scholarly journals Screening of Tomato Seed Bacterial Endophytes for Antifungal Activity Reveals Lipopeptide Producing Bacillus siamensis Strain NKIT9 as a Potential Bio-Control Agent

2021 ◽  
Vol 12 ◽  
Author(s):  
Ayushi Sharma ◽  
Nutan Kaushik ◽  
Abhishek Sharma ◽  
Abhay Bajaj ◽  
Mandar Rasane ◽  
...  

The current study investigates the diversity pattern and fungicidal potential of bacterial endophytes isolated from two different organic varieties of tomato plants (V1 and V2). A total of seventy-five bacterial isolates identified by 16S rRNA gene sequencing revealed a majority of genus as Bacillus and one Planococcus, which were grouped into eight different species. The Shannon diversity H’ (1.56), Simpson’s index of diversity (0.93), Magalef’ index (2.23), Evenness (0.96), and Species richness (7) indicated the high endophytic bacterial diversity in the V1 variety of the tomato. Bacterial endophytes isolated from both of the varieties were screened for their antifungal activity against five economically critical fungal pathogens (viz., Botrytis cinerea, Rhizoctonia solani, Fusarium solani, Verticillium lateritium, and Alternaria solani) of tomato crop through dual culture assay. The data revealed B. siamensis strain NKIT9 as the most potent antagonist, significantly (p < 0.05) inhibiting the mycelial growth between 75 to 90% against selected fungal pathogens. High bioactivity of lipopeptide extract of strain NKIT9 was recorded against R. solani with minimum IC50 value of 230 μg/ml. The Ultra Performance Liquid Chromatography-High Definition Mass Spectrometry (UPLC-HDMS) analysis of this lipopeptide extract revealed the presence of Surfactin and Bacillomycin D. Furthermore, in-vitro results showed that the selected bacterial strain significantly minimized the disease incidence in damping-off assay which makes this strain a promising antifungal bio-control agent. Moreover, in the pot experiment the NKIT9 increased the fruit yield by 59.2% compared with the untreated R. solani infested control.

2020 ◽  
Author(s):  
Ayushi Sharma ◽  
Nutan Kaushik ◽  
Abhishek Sharma ◽  
Abhay Bajaj ◽  
Mandar Rasane ◽  
...  

ABSTRACTThe current study investigates the diversity pattern and fungicidal potential of bacterial endophytes isolated from two different organic varieties of tomato plants (V1 and V2). A total of seventy-four bacterial isolates identified by 16S rRNA sequencing revealed a single genus Bacillus with 16 different species. The Shannon diversity H’ (1.45), Simpson’s index of diversity (0.9), Magalef’ index (2.1), Evenness (0.96), and Species richness (8) indicated the high endophytic bacterial diversity in the V1 variety of the tomato. Bacterial endophytes isolated from both the varieties were screened for their antifungal activity against five economically critical fungal pathogens (viz., Botrytis cinerea, Rhizoctonia solani, Fusarium solani, Verticillium lateritium, and Alternaria solani) of tomato crop through dual culture assay. The data revealed B. siamensis KCTC 13613(T) as the most potent antagonist significantly (p < 0.05), inhibiting the mycelial growth between 75 to 90% against selected fungal pathogens. High bioactivity of lipopeptide extract of B. siamensis was recorded against R. solani with IC50 value of 72 ppm. The UPLC-HDMS analysis of this lipopeptide extract revealed the presence of, Surfactin and Bacillomycin D.


Horticulturae ◽  
2021 ◽  
Vol 7 (7) ◽  
pp. 198
Author(s):  
Hongbo Yuan ◽  
Bingke Shi ◽  
Tianxiang Huang ◽  
Zengqiang Zhou ◽  
Li Wang ◽  
...  

Valsa canker caused by Valsa pyri is one of the most destructive diseases of commercial pear. For the present analysis, 29 different endophytic fungal strains were isolated from the branches of a healthy pear tree. In dual culture assays, strain ZZ1 exhibited robust antifungal activity against all tested pathogens including Valsa pyri. Microscopic analyses suggested that following co-culture with ZZ1, the hyphae of V. pyri were ragged, thin, and ruptured. ZZ1 also induced significant decreases in lesion length and disease incidence on detached pear branches inoculated with V. pyri. ZZ1 isolate-derived culture filtrates also exhibited antifungal activity against V. pyri, decreasing mycelial growth and conidium germination and inhibiting V. pyri-associated lesion development on pear branches. These results suggest that the ZZ1 isolate has the potential for use as a biological control agent against V. pyri. The strain was further identified as Penicillium citrinum based on its morphological characteristics and molecular analyses. Overall, these data highlight a potentially valuable new biocontrol resource for combating pear Valsa canker.


Author(s):  
R. C. Osaro-Matthew ◽  
F. S. Ire ◽  
N. Frank-Peterside

Aim: The present study was carried out to isolate actinomycetes from rhizospheric soil of Curcuma longa and Zingiber officinale and evaluate their antifungal potential. Methods: Actinomycetes were isolated from the rhizosphere soil of two Zingiberaceae plants (Curcuma longa and Zingiber officinale), using four different culture media. Isolates were screened for antifungal activity using dual culture technique against two reference strains Colletotrichum coccodes (DSM 2492) and Alternaria pimpriana (DSM 62023). The most potent strain was identified based on 16S rRNA gene sequence and the bioactive components of the strain were identified using GC-MS chromatography. Results: Fifteen strains of actinomycetes were isolated, SCA (starch casein agar) was found best for cultivation of actinomycetes. The 15 strains were grouped into three genera Norcadia 8(54%), Streptomyces 5(33%) and Janibacter 2(13%) based on morphological, biochemical and physiological identification. Among the 15 isolates 6(40%) strains showed activity against either of the test organisms, while 1(6.7%) was active against the both test organisms. Comparative analysis of the 16S rRNA gene sequences identified the potent isolates as Janibacter sp. strain RC18. GC-MS analysis revealed the presence of 20 compounds with 10 identified as potent antimicrobial metabolites. Conclusion: This study has revealed that rhizosphere of ginger and turmeric harbours rare actinomycetes that are valuable tool for sustainable agriculture.


2021 ◽  
Vol 31 (1) ◽  
Author(s):  
H. H. Al-Nadabi ◽  
N. S. Al-Buraiki ◽  
A. A. Al-Nabhani ◽  
S. N. Maharachchikumbura ◽  
R. Velazhahan ◽  
...  

Abstract Background Date palm ((Phoenix doctylifera L.) suffers from several fungal diseases. The endophytic microorganisms present in higher plants generally offer protection to their host plants against invading phytopathogenic fungi and bacteria. In the present study, endophytic bacteria associated with date palm leaves were isolated and their in vitro antagonistic potential against fungal pathogens causing leaf spots in date palm was demonstrated. Results Endophytic bacteria were isolated from date palm leaves of 3 different cultivars viz., Nighal, Khalas and Khinaizi and evaluated for their inhibitory activity against leaf spot pathogens of date palm viz., Fusarium solani, Alternaria sp., Nigrospora sp., Thieloviopsis sp., Curvularia subpapendrofii and Tilletiopsis minor using an in vitro dual culture assay. Of the 24 endophytic bacterial strains tested, the endophytes designated B1, B7, B8 and B9 obtained from cv. Nighal showed inhibitory activities (more than 55% mycelial growth inhibition) against F. solani and Alternaria sp. None of the bacterial endophytes inhibited the growth of other fungal pathogens tested. These antagonistic bacterial strains were identified as Pantoea septica on the basis of 16S rRNA gene sequence analysis. The hyphae of F. solani and Alternaria sp. exhibited morphological abnormalities such as shrinkage and disintegration when grown in the presence of antagonistic bacterial endophytes. The cell-free culture filtrates of the bacterial endophytes caused inhibition of mycelial growth and induced leakage of electrolytes from the mycelia of F. solani and Alternaria sp. This is the first study that describes inhibition of the date palm pathogens F. solani and Alternaria sp. by P. septica. Conclusion Endophytic Pantoea septica strains isolated from date palm leaves inhibited the mycelial growth of F. solani and Alternaria sp. and induced morphological changes in their mycelia. The culture filtrates of these bacterial strains also inhibited the mycelial growth and caused leakage of electrolytes from the mycelia of F. solani and Alternaria sp. These promising bacterial strains can be exploited as biocontrol agents to control F. solani and Alternaria sp.-induced leaf spot diseases of date palm.


2020 ◽  
Vol 24 (10) ◽  
pp. 1811-1814
Author(s):  
J.O. Nwogwugwu ◽  
A.A. Batcho ◽  
I Time

The Impact of fungal pathogens on man is enormous, stretching from infecting humans to destroying a third of all food crops annually thereby causing economic losses and impacting global poverty. As a result, fungal disease incidence was assessed under natural field infection and data were collected randomly from 12 months old jatropha seedlings with cankers. Colletotrichum spp. isolated from the diseased sampled were interacted with known fungi and bacteria biocontrol agents. The experiment was carried out on completely randomized design (CRD) in the laboratory. Data were analyzed using ANOVA and statistically different means were separated using LSD (p≤0.05). The dual culture interaction proved that all the BCA used, possessed antagonistic potential, and could serve best for prophylactic protection. Plant diseases may be suppressed by the activities of one or more plant-associated microbes hence keeping them in check below threshold levels. This will help to maintain the quality and abundance of food, thereby mitigating hunger. Negative environmental consequences caused by excessive use and misuse of agrochemicals, requires that healthier and costeffective approaches to disease management should be adopted. Keywords: Biodiesel; Agrochemicals; Biocontrol agent; Dual culture; Pathogens


2020 ◽  
Vol 18 (1) ◽  
pp. 103-112
Author(s):  
R. A. OLOYEDE ◽  
A. A. ILUPEJU ◽  
O. O. OYELAKIN ◽  
W. R. AJIJOLA

Fusarium wilt is one of the important diseases of cucumber and causes economic loss to farmers. The present study was undertaken to evaluate the potential of rhizosphere lactic acid bacteria as biocontrol agents of Fusarium wilt of cucumber. Lactic acid bacteria (LAB) were isolated and identified from the rhizosphere of five medicinal plants. The in-vitro antagonistic activity of LAB strains on F. oxysporum f.sp. cucumerinum was evaluated by dual culture method. The screen house experiment was then conducted to assess the effect of antagonistic LAB isolates on Fusarium wilt disease incidence in cucumber plants. The antagonistic LAB strains were further characterized using 16S rRNA gene sequencing technique. The total LAB counts of rhizospheric soil samples ranged from 7.0×105 cfu/g to 15.0×105 cfu/g. The LAB isolates were identified as strains of Lactobacillus acidophilus (21.4%), L. plantarum (35.7%), L. fermentum (28.6%), L. alimentarius (7.1%) and L. brevis (7.1%). Treatment of cucumber seeds with antagonistic LAB strains significantly reduced Fusarium wilt of cucumber incidence from 95% to 48%. Lactobacillus fermentum isolated from the rhizosphere of A. indica exhibited strong disease suppression (49.5%). The study therefore revealed that the rhizospheric-LAB could be applied to reduce the manifestation of Fusarium wilt in cucumber.    


2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Chao Xue ◽  
C. Ryan Penton ◽  
Zongzhuan Shen ◽  
Ruifu Zhang ◽  
Qiwei Huang ◽  
...  

Abstract Panama disease caused by Fusarium oxysporum f. sp. cubense infection on banana is devastating banana plantations worldwide. Biological control has been proposed to suppress Panama disease, though the stability and survival of bio-control microorganisms in field setting is largely unknown. In order to develop a bio-control strategy for this disease, 16S rRNA gene sequencing was used to assess the microbial community of a disease-suppressive soil. Bacillus was identified as the dominant bacterial group in the suppressive soil. For this reason, B. amyloliquefaciens NJN-6 isolated from the suppressive soil was selected as a potential bio-control agent. A bioorganic fertilizer (BIO), formulated by combining this isolate with compost, was applied in nursery pots to assess the bio-control of Panama disease. Results showed that BIO significantly decreased disease incidence by 68.5%, resulting in a doubled yield. Moreover, bacterial community structure was significantly correlated to disease incidence and yield and Bacillus colonization was negatively correlated with pathogen abundance and disease incidence, but positively correlated to yield. In total, the application of BIO altered the rhizo-bacterial community by establishing beneficial strains that dominated the microbial community and decreased pathogen colonization in the banana rhizosphere, which plays an important role in the management of Panama disease.


2018 ◽  
Vol 33 (3-4) ◽  
pp. 197-204 ◽  
Author(s):  
Svetlana Zivkovic ◽  
Milos Stevanovic ◽  
Sanja Djurovic ◽  
Danijela Ristic ◽  
Stefan Stosic

Chitosan and its derivatives have been reported as a promising alternative for control of postharvest fungal pathogens. The objective of this study was to evaluate in vitro and in situ antifungal activity of chitosan against Alternaria alternata and Colletotrichum gloeosporioides isolated from decayed apple fruits. The fungi were tested in vitro using PDA medium with three concentrations of chitosan (1, 2 and 3 mg/ml). Fungal growth of the test pathogens was significantly affected by all chitosan doses (P<0.05) after 7 days of incubation at 25?C. Water solution of 3 mg/ml of chitosan inhibited completely the conidial germination of A. alternata and C. gloeosporioides after 18 h incubation at 25?C. The results obtained from biocontrol assay indicate that the inhibition of postharvest decay of A. alternata and C. gloeosporioides was significantly influenced by chitosan concentrations. Disease incidence in chitosan-treated fruit after 7 days incubation at 25?C was significantly lower than in the positive control for both fungi tested (P<0.05). A. alternata and C. gloeosporioides used in this study were progressively inhibited in vitro and in situ with increasing concentrations of chitosan from 1 to 3 mg/ml.


Agronomy ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 373
Author(s):  
Siti Fairuz Yusoff ◽  
Farah Farhanah Haron ◽  
Norhayu Asib ◽  
Mahmud Tengku Muda Mohamed ◽  
Siti Izera Ismail

Postharvest fruits including tomatoes are commonly infected by gray mold disease resulting in significant economic losses in the fruit industry. Therefore, this study aimed to develop botanical fungicide derived from Vernonia amygdalina leaf extract to control gray mold on tomato. The emulsion formulation containing surfactant, oil carrier and water was optimized at different non-ionic alkyl polyglucoside surfactants through eleven combinations of oil to surfactant ratio (0:10, 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, 9:1 and 10:0 w/w). From eight selected formulations, two formulations, F5 and F7 showed stable in storage, remarkable thermodynamic stability, smaller particle size (66.44 and 139.63 nm), highly stable in zeta potential (−32.70 and −31.70 mV), low in polydispersity index (0.41 and 0.40 PdI), low in viscosity (4.20 and 4.37 cP) and low in surface tension (27.62 and 26.41 mN/m) as compared to other formulations. In situ antifungal activity on tomato fruits showed F5 formulation had a fungicidal activity against B. cinerea with zero disease incidence and severity, whereas F7 formulation reduced 62.5% disease incidence compared to a positive control with scale 1. Based on these findings, F5 formulation exhibited pronounced antifungal activity and may contribute to the development of new and safe antifungal product against gray mold on tomato.


2021 ◽  
Vol 22 (14) ◽  
pp. 7715
Author(s):  
Grzegorz Czernel ◽  
Dominika Bloch ◽  
Arkadiusz Matwijczuk ◽  
Jolanta Cieśla ◽  
Monika Kędzierska-Matysek ◽  
...  

Silver nanoparticles (AgNPs) were synthesized using aqueous honey solutions with a concentration of 2%, 10%, and 20%—AgNPs-H2, AgNPs-H10, and AgNPs-H20. The reaction was conducted at 35 °C and 70 °C. Additionally, nanoparticles obtained with the citrate method (AgNPs-C), while amphotericin B (AmB) and fluconazole were used as controls. The presence and physicochemical properties of AgNPs was affirmed by analyzing the sample with ultraviolet–visible (UV–Vis) and fluorescence spectroscopy, scanning electron microscopy (SEM), and dynamic light scattering (DLS). The 20% honey solution caused an inhibition of the synthesis of nanoparticles at 35 °C. The antifungal activity of the AgNPs was evaluated using opportunistic human fungal pathogens Candida albicans and Candida parapsilosis. The antifungal effect was determined by the minimum inhibitory concentration (MIC) and disc diffusion assay. The highest activity in the MIC tests was observed in the AgNPs-H2 variant. AgNPs-H10 and AgNPs-H20 showed no activity or even stimulated fungal growth. The results of the Kirby–Bauer disc diffusion susceptibility test for C. parapsilosis strains indicated stronger antifungal activity of AgNPs-H than fluconazole. The study demonstrated that the antifungal activity of AgNPs is closely related to the concentration of honey used for the synthesis thereof.


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