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Control Effect and Mechanism of Trichoderma asperellum TM11 against Blueberry Root Rot

Si Li
Si Li
, 
Fu-Mei Zhang
Fu-Mei Zhang
, 
Xiao-Jing Shang
Xiao-Jing Shang
 und 
Rui Hou
Rui Hou
   | 20. Sept. 2023
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Article Category: Original Paper
Online veröffentlicht: 20. Sept. 2023
Seitenbereich: 325 - 337
Eingereicht: 25. Mai 2023
Akzeptiert: 02. Aug. 2023
DOI: https://doi.org/10.33073/pjm-2023-034
Schlüsselwörter
© 2023 Si Li et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Inhibitory activity of strain TM11 against Fusarium commune and Fusarium oxysporum.CK is F. commune and F. oxysporum alone; on the left side of each plate is the TM11 colony, and on the right side is the colony of F. commune or F. oxysporum.
Inhibitory activity of strain TM11 against Fusarium commune and Fusarium oxysporum.CK is F. commune and F. oxysporum alone; on the left side of each plate is the TM11 colony, and on the right side is the colony of F. commune or F. oxysporum.

Fig. 2.

Morphological characteristics of strain TM11.A–B) colony of TM11 surface and base, C) conidiophores, D) conidia (scale 20 microns).
Morphological characteristics of strain TM11.A–B) colony of TM11 surface and base, C) conidiophores, D) conidia (scale 20 microns).

Fig. 3.

Phylogenetic trees based on ITS (A) and SSU (B) sequences using the neighbor-joining method.The outgroup was Schizophyllum commune and Nectria berolinensis; numerical values above the branches are bootstrap percentiles from 1,000 replicates. Parsimony bootstrap values of more than 50% are shown at the nodes.
Phylogenetic trees based on ITS (A) and SSU (B) sequences using the neighbor-joining method.The outgroup was Schizophyllum commune and Nectria berolinensis; numerical values above the branches are bootstrap percentiles from 1,000 replicates. Parsimony bootstrap values of more than 50% are shown at the nodes.

Fig. 4.

Inhibition of Fusarium oxysporum and Fusarium commune hyphal growth by Trichoderma asperellum strain TM11 under a microscope.A) Strain TM11 causes lysis of F. oxysporum lysis of the hyphae (as shown by arrow), B) strain TM11 causes F. oxysporum lysis of the hyphae, and the cytoplasm has leaked out of that hyphal (as shown by arrow), C) strain TM11 to coil around the hyphae of F. commune (as shown by arrow), D) strain TM11 causes F. commune lysis of the hyphae, and the cytoplasm has leaked out of that hyphal (as shown by arrow).
Inhibition of Fusarium oxysporum and Fusarium commune hyphal growth by Trichoderma asperellum strain TM11 under a microscope.A) Strain TM11 causes lysis of F. oxysporum lysis of the hyphae (as shown by arrow), B) strain TM11 causes F. oxysporum lysis of the hyphae, and the cytoplasm has leaked out of that hyphal (as shown by arrow), C) strain TM11 to coil around the hyphae of F. commune (as shown by arrow), D) strain TM11 causes F. commune lysis of the hyphae, and the cytoplasm has leaked out of that hyphal (as shown by arrow).

Fig. 5.

Inhibitory effects of volatile metabolites and fermentation metabolites of Trichoderma asperellum strain TM11 on the growth of Fusarium commune and Fusarum oxysporum.CK1 and CK2 represent the growth of F. commune and F. oxysporum on PDA; volatile metabolites mean volatile organic compounds (VOCs) and fermentation metabolites mean secondary metabolites, including alkaloids, ketones, esters, phenols and organic acids, such as erucamide, dibutyl phthalate and benzophenone.
Inhibitory effects of volatile metabolites and fermentation metabolites of Trichoderma asperellum strain TM11 on the growth of Fusarium commune and Fusarum oxysporum.CK1 and CK2 represent the growth of F. commune and F. oxysporum on PDA; volatile metabolites mean volatile organic compounds (VOCs) and fermentation metabolites mean secondary metabolites, including alkaloids, ketones, esters, phenols and organic acids, such as erucamide, dibutyl phthalate and benzophenone.

Fig. 6.

Effect of blueberry seedling growth inoculated with Fusarium oxysporum or Fusarium commune.A) Control group: inoculated with F. commune alone, B) treatment 1: F. commune inoculated first, and TM11 inoculated 48 hours later, C) treatment 2: TM11 inoculated first, then F. commune inoculated 48 hours later, D) control group: inoculated with F. oxysporum alone, E) treatment 1: F. oxysporum inoculated first, and TM11 inoculated 48 hours later, F) treatment 2: TM11 inoculated first, then F. oxysporum inoculated 48 hours later.
Effect of blueberry seedling growth inoculated with Fusarium oxysporum or Fusarium commune.A) Control group: inoculated with F. commune alone, B) treatment 1: F. commune inoculated first, and TM11 inoculated 48 hours later, C) treatment 2: TM11 inoculated first, then F. commune inoculated 48 hours later, D) control group: inoculated with F. oxysporum alone, E) treatment 1: F. oxysporum inoculated first, and TM11 inoculated 48 hours later, F) treatment 2: TM11 inoculated first, then F. oxysporum inoculated 48 hours later.

Inhibitory effects of volatile and fermentation metabolites of Trichoderma asperellum strain TM11 on the growth of Fusarium commune and Fusarium oxysporum.

Treatment Fusarium commune Fusarium oxysporum
Diameter of pathogen colony (cm) Percentage inhibition (%) Diameter of pathogen colony (cm) Percentage inhibition (%)
CK1 5.77 ± 0.05a n.d. 5.97 ± 0.05a n.d.
Volatile metabolites 3.83 ± 0.04b 33.53 ± 0.72a 3.77 ± 0.17b 36.87 ± 2.74a
CK2 7.00 ± 0.22a n.d. 8.87 ± 0.05a n.d.
Fermentation metabolites 5.83 ± 0.05b 16.67 ± 2.81b 5.60 ± 0.08b 36.84 ± 0.98a

Types and relative abundances of antimicrobial metabolites produced in culture filtrates by Trichoderma asperellum strain TM11 based on HPLC–MS analysis of methanol extracts of fermentation broth.

Chemical compound Peak area Retention time (min) m/z
Erucamide 221862.78 7.22 338
Dibutyl phthalate 158594.53 6.73 279
Benzophenone 24466.55 6.00 183
Benzothiazole 49711.09 7.09 136
Citric acid 49252.02 1.11 193
Betainel 6165.18 0.89 118
Dipropyl phthalate 5286.64 6.07 251
alpha-Curcumene 3619.38 5.43 203
Phenylalanine 2535.44 2.52 166
3-Hydroxycinnamic acid 2500.47 2.90 165
Chlorogenic acid 1903.63 3.02 355
4-Hydroxybenzoic acid 517.80 3.16 139

Effects of inoculation with Trichoderma asperellum strain TM11 and Fusarium commune or Fusarium oxysporum on SOD, POD, and CAT activities in leaf and root tissue of blueberry seedlings.

Treatment Leaf Root
CAT (U/g fresh weight) SOD (U/g fresh weight) POD (U/g fresh weight) CAT (U/g fresh weight) SOD (U/g fresh weight) POD (U/g fresh weight)
CK 128.14 ± 5.86b 208.01 ± 4.05c 142.08 ± 4.05d 140.96 ± 3.30c 406.44 ± 2.02b 150.86 ± 3.60d
TM11 224.59 ± 12.19a 446.95 ± 3.21a 353.22 ± 3.37a 279.22 ± 1.36a 490.13 ± 1.77a 363.61 ± 11.14a
TM11 + F. commune 217.09 ± 7.41a 352.75 ± 4.79b 309.32 ± 6.20b 228.79 ± 3.43b 469.36 ± 4.60a 313.66 ± 4.75b
TM11 + F. oxysporum 213.37 ± 9.78a 349.54 ± 4.29b 205.23 ± 0.30c 274.70 ± 1.65a 490.39 ± 12.74a 306.37 ± 9.38b

Relative control effect of Trichoderma asperellum strain TM11 against Fusarium commune and Fusarium oxysporum disease in blueberry plants in a pot test after four weeks.

Treatment Fusarium commune Fusarium oxysporum
Disease index Percentage disease (%) Disease index Percentage disease (%)
Control 96.00 ± 3.27a n.d. 97.33 ± 1.88a n.d.
Inoculation with TM11 first 6.67 ± 1.89c 93.05 ± 1.99a 14.67 ± 1.89c 84.76 ± 1.57a
Inoculation with pathogens first 28.00 ± 3.27b 70.92 ± 2.41b 36.00 ± 3.27b 62.46 ± 1.28b

j.pjm-2023-034.tab.006

Grade 0: no disease;
Grade 1: diseased spots on the root of blueberry seedlings less than 1.0 cm in size, but the plants were healthy;
Grade 2: blueberry seedling root lesions of 1.0–2.0 cm, seedling leaves slightly withered, and lower leaves rarely shed;
Grade 3: root lesions of blueberry seedlings were more than 2.0 cm, and seedling leaves were withered or fell off;
Grade 4: blueberry seedling showed brown root rot, or the whole plant withered;
Grade 5: blueberry seedling was dead.

Percentage inhibition of the mycelial colonies of the blueberry root rot pathogens Fusarium commune and Fusarium oxysporum by TM11 compared with the untreated control (CK) assessed in dual culture plates after 5 days of incubation.

Strain Fusarium commune Fusarium oxysporum
Treatment colony radius (cm) Percentage inhibition (%) Treatment colony radius (cm) Percentage inhibition (%)
CK 5.42 ± 0.08a n.d. 5.53 ± 0.05a n.d.
TM11 1.73 ± 0.09b 68.01 ± 1.45 1.90 ± 0.06b 65.65 ± 1.22
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