Comparative study of mycelia growth and sporophore yield of Auricularia polytricha (Mont.) Sacc on selected palm oil wastes as fruiting substrate

The potential for using agricultural and industrial by-products as substrate for the production of the edible mushroom, Auricularia polytricha, was evaluated using several formulations of selected palm oil wastes mixed with sawdust and further supplemented with selected nitrogen sources. The best substrate formulations were sawdust (SD) mixed with oil palm frond (OPF; 90:10) added with 15 % spent grain (SG) and sawdust mixed with empty fruit bunch (EFB; 50:50) added with 10 % spent grain (SG) with mycelia growth rate of 8 mm/day and 7 mm/day respectively. These two substrate formulations were then subjected to different moisture content levels (65 %, 75 % and 85 %). Highest total fresh sporophore yield at 0.43 % was obtained on SD?+?OPF (90:10)?+?15 % SG at 85 % moisture content, followed closely by SD?+?EFB (50:50)?+?10 % SG with 0.40 % total yield, also at 85 % moisture content. Each of the substrate formulations at 85 % moisture content gave the highest biological efficiency (BE) at 288.9 % and 260.7 %, respectively. Both yield and biological efficiency of A. polytricha on these two formulations were almost three times higher when compared to sawdust substrate alone, thus proving the potential of these formulations to improve yield of this mushroom.     

Antagonistic effects of Streptomyces violaceusniger strain G10 on Fusarium oxysporum f.sp. cubense race 4: Indirect evidence for the role of antibiosis in the antagonistic process

Fusarium oxysporum f.sp. cubense is the causal pathogen of wilt disease of banana. A cost-effective measure of control for this disease is still not available. Streptomyces violaceusniger strain G10 acts as an antifungal agent antagonistic towards many different phytopathogenic fungi, including different pathogenic races of the Fusarium wilt pathogen. In an attempt to understand the mode of action of this antagonist in nature, the interaction between S. violaceusniger strain G10 and F. oxysporum f.sp. cubense was first studied by paired incubation on agar plates. Evidence for the in vitro antibiosis of strain G10 was demonstrated by inhibition zones in the “cross-plug” assay plates. Microscopic observations showed lysis of hyphal ends in the inhibited fungal colonies. Culture of strain G10 in liquid media produces antifungal metabolites, which showed in vitro antagonistic effects against F. oxysporum f.sp. cubense such as swelling, distortion and excessive branching of hyphae, and inhibition of spore germination. An indirect method was used to show that antibiosis is one of the mechanisms of antagonism by which strain G10 acts against F. oxysporun f.sp. cubense in soil. This study suggests the potential of developing strain G10 for the biological control of Fusarium wilt disease of banana. Journal of Industrial Microbiology & Biotechnology (2002) 28, 303–310 DOI: 10.1038/sj/jim/7000247 Received 08 August 2001/ Accepted in revised form 16 February 2002     

Laccase production from oil palm industry solid waste: Statistical optimization of selected process parameters

Oil palm frond parenchyma tissue was used as a solid substrate for the production of laccase via solid‐state fermentation using the white rot fungus Pycnoporus sanguineus. With a rectangular aluminium tray as solid‐state fermentation bioreactor, process parameters such as bed height, moisture and supplemented nitrogen (as urea solution) levels were studied and optimized using a statistical design of experiment. The moisture level exerted a significant effect on the process. The interaction effect observed between bed height and supplemented nitrogen level suggested that uniform distribution of supplemented nitrogen into the substrate bed was important. The proposed regression model sufficiently predicted the process response over the experimental range tested. The optimum parameter combination for laccase production was a 3‐cm bed height, 72% w/w moisture and 0.21% w/v supplemented nitrogen. Laccase productivity remained constant when the tray size was increased from 1.4 to 3.4‐fold.     

Enzymes from spent mushroom substrate of <Emphasis Type="Italic">Pleurotus sajor</Emphasis>-<Emphasis Type="Italic">caju</Emphasis> for the decolourisation and detoxification of textile dyes

The potential of ligninolytic enzymes, including lignin peroxidase (LiP) as the main enzyme from the spent mushroom substrate of Pleurotus sajor-caju was evaluated for the decolourisation of five dyes from azo and anthraquinone dye groups. Among the azo dyes, reactive black 5 and reactive orange 16 were 84.0 and 80.9% decolourised respectively, after 4 h of incubation with 45 U of LiP as compared to 32.1% decolourisation of disperse blue 79. Among the anthraquinone dyes, disperse red 60 was decolourised to 47.2% after 4 h of incubation with 45 U of LiP as compared to 5.9% decolourisation of disperse blue 56. Increasing the LiP concentration and incubation time had a positive effect on the decolourisation of anthraquinone dyes as compared to azo dyes. A 67.9% decolourisation of synthetic textile waste-water was achieved after 4 h of incubation with 25 U of LiP. Increasing the incubation time significantly increased (P < 0.05) the decolourisation of synthetic textile waste-water. Further, there was a 52.4% reduction in the toxicity of synthetic textile waste-water treated with 55 U of LiP for 4 h. However, only 35.7% reduction in toxicity was achieved when the synthetic textile waste-water was treated with 55 U of LiP for 24 h. In this study, it was shown that the spent mushroom substrate of P. sajor-caju could be a cheap source of ligninolytic enzymes for the decolourisation of dyes in textile industry wastewaters.     

Evaluation of Streptomyces sp. strain g10 for suppression of Fusarium wilt and rhizosphere colonization in pot-grown banana plantlets

Streptomyces sp. strain g10 exhibited strong antagonism towards Fusarium oxysporum f.sp. cubense (Foc) races 1, 2 and 4 in plate assays by producing extracellular antifungal metabolites. Treating the planting hole and roots of 4-week-old tissue-culture-derived Novaria banana plantlets with strain g10 suspension (10⁸ cfu/ml), significantly (P<0.05) reduced wilt severity when the plantlets were inoculated with 10⁴ spores/ml Foc race 4. The final disease severity index for leaf symptom (LSI) and rhizome discoloration (RDI) was reduced about 47 and 53%, respectively, in strain g10-treated plantlets compared to untreated plantlets. Reduction in disease incidence was not significant (P<0.05) when plantlets were inoculated with a higher concentration (10⁶ spores/ml) of Foc race 4. Rhizosphere population of strain g10 showed significant (P=0.05) increase of more than 2-fold at the end of the 3rd week compared to the 2nd week after soil amendment with the antagonist. Although the level dropped, the rhizosphere population at the end of the 6th week was still nearly 2-fold higher than the level detected after 2 weeks. In contrast, the root-free population declined significantly (P=0.05), nearly 4-fold after 6 weeks when compared to the level detected after 2 weeks. Neither growth-inhibiting nor growth-stimulating effects were observed in plantlets grown in strain g10-amended soil.     

Escaping introns in COI through cDNA barcoding of mushrooms: Pleurotus as a test case

DNA barcoding involves the use of one or more short, standardized DNA fragments for the rapid identification of species. A 648‐bp segment near the 5′ terminus of the mitochondrial cytochrome c oxidase subunit I (COI) gene has been adopted as the universal DNA barcode for members of the animal kingdom, but its utility in mushrooms is complicated by the frequent occurrence of large introns. As a consequence, ITS has been adopted as the standard DNA barcode marker for mushrooms despite several shortcomings. This study employed newly designed primers coupled with cDNA analysis to examine COI sequence diversity in six species of Pleurotus and compared these results with those for ITS. The ability of the COI gene to discriminate six species of Pleurotus, the commonly cultivated oyster mushroom, was examined by analysis of cDNA. The amplification success, sequence variation within and among species, and the ability to design effective primers was tested. We compared ITS sequences to their COI cDNA counterparts for all isolates. ITS discriminated between all six species, but some sequence results were uninterpretable, because of length variation among ITS copies. By comparison, a complete COI sequences were recovered from all but three individuals of Pleurotus giganteus where only the 5′ region was obtained. The COI sequences permitted the resolution of all species when partial data was excluded for P. giganteus. Our results suggest that COI can be a useful barcode marker for mushrooms when cDNA analysis is adopted, permitting identifications in cases where ITS cannot be recovered or where it offers higher resolution when fresh tissue is. The suitability of this approach remains to be confirmed for other mushrooms.     

Fructose formation in stored blood

At the high glucose concentrations used in the collection and storage of donor blood the activity of the fructose-forming polyol pathway (Reaction I and II) could act to deplete NADPH and thus GSH, thereby exposing the cells to oxidative stress. Fructose levels were found to be high in red cells and the supernatant plasma of blood collected into CP2D, which contains 258 mM glucose. Elevated fructose was not produced by the polyol pathway, but was formed by the autoclaving process. A high fructose concentration sufficient to account for the fructose in donor red cells was also found in the CP2D anticoagulant and in samples of autoclaved glucose.