Antifungal Activity and Secondary Metabolites of P. protegens ML15

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Questions and Answers

What method was used to isolate the bacterial genomic DNA?

Bacterial & Yeast Genomic DNA Purification Kit (EURx)

How was the quality of the extracted DNA measured and verified?

NanoDrop 2000 spectrophotometer and gel electrophoresis in ChemiDoc XRS+

Which platform was used for whole-genome sequencing?

Illumina HiSeq platform

What software was used for quality filtering and trimming of raw sequencing reads?

fastp software Signup and view all the answers

What tool was used to assign COG functional categories to the identified gene sequences?

ReCOGnizer Signup and view all the answers

What was the impact of treatments T1 and T2 on the total phenolic content of tomatoes?

Slightly lower total phenolic content compared to T3 and T4 Signup and view all the answers

How did the ascorbic acid content change in tomatoes infected with fungi and inoculated with bacteria?

T1 caused a smaller reduction in ascorbic acid than T2 Signup and view all the answers

What was the trend observed in the titratable acid content of tomato juice samples?

Treatments with B. cinerea infection generally had a higher rate of citric acid than those without B. cinerea infection Signup and view all the answers

Which treatment exhibited the lowest total phenolic content?

CWB Signup and view all the answers

What impact did the application of P. protegens ML15 have on tomato quality and nutritional value?

Altering the levels of important phytochemicals such as antioxidant activity, total phenolic content, and ascorbic acid Signup and view all the answers

What method was used to identify potential phloroglucinol production?

TLC method Signup and view all the answers

How many bioactive metabolites were revealed by GC-MS analysis of the ethyl acetate extract?

29 Signup and view all the answers

What was the total assembly length of the genome assembly of P. protegens ML15?

6,972,288 bp Signup and view all the answers

How many protein coding sequences were contained in the genome of P. protegens ML15?

6,358 Signup and view all the answers

What genes did the genome of P. protegens ML15 contain for biosynthesis?

HCN, EPS matrix, bacillibactin biosynthesis, and enterobactin uptake Signup and view all the answers

How was the total antioxidants measured in cherry tomatoes?

Using the DPPH assay Signup and view all the answers

What method was used to determine the total phenolic content in the cherry tomatoes?

Folin-Ciocalteu method Signup and view all the answers

How was the ascorbic acid content determined in the cherry tomatoes?

By titration with iodine and potassium iodide solution Signup and view all the answers

What method was used to measure the titratable acidity in the cherry tomatoes?

Titrating with NaOH and using phenolphthalein as an indicator Signup and view all the answers

What specific factors contributed to the ability of P. protegens ML15 to effectively inhibit the growth of B. cinerea BC21?

Production of siderophores, HCN, lipase, EPS, ammonia, and VOCs Signup and view all the answers

What are the biocontrol properties of Pseudomonas protegens ML15 against Botrytis cinerea?

Inhibited spore germination and growth of the pathogen, reducing disease lesion Signup and view all the answers

What genes does Pseudomonas protegens ML15 encode for related to ammonia assimilation?

nirK, ureB, and the gltBDSPT operon Signup and view all the answers

How did the application of Pseudomonas protegens ML15 affect the antioxidant activity in tomatoes?

Did not significantly affect antioxidant activity, but improved it to some extent Signup and view all the answers

What are the plant growth-promoting abilities of Pseudomonas protegens ML15?

Production of gamma-aminobutyric acid (GABA) and indole-3-acetic acid (IAA) Signup and view all the answers

How did the antifungal activity of Pseudomonas protegens ML15 compare to phloroglucinol?

Surpassed the performance of phloroglucinol Signup and view all the answers

Which genes were inspected in the bacterial strain for biocontrol and plant growth promotion?

ACC deamination (acdS), acetoin and 2,3-butanediol synthesis (budA, budB, budC), achromobactin biosynthesis and transport (acsA, acsB, acsC, acsD, cbrA, cbrB, cbrC, cbrD), alkaline protease biosynthesis (aprA), and many others. Signup and view all the answers

How were proteomes compared against reference protein sequences?

Proteomes were compared using BLASTp with thresholds for e-value, sequence identity, and sequence query coverage. Signup and view all the answers

What experiment was performed to evaluate the antagonist effect of the selected isolate?

A biocontrol experiment on cherry tomatoes (Solanum lycopersicum var. cerasiforme) was performed to assess the antagonist effect of the selected isolate against Botrytis cinerea. Signup and view all the answers

How was the inhibition rate calculated in the experiment?

The inhibition rate was calculated based on the average area of lesions infected by Botrytis cinerea on fruit inoculated with distilled water or bacterial culture/CFS/phloroglucinol. Signup and view all the answers

What was the purpose of the screening of 24 Pseudomonas sp.?

The screening aimed to check for the presence of genes involved in biocontrol and plant growth promotion. Signup and view all the answers

Study Notes

Researchers inspected the genome of a bacterial strain for presence of genes involved in biocontrol and plant growth promotion, including: ACC deamination (acdS), acetoin and 2,3-butanediol synthesis (budA, budB, budC), achromobactin biosynthesis and transport (acsA, acsB, acsC, acsD, cbrA, cbrB, cbrC, cbrD), alkaline protease biosynthesis (aprA), ammonia assimilation (gltB, gltD, gltP, gltS, gltT), auxin biosynthesis (ipdC), bacillibactin biosynthesis (dhbA, dhbB, dhbC, dhbE, dhbF), biofilm formation (efp, flgB, flgC, flgD, flgE, flgF, flgG, flgH, flgI, hfq, motA, motB), chitinase biosynthesis (ChiC), elastase biosynthesis (lasA, lasB), enterobactin biosynthesis (entD, fepA, fes), exopolysaccharides biosynthesis (algA, algB, algD, algE, algG, algK, algL, algX), fengycin synthesis (fenA, fenB, fenC, fenD, fenE), GABA biosynthesis (gabD, gabT), HCN biosynthesis (hcnA, hcnB, hcnC), IAA biosynthesis (trpA, trpB, trpC, trpD, trpE, trpF, trpG), lipase biosynthesis (lipA, lipB), nitric oxide synthesis (nirK), phenazine biosynthesis (phzA, phzB, phzC, phzD, phzE, phzF, phzG, phzH, phzM, phzS), phosphate solubilization (pqqA, pqqB, pqqC, pqqD, pqqE, pqqF, pqqG, pstA, pstB, pstC, pstS), protease IV biosynthesis (prpL), pyochelin receptor (fptA), pyoverdine receptor (fpvA), pyoverdine biosynthesis (pvdA, pvdD, pvdE, pvdF, pvdG, pvdH, pvdI, pvdJ, pvdL, pvdM, pvdN, pvdO, pvdP, pvdQ, pvdS, pvdY), rhamnolipid biosynthesis (rhlA, rhlB, rhlC), surfactin biosynthesis (srfAA, srfAB, srfAC, srfAD), urease biosynthesis (ureA, ureB), vibrioferrin synthesis (pvsA, pvsB, pvsC, pvsD, pvsE).
24 Pseudomonas sp. were screened for the presence of the aforementioned genes: Pseudomonas aeruginosa B18, Pseudomonas aeruginosa FG106, Pseudomonas aeruginosa L10, Pseudomonas aeruginosa M18, Pseudomonas aeruginosa PAO1, Pseudomonas chloropsis JD37, Pseudomonas chlororaphis HT66, Pseudomonas chlororaphis PA23, Pseudomonas chlororaphis subsp. aurantiaca, Pseudomonas fluorescens CHA0, Pseudomonas fluorescens SBW25, Pseudomonas fluorescens UM270, Pseudomonas protegens FD6, Pseudomonas protegens Pf-5, Pseudomonas psychrotolerans CS51, Pseudomonas putida BIRD-1, Pseudomonas putida LWPZF, Pseudomonas seleniipraecipitans D1-6, Pseudomonas simiae WCS417, Pseudomonas sp. ANT_H12B, Pseudomonas sp. B10, Pseudomonas sp. UW4, Pseudomonas sp. VI4.1, Pseudomonas syringae GR12-2.
Proteomes were compared against reference protein sequences using BLASTp with the following thresholds for further analysis: e-value not exceeding 1e-5, minimum sequence identity of at least 50% and sequence query coverage per HSP higher than 70%.
Biocontrol experiment was performed on cherry tomatoes (Solanum lycopersicum var. cerasiforme) to evaluate the antagonist effect of the selected isolate against Botrytis cinerea, a post-harvest pathogen.
Fruit were surface sterilized and inoculated with mycelial disks of B. cinerea with or without bacterial culture, CFS, phloroglucinol, distilled water, or sterile distilled water as controls.
The inhibition rate was calculated based on the average area of lesions infected by B. cinerea on fruit inoculated with distilled water or bacterial culture/CFS/phloroglucinol.
Different postharvest quality parameters of cherry tomatoes were evaluated, but the text did not provide sufficient context for a summary.

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Description

Explore the antifungal activity and identification of secondary metabolites produced by P. protegens ML15 through a three-day incubation period and TLC method. Analyze the compounds observed in the cell-free extract of P.protegens ML15 and their comparison with standard on TLC plate.