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A convenient culture method for observing filamentous fungi
Abstract
It is important to observe and identify the morphology of individual form and group (colony) during the cultivation of fungi. In this study, half medium combined with micromanipulation technique was used to observe the morphological changes in individual and group of filamentous fungi. The results showed that this novel method could be used to make real-time measurements of the morphology of filamentous fungi without dyeing and slicing, and does not affect the normal growth of mycelia. In addition, it can be applied to many strains for observing the mycelium undergoing natural growth and the colony characteristics simultaneously. Overall, in this experiment, a simple and rapid method is designed for the observation of individual and colony morphology of filamentous fungi, which needs a low cost and workload.
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The effect of addition of autoclaved and filter-sterilized culture filtrate of Piriformospora indica (a root endophytic fungus) to the growing Linum album hairy root cultures on growth and lignan production was investigated. The addition resulted in a significant enhancement in lignan production and growth. The podophyllotoxin and 6-methoxypodophyllotoxin (the lignans) concentrations were maximally improved by 3.8 times (233.8mg/L) and 4.4 times (131.9mg/L) in comparison to control cultures, respectively, upon addition of 3.0% (v/v) filter-sterilized culture filtrate of P. indica to the hairy root cultures of L. album for exposure time of 48h. This increase in the lignan content also coincided with the increase in phenylalanine ammonia lyase activity, which was 3.1-fold (371.4μkat/kg protein) higher compared to control cultures under the same conditions. The maximal increase in hairy root biomass was, however, obtained under different conditions; it was enhanced by 1.4 times (21.8g/L) in comparison to control cultures, when 2% (v/v) filter-sterilized culture filtrate was in contact with L. album cultures for 96h.
The aim of this study was to isolate protoplasts from endophytic fungi and to carry out self-fusion of protoplasts for their enhancement of metal resistance. Self-fusant CBRF59T3 with resistance to 25mM Cd(II) was constructed by self-fusion of inactivated protoplasts from Mucor sp. CBRF59. The inoculation of CBRF59 and CBRF59T3 improved significantly the availability of Cd(II) and Pb(II) in the soil. Compared with CBRF59, CBRF59T3 inoculation increased the content of water-soluble Cd(II) by 24%. The dry weight of rape inoculated with CBRF59 and CBRF59T3 was both higher than that of the uninoculation rape. Inoculation of CBRF59T3 further increased the dry weight of rape by 62% than CBRF59 in the higher Cd(II)-+Pb(II)-contaminated soil. Compared with CBRF59, CBRF59T3 inoculation increased the concentration of Cd(II) in rape shoots by 35-189% in Cd(II)- and Cd(II)-+Pb(II)-contaminated soils. The inoculation of CBRF59T3 also enhanced the translocation of Cd(II) from roots to shoots and increased the amount of extracted Cd(II) by rape. The results indicated that the mutants constructed by protoplast fusion is a feasible and efficient method to improve stress tolerance of uncharacterized fungi for phytoremediation of soils contaminated by heavy metals.
