebrodensis is an edible and commercially available mushroom in Korea. This study was conducted in order to evaluate the anticancer and immunopotentiating activities of crude polysaccharides, extracted in methanol, neutral saline, and hot water (hereafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of P. nebrodensis. beta-Glucan and protein contents in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. nebrodensis ranged from 23.79~36.63 g/100 g and 4.45~6.12 g/100 g, respectively. Crude polysaccharides were not cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at a range of 10~2,000 microg/mL. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 11.76~27.06% in mice previously inoculated with sarcoma 180. Treatment with Fr. NaCl resulted in an increase in the numbers of spleen cells by 1.49 fold at the concentration of 50 microg/mL, compared with control. Fr. HW improved the immuno-potentiating activity of B lymphocytes through an increase in alkaline phosphatase activity by 1.65 fold, compared with control at 200 microg/mL. Maximum production of nitric oxide (14.3 microM) was recorded in the Fr. NaCl fraction at 200 microg/mL. Production of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6) was significantly higher, compared to control, and IL-6 production was highest, in contrast to TNF-alpha, IL-1beta, and positive control, concanavalin at the tested concentration of the various fractions. Results of the current study suggest that polysaccharides extracted from P. nebrodensis have a strong anticancer effect and may be useful as an ingredient of biopharmaceutical products for treatment of cancer.

Asunto(s)

Animales , Ratones , Agaricales , Fosfatasa Alcalina , Linfocitos B , Línea Celular , Frutas , Inmunomodulación , Inyecciones Intraperitoneales , Interleucina-1beta , Interleucina-6 , Corea (Geográfico) , Cuidados para Prolongación de la Vida , Metanol , Óxido Nítrico , Pleurotus , Polisacáridos , Sarcoma , Sarcoma 180 , Bazo , Factor de Necrosis Tumoral alfa , Agua

RESUMEN

The molecular phylogeny in nine different commercial cultivated strains of Pleurotus nebrodensis was studied based on their internal transcribed spacer (ITS) region and RAPD. In the sequence of ITS region of selected strains, it was revealed that the total length ranged from 592 to 614 bp. The size of ITS1 and ITS2 regions varied among the strains from 219 to 228 bp and 211 to 229 bp, respectively. The sequence of ITS2 was more variable than ITS1 and the region of 5.8S sequences were identical. Phylogenetic tree of the ITS region sequences indicated that selected strains were classified into five clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by RAPD with 20 arbitrary primers. Twelve primers were efficient to applying amplification of the genomic DNA. The sizes of the polymorphic fragments obtained were in the range of 200 to 2000 bp. RAPD and ITS analysis techniques were able to detect genetic variation among the tested strains. Experimental results suggested that IUM-1381, IUM-3914, IUM-1495 and AY-581431 strains were genetically very similar. Therefore, all IUM and NCBI gene bank strains of P. nebrodensis were genetically same with some variations.

RESUMEN

The molecular phylogeny in nine different commercial cultivated strains of Pleurotus nebrodensis was studied based on their internal transcribed spacer (ITS) region and RAPD. In the sequence of ITS region of selected strains, it was revealed that the total length ranged from 592 to 614 bp. The size of ITS1 and ITS2 regions varied among the strains from 219 to 228 bp and 211 to 229 bp, respectively. The sequence of ITS2 was more variable than ITS1 and the region of 5.8S sequences were identical. Phylogenetic tree of the ITS region sequences indicated that selected strains were classified into five clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by RAPD with 20 arbitrary primers. Twelve primers were efficient to applying amplification of the genomic DNA. The sizes of the polymorphic fragments obtained were in the range of 200 to 2000 bp. RAPD and ITS analysis techniques were able to detect genetic variation among the tested strains. Experimental results suggested that IUM-1381, IUM-3914, IUM-1495 and AY-581431 strains were genetically very similar. Therefore, all IUM and NCBI gene bank strains of P. nebrodensis were genetically same with some variations.

Asunto(s)

ADN , Variación Genética , Filogenia , Pleurotus