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1. Mass multiplication of Pseudomonas fluorescens By, A.Vikas (2022002204) AC&RI - Madurai

2. Introduction Pseudomonas fluorescens is a common Gram-negative, rod-shaped bacterium. The word Pseudomonas means false unit, being derived from the Greek words 'pseud ē s' and 'monas', meaning 'false' and 'a single unit' respectively.

3. T he word was used early in the history of microbiology to refer to germs. The specific name fluorescen s refers to the microbes secretion of a soluble fluorescent pigment called pyoverdin , which is a type of siderophore .

4. Biological properties Some P. fluorescens strains present have biocontrol properties, protecting the roots of some plant species against parasitic fungi such as F usarium or the o omycete Pythium , as well as some phytophagous n ematodes .

5. Medical implications By culturing P. fluorescens , mupirocin can be produced, which has been found to be useful in treating skin, ear and eye disorders . Pseudomonas fluorescens demonstrates hemolytic activity, and as a result, has been known to infect blood transfusions. Pseudomonas fluorescens produces the antibiotic Obafluorin .

6. Agricultural research Pseudomonas fluorescens is increasingly recognized for its biocontrol properties in agriculture. The bacterium’s ability to produce secondary metabolites not only inhibit the growth of pathogens but also induce systemic resistance in plants, enhancing their natural defense mechanisms.
Moreover, the application of P.fluorescens as a biocontrol agent has been shown to be a sustainable alternative to chemical pesticides, promoting environmental health and reducing the ecological footprint of agricultural practices.

7. Mass multiplication by king’s b medium The Ki ng ’ s B medium is prepared and poured into the fermentor and sterilized at 15 lb pressure for 15 minutes. After the broth has cooled below the mother culture of P. fluorescens is added to the K ing’s B medium in the fermentor at the rate of 3 lit for 40 lit of the broth. Then it is incubated in the fermentor for 2 days with frequent mixing of the broth by operating the stirrer. Then the broth containing the bacterial growth is collected in plastic buckets and used for mixing with talc powder for commercial formulation.

8. PREPARATION OF MOTHER CULTURE Mother culture is prepared by using the K ing’s B medium Peptone : 20.0 g K2HPO4 : 1.5 g Mg SO4 : 1.5 g Glycerol : 10 ml Distilled water : 1000 ml The above broth is dispersed into conical flasks and autoclaved at 15 lb pressure for 15 minutes and cooled and inoculated with a loop of P.fluorescens a nd incubated for 2 days.

9. Mass multiplication by Boiled coconut water An easy, farmer-friendly field level multiplication method for Pseudomonas fluorescens . I t was multiplied in boiled coconut water under non-sterile conditions and the population reached to 109 cfu /ml after 48 hours. Though the inoculation of the growth medium was done without complete sterilization, there was no detectable growth of contaminants in the inoculated boiled coconut water. Since the population level of the bacterial strain in the inoculated bottles had gone up to 108-9 cfu /ml, the same preparation could be further diluted up to 10 to 50 times and could be applied to plants in the form of spray or soil drench.

10. Mass multiplication by liquid and solid fermentation Prepare a mother culture of P. fluorescens in King's B broth, then use it to liquid ferment a substrate that includes pongamia and neem cake extracts, sugarcane molasses, and King's B broth.  Next, use the liquid fermentation product to solid ferment a substrate that includes sterile pongamia and neem deoiled cake, as well as wheat bran.

11. McQs 1. As per the gram stain classification, P.fluorescens comes under which category? A) gram negative B) gram positive C) gram neutral D) unclassified 2. What is the shape of P.fluorescens ? A) comma shaped B) globular C) rod shaped D) Irregularly shaped 3. The name is derived from........ A) Spanish B) Latin C) Greek D) French 4. What does fluorescens refer to? A) light B) bright C) pigment D)dye

12. McQs 5. What is the name of the fluorescent pigment? A) xanthophyll B) carotene C) chlorophyll D) pyoverdin 6) Which multiplication method is farmer friendly? A) liquid and solid fermentation B) King’s B medium C) boiled coconut water D) none of the given options 7. P.fluorescens in agriculture can be used as........ A) biocontrol agent B) fertilizer C) organic manure D) PGR

13. McQs 8. What is/are the useful product(s) obtained from P.fluorescens ? A) Mupirocin B) Obafluorin C) Dye D) both options a and b 9. What is the sterilization condition of King’s B medium? A) 60°C for 30 mins B) 72°C for 15s C) 105°C overnight D) 15lb pressure for 15mins 10. In what form can P.fluorescens be applied to the plants? A) granular B) spray C) soil drench D) both options B and C

14. Bibliography http://jtropag.kau.in https://agritech.trau.ac.in http://celkau.in https://www.researhgate.net https://patents.google.com