Physiological and biochemical characterization of Trichoderma harzianum, a biological control agent against soilborne fungal plant pathogens

Grondona, Isabel; Hermosa, Rosa; Tejada, Margarita; Gomis, M; Mateos, Pedro F.; Bridge, P. D.; Monte, Enrique; García-Acha, Isabel

doi:10.1128/aem.63.8.3189-3198.1997

Cited by 134 publications

(68 citation statements)

References 48 publications

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“…Induction of BGN16.3 at a different pH or by nitrogen starvation was also tested, with negative results (not shown). (2), 0.5% chitin (3), 0.5% pustulan (4), 0.5% B. cinerea cell walls (5) or no carbon source (6). The purified BGN16.3 was used as positive control (7).…”

Section: Regulation Of the Bgn163 Productionmentioning

confidence: 99%

“…The antagonism by T. harzianum has been explained by different mechanisms [3]. One of them, mycoparasitism, involves the production of several hydrolytic enzymes for the local degradation of the host fungal cell wall and further penetration inside its hyphae as main steps [1].Several mycoparasitic strains included in different taxonomic groups in the Trichoderma genus [4,5] secrete complex sets of enzymes [6]. Within these enzymes we can find hydrolytic activities able to degrade most components of fungal cell walls (chitinases, glucanases, proteases, lipases, etc.).…”

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confidence: 99%

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BGN16.3, a novel acidic β‐1,6‐glucanase from mycoparasitic fungus Trichoderma harzianum CECT 2413

et al. 2005

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Trichoderma harzianum is a filamentous fungus that has been proposed as a potential biocontrol agent against phytopathogenic fungi [1] and more recently as opportunistic, avirulent plant symbiont [2]. The antagonism by T. harzianum has been explained by different mechanisms [3]. One of them, mycoparasitism, involves the production of several hydrolytic enzymes for the local degradation of the host fungal cell wall and further penetration inside its hyphae as main steps [1].Several mycoparasitic strains included in different taxonomic groups in the Trichoderma genus [4,5] secrete complex sets of enzymes [6]. Within these enzymes we can find hydrolytic activities able to degrade most components of fungal cell walls (chitinases, glucanases, proteases, lipases, etc.). These are usually present as isozyme groups composed by proteins with the same activity but different catalytic and molecular properties [7][8][9][10][11][12].Chitinases and b-1,3-glucanases are considered the main enzymes responsible for the degradation of the host cell walls, as chitin and b-1,3-glucan are their two major components. However, other enzymes hydrolyzing less abundant, but structurally important components (as b-1,6-glucan), can contribute to the efficient disorganization and further degradation of the cell wall by Trichoderma. b-1,6-glucan has been described in budding yeasts as the link between cell wall proteins and the main b-1,3-glucan ⁄ chitin polysaccharide [13] supporting an important role for this polymer in the structure of the fungal cell wall.Although b-1,6-glucanases are widely distributed among filamentous fungi, few of them have been purified and characterized [10,[14][15][16][17] and few gene sequences have been published [18][19][20][21][22].We have previously described two b-1,6-glucanases in T. harzianum CECT 2413: BGN16.1 and BGN16.2 A new component of the b-1,6-glucanase (EC 3.2.1.75) multienzymatic complex secreted by Trichoderma harzianum has been identified and fully characterized. The protein, namely BGN16.3, is the third isozyme displaying endo-b-1,6-glucanase activity described up to now in T. harzianum CECT 2413. BGN16.3 is an acidic b-1,6-glucanase that is specifically induced by the presence of fungal cell walls in T. harzianum growth media. The protein was purified to electrophoretical homogenity using its affinity to b-1,6-glucan as first purification step, followed by chomatofocusing and gel filtration. BGN16.3 has a molecular mass of 46 kDa in SDS ⁄ PAGE and a pI of 4.5. The enzyme only showed activity against substrates with b-1,6-glycosidic linkages, and it has an endohydrolytic mode of action as shown by HPLC analysis of the products of pustulan hydrolysis. The expression profile analysis of BGN16.3 showed a carbon source control of the accumulation of the enzyme, which is fast and strongly induced by fungal cell walls, a condition often regarded as mycoparasitic simulation. The likely involvement b-1,6-glucanases in this process is discussed.

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Section: Regulation Of the Bgn163 Productionmentioning

confidence: 99%

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confidence: 99%

BGN16.3, a novel acidic β‐1,6‐glucanase from mycoparasitic fungus Trichoderma harzianum CECT 2413

et al. 2005

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“…Fungi of this genus commonly occur in the natural environment. They are present in soil and are responsible for the decomposition of dead organic tissue [39]. Trichoderma spp.…”

Section: Discussionmentioning

confidence: 99%

“…are ubiquitous and colonize dead wood very quickly, although they are unable to colonize living trees. They are frequently isolated from dead spruce wood [39][40][41]. Interestingly, fungi belonging to this genus (e.g., T. harzianum) may be antagonistic to insects colonizing spruce trees, an example of such an insect being the bark beetle [9].…”

Section: Discussionmentioning

confidence: 99%

Communities of Microscopic Fungi in Dead Spruce in Relation to the Season and Degree of Wood Decay in Poland’s Karkonosze Mountains

Pusz¹,

Ogórek²

2020

Pol. J. Environ. Stud.

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By the end of the 20 th century the Karkonosze Mountains in southwestern Poland were largely associated with vast swathes of land covered by dead or dying spruce forests [1]. Their condition was dire enough to be regarded as an ecological disaster, and the blame was placed entirely on air pollution and acid rain. It is now a well-established fact that this was a multi-stage and multi-component process, and that these stages and components comprised, inter alia, herbivorous insect gradation, the presence of spruce monocultures and atmospheric conditions [2]. According to Manion [3], the phenomenon was also a consequence of the so-called 'decline disease spiral', which is a group of factors that Pol. AbstractTowards the end of the 20 th century the Karkonosze Mountains in southwestern Poland were characterized by vast swathes of dead and dying spruce forests whose condition at the time was described as an ecological disaster. Wood-decaying fungi play an important role in global carbon and nitrogen circulation by promoting the bioconversion of organic matter. The aim of our study was to determine the composition of the fungal community in the 'post-disturbance' dead spruce wood in relation to the season and degree of wood decay in the Karkonosze Mountains. Mycological analyses were carried out in 2015 and 2016 in the submontane and mountain forest zone, as well as in the subalpine zone of Karkonoski National Park (KNP). The findings reveal that the prevalent fungi colonising the internal tissues of dead wood belonged to the genus Trichoderma. Their proportion in the material under study increased in advanced stages of wood decay. However, the mycobiota of dead spruce at lower levels of decay were characterized by the highest diversity. The analysis also showed seasonal variations in the composition of the fungal communities colonizing dead wood.

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“…Soil-borne fungi from genus Trichoderma have been studied for their capacity to produce complex mixtures of extracellular enzymes (Grondona et al 1997), specially cellulases, and also for their antagonism against other fungi (Papavizas 1985). Strains of different taxons from the Trichoderma genus (Hermosa et al 2000 have invited agronomic interest not only because of their potential use as biocontrol agents against several fungi (Howell 2003), but also as plant avirulent symbionts that can trigger plant induced resistance and promote plant growth (Harman et al 2004).…”

Section: Introductionmentioning

confidence: 99%

Cloning and characterization ofbgn16·3, coding for a β-1,6-glucanase expressed duringTrichoderma harzianummycoparasitism

Montero

Sanz

Rey

et al. 2007

Journal of Applied Microbiology

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Aims: To clone and characterize the gene coding for BGN16·3, a β‐1,6‐glucanase putatively implicated in mycoparasitism by Trichoderma harzianum, a biocontrol agent used against plant pathogenic fungi. Methods and Results: Using degenerate primed PCR and cDNA library screening, we have cloned the cDNA coding BGN16·3. bgn16·3 showed a significant sequence identity (50%) to bgn16·1; however, they both have low identity to the previously cloned bgn16·2, allowing the identification of amino acid sequences putatively involved in the common catalytic activity of the three proteins. bgn16·3 is a single‐copy gene and highly homologous sequences are present in all tested Trichoderma species. bgn16·3 expression pattern is analysed by Northern blot, finding that it is expressed during the interaction of T. harzianum CECT 2413 with Botrytis cinerea, supporting the implication of the enzyme in the mycoparasitic process. Conclusions: The cloned bgn16·3 completes the knowledge on the β‐1,6‐glucanase isozyme system from T. harzianum CECT 2413. A highly homologous gene is present in all analysed Trichoderma strains. bgn16·3 is expressed under few specific conditions, including the mycoparasitic process. Significance and Impact of the Study: This study contributes to the knowledge of β‐1,6‐glucanases. It implicates this group of enzymes in the mycoparasitism by some biocontrol agents such as T. harzianum.

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Physiological and biochemical characterization of Trichoderma harzianum, a biological control agent against soilborne fungal plant pathogens

Cited by 134 publications

References 48 publications

BGN16.3, a novel acidic β‐1,6‐glucanase from mycoparasitic fungus Trichoderma harzianum CECT 2413

BGN16.3, a novel acidic β‐1,6‐glucanase from mycoparasitic fungus Trichoderma harzianum CECT 2413

Communities of Microscopic Fungi in Dead Spruce in Relation to the Season and Degree of Wood Decay in Poland’s Karkonosze Mountains

Cloning and characterization ofbgn16·3, coding for a β-1,6-glucanase expressed duringTrichoderma harzianummycoparasitism

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