Construction of protoplast genetic transformation system for Mycena--symbiont of Gastrodia elata / 中国中药杂志

Mycena, a symbiont of Gastrodia elata, promotes seed germination of G. elata and plays a crucial role in the sexual reproduction of G. elata. However, the lack of genetic transformation system of Mycena blocks the research on the interaction mechanism of the two. In order to establish the protoplast transformation system of Mycena, this study analyzed the protoplast enzymatic hydrolysis system, screened the resistance markers and regeneration medium, and explored the transient transformation. After hydrolysis of Mycena hyphae with complexes enzymes for 8 h and centrifugation at 4 000 r·min~(-1), high-concentration and quality protoplast was obtained. The optimum regeneration medium for Mycena was RMV, and the optimum resistance marker was 50 mg·mL~(-1) hygromycin. The pLH-HygB-HuSHXG-GFP-HdSHXG was transformed into the protoplast of Mycena which then expressed GFP. The established protoplast transformation system of Mycena laid a foundation for analyzing the functional genes of Mycena and the molecular mechanism of the symbiosis of Mycena and G. elata.

Isolation and Identification of Mycena of Pleurotus in Wumeng Mountain Area and Optimization of Its Production Conditions / 中国实验方剂学杂志

Objective:Isolate and identify Mycena, expand the resources of geminating fungus of Gastrodia elata and optimize the culture conditions of Mycena，in order to provide information and guidance for the production of geminating fungus of G. elata. Method:Juvenile tuber tissue mass transfer separation and purification technology was used for the separation and purification of strains，traditional morphology microscopy was used to isolate the colony mycelia spores morphological characteristics, such as identification，polymerase chain reaction（PCR） amplification rDNA （Ribosomal DNA） internal transcribed spacer（ITS） was used for sequencing analysis and further homology with NCBI database retrieval，MEGA6 software was used to establish Phylogenetic tree by the Maximum likelihood method （MaximumLikelihood，M-L）, so as to classify and identify isolated strains. At the same time，orthogonal test was used to optimize the optimal growth conditions of Mycena. Result:A total of 86 strains were isolated, which belong to 21 species in 12 genera. WMMFJ，SHXG，WMM-21 and MFJ8103 were identified as M. purpureofusca, and ZT01-6 and ZT01-8 were identified as M. cf. purpureofusca. The growth rate of Mycena in wheat bran medium was significantly higher than in PDA medium. The optimal medium composition for the growth of germinating bacteria was 100 g potato，150 g wheat bran and 20 g corn flour，100 g glucose. And 1，3，5-Trihydroxybenzene significantly promoted the growth of WMMFJ，and played a role in promoting the growth of WMM-21 and ZT01-6，and 2-Methoxyphenol promoted the growth of WMMFJ. Conclusion:Six strains of Mycena were isolated and identified，four of them are M. purpureofusca，and two of them are M. cf. Purpureofusca. The separation method improved the separation effect of germinating bacteria.

Mycorrhizal Cultivation of Dendrobium officinale / 中国药学杂志

OBJECTIVE: To provide the scientific basis for the application of mycorrhizal fungi in Dendrobium officinale artificial cultivation. METHODS: The dominant strain was screened in a planting base by using biomass as the index. The effects of initial inoculums and supplementary inoculation on the growth and the chemical composition of D. officinale were further investigated. RESULTS: Mycorrhizal fungi M2(Mycena sp.) was found to be the advantageous strain. After 16 months of growth, the stem dry weight(SDW) and plant dry weight(PDW) of the three M2-treatment groups were significantly higher than those of CK, and the SDW of group M(M2 initial inoculums of 1.0 g·clump-1) was the highest, which increased by 39.2% and 11.1% compared with CK and group L(M2 initial inoculums of 1.0 g·clump-1)(P<0.05), respectively. Furthermore, M2 had no effect on the chemical properties of stem polysaccharide, and could improve the contents of naringenin and bibenzyls in the stems. The supplementary inoculation of M2 enhanced the plant-growth-promoting effects of M2. The SDW of the supplementary group increased by 18.0%(P<0.05) compared with the non-supplementary group, while supplementary inoculation had a certain effect on the chemical properties of polysaccharide. CONCLUSION: Mycorrhizal fungus M2 has good application prospects for D. officinale artificial cultivation.

Effects of mycorrhizal fungi on the growth and dendrobine and polysaccharide accumulation of Dendrobium nobile lindl seedlings / 中国药学杂志

OBJECTIVE: To study the effect of mycorrhizal fungi on the growth and the main chemical composition accumulation of Dendrobium nobile seedlings. METHODS: Seedlings which inoculated mycorrhizal fungus MF23 grew for 18 months, and were sampled regularly. The stem height and the dry weight of plant organs were measured and the content of dendrobine and polysaccharide were determined to investigate the effect of MF23 on the plant growth and the main chemical composition accumulation of stem. RESULTS: After 18 months growth, the stem height and the dry weight of root, stem and leaf of inoculated seedlings were significantly higher than those of the control, increased by 25.34%, 29.28%, 34.64% and 47.12%, respectively. MF23 increased the highest amount of dendrobine and polysaccharide in stem and also prolonged the accumulation process of polysaccharide. After 12 months growth, the content of dendrobine in both the treatment and the control group reached the highest levels, which were 0.59% and 0.51%, respectively. The content of polysaccharide in the treatment and the control group reached the highest levels after 15 (20.05%) and 12 (17.89%) months growth, respectively. CONCLUSION: Mycorrhizal fungus MF23 significantly promotes the growth of D. nobile seedlings, and had a good application prospects on the cultivation and production of D. nobile.

Distribution of bioluminescent fungi across old-growth and secondary tropical rain forest in Costa Rica

Most research on bioluminescent fungi is concentrated on their taxonomic relationships, while the basics of their natural history and ecological relationships are poorly understood. In this study, we compared the distribution of bioluminescent fungi between old-growth and secondary forest as related to four different soil types at the tropical rainforest of La Selva Biological Station in Costa Rica. The study was conducted during the wet season of 2009. Bioluminescent fungi were sought following eight different transects distributed evenly in old-growth and secondary forests across four different soil types, covering an area of 9 420m². We found fungi in four different substrates: litter, fallen branches, dead trunks, and roots, for a total of 61 samples. Correspondence analysis showed that the occurrence of fungi and soil types were related (inertia=0.21, p=0.071). We found a significant relationship between the presence of fungi and the distribution of soil types (X²=18.89, df=9, p=0.026). We found only three samples with fruiting bodies, two of which had Mycena and the other had one fungus of the order Xylariales (possibly Hypoxylon sp., Kretzschmariella sp., Xylaria sp.). Future work will concentrate on exploring other aspects of their ecology, such as their dispersal and substrate preference. This information will facilitate field identification and will foster more research on the distribution, seasonality, reproductive phenology and ecological requirements of this group of Fungi.

La mayoría de las investigaciones sobre los hongos bioluminiscentes se ha centrado en relaciones taxonómicas. Los aspectos básicos de la historia natural y relaciones ecológicas de este grupo son poco conocidos. En este estudio, comparamos la distribución de hongos bioluminiscentes entre el bosque primario y el secundario en la Estación Biológica La Selva, Costa Rica en relación con cuatro tipos de suelo. El estudio se realizó durante la estación lluviosa del 2009. Se buscaron hongos bioluminiscentes en ocho transectos distribuidos de manera uniforme en el bosque primario y secundario y en cuatro tipos de suelo, cubriendo un área de 9 420m². Se encontraron hongos en cuatro sustratos: arena, ramas caídas, troncos muertos, y raíces muertas, para un total de 61 muestras. Se encontró una relación significativa entre la presencia de hongos y la distribución de los tipos de suelo (x²=18.89, gl=9, p=0.026). Sólo se encontraron tres muestras con cuerpos fructíferos, dos de ellos del género Mycena, mientras que el otro presentaba un hongo del orden Xylariales (posiblemente Hypoxylon sp. Kretzschmariella sp. Xylaria sp.). Futuras líneas de investigación deben concentrarse en aspectos básicos de la ecología del grupo, tales como su dispersión y preferencia de sustrato. Esta información fomentará una mayor investigación sobre la distribución, la estacionalidad y fenología reproductiva, y los requerimientos ecológicos de este grupo de hongos.

Seed Germination of Gastrodia elata Using Symbiotic Fungi, Mycena osmundicola

The germination rate and longevity of seeds of Gastrodia elata Blume have been observed for 48 weeks using Mycena osmundicola strain H-21, one of fungi stimulating seed germination. Storage condition of post-harvest seeds was observed in the different temperature ranges of -30degrees C, -5degrees C, 5degrees C and 30degrees C for 48 weeks. After storage period of 48 weeks, the germination rate of G. elata was 65.7% at 5degrees C and 71.6% at -5degrees C, respectively. Although the germination rate of G. elata was 77.3% for 11 weeks at 25degrees C, the germination rate had been decreased gradually to 49.3% at 13 weeks, 0.3% at 23 weeks and then 0% at 25 weeks. The germination rate was reached to the level of 10% for 2 weeks at -30degrees C and then decreased to 0%.