Pengoptimuman Campuran Enzim Selulase Rekombinan Untuk Pencuraian Tandan Kosong Kelapa Sawit

Penggunaan enzim selulase untuk penguraian biojisim pertanian lignoselulosa telah lama dikaji dan pelbagai usaha telah dilakukan untuk meningkatkan kecekapan proses hidrolisis. Keberkesanan penguraian biojisim pertanian kepada gula ringkas memerlukan satu campuran enzim yang mengandungi pelbagai jenis aktiviti selulolitik. Dalam kajian ini, satu campuran multi-enzim rekombinan yang terdiri daripada tiga komponen asas selulase iaitu endoglukanase (EglB) dan β-glukosidase (BglA) daripada Aspergillus niger serta selobiohidrolase (CbhII) daripada Trichoderma virens telah dibentuk khusus untuk hidrolisis tandan kosong kelapa sawit (TKKS). Penghasilan enzim selulase rekombinan telah dilakukan menggunakan hos pengekspresan Pichia pastoris. Pengoptimuman nisbah enzim untuk tindak balas ditentukan menggunakan Kaedah Gerak Balas Permukaan (RSM). Hasil menunjukkan hidrolisis TKKS pada suhu 50 °C dan pH 5.0 menggunakan enzim pada nisbah 641.4 unit CMCase: 10.14 unit Avicelase: 93.8 unit β-glukosidase, menghasilkan gula terturun dan glukosa tertinggi, masing-masing sebanyak 63 mg dan 40 mg per gram substrat TKKS. Hasil hidrolisis TKKS oleh campuran multi enzim yang telah dibentuk dalam kajian ini menunjukkan ketiga-tiga gabungan enzim rekombinan ini berpotensi untuk digunakan bagi penguraian TKKS.

EFECTO DE LA INTERACCIÓN DUAL DE ESPECIES DE Trichoderma EN EL CRECIMIENTO DE Capsicum chinense Jacq.

La interacción de especies bioestimulantes de Trichoderma puede tener beneficios para aumentar la producción de metabolitos involucrados en la promoción de crecimiento vegetal. La combinación de dos organismos con mayor grado de antagonismo podría potenciar los efectos promotores del crecimiento en plantas. El objetivo del estudio fue seleccionar in vitro especies nativas de Trichoderma cultivadas de manera dual y evaluar su efecto promotor del crecimiento en Capsicum chinense. La selección de aislados in vitro se realizó por medio de enfrentamientos duales con las siguientes características: crecimiento proporcional de ambos aislados en la caja Petri, presencia de barrera micelial y coloración del medio de cultivo. Los enfrentamientos seleccionados y sus aislados individuales se evaluaron en plantas de C. chinense cv. Mayapan. Además, dos testigos sin inoculación fúngica, uno como testigo al 100% (250N-200P-300K) de la fertilización química recomendada para el cultivo; y el segundo como testigo al 50%, con la mitad de la fertilización indicada. Tres aplicaciones de suspensiones de 106 conidios mL-1 se realizaron en la etapa de almácigo y otras tres durante el crecimiento del cultivo. El efecto de los tratamientos se evaluó a través del crecimiento vegetal y la producción del cultivo a los 145 días después de la siembra. La interacción de Trichoderma virens (Th05-02)/Trichoderma koningiopsis (Th41-11) fue igual (p>0.05) al testigo fertilizado al 100% en las variables evaluadas. Además, se obtuvieron incrementos del 14.53% en altura de la planta, 11.20% en diámetro del tallo, 28.54% en la biomasa seca aérea, 12.97% en biomasa seca de raíz, 33.13% en número de frutos y 25.77% en el peso de frutos, respecto al testigo con 50% de fertilización química. La interacción de las especies T. virens (Th05-02)/T. koningiopsis (Th41-11) ejerció un efecto promotor de crecimiento vegetal con la reducción del 50% de la fertilización química recomendada en el cultivo de chile habanero.

Complete Genome Sequences and Genome-Wide Characterization of Trichoderma Biocontrol Agents Provide New Insights into their Evolution and Variation in Genome Organization, Sexual Development, and Fungal-Plant Interactions

Telomere-to-telomere and gapless reference genome assemblies are necessary to ensure that all genomic variants are studied and discovered, including centromeres, telomeres, AT-rich blocks, mating type loci, biosynthetic, and metabolic gene clusters. Here, we applied long-range sequencing technologies to determine the near-completed genome sequences of four widely used biocontrol agents or biofertilizers: Trichoderma virens Gv29-8 and FT-333, Trichoderma asperellum FT101, and Trichoderma atroviride P1.

Purification, Identification And Characterization of Nag2 N-Acetylglucosaminidase From Trichoderma Virens Strain Mango

Background: N -acetylglucosaminidase (NAGase) could liberate N -acetylglucosamine (GlcNAc) from GlcNAc-containing oligosaccharides. Trichoderma spp. is an important source of chitinase, particularly NAGase for industrial use. nag1 and nag2 genes encoding NAGase , are found in the genome in Trichoderma spp. The deduced Nag1 and Nag2 shares ~55% homology in Trichoderma virens. Most studies were focus on Nag1 and nag1 previously. Results: The native NAGase (TvmNAG2) was purified to homogeneity with molecular mass of ~68 kDa on SDS-PAGE analysis, and identified as Nag2 by MALDI/MS analysis from an isolate T. virens strain mango. RT-PCR analyses revealed that only nag2 gene was expressed in liquid culture of T. virens , while both of nag1 and nag2 were expressed in T. virens cultured on the plates. TvmNAG2 was thermally stable up to 60 o C for 2 h, and the optimal pH and temperature were 5.0 and 60-65 o C, respectively, using p -nitrophenyl- N -acetyl- β -D-glucosaminide ( p NP-NAG) as substrate. Using colloidal chitin as substrate, the end product catalyzed by TvmNAG2 was GlcNAc, based on HPLC and TLC analyses. The optimal temperature for TvmNAG2 to produce GlcNAc was 40 o C. TvmNAG2 possesses antifungal activity, inhibiting the mycelium growth of Sclerotium rolfsii . And it was resistant to the proteolysis by papain and trypsin. Conclusions: The native Nag2, TvmNAG2 was purified and identified from T. virens strain mango, as well as enzymatic properties. To our knowledge, it is the first report with the properties of native Trichoderma Nag2.

Synergistic Effect between Trichoderma virens and Bacillus velezensis on the Control of Tomato Bacterial Wilt Disease

(1) Background: Ralstonia solanacearum causes tomato bacterial wilt disease, one of the most serious tomato diseases. As the combination of Trichoderma virens (Tvien6) and Bacillus velezensis (X5) was more effective at controlling tomato bacterial wilt disease than a single agent, we investigated the synergistic effect of Tvien6 and X5 in controlling this disease; (2) Methods: The disease incidence, plant heights and weights, relative chlorophyll content (SPAD values), defensive enzymes (PPO, POD, and SOD) activities, and metabolome were estimated among four treatment groups (BR treatment, X5 + R. solanacearum (RS-15); TR treatment, Tvien6+ RS-15; TBR treatment, Tvien6 + X5 + RS-15; and R treatment, RS-15); (3) Results: The R treatment group had the highest disease incidence and lowest plant heights, plant weights, SPAD values, defensive enzyme activities, and D-fructose and D-glucose contents; the TBR treatment group had the lowest disease incidence and highest plant heights, plant weights, SPAD values, defensive enzyme activities, and D-fructose and D-glucose contents; (4) Conclusions: The results revealed that Tvien6 and X5 can both individually promote tomato plant growth, increase leaf chlorophyll content, enhance defensive enzyme activities, and induce the accumulation of D-fructose and D-glucose; however, they were more effective when combined.

Characterization of Fungal FAD-Dependent AA3_2 Glucose Oxidoreductases from Hitherto Unexplored Phylogenetic Clades

The CAZy auxiliary activity family 3 (AA3) comprises FAD-dependent enzymes belonging to the superfamily of glucose-methanol-choline (GMC) oxidoreductases. Glucose oxidase (GOx; EC 1.1.3.4) and glucose dehydrogenase (GDH; EC 1.1.5.9) are part of subfamily AA3_2 and catalyze the oxidation of β-D-glucose at its anomeric carbon to D-glucono-1,5-lactone. Recent phylogenetic analysis showed that AA3_2 glucose oxidoreductases can be grouped into four major clades, GOx I and GDH I–III, and in minor clades such as GOx II or distinct subclades. This wide sequence space of AA3_2 glucose oxidoreductases has, however, not been studied in detail, with mainly members of GOx I and GDH I studied biochemically or structurally. Here, we report the biochemical characterization of four fungal glucose oxidoreductases from distinct, hitherto unexplored clades or subclades. The enzyme from Aureobasidium subglaciale, belonging to the minor GOx II clade, showed a typical preference for oxygen and glucose, confirming the correct annotation of this clade. The other three enzymes exhibited strict dehydrogenase activity with different substrate specificities. GDH II from Trichoderma virens showed an almost six-fold higher catalytic efficiency for maltose compared to glucose. The preferred substrate for the two GDH III enzymes from Rhizoctonia solani and Ustilago maydis was gentiobiose, a β(1→6) disaccharide, as judged from the catalytic efficiency. Overall, the newly studied AA3_2 glucose oxidoreductases showed a much broader substrate spectrum than the archetypal GOx from Aspergillus niger, which belongs to clade GOx I.

UJI FORMULA BIOFUNGISIDA TABLET BERBAHAN AKTIF KONSORSIUM Trichoderma virens ENDOFIT DAN MIKORIZA INDIGENOUS TERHADAP PENYAKIT JAP PADA BIBIT KARET

The objective of this research is to determine the effect of biofungicide tablets with active ingredient of the consortium consisted Trichoderma virens endophyte and mycorrhiza indigenous and to get the best formula to control the JAP disease on rubber plant. The research was carried out experimentally by using a completely randomized design (RAL), consisting of 6 treatments and 4 replications in order to obtain 24 experimental units. Each experimental unit consisted of two seeds, so there were 48 rubber seeds. The treatments seed were: F1 (15 ml T. virens endophyte), F2 (15 gr inokulan mychorrhiza), F3 (15 ml T. virens endophyte + 15 gr inokulan mychorrhiza + 100 gr palm fronds powder + 25 gr zeolite + 25 gr tapioca powder), F4 (15 ml T. virens endophyte + 15 gr inokulan mychorrhiza + 100 gr sago powder + 25 gr sago dregs + 25 gr tapioca powder), F5 (15 gr inokulan mychorrhiza + 100 gr solid + 25 gr zeolite + 25 gr tapioca powder) and F6 (15 ml T. virens endophyte + 100 gr cocoa shells powder + 25 gr sago dregs + 25 gr tapioca powder). Parameter observerd were initial symptoms on seedlings, intensity of disease, addition of plant height, stem diameter, and weight of dry seedlings. The data obtained were analyzed for variance followed by further tests of honest real difference (BNJ) at the 5% level. The results indicated that F3 (15 ml T. virens endophyte + 15 gr inokulan mychorrhiza + 100 gr palm fronds powder + 25 gr zeolite + 25 gr tapioca powder) was the best formula in controlling JAP with initial symptoms of infection of 24,50 days with the seedling height was 69,46 cm, stem diameter and dry weight were 6,60 cm and 103,62 gr respectively.