scholarly journals Molecular Characterization of Trichoderma spp. Isolates by Internal Transcribed Spacer (ITS) Region Sequencing Technique and its Use as a Biocontrol Agent

2020 ◽  
Vol 14 (1) ◽  
pp. 70-77
Author(s):  
Ziyaul Haque ◽  
Mohammed S. Iqbal ◽  
Ausaf Ahmad ◽  
Mohd S. Khan ◽  
Jyoti Prakash

Objective: In the present investigation, Trichoderma spp., isolated from rhizospheric soil, has been identified by Internal Transcribed Spacer (ITS) region sequencing technique and its antagonistic activity was evaluated against A. niger. Methods: The sequencing analysis was done with its ITS1 region of the rRNA gene. Using the ITS1 amplified products for all isolated fungi, a bi-directional DNA sequencing was done with high quality bases (>98% - 100%). Antagonistic activity was done using dual culture technique. Results: All of the ITS1 nucleotide sequences obtained in this study matched 97% - 100% with the published sequence of Trichoderma spp. The results confirmed the strains as T. asperellum and T. viride with gene bank accession no. (ZTa); MK937669 and (ZTv); MK503705, respectively. When phylogenetic analysis was done for the isolates, the optimal tree with the sum of branch length = 0.69585023 and 0.10077756 for T. asperellum and T. viride, respectively, was observed. There were a total of 678 and 767 for T. asperellum and T. viride positions in the final dataset, respectively. Antagonistic activity was done for the isolated strains of Trichoderma spp. against A. niger, and it was found that T. asperellum showed maximum antagonistic activity (79.33±7.09%). Conclusion: The findings prolong the genome availability for relative investigations pointing out phenotypic variances to compare with Trichoderma genetic diversity. The present investigation delivered the Bases of future studies for better knowledge in understanding the complicated connections of Trichoderma spp. to be used as an effective biocontrol agent.

2020 ◽  
Vol 13 (2) ◽  
pp. 54-65 ◽  
Author(s):  
M.E.A. Bendaha ◽  
H.A. Belaouni

SummaryThis study aims to develop a biocontrol agent against Fusarium oxysporum f.sp. radicis-lycopersici (FORL) in tomato. For this, a set of 23 bacterial endophytic isolates has been screened for their ability to inhibit in vitro the growth of FORL using the dual plate assay. Three isolates with the most sound antagonistic activity to FORL have been qualitatively screened for siderophore production, phosphates solubilization and indolic acetic acid (IAA) synthesis as growth promotion traits. Antagonistic values of the three candidates against FORL were respectively: 51.51 % (EB4B), 51.18 % (EB22K) and 41.40 % (EB2A). Based on 16S rRNA gene sequence analysis, the isolates EB4B and EB22K were closely related to Enterobacter ludwigii EN-119, while the strain EB2A has been assigned to Leclercia adecarboxylata NBRC 102595. The promotion of tomato growth has been assessed in vitro using the strains EB2A, EB4B and EB22K in presence of the phytopathogen FORL. The treatments with the selected isolates increased significantly the root length and dry weight. Best results were observed in isolate EB4B in terms of growth promotion in the absence of FORL, improving 326.60 % of the root length and 142.70 % of plant dry weight if compared with untreated controls. In the presence of FORL, the strain EB4B improved both root length (180.81 %) and plant dry weight (202.15 %). These results encourage further characterization of the observed beneficial effect of Enterobacter sp. EB4B for a possible use as biofertilizer and biocontrol agent against FORL.


2016 ◽  
Vol 137 (2) ◽  
pp. AB181
Author(s):  
Brett J. Green ◽  
Angela R. Lemons ◽  
Yeonmi Park ◽  
Jean M. Cox-Ganser ◽  
Ju-Hyeong Park

Author(s):  
H.V. Parmar ◽  
N.M. Gohel

Background: Chickpea wilt complex caused by several soil-borne pathogens is the major yield-reducing malady worldwide. Biological control is one of the best, low-cost and ecologically sustainable method for managing plant diseases caused by soil-borne pathogens. Methods: In this present investigation Panchagavya and Trichoderma spp. were evaluated by following poisoned food technique and dual culture technique against wilt complex causing pathogens i.e. Fusarium oxysporum f. sp. ciceri, Fusarium solani and Macrophomina phaseolina. Result: Among the different isolates of Trichoderma spp. evaluated, Trichoderma viride (AAU isolate) was highly antagonistic to F. oxysporum f. sp. ciceri (52.78%) and F. solani (65.37%) whereas, Trichoderma asperellum (AAU isolate) was highly antagonistic to M. phaseolina (65.93%). Panchagavya at the highest concentration (50%) showed significantly higher efficacy (80.74, 66.62 and 49.67%) in inhibiting the mycelial growth of all three pathogens and at the lowest concentration it was moderately effective.


Author(s):  
Jaygendra Kumar ◽  
Mukesh Kumar ◽  
Akash Tomar ◽  
. Vaishali ◽  
Pushpendra Kumar ◽  
...  

Trichoderma species are well known for their biocontrol activity which colonize many soil and tuber-borne and foliage plant pathogens. In this study, 12 native isolates of Trichiderma spp were collected from various crop rhizosphere soil samples and characterized them phenotypically based on morphological and cultural features and genotypically based on sequence analysis of internal transcribed spacer (ITS) region-PCR amplification. The results obtained from phenotypic and genotypic observation revealed that isolates were belonged to five different species namely T. asperellum, T. harzianum, T. longibrachiatum, T. koningii and T. koningiopsis. All Trichoderma isolates produced ~600 bp amplicon and phylogenetic analysis revealed that all isolates were grouped with respective species. Further, the antagonistic potential of all the isolates was evaluated against Fusarium spp. following in vitro dual culture method. The results showed that isolates of T. harzianum exhibited maximum growth inhibition activity. The highest rate of inhibition was recorded with T. harzianum isolate TBT6 (87.1%) followed by TBT7 (82.2%), while the least inhibition was observed in T. longibrachiatum isolate TBT10 (59.7%) after 7 days of incubation. The antagonistic T. harzianum isolate TBT6 can be used for development of Trichoderma based bio-formulation and served as bio-control agent against Fusaium spp. under field conditions.


Agronomy ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 891
Author(s):  
Mila Santos ◽  
Fernando Diánez ◽  
Alejandro Moreno-Gavíra ◽  
Brenda Sánchez-Montesinos ◽  
Francisco J. Gea

A study was conducted to explore the efficacy of potential biocontrol agent Cladobotryum mycophilum against different phytopathogenic fungi. The growth rates of 24 isolates of C. mycophilum were determined, and their antagonistic activity was analysed in vitro and in vivo against Botrytis cinerea, Fusarium oxysporum f. sp. radicis-lycopersici, Fusarium oxysporum f.sp. cucumerinum, Fusarium solani, Phytophthora parasitica, Phytophthora capsici, Pythium aphanidermatum and Mycosphaerella melonis. Most isolates grow rapidly, reaching the opposite end of the Petri dish within 72–96 h. Under dual-culture assays, C. mycophilum showed antagonistic activity in vitro against all phytopathogenic fungi tested, with mycelial growth inhibition ranging from 30 to 90% against all the different phytopathogens tested. Similarly, of all the selected isolates, CL60A, CL17A and CL18A significantly (p < 0.05) reduced the disease incidence and severity in the plant assays compared to the controls for the different pathosystems studied. Based on these results, we conclude that C. mycophilum can be considered as a potential biological control agent in agriculture. This is the first study of Cladobotryum mycophilum as a biological control agent for different diseases caused by highly relevant phytopathogens in horticulture.


2015 ◽  
Vol 45 (6) ◽  
pp. 951-957 ◽  
Author(s):  
Poholl Adan Sagratzki Cavero ◽  
Rogério Eiji Hanada ◽  
Luadir Gasparotto ◽  
Rosalee Albuquerque Coelho Neto ◽  
Jorge Teodoro de Souza

Black Sigatoka disease caused by Mycosphaerella fijiensis is the most severe banana disease worldwide. The pathogen is in an invasive phase in Brazil and is already present in most States of the country. The potential of 29 isolates of Trichoderma spp. was studied for the control of black Sigatoka disease under field conditions. Four isolates were able to significantly reduce disease severity and were further tested in a second field experiment. Isolate 2.047 showed the best results in both field experiments and was selected for fungicide sensitivity tests and mass production. This isolate was identified as Trichoderma atroviride by sequencing fragments of the ITS region of the rDNA and tef-1α of the RNA polymerase. Trichoderma atroviride was as effective as the fungicide Azoxystrobin, which is recommended for controlling black Sigatoka. This biocontrol agent has potential to control the disease and may be scaled-up for field applications on rice-based solid fermentation


2014 ◽  
Vol 16 (1) ◽  
pp. 33-43 ◽  
Author(s):  
William R. Rittenour ◽  
Christina E. Ciaccio ◽  
Charles S. Barnes ◽  
Michael L. Kashon ◽  
Angela R. Lemons ◽  
...  

2015 ◽  
Vol 65 (Pt_8) ◽  
pp. 2466-2471 ◽  
Author(s):  
Melissa Fontes Landell ◽  
Luciana R. Brandão ◽  
Silvana V. B. Safar ◽  
Fatima C. O. Gomes ◽  
Ciro R. Félix ◽  
...  

Two independent surveys of yeasts associated with different bromeliads in different Brazilian regions led to the proposal of a novel yeast species, Bullera vrieseae sp. nov., belonging to the Tremellales clade (Agaricomycotina, Basidiomycota). Analysis of the sequences in the internal transcribed spacer (ITS) region and D1/D2 domain of the LSU rRNA gene suggested affinity to a phylogenetic lineage that includes Bullera miyagiana and Bullera sakaeratica. Six isolates of the novel species were obtained from different bromeliads and regions in Brazil. Sequence analysis of the D1/D2 domains of the large subunit of the rRNA gene showed that the novel species differs from B. miyagiana and B. sakaeratica by 85 and 64 nt substitutions, respectively and by more than 75 nt substitutions in the ITS region. Phenotypically, Bullera vrieseae sp. nov. can be distinguished from both species based on the assimilation of meso-erythritol, which was negative for B. vrieseae sp. nov. but positive for the others, assimilation of d-glucosamine, which was positive for B. vrieseae sp. nov. but negative for B. miyagiana and of l-sorbose, which was negative for B. vrieseae sp. nov. but positive for B. sakaeratica. The novel species Bullera vrieseae sp. nov. is proposed to accommodate these isolates. The type strain of Bullera vrieseae sp. nov. is UFMG-CM-Y379T (BRO443T; ex-type CBS 13870T).


Sign in / Sign up

Export Citation Format

Share Document