Monitoring of Trichoderma species in agricultural soil in response to application of biopreparations

Oskiera, Michał; Szczech, Magdalena; Stępowska, A.; Smolińska, U.; Bartoszewski, Grzegorz

doi:10.1016/j.biocontrol.2017.07.005

Cited by 29 publications

(24 citation statements)

References 43 publications

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“…Assuming 100% DNA recovery, the protocol quantified up to 2 × 10 3 and 4 × 10 4 conidia g -1 in plants and soil, respectively. These results agree with those reported for fungi other than Trichoderma ( Selma et al, 2008 ; Garrido et al, 2009 ) and for other Trichoderma species, such as T. virens ( Dodd et al, 2004 ; Oskiera et al, 2017 ).…”

Section: Discussionsupporting

confidence: 92%

A Ready-to-Use Single- and Duplex-TaqMan-qPCR Assay to Detect and Quantify the Biocontrol Agents Trichoderma asperellum and Trichoderma gamsii

et al. 2018

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Trichoderma asperellum strain icc012 and Trichoderma gamsii strain icc080, the microbial active ingredients of RemedierTM (ISAGRO, Novara, Italy), are biocontrol agents (BCAs) employable for crop protection against a wide range of fungal pathogens, including soil-borne pathogens and fungi involved in grapevine trunk disease. In this study, single and duplex real-time quantitative PCR (qPCR) methods to detect and quantify T. asperellum and T. gamsii were developed. Primers/probe sets were designed on the T. asperellum and T. gamsii rpb2 genes and tested for specificity on a panel of microorganisms commonly associated with grape wood and soil. No differences were observed comparing single- and duplex-qPCR assays on different BCAs, 1 pg of target DNA was detected approximately at Cq = 34. R2-values and the efficiency were always equal to 0.99 and >80%, respectively. The detection limit of the duplex-qPCR assay on artificially inoculated samples was 2 × 103 and 4 × 104 conidia g-1 of grape wood tissue and soil, respectively. The methods will be useful to better schedule BCA application in the field and in grapevine nurseries, as well as for investigating the dynamic of BCA populations.

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Section: Discussionsupporting

confidence: 92%

A Ready-to-Use Single- and Duplex-TaqMan-qPCR Assay to Detect and Quantify the Biocontrol Agents Trichoderma asperellum and Trichoderma gamsii

et al. 2018

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“…Field experiments with strain SCI (Longa et al 2009) also reported greater colonization (1 9 10 6 CFU per gram soil) in the upper soil profile of the grapevine rhizosphere for up to 18 weeks after inoculation. Similarly, T. atroviride and T. harzianum inoculants in open-field lettuce production persisted for up to 2 years at levels of ≥10 4 CFU per gram of soil (Oskiera et al 2017). These studies however, only provide information on Trichoderma soil colonization and not the interactions with plant roots.…”

Section: Discussionmentioning

confidence: 96%

Quantification of Trichoderma afroharzianum, Trichoderma harzianum and Trichoderma gamsii inoculants in soil, the wheat rhizosphere and in planta suppression of the crown rot pathogen Fusarium pseudograminearum

Stummer

Zhang

et al. 2020

J. Appl. Microbiol.

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Aims Develop quantitative assays (qPCR) to determine the detection threshold limits, colonization and persistence of Trichoderma gamsii, Trichoderma afroharzianum and T. harzianum inoculants in cropping soils, the wheat rhizosphere and their in planta suppressive efficacy against the crown rot pathogen Fusarium pseudograminearum. Methods and Results Trichoderma qPCR primers were designed from the internal transcribed spacer region of 5.8S rDNA and from sequences of DNA fragments diagnostic for each inoculant genotype. The minimum detection thresholds of qPCR assays varied between 1 × 103 (log 3) and 8 × 104 (log 4·9) conidia (genome) equivalents per gram of soil for multi‐ and single‐copy target sequences, respectively and were independent of soil type. There was a strong correlation (r > 0·974) between culture‐dependent and culture‐independent (qPCR) quantification methods. In wheat bioassays, Trichoderma inoculants colonized rhizosphere soils and wheat roots at 56–112 days postemergence to a depth of 20 cm but were more abundant (P < 0·001) at 0–10 cm root depth, means ranging from 2 × 102 (log 2·3) to 4 × 105 (log 5·6) conidia equivalents per gram of rhizosphere soil or root tissue. Inoculants reduced (P < 0·001) F. pseudograminearum biomass in wheat crown and root tissue by up to 5754‐fold and increased (P = 0·008) plant biomass, relative to the pathogen control. Conclusions The qPCR assays provided sensitive and accurate assessment of wheat root and rhizosphere soil colonization of Trichoderma inoculants. Strains persisted through to grain maturity at levels shown to significantly suppress F. pseudograminearum in planta. Significance and Impact of the Study The qPCR assays developed here were used to determine the wheat rhizosphere dynamics of T. harzianum, T. afroharzianum and T. gamsii inoculants and their suppressive efficacy against F. pseudograminearum in planta. These assays can be applied to monitor inoculant dynamics in suppressing crown rot and other wheat root diseases in the field.

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“…Recently, Oskiera et al ( 2017 ) described a multiplex PCR assay for monitoring the population levels of T. atroviride and T. harzianum applied as biopreparations in the field. For the detection in soil, a maximum of 3 primer pairs targeting markers TaQ (Q01_3F/Q01_3R) or TaX (X18_1F/X18_3R) (Table 1 ), the Trichoderma chitinase-specific chit42_1af/chit42_2ar (Kullnig-Gradinger et al, 2002 ) and ITS primers (5.8SR/LR6) specific to fungi were combined for the detection of T. atroviride , while two markers, TVD (TVD_If/TVD_Ir) (Bhat, 2007 ) and ThQ (QTh_5F/QTh_4R) (Table 1 ) were combined in multiplex-PCR for T. harzianum detection.…”

Section: Species-specific Pcrmentioning

confidence: 99%

Molecular Tools for Monitoring Trichoderma in Agricultural Environments

et al. 2018

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Various Trichoderma species possess significance in agricultural systems as biofertilizers or biocontrol agents (BCAs). Besides these beneficial features, certain Trichoderma species can also act as agricultural pests, causing the green mold disease of cultivated mushrooms. This double-faced nature of the genus in agricultural environments points at the importance of proper monitoring tools, which can be used to follow the presence and performance of candidate as well as patented and/or registered biocontrol strains, to assess the possible risks arising from their application, but also to track harmful, unwanted Trichoderma species like the green molds in mushroom growing facilities. The objective of this review is to discuss the molecular tools available for the species- and strain-specific monitoring of Trichoderma, ranging from immunological approaches and fingerprinting tools to exogenous markers, specific primers used in polymerase chain reaction (PCR) as well as “omics” approaches.

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Monitoring of Trichoderma species in agricultural soil in response to application of biopreparations

Cited by 29 publications

References 43 publications

A Ready-to-Use Single- and Duplex-TaqMan-qPCR Assay to Detect and Quantify the Biocontrol Agents Trichoderma asperellum and Trichoderma gamsii

A Ready-to-Use Single- and Duplex-TaqMan-qPCR Assay to Detect and Quantify the Biocontrol Agents Trichoderma asperellum and Trichoderma gamsii

Quantification of Trichoderma afroharzianum, Trichoderma harzianum and Trichoderma gamsii inoculants in soil, the wheat rhizosphere and in planta suppression of the crown rot pathogen Fusarium pseudograminearum

Molecular Tools for Monitoring Trichoderma in Agricultural Environments

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