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Aims@#This study was aimed to characterize laccase from a selected fungal strain and examine the enzyme’s ability to remove lignin from paper pulp. @*Methodology and results@#Twelve fungal strains were screened for laccase production, resulting in the selection of Trichoderma sp. UBDFT12. The highest laccase activity (103 U/L) was observed from the culture filtrate on the fourth day of incubation. The optimum temperature and pH for the enzyme were 40 °C and pH 4, respectively. However, the enzyme stability was found to be reduced with time after 1 h incubation. At 1 mM, it was found that AgNO3, CaCO3, CuSO4, KCl, MgSO4, MnSO4 and ZnSO4 increased the laccase activity to 107, 107, 111, 112, 106, 105 and 107%, respectively, whereas FeSO4 and NH4Cl reduced the activity to 84 and 99%, respectively. The addition of 1% H2O2, 1% NaCl, 1% sodium dodecyl sulphate (SDS), 1 mM ethylenediaminetetraacetic acid (EDTA), 10 mM EDTA, 1 mM phenanthroline and 10 mM phenanthroline reduced the activity to 95, 73, 0, 79, 79, 73 and 37%, respectively. The culture filtrate was partially purified via ammonium sulphate precipitation and the recovered enzyme had a specific activity of 0.176 U/mg. Using sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE), the molecular weight of the enzyme was approximately 65 kDa and its activity was confirmed by zymography. The culture filtrate was also found to be able to remove lignin from different types of paper pulp.@*Conclusion, significance and impact of study@#Laccase produced by Trichoderma sp. UBDFT12 was found to have the ability to remove lignin from paper pulp.
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The banana Fusarium wilt (BFW) caused by Fusarium oxysporum f. sp. cubense tropical race4 (FocTR4) is difficult to control worldwide, which causes a huge economic losse to banana industry. The purpose of this study was to screen Trichoderma strains with antagonistic activity against FocTR4, to isolate and purify the active compound from the fermentation broth, so as to provide important biocontrol strains and active compound resources. In this work, Trichoderma strains were isolated and screened from the rhizosphere soil of crops, and the strains capable of efficiently inhibiting FocTR4 were screened by plate confrontation, and further confirmed by testing inhibition for the conidial germination and mycelial growth of FocTR4. The phylogenetic tree clarified the taxonomic status of the biocontrol strains. Moreover, the active components in the fermentation broth of the strains were separated and purified by column chromatography, the structure of the most active component was analyzed by nuclear magnetic resonance spectroscopy (NMR), the BFW control effect was tested by pot experiments. We obtained a strain JSHA-CD-1003 with antagonistic activity against FocTR4, and the inhibition rate from plate confrontation was 60.6%. The fermentation broth of JSHA-CD-1003 completely inhibited the germination of FocTR4 conidia within 24 hours. The inhibition rate of FocTR4 hyphae growth was 52.6% within 7 d. A phylogenetic tree was constructed based on the ITS and tef1-α gene tandem sequences, and JSHA-CD-1003 was identified as Trichoderma brevicompactum. Purification and NMR identification showed that the single active compound was trichodermin, and the minimum inhibitory concentration (MIC) was 25 μg/mL. Pot experiments showed that the fermentation broth of strain JSHA-CD-1003 was effective against BFW. The control rate of leaf yellowing was 47.4%, and the rate of bulb browning was 52.0%. Therefore, JSHA-CD-1003 effectively inhibited FocTR4 conidial germination and mycelium growth through producing trichodermin, and showed biocontrol effect on banana wilt caused by FocTR4, thus is a potential biocontrol strain.
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Fusarium , Musa , Filogenia , Tricodermina , HypocrealesRESUMEN
ObjectiveRoot rot is one of the most serious diseases in the cultivation and production of Atractylodes lancea. Trichoderma spp. are effective in the biocontrol of root rot without causing environmental pollution. This study aims to isolate and study a Trichoderma strain capable preventing and controlling root rot from the rhizosphere of A. lancea and to solve the problem of disease prevention and control in the planting and production of A. lancea. MethodTrichoderma T2204 was isolated by the dilution-coating method and identified by ITS sequencing. The inhibitory activities of T2204 and its volatiles against two pathogenic fungal strains were examined by dual-culture and co-culture experiments. The biocontrol potential of T2204 on root rot of A. lancea and the effect of T2204 on the accumulation of medicinal compounds in the rhizosphere of A. lancea were investigated by pot experiments and GC-MS， respectively. In addition， the optimal medium， photoperiod， temperature， pH， and carbon and nitrogen sources for the culture of T2204 were explored. ResultThe Trichoderma isolate T2204 was identified as T. citrinoviride and had direct inhibitory effects on two highly pathogenic strains causing root rot. In the dual-culture experiments with the two pathogenic strains， T2204 showcased the inhibition rates of 77.90% and 76.80%， respectively. In the co-culture experiments with the two pathogenic strains， the volatile organic compounds produced by T2204 showed the inhibition rates of 57.11% and 81.11%， respectively. The pot experiments showed that the survival rate of A. lancea seedlings infected by root rot reached 100% after inoculation with T2204 and was only 50% in the case without inoculation of T2204. After 150 days of cultivation， the dry weight and atractylodin content of the rhizome of A. lancea plants treated with T2204 increased by 32% （P<0.05） and 11%， respectively， compared with the untreated group. The optimal conditions for the growth of T2204 were PDA or PSA medium， photoperiod of 12 h dark/12 h light， 25-30 °C， pH 5-6， carbon sources of glucose， D-fructose， soluble starch， and maltose， and the nitrogen sources of ammonium sulfate and ammonium dihydrogen phosphate. The optimal conditions for the sporulation of T2204 were PSA or CMA medium， photoperiod of 12 h dark/12 h light， 20-30 °C， pH 8， carbon source of sucrose， and nitrogen source of sodium nitrate. ConclusionT2204 could improve the growth and root rot resistance of A. lancea and promote the accumulation of medicinal compounds. The findings laid a foundation for the industrialized production and application of T2204 in the production of A. lancea in the future.
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This study evaluated the initial development of paricá (Schizolobium parahyba var. amazonicum) seedlings under different nitrogen rates with the application of Trichoderma spp., using a randomized complete block design in a 4x5 factorial scheme (strains and rates) with seven replications. The evaluated traits were plant height, stem diameter, leaf and stem fresh weights, leaf and stem dry weights, and aerial part dry and fresh weights. Trichoderma spp. strains did not satisfactorily promote paricá seedlings (Schizolobium parahyba var. amazonicum) under high nitrogen rates. However, the Trichoderma harzianum IBLF 006 WP strain was efficient only under low nitrogen availability.
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Tomato (Solanum lycopersicum) is a widely cultivated and versatile fruit. Tomato production faces various constraints, both biotic and abiotic stresses. Fusarium wilt, caused by the soil-borne fungus Fusarium oxysporum f. sp. lycopersici, is a devastating disease that affects tomato plants worldwide, causing significant economic losses in tomato cultivation. Inthis study the potential use of biofortified vermicompost for wilt management was investigated. The biological control agents viz. Trichoderma harzianum, Pseudomonas fluorescens and Bacillus subtilis were used to fortified the vermicpompost. The study monitored various antioxidants, plant growth indicators, and disease rates across designated treatments at different intervals. The findings revealed substantial differences: tomato plants treated with biofortified vermicompost showed notable reductions in disease occurrence, improved growth, increased yields, and heightened antioxidant activity compared to the control. Among these treatments, the most remarkable outcomes were observed in plants treated with T. harzianum fortified vermicompost, exhibiting the highest recorded values across the parameters studied. Data revealed that the Tomato plants treated with vermicompost fortified with Trichoderma showed maximum root length (14.95 cm) after 15 days of sowing followed by T-2 (11.25 cm) and T-3 (9.85 cm). Maximum dry weight was observed in plants treated with vermicompost fortified with Trichoderma. Maximum PAL, PO and PPO activity was recorded in leaves from plant grown in vermicompost fortified with T. harzianum (T1) at 48 h followed by T2, T3 and T4. The findings revealed tomato plants treated with biofortified vermicompost showed notable reductions in disease occurrence, improved growth, increased yields, and higher accumulation of defense enzyme activity compared to the control.
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In the present study Trichoderma asperellum (ECK), Trichoderma asperellum (TNAU), Streptomyces sp (native) and Pseudomonas fluorescens (TNAU) were tested in vitro by dual culture technique. Among the six treatments Trichoderma asperellum (ECK) and Streptomyces sp (native) was found to be more effective as compared to other bio-control agents and inhibited maximum fungal growth (100 %) of Macrophomina phaseolina followed by Trichoderma asperellum (TNAU) (81.6 %). The disease incidence controlled by all other isolates was ranged from 67.72 to 81.66%. The growth promoting activity of various bio-control agents and chemical was assessed based on seedling vigour index by standard Roll Towel Method (ISTA,1993). The treated seeds and controls are placed on the pre-soaked germination paper. After one-week various growth parameters like root length, shoot length, vigour index are determined by using the formulae. The seeds treated with Trichoderma asperellum (ECK) and Streptomyces sp (native) exhibited increased yield parameters like increased germination percentage, root length, shoot length, and Seedling vigour index 2562.0. Under field conditions the seeds are treated with different biocontrol agents and carbendazim. The various growth parameters like root length, shoot length and seedling vigour index observed under field conditions. Streptomyces sp (native) was found to be most effective in reducing pre and post emergence mortality and also it enhances the yield than other treatments. In field trail among the six treatments, treatment containing seed treatment of Streptomyces sp (native) was found to be most effective in reducing pre and post emergence mortality with germination of 87.0 %. Streptomyces sp (native) was found to be more effective as compared to other bio-control agents in increasing the yield parameters like root length, shoot length, and seedling vigour and also it enhances the yield (322.8 Kg/ha) compared to other treatments. Streptomyces sp (native) needs further species level characterization and other biotrophic and tritrophic interaction studies for developing commercial formulation.
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Chilli is an important spice cum vegetable crop from the Solanaceae family grown around the world for its pungent flavor. Fusarium wilt of chilli is one of the most important diseases as it drastically reduces the yield. In the present experiment a total of 4 different botanical extracts and 4 isolates of Trichoderma were evaluated against the growth of wilt pathogen Fusarium solani. Among them neem oil and Tr1 isolate of Trichoderma recorded better results hence they were selected and further studied under pot culture conditions alone in combinations with panchagavya on the incidence of Fusarium wilt of chilli in local variety of chilli Out of 7 different treatments tested, highest germination (93.24%) was recorded from T7. And, highest percent disease inhibition of Fusarium wilt (69.56%) was recorded from T7. On a similar note, plant growth parameters such as improved plant height, dry weight, no. of flowers/ fruits per plant and fruit length were recorded from plants treated with T7. It was clear the IDM strategies perform better in reducing disease as compared to that of chemical control besides improving yield and growth of treated plants.
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Okra (Abelmoschus esculentus L.) also known as bhendi is one of the most common vegetable preferred in every household of India. Cercospora leaf spot incited by Cercospora abelmoschi. is one of the emerging disease in Uttar Pradesh Region. An experiment was conducted in Central Research Farm ,SHUATS , Prayagraj in Kharif season of 2022 to evaluate the efficacy of bioagents and chemicals viz., T0 – Untreated control,T1 Mancozeb (1%) + Trichoderma(4%) , T2 - Mancozeb (1%) + Pseudomonas(4%) , T3 Mancozeb (1%) + Bacillus subtilis(4%), T4 - Mancozeb (1%) + Trichoderma (2%) + Pseudomonas(2%) ,T5 - Mancozeb (1%) + Pseudomonas(2%) + Bacillus subtilis(2%), T6 Mancozeb (1%) + Bacillus subtilis(2%) + Trichoderma(2%), T7 - Mancozeb (1%) against Cercospora leaf spot of okra. C. abelmoschi initiates with sooty black, angular spots and cause heavy defoliation Studies revealed that minimum disease intensity was observed in T4 - Mancozeb (1%) + Trichoderma (2%) + Pseudomonas (2%) and is hereby considered as the best treatment out of all the treatments.
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Okra (Abelmoschus esculentus L.) is one of the fore most vegetable crop grown during kharif as well as summer seasons. Cercospora leaf spot incited by Cercospora spp. is one of the emerging disease in all regions wherever okra is grown. C. abelmoschi causes sooty black, angular spots and cause severe defoliation common during humid seasons. An experiment was conducted to evaluate the efficacy of bioagents and chemicals viz., T0 – Untreated control,T1 Mancozeb (1%) + Trichoderma (4%), T2 - Mancozeb (1%) + Pseudomonas (4%), T3 Mancozeb (1%) + Bacillus subtilis (4%), T4 - Mancozeb (1%) + Trichoderma(2%) + Pseudomonas(2%) ,T5 - Mancozeb (1%) + Pseudomonas (2%) + Bacillus subtilis (2%), T6 Mancozeb (1%) + Bacillus subtilis (2%) + Trichoderma (2%), T7 - Mancozeb (1%) against Cercospora leaf spot of okra. Studies revealed that minimum disease intensity , Maximum plant height , maximum no. of branches per plant and Maximum no. of fruits was observed in T4 - Mancozeb (1%) + Trichoderma (2%) + Pseudomonas (2%) and is hereby considered as the best treatment.
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The use of chemical pesticides in agricultural practices has led to significant concerns related to health and the environment in the last few decades. Consequently, there has been a growing interest in finding alternative approaches for controlling pests and diseases that are effective and environmentally friendly. Among these alternatives, Trichoderma harzianum has gained attention due to its remarkable ability to combat various crop pathogens in sustainable form. Biocontrol agents have gained prominence as eco-friendly substitutes for conventional chemical pesticides in disease management and crop improvement. T. harzianum, a common filamentous fungus, has emerged as a promising biocontrol agent due to its multifaceted strategies for suppressing diseases and promoting crop growth. T. harzianum fosters plant growth by aiding nutrient absorption, improving soil structure, and generating growth-promoting substances like auxins and cytokinins. Field studies have substantiated the efficacy of T. harzianum in managing a wide array of plant diseases caused by fungi, bacteria, and nematodes whether used as a seed treatment, soil application, or foliar spray, T. harzianum establishes a beneficial presence early in the plant's life cycle, providing enduring protection. In conclusion, T. harzianum holds immense potential as a biocontrol agent to sustainably safeguard crops. Trichoderma species play a pivotal role in managing plant diseases due to their versatile mechanisms. As global agriculture seeks alternatives to chemical pesticides, leveraging T. harzianum 's potential offers a valuable avenue toward resilient and environmentally safe crop production.
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Leaf blight of the medicinal plants (Piper longum, Tylophora indica and Hibiscus subdariffa) is one of the most significant diseases that severely damage these crops throughout the year in West Bengal .So, management of this disease is essential at now. A few chemical fungicides and trichoderma isolates were tested under in vitro against the disease-associated pathogens in order to identify efficient chemical fungicides and bio agents.It was found that the trichoderma isolates T-2 and T-3 were effective against Colletotrichum gloeosporioides, whereas the isolates T-1 and T-2 were efficient in inhibiting the growth of the test pathogen better against Fusarium sp. and Sclerotium rolfsii.With a higher percentage of inhibition (77.60%), copper oxychloride was the most effective fungicide against Fusarium sp. Conversely, carbendazim and dithane m-45 were more effective against Sclerotium rolfsii and Colletotrichum gloeosporioides, showing 78.33% and 77.93% of inhibition, respectively, and less effective against Fusarium sp at higher concentrations (400 ppm). To determine whether or not these chemical fungicides and trichoderma isolates are effective under in vivo, they will be tested in the field against the corresponding pathogens associated with the disease.
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The high cost of raw materials has long hindered the large-scale commercialization of Trichoderma sp. conidiophore formulations. This study addresses the need for affordable, nutrient-rich alternatives by exploring the use of lignocellulosic residues, comprising lignin, cellulose, and hemicellulose, as a carbon source for cultivating Trichoderma. These fungi possess unique attributes, such as filamentous growth and prolific spore production, which make them valuable for composting lignocellulosic waste. Using different substrates with a combination of straws effectively helps in the decomposition of straw and enhances the mass multiplication of Trichoderma lexii (T-94a). Different substrates such as broken rice, grounded maize, gram flour, sorghum & broken wheat were used in combination with paddy straws. A combination of Maize + straw induced an early response in the mycelial growth of Trichoderma lexii (T-94a). After 5 and 10 days of inoculation, physical parameters such as growth and sporulation characteristics of T. lexii (T-94a) show highly uniform growth and profuse sporulation on maize and straw combination as compared to broken rice & straw, sorghum, and straw combinations. This finding has promising implications for improving agricultural practices and waste management. Further research can fine-tune the optimal substrate ratios to enhance the efficiency of biocontrol agents and waste decomposition. Scaling up the application of these substrate combinations in real-world farming scenarios and assessing their long-term environmental impact will be essential for realizing the full potential of these sustainable agricultural practices.
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Rapeseed-mustard is one of the major oilseed crops cultivated in India. Alternaria brassicae is the most destructive pathogen of oilseeds. The present study aims to evaluate the effect of bio-agents and elicitors on Alternaria brassicae of mustard in vitro using dual culture technique and poison food technique in completely randomized design (CRD). The study was conducted at the Department of Plant Pathology Laboratory, Sam Higginbottom University of Agriculture, Technology And Sciences, Prayagraj. The results revealed that among bio-agents, Trichoderma viride + Pseudomonas fluorescens (87.70mm) recorded the maximum per cent inhibition of mycelial growth of Alternaria brassicae, followed by Trichoderma viride (82.60mm). Among elicitors, the maximum per cent inhibition of the pathogen was recorded by salicylic acid at 150 ppm (75.33%), followed by salicylic acid at 100 ppm (58.69%).
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The efficacy of four systemic and combination fungicides, namely Carbendazim 12% + Mancozeb 63% WP, Tebuconazole 25% WG, Hexaconazole 4% + Zineb 68% WG, and Tebuconazole 50% + Trifloxystrobin 25% WG, against Rhizoctonia bataticola, the causal agent of dry root rot in groundnut was evaluated using the poison food technique. Among the tested fungicides, Hexaconazole 4% + Zineb 68% WG exhibited the highest efficacy, with 100% inhibition of mycelial growth at all concentrations tested. Tests of compatibility of these fungicides with Trichoderma asperellum revealed that Hexaconazole 4% + Zineb 68% WG caused 38.88% of growth inhibition in T. asperellum while the other fungicides tested were totally incompatible with100 percent inhibition.
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Biocontrol agents (BCA) are an important tool for preventing crop losses and increasing plant food production in agriculture. Providing an eco-friendly and more sustainable alternative to chemical pesticides and fertilizers. A number of biocontrol applications are often conducted on the fungus Trichoderma, which is one of the most widely used and studied organisms in the field due to its numerous biocontrol traits, including parasitism, secondary metabolite production, antibiosis which induced plant defense mechanisms. It is well known that several Trichoderma species act as mycoparasites. Certain species within this group also possess the ability to oppose many organisms, including nematodes and plant pests, hence exhibiting their versatility as a biological control agent. The utilization of bioformulations incorporating Trichoderma species or other beneficial microbes for plants, such as plant growth-promoting bacteria (PGPB), has gained significant traction in the agricultural industry. The present review provides a comprehensive summary of the biocontrol studies conducted on six commonly utilized Trichoderma species. It focuses on elucidating their biocontrol attributes and explores the potential of incorporating these fungal genera into Trichoderma-based formulations for safeguarding against plant diseases. Furthermore, it discusses the viability of Trichoderma as a substitute for chemical pesticides and fertilizers.
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A survey was conducted during Rabi 2020 - 2021 to know the severity of Anthracnose of chilli in fields ofeight villages in Krishna district of Andhra Pradesh. The disease severity ranged from 13.30 to 29.65 per cent irrespective of location surveyed. The disease severity was least in Tadikilapudi village (13.3%) and highest in kamavarapukota village (29.65%) during the month of December 2020. To manage the disease severity evaluate the efficacy of bio-agents viz.., Seed Treatment with Pseudomonas fluorescens, Trichodermaviride and Microalgae.The foliar application of Neem oil is given to all the treatments. Among the Treatments disease intensity (%) was significantly decreased in treatment T6 - Pseudomonas fluorescens + Trichodermaviride + microalgae (16.03%). The plant height (cm) was significantly increased in treatment T6 – Pseudomonas fluorescens + Trichodermaviride + microalgae (62.2 cm) respectively, fruit length (cm) was significantly increased (15.96cm), yield was significantly increased in treatment T6 (128.33 q/ha) as compared to control T0.
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Mango gummosis caused by Lasiodiplodia theobromae (Pat.) Griffon & Moube [synonym: Botryodiplodia theobromae] is a serious disease in India especially on popular varieties of mango during monsoon and post-monsoon periods. Severe infection with pathogen causes up to 30- 100 % yield losses in mango. Gummosis infected orchards shows abundant gum secretion from branches, stem and main trunk and also Vascular discoloration. In severe cases infected mango trees may die. The pathogen produces grey-brown to black colonies with dense aerial mycelia on the PDA medium. Pycnidia were separate or aggregated, dark brown, thick or thin-walled. Conidiophores were hyaline, cylindrical to sub-obpyriform, with oblong, straight and hyaline single celled conidia andinitially. Gradually the conidia became dark brown and produced one septum with longitudinal striations. The pathogen has wide host range so difficult to manage the disease at field level. There are sevral Management strategies for mango gummosis like resistant or tolerant varieties, effective fungicides, botanicals and effective biological control agents role in disease management. This review attempts to summarize the Knowledge on mango gummosis, symptomotology, pathogen host range, morphological and cultural characters of Lasiodiplodia and management of the disease.
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The betelvine (Piper betle L.) is a perennial climbing evergreen shrub belonging to the Piperaceae, also known as the pepper family. As leaves has economic part of betelvine crop and consumed freshly harvested leaves. The present study were carried out at farmer’s field in collaboration with Department of Plant Pathology, Sam Higginbottom University of Agriculture Technology and Sciences Prayagraj to find out the ecological, environmentally and biological safe treatment to manage the disease as well increases the plant height and leaves length. Among all the treatment Trichoderma harzianum found to increase maximum plant height at different days after planting. While, Trichoderma viride increases the maximum leaves length and breadth during both the year.
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A field experiment entitled, Influence of different organic sources of nutrients on yield and quality of pomegranate (Punica granatum L.) cv. Bhagwa. The current study was designed in RBD with three replications consisting of twenty-two treatments with four different organic manures viz., farmyard manure, vermicompost, poultry manure and neem cake as a source of nitrogen and recommended dose of manure and fertilizers based on plant age with or without biofertilizers (Azotobacter, PSB & KMB) and biopesticides (Trichoderma viride and and Paecilomyces lilacinus). The findings based on pooled data discovered that maximum number of fruits per plant (59.50), yield per plant (12.41 kg), yield per hectare (19.86 tonne), fruit weight (239.39 g), fruit diameter (7.55 cm), aril weight per fruit (147.70 g), juice content per fruit (97.55 ml), peel weight per fruit (91.69 g) along with peel: aril ratio (0.62) was significantly noted under treatment 100 % RDN through poultry manure + 50 ml PSB + 25 ml KMB + 5 g Trichoderma viride + 5 ml Paecilomyces lilacinus. The maximum net return (4,12,840/ha) and BCR (3.26) were also obtained in same treatment. Significantly maximum TSS (16.15°Brix), reducing sugar (14.44 %), non-reducing (2.04 %) and total sugar (16.48 %) along with minimum titrable acidity (0.41 %) were recorded with application of 37.5 % RDN through FYM + 37.5 % RDN through neem cake + 50 ml Azotobacter culture + 50 ml PSB + 25 ml KMB + 5 g Trichoderma viride + 5 ml Paecilomyces lilacinus treatment. Whereas, maximum ascorbic acid content (16.72 mg/100 ml juice) was significantly found under treatment 100 % RDN through FYM + 50 ml PSB + 25 ml KMB + 5 g Trichoderma viride + 5 ml Paecilomyces lilacinus.
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Nine compounds were isolated from the crude extract of the solid culture of endophyte Trichoderma atroviride B7 of Colquhounia coccinea var. mollis by silica gel column chromatography, Sephadex LH-20 gel column chromatography, and HPLC. They were identified as atroviridanol (1), 3-oxo-3-[(2-phenylethyl) amino]-propanoic acid (2), N-(2′-phenylethyl)-acetamide (3), neoechinulin A (4), echinulin (5), gancidin W (6), N-isobutyl-3-methylbutanamide (7), 5-acetamido-1-pentanol (8), and N-2-methylpropyl-2-methylbutenamide (9) by NMR, HR-MS, and so on. Among them, compound 1 is a new compound, and compounds 2-9 are firstly isolated from Trichoderma spp.