Trichoderma viride Strains against Vegetable Grey Mold in Greenhouse

[Objective] The paper aimed to study the control effects of live spore preparations of Trichoderma viride strains against vegetable grey mold in greenhouse. [Method] Trichoderma viride strains NW-411 live spore preparations were prepared by solid-state fermentation,106-107 spore/g diluent was made to conduct field control experiment,traits change of cucumber and tomato plants inoculated grey mold were investigated,control effect was calculated. [Result] Cucumber and tomato plants without dilution treatment of T. viride spores could be infected with different changes in trait. T. viride spore preparations had a significant preventive effect on greenhouse cucumber and tomato gray mold,the optimal concentration of spores was in the range of 2.3×10^6-2.3×10^7 spore/g. The incidence of cucumber and tomato plants were reduced to 4.2% and 3.1%,the incidence rate decreased 29.8% and 39.1% compared with plants without treatment,biological control effect was over 87%,and the plant growth can be enhanced obviously. [Conclusion] Live spores preparation of T. viride not only had a significant effect on grey mold,but also significantly enhanced the plants growth in greenhouse,which is a safety and environmental protection biological agent,and worthy to be widely spread in large-scale green vegetable production.

Physicochemical properties and release characteristics of calcium alginate microspheres loaded with Trichoderma viride spores

Novel agroformulations for simultaneous delivery of chemical and biologically active agents to the plants were prepared by encapsulation of Trichoderma viride spores in calcium alginate microspheres.The impact of calcium ions concentration on the viability and sporulation of T.viride spores as well as on the microsphere important physicochemical properties were investigated.Intermolecular interactions in microspheres are complex including mainly hydrogen bonds and electrostatic interactions.T.viride germination inside matrix and germ tubes penetration out of microspheres revealed calcium alginate microspheres provide a supportive environment for T.viride growth.Differences in physicochemical properties and bioactive agents release behaviour from microspheres were ascribed to the changes in microsphere structure.Fitting to Korsmeyer-Peppas empirical model revealed the underlying T.viride release mechanism as anomalous transport kinetics(a combination of two diffusion mechanisms and the Type II transport(polymer swelling and relaxation of the polymeric matrix)).The increasing amount of T.viride spores in the surrounding medium is closely related to the release from microspheres and germination.The rate controlling mechanism of calcium release is Fickian diffusion.A decrease in the release rate with increasing calcium ion concentrations is in accordance with the calcium ions effect on the strength of the alginate network structure.T.viride germination inside microsphere diminished the amount of released calcium ions and slowed release kinetics in comparison with microspheres prepared without T.viride.The results indicated investigated agroformulations have a great potential to be used for plant protection and nutrition.

Effect of Trichoderma viride biofertilizer on ammonia volatilization from an alkaline soil in Northern China

Ammonia（NH3） volatilization is one of the primary pathways of nitrogen（N） loss from soils after chemical fertilizer is applied, especially from the alkaline soils in Northern China, which results in lower efficiency for chemical fertilizers. Therefore, we conducted an incubation experiment using an alkaline soil from Tianjin（p H 8.37–8.43） to evaluate the suppression effect of Trichoderma viride（T. viride） biofertilizer on NH3 volatilization, and compared the differences in microbial community structure among all samples. The results showed that viable T. viride biofertilizer（T） decreased NH3 volatilization by 42.21% compared with conventional fertilizer（（CK）, urea）, while nonviable T. viride biofertilizer（TS） decreased NH3 volatilization by 32.42%. NH3 volatilization was significantly higher in CK and sweet potato starch wastewater（SPSW） treatments during the peak period. T. viride biofertilizer also improved the transfer of ammonium from soil to sweet sorghum. Plant dry weights increased 91.23% and 61.08% for T and TS, respectively, compared to CK. Moreover, T. viride biofertilizer enhanced nitrification by increasing the abundance of ammonium-oxidizing archaea（AOA） and ammonium-oxidizing bacteria（AOB）. The results of high-throughput sequencing indicated that the microbial community structure and composition were significantly changed by the application of T. viride biofertilizer. This study demonstrated the immense potential of T. viride biofertilizer in reducing NH3 volatilization from alkaline soil and simultaneously improving the utilization of fertilizer N by sweet sorghum.

Cloning of Endo-β-Glucanase Ⅲ and Expression in Eerevisiae Fermentum

Objective The aim was to construct bioengineering strains that could degrade the cellulosic solid waste. Method The cDNA of endo-β-glucanase III of Trichoderma vi ride AS313711 was cloned by RT-PCR method. After sequenced, this gene was constructed to expression vector pESP-2, and then the plasmid was transformed into competent cell of cerevisiae fermentum by electric shock, the transformant was then obtained. The enzyme activity of this transformant at the different temperatures and pH was measured by DNS method. Result The length of ORF of EG III was 1 257 bp, encoding 418 amino acids, while the deduced molecular weight was 44.1 × 103 kD. Conclusion The enzyme activity of EG III was the highest when it was at PH 4.9 and tempeture was of 60℃. Then the corresponding enzyme activity was about 100%.

Improvement of Feeding Value of Rapeseed Meal by Mixed Solid State Fermentation

[Objective] To optimize solid state fermentation conditions of rapeseed meal and thus to reduce glucosinolates and neutral detergent fibers by mixed cultures of Aspergillus oryzae and Tnchoderrna viride. [ Method ] The optimal fermentation conditions were determined by single factor test and orthogonal design. [ Result J The optimum fermentation conditions are as following: inoculum weight ratio (Aspergillus oryzae vs Trichoderma vinde), 1:1 ; inoculum size, 30% ; water content, 40% ; fermentation time, 96 h; and fermentation temperature, 30℃. Under these conditions, glucosinolates were reduced by 90.71% and neutral detergent fibers were degraded by 20.65%. [ Condusion] In laboratory, solid state fermentation with Aspergillus oryzae and Trichoderrna viride can be used to produce high-quality rapeseed meal.

Mutagenesis of T richoderma V iride by Ultraviolet and Plasma

Considering the importance of a microbial strain capable of increased cellulase production, a mutant strain UP4 of Trichoderma viride was developed by ultraviolet （UV） and plasma mutation. The mutant produced a 21.0 IU/mL FPase which was 98.1% higher than that of the parent strain Trichoderma viride ZY-1. In addition, the effect of ultraviolet and plasma mutagenesis was not merely simple superimposition of single ultraviolet mutation and single plasma mutation. Meanwhile, there appeared a capsule around some of the spores after the ultraviolet and plasma treatment, namely, the spore surface of the strain became fuzzy after ultraviolet or ultraviolet and plasma mutagenesis.

Characterization of Humic System in Fertilizer Raw Materials

The aim of the study was to characterize humus system of natural and artificial products. Humus systems from leonardite, lignite biotransformed with Trichoderma sp. （Plantagra）, plant materials after pyrolisis （charcoal） and composts are compared. Humus systems are characterized by Kononova-Belchikova＇s method, and heavy metals content was measured by atomic absorption spectroscopy （AAS）. Humic acids from the International Humic Substances Society （IHSS） collection are the standards for humus substances quality of compared products. Data obtained for leonardite indicate that the studied substances from factory, Izmir, Turkey contain humic acids over 94%. Compared to the standard, heavy metals content in these materials demonstrate high amounts. Organic carbon content in the composts is very low compared to the leonardite materials and IHSS collection, where the heavy metals content is lower. Biotransformed lignite is characterized with lower content of organic carbon, but humic acids are with high degree of humification. Results obtained show that the fourth studied humus systems may be used in agriculture on base of the high humic acids content. It is recommended to measure heavy metals content before applying the materials in agriculture.

Isolation of a salt tolerant laccase secreting strain of Trichoderma sp. NFCCI-2745 and optimization of culture conditions and assessing its efectiveness in treating saline phenolic efuents

Most of the hazardous pollutants are phenolic in nature and persists in the environment. The ability of laccases to oxidize phenolic compounds and reduce molecular oxygen to water has led to intensive studies of these enzymes. Therefore the fungal strains with high laccase activity and substrate affinity that can tolerate harsh environmental conditions have a potential for biotechnological applications. Salt tolerant laccase secreting fungi can be utilized in treatment of saline and phenolic rich industrial effluents such as coir effluent and textile effluent that needed to be diluted several fold before microbial treatment. This is the first study describing the isolation and optimization of a salt tolerant strain of Trichoderma sp. potential for industrial applications. The fungus was identified based on morphological characteristics and was subsequently confirmed with molecular techniques and deposited at National Fungal Culture Collections of India （NFCCI） under the Accession No. Trichoderma viride NFCCI 2745. In contrast to other laccase secreting fungi, light conditions did not exert much influence on laccase production of this strain and salinity enhanced its laccase secretion. The fungus effectively removed the phenolic content of the textile effluent, coir-ret liquor and wood processing effluent within 96 hr of incubation. The tolerance of the fungus to high salinity and phenolic compounds makes this strain ideal for treating saline and phenolic rich industrial effluents.

Enhancement of Essential Oil Yield from Melaleuca Leucadendra L.Leaves by Lignocellulose Degradation Pre-Treatment Using Filamentous Fungi

The essential oil from Melaleuca leucadendra L.leaves has been widely used as a perfume and traditional remedy,cosmetics and pharmaceutical products ingredient since many years ago.The common technology to recover the oil is hydro-distillation and steam-distillation.However,all oil can not be fully extracted from the leaves by this method due to the recalcitrant structure of leaves that hindrance the access of the solvent.Adding a submerged fermentation as a pretreatment step prior to the extraction process helped to loosen the lignocellulose structure and enhance oil release in the extraction process.In this study,the raw materials were collected from the natural forest in Buru Island,Maluku,Indonesia.The biological agents applied in these processes were Phanerochaete chrysosporium ITBCC136 and Trichoderma viride ITBCC143.The oil extraction process was conducted by method of steam-distillation,the oil was analysed using gas chromatography-mass spectroscopy(GC-MS),and the lignocellulose content in the biomass was measured by the fractionation method.The treatment using T.viride provided the highest increase in yield percentage up to 3.47%as compared with control of 1.45%,with the lowest percentages of the remained cellulose,while the fermentation with the presence of P.chrysosporium did not affect the oil yield even the lignin content was decrease as much as 21%.The percentages of 1,8-cineole in the oil were almost unchanged,which was about 20%of the oil.