A simple method for the determination of bioethanol from lignocellulosic materials using gas chromatography-flame ionisation detector (GC-FID)

Aims@#The utilisation of lignocellulosic biomass for bioethanol production reduces the dependency on fossil fuels as a source of energy and emission of greenhouse gas (GHG). However, studies in this emerging field are hampered by the cost of ethanol quantification methods. Due to the volatile nature of ethanol, the method for the quantification of bioethanol production should be reproducible and rapid to avoid any evaporation loss to the surroundings. Therefore, this study aimed to develop a simple, rapid and precise bioethanol quantification method using a gas chromatographyflame ionisation detector (GC-FID) without having to go through distillation process for ethanol purification.@*Methodology and results@#The bioethanol was produced via consolidated bioprocessing (CBP) using Trichoderma asperellum B1581 and paddy straw. The peak corresponding to ethanol was obtained at 2.347 min with a peak area of 189.66, equating to 0.159% (v/v) or 1.25 g/L ethanol. A comparison between the quantity of ethanol detected by GC-FID and spectrophotometric analysis (340 nm) showed no significant difference (p>0.05) in the amount of ethanol detected by GC analysis, thus validating the accuracy of the GC method.@*Conclusion, significance and impact of study@#This work presents a simple, precise and reliable method to determine the amount of bioethanol in the sample using a GC-FID. Currently, there are many GC-FID methods available for the determination of ethanol/alcohol in a human blood samples or in beverages but not in bioethanol samples. Thus, this method was developed to facilitate the determination of bioethanol in the samples produced from lignocellulosic materials.

Morphological and molecular characterization of Trichoderma species isolated from rhizosphere soils in Malaysia

Aims@#Knowledge of the Trichoderma taxa is important for both control efficiency and environmental conservation. Therefore, the objective of this study is to isolate and identify Trichoderma species from various rhizosphere soil samples using phenotypic and molecular characterization.@*Methodology and results@#Native Trichoderma spp. were isolated from agricultural fields in 17 sites from seven states of Malaysia. These isolates were characterized via morphological observation and molecular phylogenetic analysis based on the translation elongation factor-1α (tef1-α) gene. About 42 isolates were classified into eight Trichoderma species, which are Trichoderma asperellum, T. hamatum, T. harzianum, T. koningiopsis, T. rodmanii, T. spirale, T. viride and T. virens. Comparison of DNA sequences of tef1-α showed that the isolates were 98-100% similar to respective Trichoderma species from GenBank, thus confirming the fungal identity. Phylogenetic trees of maximum likelihood (ML) dataset of tef1-α inferred that the isolates were clustered according to species.@*Conclusion, significance and impact of study@#Findings in the present study will be beneficial for the purposes of biodiversity conservation and plant disease management using biocontrol agents.

Screening of lignocellulolytic fungi for hydrolyzation of lignocellulosic materials in paddy straw for bioethanol production

Aims@#Paddy straw is known to have lignocellulosic materials such as cellulose and hemicellulose which can be readily converted into fermentable sugar for production of bioethanol via simultaneous saccharification and fermentation (SSF). In order to produce ethanol competently, the degradation of biomass by cellulase and highly ethanol-producing microorganism in fermentation process are necessarily needed. However, there is lacking in cellulose degrading organism in producing adequate amount of lignocellulosic enzyme. Therefore, the screening and selection for the best fungi to hydrolyze the lignocellulosic materials as well as forming consortium between two species of fungi has become the main focus. @*Methodology and results@#Thirteen strains of fast-growing fungi were tested qualitatively for cellulase (congo red staining) and polyphenol oxidase (Bavendamm test). All tested strains displayed lignocellulolytic fungi characteristics. The selection was narrowed down by quantitative assay on endoglucanase, exoglucanase, β-glucosidase and xylanase and the highest cellulases enzyme producer were Trichoderma asperellum B1581 (3.93 U/mL endoglucanase; 2.37 U/mL exoglucanase; 3.00 IU/mL β-glucosidase; 54.87 U/mL xylanase), followed by Aspergillus niger B2484 (5.60 U/mL endoglucanase; 1.08 U/mL exoglucanase; 1.57 IU/mL β-glucosidase; 56.85 U/mL xylanase). In compatibility test, both T. asperellum B1581 and A. niger B2484 were inoculated on the same Petri dish for 4 days and the interaction showed by the two species was mutual intermingling. @*Conclusions, significance and impact of study@#Both T. asperellum B1581 and A. niger B2484 produced the highest cellulase enzyme. Since both strains can co-exist and produce enzymes that complete each other, a fungal consortium was suggested to increase the yield of sugars in saccharification process.

Volatile organic compounds profiles emitted by Cochliobolus miyabeanus, a causal agent of brown spot disease of rice

Aims@#Brown spot disease is among the important crop diseases of rice caused by the infection of a pathogenic fungus, Cochliobolus miyabeanus that results in yield losses. Nowadays, limited studies on volatile organic compounds (VOCs) have been carried out using pathogenic fungal isolate. Hence, this study was conducted to identify VOCs produced by C. miyabeanus wild-type isolate, WK1C, a causal agent of brown spot disease using gas chromatography-mass spectrometry (GC-MS). @*Methodology and results@#Fungal isolate WK1C was cultured on potato dextrose agar (PDA) and in potato dextrose broth (PDB) for extraction. The extracts were analysed using GC-MS and the profiles of VOCs were obtained. Cochliobolus miyabeanus WK1C isolate showed a significant presence of various types of organic compound including ester, alcohol, phenol, alkane, alkene, ketone, carboxylic acid, amide and aldehyde. @*Conclusion, significance and impact of study@#This study important for a preliminary assessment of VOCs profiles of C. miyabeanus, a causal agent of brown spot disease. In order to identify the compounds contribute to pathogenicity, further study can be conducted to identify the virulence factor of brown spot disease using different approaches

Secondary metabolite profiles and mating populations of Fusarium species in section Liseola associated with bakanae disease of rice

A total of 25 strains of Fusarium species that belong to F. fujikuroi (a pathogen of bakanae disease), F. proliferatum, F.sacchari, F. subglutinans and F. verticillioides were isolated from rice plants showing typical bakanae symptoms in Malaysia and Indonesia and screened for their secondary metabolites. The objectives of the studies were to determine the physiological variability based on production of moniliformin (MON), fumonisin (FB1), gibberellic acid (GA3) and fusaric acid (FA) as well as to ascertain the mating populations (MPs) within the Gibberella fujikuroi species complex based on their ability to produce perithecia and viable ascospores. Production of GA3 could be used to separate F. fujikuroi that belongs to MP-C from other species. In crosses with seven standard testers of MPs, 76% of strains could be assigned to at least one of the G. fujikuroi species complex namely MP-A (G. moniliformis), MP-B (G. sacchari), MP-C (G. fujikuroi) and MP-D (G. intermedia). Single strain (M3237P) that was assigned as MP-C, and has also been identified morphologically as F. fujikuroi was also crossed-fertile with MP-D tester. The secondary metabolites profiles and the presence of MP-A, MP-B, MP-C and MP-D strains on samples of bakanae-infected rice plants are new records in Malaysia.