Quantification of the biocontrol agent Trichoderma harzianum with real-time TaqMan PCR and its potential extrapolation to the hyphal biomass

López-Mondéjar, Rubén; Antón, Anabel; Raidl, Stefan; Ros, Margarita; Pascual, José Antonio

doi:10.1016/j.biortech.2009.10.019

Cited by 65 publications

(40 citation statements)

References 18 publications

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“…Techniques used so far include plating out of aggregates to obtain colony forming units, ergosterol essay (Feeney et al, 2006), MAb-ELISAfor specific species or quantification by PCR (Lopez-Mondejar et al, 2009). These techniques however only enable a single snap shot in time, and, perhaps more importantly, require the destruction of the physical environment which contributed to the growth dynamics.…”

Section: Discussionmentioning

confidence: 99%

Modelling and quantifying the effect of heterogeneity in soil physical conditions on fungal growth

et al. 2010

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Abstract. Despite the importance of fungi in soil ecosystem services, a theoretical framework that links soil management strategies with fungal ecology is still lacking. One of the key challenges is to understand how the complex geometrical shape of pores in soil affects fungal spread and species interaction. Progress in this area has long been hampered by a lack of experimental techniques for quantification. In this paper we use X-ray computed tomography to quantify and characterize the pore geometry at microscopic scales (30 µm) that are relevant for fungal spread in soil. We analysed the pore geometry for replicated samples with bulk-densities ranging from 1.2-1.6 g/cm 3 . The bulk-density of soils significantly affected the total volume, mean pore diameter and connectivity of the pore volume. A previously described fungal growth model comprising a minimal set of physiological processes required to produce a range of phenotypic responses was used to analyse the effect of these geometric descriptors on fungal invasion, and we showed that the degree and rate of fungal invasion was affected mainly by pore volume and pore connectivity. The presented experimental and theoretical framework is a significant first step towards understanding how environmental change and soil management impact on fungal diversity in soils.

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Section: Discussionmentioning

confidence: 99%

Modelling and quantifying the effect of heterogeneity in soil physical conditions on fungal growth

et al. 2010

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“…The standard curves were developed according to López-Mondéjar et al (2010). The approximately 242 bp fragment from the selected sequence characterized amplified region (SCAR) of FOC was cloned into a PMD-19 vector (TaKaRa Bio Inc., Dalian, China).…”

Section: Real-time Pcr Quantification Of Soil Focmentioning

confidence: 99%

Control of soil-borne pathogen Fusarium oxysporum by biological soil disinfestation with incorporation of various organic matters

et al. 2015

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Banana Fusarium wilt caused by Fusarium oxysporum f. sp. cubense (FOC) is a worldwide soilborne disease that causes serious economic losses every year. Effective ways of preventing the disease are still not available. In this study, biological soil disinfestation (BSD) with incorporation of various organic matters under flooding conditions combined with plastic mulching was applied to suppress FOC and prevent banana Fusarium wilt disease. The biological properties of the soil during the BSD process were investigated using quantitative real-time PCR and denaturing gradient gel electrophoresis. The results showed that the values of soil redox potential significantly decreased by >800 mV in the treatments that incorporated rice straw and bagasse under flooding conditions compared with untreated soil. The lowest soil pH (5.36) and a significant decline in the FOC population to 2.79 % of untreated soil were obtained in flooded soil with the highest amount of rice straw. Incorporation of bagasse, pig manure, and lime also significantly decreased the populations of FOC, but were less effective than rice straw. Application of rice straw under flooding conditions significantly increased soil bacterial diversity, and the increased bacteria were considered to have an antagonistic effect on FOC. In 2013 field experiment, a wilt disease control efficiency of 82.3 % was obtained in flooded soil incorporated with 0.5 % (w/w) rice straw compared with untreated soil, and the control efficiency of BSD was further confirmed in 2014 field experiment. These results indicated that BSD can reduce FOC populations in soil, ameliorate soil microbial communities, and reduce the occurrence of banana Fusarium wilt disease.

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“…Next, 3 μl total RNA was reverse-transcribed in a 20 μl reaction volume with the PrimeScript 1st Strand cDNA Synthesis Kit (TaKaRa), using the specific primer according to the manufacturer's instructions. The internal transcribed spacer (ITS) regions of SQR-T037 were amplified in a realtime PCR assay using primers (ITS1 S and ITS1 R) designed by López-Mondéjar et al (2010) and Beaulieu et al (2011). The strain-specific TaqMan probe ITS TM-037 Fam: 5′-FAM-AACTCTTTTTGTATACCCCCTC GCGGGT-TAMRA-3′ (FAM: 6-carboxyfluorescein, TAMRA: 6-carboxy-tetramethylrhodamine) (TaKaRa) was modified from that described by López-Mondéjar et al (2010), which was designed on one nucleotide mismatch of ITS where no sequence in the database (http://www.ncbi.nlm.nih.gov/BLAST) was identical to our isolate.…”

Section: Soil Rna Extraction and Rt-qpcrmentioning

confidence: 99%

Colonization of Trichoderma harzianum strain SQR-T037 on tomato roots and its relationship to plant growth, nutrient availability and soil microflora

et al. 2014

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Aims This research was to study the capacity of Trichoderma harzianum strain SQR-T037 to colonize tomato roots and how this strain and its bio-organic fertilizer stimulate plant growth and increase yields under field conditions. Methods Field trials were conducted with a reduced application of chemical fertilizer (75 % of the recommended application) plus Trichoderma-enriched bioorganic fertilizer (BF) or organic fertilizer (OF) or Trichoderma spore suspension (SS), while 100 % of the recommended chemical fertilizer (CF) was used as control.Results Trichoderma SQR-T037 could efficiently colonize tomato roots and soils based on the reverse transcription-quantitative PCR analysis, and significantly stimulate biomass accumulation at an early stage. The 75 % rates of chemical fertilizer coupled with bioorganic fertilizer (BF) produced tomato yields equivalent to those obtained using the 100 % rates of chemical fertilizer (CF), while inoculation with the Trichoderma alone (SS) or supplement with organic fertilizer alone (OF) would cause 11 and 13 % decreases in yield over the control (CF). The efficacy of BF for maintaining a stable tomato yield may be due to the enhanced soil nutrients availability and the increased abundance of soil microflora, including bacteria, fungi, actinomycetes and Trichoderma communities, which had a positive linear correlation in most of the cases revealed by the Pearson correlation analysis. Plant SoilConclusions The results of this study imply that the reason by which the bio-organic fertilizer promotes plant growth and increases the tomato yields can be attributed to the enhancement of the rhizospheric microflora, which promotes nutrient activation. Therefore, T. harzianum could be employed in a combination with composts. In this way, the application rate of chemical fertilizers can be practically decreased by 25 % to obtain maximum benefits.

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Quantification of the biocontrol agent Trichoderma harzianum with real-time TaqMan PCR and its potential extrapolation to the hyphal biomass

Cited by 65 publications

References 18 publications

Modelling and quantifying the effect of heterogeneity in soil physical conditions on fungal growth

Modelling and quantifying the effect of heterogeneity in soil physical conditions on fungal growth

Control of soil-borne pathogen Fusarium oxysporum by biological soil disinfestation with incorporation of various organic matters

Colonization of Trichoderma harzianum strain SQR-T037 on tomato roots and its relationship to plant growth, nutrient availability and soil microflora

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