Isolation, production and <i>in vitro</i> effects of the major secondary metabolite produced by <i>Trichoderma</i> species used for the control of grapevine trunk diseases

Mutawila, Cheusi; Vinale, Francesco; Halleen, F.; Lorito, Matteo; Mostert, Lizel

doi:10.1111/ppa.12385

Cited by 56 publications

(42 citation statements)

References 33 publications

(43 reference statements)

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“…After selection of endophytic fungi by the in vitro diffusible metabolite test according to the methodology to Mutawila et al (2015), only one sample was chosen for synergism evaluation. The extract of endophytic fungus was used together with extract of L. sidoides.…”

Section: Synergism Evaluationmentioning

confidence: 99%

Secondary metabolites from endophytic fungus from Lippia sidoides Cham.

Ferreira¹,

Santos²,

Soares³

et al. 2017

J. Med. Plants Res.

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Lippia sidoidesCham. (Verbenaceae) is a species native to the Brazilian northeast, widely used in popular medicine. Its leaves were used for the isolation of endophytic fungi and extraction of metabolites. Among them, three were selected according to fungitoxicity tests against the maize phytopathogenic fungus, Curvularia lunata (Wakker). However, the objective of this study was to identify the role of L. sidoides extracts associated with their endophytic fungi, necessary to reduce excess of fungicides applied on the maize crop. Metabolites were evaluated for antioxidant activity by 2.2-diphenyl-1-picrylhydrazyl (DPPH), phenols, total flavonoids and one of it endophytic fungus were evaluated for synergism (Verticilium sp. and plant extracts). The endophytic fungi and plant extracts evaluated for phenolic content ranged from 0.29 ± 0.05 to 96.94 ± 11.86 mgEAG/g, the content of flavonoids from 14.31 ± 1.56 to 192.33 ± 4.58 mgER/g, and antioxidant activity could only be observed for the plant extract with EC 50 81 ± 0.3%. The secondary metabolites identified by HPLC in the plant extract were catechin, quercetin, gallic acid and naringin. Naringenin, catechin, epigallocatechin gallate and quercetin were identified in the extract of the fungi viz. Verticillium sp. and Fusarium sp. Synergistic analysis between a 1:1 proportion of plant and fungal extracts has shown more efficient (79.0%) inhibition of C. lunata. Thus, alternative control of phytopathogenic fungi can be accomplished using plant extracts associated with their endophytic fungi, reducing the excess of fungicides applied on the maize crop.

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Section: Synergism Evaluationmentioning

confidence: 99%

Secondary metabolites from endophytic fungus from Lippia sidoides Cham.

Ferreira¹,

Santos²,

Soares³

et al. 2017

J. Med. Plants Res.

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“…To evaluate the capacity of TaWT, Tr, and Tv of growing using Phi as the P source, we used a modified Vogel's Minimal Medium (VMM) recipe [34], in which the source of P, monopotassium phosphate, was replaced by monopotassium phosphite. VMM is commonly used to cultivate Neurospora crassa, however, it has been also successfully used to grow Trichoderma species including TaWT, Tr, Tv, and T. asperellum [35,36,37,38]. VMM was prepared as a 50X solution and used as a 1X solution and supplemented with glucose 2% and Bacto Agar (DIFCOÔ, USA) 1.5% when needed.…”

Section: Evaluation Of the Capacity Of Trichoderma Wild Type Strains mentioning

confidence: 99%

Assessment of the ptxD gene as a growth and selective marker in Trichoderma atroviride using Pccg6, a novel constitutive promoter

Carreras-Villaseñor

Rico-Ruiz

Lopez-Hernandez

et al. 2019

Preprint

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BackgroundTrichoderma species are among the most effective cell factories to produce recombinant proteins, whose productivity relies on the molecular toolkit and promoters available for the expression of the target protein. Although inducible promoter systems have been developed for producing recombinant proteins in Trichoderma, constitutive promoters are often a desirable alternative. Constitutive promoters are simple to use, do not require external stimuli or chemical inducers to be activated, and lead to purer enzyme preparations. Moreover, most of the promoters for homologous and heterologous expression reported in Trichoderma have been commonly evaluated by directly assessing production of industrial enzymes, requiring optimization of laborious protocols. ResultsHere we report the identification of Pccg6, a novel Trichoderma atroviride constitutive promoter, that has similar transcriptional strength as that of the commonly used pki1 promoter. Pccg6 displayed conserved arrangements of transcription factor binding sites between promoter sequences of Trichoderma ccg6 orthologues genes, potentially involved in their regulatory properties. The predicted ccg6-encoded protein potentially belongs to the SPE1/SPI1 protein family and shares high identity with CCG6 orthologue sequences from other fungal species including Trichoderma reesei, Trichoderma virens, Trichoderma asperellum, and lo a lesser extent to that of Neurosposa crassa. We also report the use of the Pccg6 promoter to drive the expression of PTXD, a phosphite oxidoreductase of bacterial origin, which allowed T. atroviride to utilize phosphite as a sole source of phosphorus. We propose ptxD as a growth reporter gene that allows real-time comparison of the functionality of different promoters by monitoring growth of Trichoderma transgenic lines and enzymatic activity of PTXD. Finally, we show that constitutive expression of ptxD provided T. atroviride a competitive advantage to outgrow bacterial contaminants when supplied with phosphite as a sole source of phosphorus. ConclusionsA new constitutive promoter, ccg6, for expression of homologous and heterologous proteins has been 3 identified and tested in T. atroviride by expressing PTXD, which resulted in an effective and visible phenotype to evaluate transcriptional activity of sequence promoters. Use of PTXD as a growth marker holds great potential for assessing activity of other promoters and for biotechnological applications as a contamination control system. BackgroundTrichoderma is a fungal genus with high level of genetic diversity and great adaptability to different environmental conditions. These characteristics, together with factors such as their high capacity to produce and secrete proteins and bioactive compounds, as well as the availability of protocols and molecular tools for their genetic manipulation, have posed Trichoderma as a model organism to study fungal biology and to develop biotechnological applications for different industries [1].Some examples of the industrial applications of Tric...

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“…Mutawila et al (2016a) recommend the application of Trichoderma after six hours from the end of sap exudation from cuts and Kotze et al (2011) observed that the use of Trichoderma showed better results than the use of chemicals. Mutawila et al (2016b) found that the 6-pentyl-a-pyrone (6PP) component produced by Trichoderma harzianum and Trichoderma atroviride was shown to inhibit trunk pathogens. In addition, the cleaning of tools is another practice that significantly reduces the infection of plant pathogens in mother plants.…”

Section: Advances Observed In the Management Of New Diseasesmentioning

confidence: 99%

Advances in Propagation of Grapevine in the World

et al. 2017

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-Grapevine production by classical grafting methods and in commercial scale emerged over 130 years. This system remained handmade until the mid-1950s, when emerged the first international certification programs aimed at obtaining mother plants with high viral sanity. The necessity to increase the scale of production on industrial model and plant material production based on minimum morphological standards appeared at the end of the 1960s. Along the 1970s, research unlocked knowledge on semi-automated grafting, process hygiene, use of plant growth regulators and understanding of physiological events of rootstock-scion compatibility, callus formation and rooting. So, until the mid-2000s, certification schemes and propagation processes advanced little in technical standard. However, grapevine growing areas were expanded and demands for plant material increased, and new diseases emerged from contaminated nurseries. These new diseases (new viral complexes, phytoplasmas, bacteria and grapevine trunk diseases) were discovered by high-sensitivity diagnostic methods. Today, there is a new discussion on the nursery segment worldwide. The propagation techniques have been reviewed from the perspective of reducing the incidence of new diseases and minimum physiological damage of nursery plants during the production stages. Therefore, technological innovations regarding equipment, practices and production inputs have been incorporated in new certification schemes. However, despite these advantages, these schemes have become more complex and multidisciplinary than previous ones, bringing difficulties in adaptation of nurserymen. Index-terms: Vitis grafting, nursery, certification, grapevine cuttings. AVANÇOS OBSERVADOS NA PROPAGAÇÃO DE VIDEIRAS NO MUNDORESUMO -A propagação de videiras a partir dos métodos clássicos de enxertia e em escala comercial teve origem há mais de 130 anos. Este sistema permaneceu artesanal até meados da década de 1950, quando se iniciaram os primeiros programas internacionais de certificação com foco na obtenção de plantas básicas com elevada sanidade para vírus. No fim da década de 1960, surgiu a necessidade de aumentar a escala para produção em um modelo industrial em que a muda apresentasse um padrão morfológico mínimo. Ao longo da década de 1970, aprofundaram-se as pesquisas relacionadas à automatização da prática de enxertia, à higienização do processo, ao uso de reguladores de crescimento e ao entendimento dos eventos fisiológicos da compatibilidade entre enxerto e porta-enxerto, formação de calos e enraizamentos. Assim, até meados dos anos 2000, os esquemas de certificação e o processo de propagação pouco evoluíram em termos técnicos. Porém, a medida que a área vitícola foi expandindo e a demanda por mudas aumentando, verificou-se que novas doenças se alastravam em escala global a partir de viveiros contaminados. Estas novas doenças (complexos virais, fitoplasmas, bactérias e fungos causadores de podridões vasculares) foram sendo descobertas à medida que os métodos de diagnose avançaram...

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Isolation, production and in vitro effects of the major secondary metabolite produced by Trichoderma species used for the control of grapevine trunk diseases

Cited by 56 publications

References 33 publications

Secondary metabolites from endophytic fungus from Lippia sidoides Cham.

Secondary metabolites from endophytic fungus from Lippia sidoides Cham.

Assessment of the ptxD gene as a growth and selective marker in Trichoderma atroviride using Pccg6, a novel constitutive promoter

Advances in Propagation of Grapevine in the World

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