scholarly journals Inhibitory Effect of Trichoderma asperellum Isolate against Ralstonia solanacearum Causing Brinjal Wilt

2021 ◽  
Vol 24 (2) ◽  
pp. 107-120
Author(s):  
SMN Islam ◽  
SS Siddique ◽  
MZH Chowdhury ◽  
NJ Mishu

A native Trichoderma isolate was collected from the agricultural soil of Gazipur. This isolate was identified as a Trichoderma asperellum through morphology and analysis of internal transcribed spacer (ITS) region of ribosomal RNA gene sequence and reconstruction of the phylogenetic tree. The antagonistic effects of the newly identified T. asperellum isolate were assessed against brinjal bacterial wilt caused by Ralstonia solanacearum both in vitro and in planta. Both qualitative and quantitative bioassays were conducted in vitro. For qualitative tests, dual culture and antibacterial activity were carried out, and pathogen growth was observed visually. The antagonism of T. asperellum cell free culture filtrate on the growth of R. solanacearum was conducted in a quantitative test. Successful antagonism was recorded after both in vitro qualitative tests. In addition, the lowest colony forming unit was recorded in 100% of CFC (2.4±0.51 ×103 cfu/ml) in quantitative test. The T. asperellum inoculated plant showed low disease incidence (13.33%) when seedlings were challenged with R. solanacearum in planta experiment. Disease incidence was 100% for seedlings when treated with only R. solanacearum. The results showed that the isolated and identified T. asperellum isolate suppressed R. solanacearum growth in vitro and protected the seedling from wilting in planta. Therefore, this isolate could be considered as a potential isolate. Ann. Bangladesh Agric. (2020) 24(2) : 107-120

Plant Disease ◽  
2019 ◽  
Vol 103 (11) ◽  
pp. 2733-2741 ◽  
Author(s):  
Hao Chou ◽  
Yi-Ting Xiao ◽  
Jyh-Nong Tsai ◽  
Ting-Ting Li ◽  
Hung-Yi Wu ◽  
...  

Brown root rot (BRR), caused by the white rot fungus Phellinus noxius, is an epidemic disease of diverse broadleaved and coniferous tree species in many tropical and subtropical regions. Flooding and trenching control measures are difficult to implement, and chemical controls can have an adverse impact on ecosystems. Previous studies have provided in vitro evidence for the potential use of Trichoderma spp. for biocontrol of BRR. Here, we analyzed the in vitro antagonistic and mycoparasitic abilities of four Trichoderma spp. isolates against four P. noxius isolates in dual culture and Ficus microcarpa wood blocks. A convenient inoculation system based on root inoculation of a highly susceptible loquat (Eriobotrya japonica) with P. noxius-colonized wheat-oat grains was developed to examine the effect of Trichoderma treatment in planta. Preventive application of Trichoderma asperellum TA, the isolate showing high antagonistic activity in vitro, was effective in preventing and delaying the wilting of P. noxius-inoculated loquat cuttings in greenhouse trials. To understand the specific niche in which T. asperellum TA interacts with P. noxius, KOH-aniline blue fluorescence microscopy was used to investigate the colonization of loquat roots by P. noxius and/or T. asperellum TA. Dilution plating assays were also conducted to quantify Trichoderma populations in the rhizosphere and potting mix. T. asperellum TA was able to robustly establish in the rhizosphere and potting mix but with scarce root penetration limited to the superficial layer. We discuss the timing and strategy for applying antagonistic Trichodema sp. on living trees or in BRR-infested areas for BRR management.


2021 ◽  
Vol 7 (1) ◽  
pp. 46
Author(s):  
Warin Intana ◽  
Suchawadee Kheawleng ◽  
Anurag Sunpapao

Postharvest fruit rot caused by Fusarium incarnatum is a destructive postharvest disease of muskmelon (Cucumis melo). Biocontrol by antagonistic microorganisms is considered an alternative to synthetic fungicide application. The aim of this study was to investigate the mechanisms of action involved in the biocontrol of postharvest fruit rot in muskmelons by Trichoderma species. Seven Trichoderma spp. isolates were selected for in vitro testing against F. incarnatum in potato dextrose agar (PDA) by dual culture assay. In other relevant works, Trichoderma asperellum T76-14 showed a significantly higher percentage of inhibition (81%) than other isolates. Through the sealed plate method, volatile organic compounds (VOCs) emitted from T. asperellum T76-14 proved effective at inhibiting the fungal growth of F. incarnatum by 62.5%. Solid-phase microextraction GC/MS analysis revealed several VOCs emitted from T. asperellum T76-14, whereas the dominant compound was tentatively identified as phenylethyl alcohol (PEA). We have tested commercial volatile (PEA) against in vitro growth of F. incarnatum; the result showed PEA at a concentration of 1.5 mg mL−1 suppressed fungal growth with 56% inhibition. Both VOCs and PEA caused abnormal changes in the fungal mycelia. In vivo testing showed that the lesion size of muskmelons exposed to VOCs from T. asperellum T76-14 was significantly smaller than that of the control. Muskmelons exposed to VOCs from T. asperellum T76-14 showed no fruit rot after incubation at seven days compared to fruit rot in the control. This study demonstrated the ability of T. asperellum T76-14 to produce volatile antifungal compounds, showing that it can be a major mechanism involved in and responsible for the successful inhibition of F. incarnatum and control of postharvest fruit rot in muskmelons.


PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6905 ◽  
Author(s):  
Elena Maria Colombo ◽  
Cristina Pizzatti ◽  
Andrea Kunova ◽  
Claudio Gardana ◽  
Marco Saracchi ◽  
...  

Biocontrol microorganisms are emerging as an effective alternative to pesticides. Ideally, biocontrol agents (BCAs) for the control of fungal plant pathogens should be selected by an in vitro method that is high-throughput and is predictive of in planta efficacy, possibly considering environmental factors, and the natural diversity of the pathogen. The purpose of our study was (1) to assess the effects ofFusariumstrain diversity (N= 5) and culture media (N= 6) on the identification of biological control activity ofStreptomycesstrains (N= 20) againstFusariumpathogens of wheat in vitro and (2) to verify the ability of our in vitro screening methods to simulate the activity in planta. Our results indicate that culture media,Fusariumstrain diversity, and their interactions affect the results of an in vitro selection by dual culture assay. The results obtained on the wheat-based culture media resulted in the highest correlation score (r= 0.5) with the in planta root rot (RR) inhibition, suggesting that this in vitro method was the best predictor of in planta performance of streptomycetes against Fusarium RR of wheat assessed as extension of the necrosis on the root. Contrarily, none of the in vitro plate assays using the media tested could appropriately predict the activity of the streptomycetes against Fusarium foot rot symptoms estimated as the necrosis at the crown level. Considering overall data of correlation, the activity in planta cannot be effectively predicted by dual culture plate studies, therefore improved in vitro methods are needed to better mimic the activity of biocontrol strains in natural conditions. This work contributes to setting up laboratory standards for preliminary screening assays ofStreptomycesBCAs against fungal pathogens.


Agriculture ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 1177
Author(s):  
Yasser S. Mostafa ◽  
Mohamed Hashem ◽  
Ali M. Alshehri ◽  
Saad Alamri ◽  
Ebrahem M. Eid ◽  
...  

This research evaluated the efficacy of essential oils in the management of cucumber powdery mildew. Essential oils of lemongrass, lemon, thyme, peppermint, abundance blend, purification blend, and thieves blend were tested in vitro and under greenhouse conditions in two separate experiments. The effects of essential oils were tested against powdery mildew disease at concentrations of 1.0–2.5 mL/L, and the consequent impact of the oils on plant growth was evaluated. Powdery mildew fungus, Podosphaera xanthii, was identified using sequencing of the ITS region. The essential oils significantly reduced disease incidence up to 77.3% compared with the positive control (p < 0.5). Moreover, the essential oils increased the plant length (up to 187 cm), leaf area (up to 27.5 cm2), fresh weight (up to 123 g), dry weight (up to 22.5 g), number of flowers (16.3), and metabolite content compared with the positive control (p < 0.5). Cell membrane injury decreased significantly in the oil-treated pants (p < 0.5), indicating the protective effect of essential oils. This study recommends the application of essential oils in an appropriate dose (2.5 mL/L) to protect cucumber plants against powdery mildew. Overdose of the oils (more than 2.5 mL/L) should be avoided due to adverse effects.


Agronomy ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1398
Author(s):  
Vishal Gupta ◽  
Krishna Kumar ◽  
Kausar Fatima ◽  
Vijay Kumar Razdan ◽  
Bhagwati Charan Sharma ◽  
...  

Saffron (Crocus sativus L.) is considered as one of the most expensive spices. Fusarium corm rot of saffron, caused by Fusarium oxysporum, is known to cause severe yield losses worldwide. In the present study, efficacy of biocontrol agents (Trichoderma asperellum, Pseudomonas fluorescens, Pseudomonas aeruginosa, Pseudomonas putida, Bacillus stratosphericus, Bacillus pumilus, and Bacillus subtilis) along with a chemical fungicide, carbendazim, was evaluated for managing the corm rot of saffron. Under in vitro conditions, using dual culture and poison food techniques on potato dextrose agar, T. asperellum and carbendazim significantly reduced the mycelial growth of the pathogen F. oxysporum, with the inhibition of 62.76 and 60.27%, respectively, compared with control. Under field conditions, dipping of saffron corms in carbendazim and T. asperellum exhibited maximum reduction of 82.77 and 77.84%, respectively, in the disease incidence, during the first year of experiment. However, during the second year, maximum reduction in the incidence of corm rot (68.63%) was recorded with the T. asperellum. Moreover, the population density of F. oxysporum was also significantly reduced by 60 and 80.19% while using T. asperellum after 75 and 260 days of sowing of saffron corms, compared to its population before planting of corms. In case of growth promotion traits, such as sprouting and flowering, biocontrol treatments reduced the number of days (average) of sprouting and flower emergence after sowing, compared to control.


Horticulturae ◽  
2021 ◽  
Vol 7 (11) ◽  
pp. 457
Author(s):  
Josefina Viejobueno ◽  
Natalia Rodríguez-Berbel ◽  
Luis Miranda ◽  
Berta de los Santos ◽  
María Camacho

The effect of antagonistic bacteria to control Macrophomina phaseolina was evaluated under in vitro, growth chamber, greenhouse and field conditions. A total of 177 bacteria, isolated from Athrocaulon macrostachyum rhizosphere of the Lebrija marsh, were screened for their potential against M. phaseolina (causes charcoal rot in strawberry) by dual culture assay. Of these isolates, 14 most promising strains were molecularly identified by the 16S rDNA sequencing method using the EzBioCloud database. These strains were tested for in vitro hydrolytic enzymes, HCN production, and biocontrol against M. phaseolina in strawberry plants. All the 14 strains produced, at least, one hydrolytic enzymatic activity and one of them, which belongs to Brevibacterium genus (Hvs8), showed the lowest records of disease incidence (20%) and severity (0.4). With these results, greenhouse and field trials were carried out with the Hvs8 strain, compared to non-treated control. In the greenhouse assays, Hvs8 strain increased root dry mass by 30%, over the control. In the field trials, production and fruit quality were not significantly different between Hvs8 treatment and non-treated control, but plant mortality and plant mortality associated to M. phaseolina decreased by more than 24% and 65% respectively, in Hvs8 treatment. This study suggests that Brevibacterium sp. Hvs8 strain could be a candidate for controlling charcoal rot in strawberry.


2015 ◽  
Vol 15 (1) ◽  
pp. 72
Author(s):  
Susanti Tasik ◽  
Siti Muslimah Widyastuti ◽  
Harjono .

Mechanism of parasitism of Trichoderma harzianum on Fusarium oxysporum on Acacia mangium seedlings. Fusarium oxysporum is one of the most important soil-borne fungi the causal agent of damping-off disease. Detailed information it needed to know how the pathogen can be inhibited by Trichoderma harzianum. The objective of this research was to investigate the inhibition mechanism of T. harzianum on F. oxysporum in vitro and in planta. Green Flourescent Protein (GFP) T. harzianum was used as biocontrol agent of F. oxysporum. An in vitro inhibition test of T. harzianum was performed using dual culture method. In the in planta inhibition tests, seedlings of A. mangium were applied with GFP T. harzianum two days before inoculation of F. oxysporum; GFP T. harzianum was simultaneously applied with F. oxysporum and GFP T. harzianum was applied two days after inoculation of F. oxysporum. The inhibition effect of T. harzianum GFP was observed at seven days incubation, indicated by attachment of T. harzianum to F. oxysporum hyphae. GFP T. harzianum hyphae covered the colonies of F. oxysporum at 12 days after incubation. The highest life percentage of A. mangium seedlings was found on the treatment of GFP T. harzianum two days before inoculation of F. oxysporum (82.22%), whereas the lowest life percentage was found on seedling applied with GFP T. harzianum two days after inoculation of F. oxysporum (64.44%).


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.


Author(s):  
Juan Carlos Martínez-Álvarez ◽  
Flavio Camacho-Angulo ◽  
Yolani de Jesús Bojórquez-Armenta ◽  
Bardo Sánchez-Soto ◽  
Jesús Damián Cordero-Ramírez ◽  
...  

El objetivo del presente estudio fue seleccionar cepas bacterianas de la rizósfera del cultivo de frijol y determinar su potencial para el control de <em>Sclerotium rolfsii</em> bajo condiciones <em>in vitro</em> e <em>in planta</em>. Se recolectaron muestras de suelo en los municipios de Ahome, Guasave y Angostura, Sinaloa, México, durante el ciclo agrícola 2018-2019. Los aislados se evaluaron bajo condiciones <em>in vitro</em> y se seleccionaron los mejores para la evaluación <em>in planta</em> e identificación molecular (tres aislados) con base a la región 16S del ADN ribosomal. Se evaluaron 65 aislados bacterianos <em>in vitro</em> contra <em>S. rolfsii</em> con un porcentaje de inhibición de 2.5 a 65%. <em>Acinetobacter pittii </em>(COHUI06)<em>,</em> <em>Pseudomonas putida</em> (SANMI02) y <em>Burkholderia</em> sp. (GLS06) inhibieron 55, 60 y 65% bajo condiciones <em>in vitro</em>; además resultaron no hemolíticas. Solo <em>Burkholderia</em> sp. ejerció mayor porcentaje de inhibición <em>in planta</em> para el control de <em>S. rolfsii,</em> con reducción de incidencia y severidad de la enfermedad en un 40 y 50% respectivamente, y promovió el peso seco de la planta. Para <em>A. pitti</em> o <em>P. putida</em> no fueron eficientes para el control del hongo <em>in planta</em>. Se sugiere hacer estudios con las bacterias en invernadero y campo.


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