A new naturally occurring Plant Growth Promoting Rhizobacteria.

Pluton is non pathogenic and occurrsNaturally in the soil

Pluton

What is Pluton

• Pluton is a high quality dry flowable formulation of thespores of a naturally occurring bacteria of the bacillusamyloliquefaciens family.

• Pluton is a natural product. It is not a GMO.

Contains 5 x 10 10 sporesPer gram

Pluton

A new naturally occurring Plant Growth Promoting Rhizobacteria.

What does Pluton do?

• Pluton contains live spores that germinate in the presence of root exudates. •Metabolites that are produced by Pluton stimulate plant growth , increase yield and induce plant resistance mechanisms.

Mode Of Action

Benefits of Pluton

1. Improved germination2. Improved rooting3. Improved yield4. Increased vigour5. Reduced frequency and intensity of disease

Toxicology

In trials carried out no

•Phytotoxic or• phytopathogenic effects observed

Biological Activity

•Pluton stimulates the growth of roots,Increases the tolerance against abiotic stress and inctrases the yield of plants• Trials have shown that auxins and otherPlant hormones are produced by Pluton inThe soil.• Pluton produces a bio fertiliser effect whichIs exerted by the enzyme Phytase.

Rate of Use

•Potatoes- 250 g/ha. Applied throughAn Amistar type applicator.Can also be injected with starter fertiliser•Other crops( carrots, parsnips or onions,250 g/ha. Can be appliedWith pre emergence herbicide.

Trial Results

Effect of seed treatments on % infection with Microdocium nivale

Septorianodorum

Pythium

Effect of Pluton on diseasesGrown On agar plates

Septoriatritici

Sclerotinia

Fusarium

Effect of Pluton on diseasesGrown On agar plates

Effect Of Pluton on Ht of OSR.In Modules. 28 d post drilling.

a b

Barworth Agriculture 2011

Effect of Pluton on Ht of BrassicaTransplants 21 d Post Sowing

Barworth Agriculture 2011

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After use of Bacillus (by coating, soil drenching etc.) and germination of spores the roots will be colonized and will use sugars and amino acids for their growth

bacteria

competitionMode of action

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Biofilm-like structures at root hairs most likely fixed by bacterial exopolysaccharides

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Growth promotion:• Based on formation of

growth hormones and enzymes for nutrient mobilisation

• Growth hormones(Auxines e.g.) support root formation

• Enzymes e.g. phytase transform organic phosphorous to anorganic phosphorous and in this way improve availibility of nutrients for the plant

Mode of action

CallibrachoaKern 2003

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Antibiosis - One piece in the puzzle of the mode of action :

Mode of action

Chromosome of FZB42 has a size of 3.9 Megabases

FZB42 devotes 8.5 % of its genomic capacity to synthesis of secondary metabolites useful to cope with other competing microorganisms in the rhizosphere

Chen et al. 2007, Nat. Biotechnol.

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Secondary metabolites with biocontrol and plant growth promoting activities

OH OHO

H2C-C-COO- H3C-C-COO- H3C-CH

O

H2C-C-COO- H3C-C

O

H3C-C

O

1 2

2x Pyruvat 2-Acetolactat Acetoin

CH2 COOH

N

H

IAA

CH2COOH

N

H

IAATrp 3? ?

surfactin

C10 -12 CHCH2CO=E-L-L-V-D-L-L

O

bacillomycinD

C 11-13CHCH 2CO =N -Y -N -P -E -S -T

HNfengycin

C13 -14 CHCH2CO=E-O-Y-T-E-A

OH

O

I-Y-Q-P

difficidin

O

POHO

OH

R

bacillibactin

HNGly - DHB

CH3

O O

O

H3 C

DHB - Gly -N

HO

O

CH3

N-Gly - DHB

H

O O

HO

OH

OHDHB

bacilysin

COOH

O

O

O

H 2 N N

H

OH

H3C-CH

H3C-CH

OH

2,3 -butandiol

HO

HO CH3

O

OH

O

macrolactin

O

HO

NH

O

OH

OOH

bacillaene

NH

NRS-NRPS

Bac-NRPS

Bmy-NRPS

Fen-NRPSSrf-NRPS

Dbe-NRPS

Sfp

Bae-MS

Mln-MS

Dif-MS

Chen et al. 2007: Nature Biotechnology

Antibiosis - One piece in the puzzle of the mode of action

Cocultivation of Rhizoctonia solani and FZB42Growth of R. solani in absence of FZB42

Suppression of growth of R. solani in presence of FZB42

R.solani R.solani+FZB42 R.solani+FZB42

Antibiosis - One piece in the puzzle of the mode of action

FZB 42 mutant Not produced cocultivation suppression

FZB42 ./. +

AK1 BacillomycinD -

AK2 Fengycin -

AK3 Bacillomycin DFengycin -

CH1 Surfactin +

CH2 FengycinSurfactin -

Kontrolle (R. solani) ./. ./.

Antibiosis - One piece in the puzzle of the mode of action

Beneficial bacteria can stimulate an inducedsystemic resistance (ISR) in plants, whereby itsdefensive capacities are enhanced or “primed”.

The ISR induced by beneficial bacteria ischaracterized by a weak, transient and localizeddefense response, but this priming leads to anenhanced expression of host defense related genesupon subsequent attack by a pathogen.

The rhizobacteria typically induce ISR via inductionof jasmonic acid (JA) and/or ethylene (ET)signalling pathways. However, some rhizobacteriaappear to induce an SA-dependent pathway,indicating that different signaling pathways mayoperate when ISR is elicited.

Pieterse et al. 2009, Nat. Chem Biol. 5:308-316

ISR - One piece in the puzzle of the mode of action

Several reports have shown that various Bacillusstrains are able to induce ISR in different plants.

Molecular patterns (MAMPs) produced by Bacilli thatare capable of eliciting immune responses in plants:

volatile organic compounds (Ryu et al., 2003,2004; Rudrappa et al., 2010; Farag et al.,2013)

cyclic lipopeptides like surfactin and fengycin(Ongena et al., 2005b, 2007; Jourdan et al.,2009; Henry et al., 2011; Dessoignies et al.,2013)

ISR - One piece in the puzzle of the mode of action

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Rela

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* P < 0.001 ; § P<0.005 Source: Soumitra Paul Chowdhury HGMU Munich

ISR - One piece in the puzzle of the mode of action

Q-RT-PCR-Results of lettuce plants from Pathcontrol project:

•Mild but effective Induction of plant immune response of FZB42-bacterized plant through activation of plant defence genes (PR1 & PDF1.2)•Lettuce plants respond to R. solani through JA pathway•Induction of ISR in the bacterized plants in response to pathogens challenge through expression of JA/ET pathway (defensin PDF1.2)

...pathogenesis-related protein-1 (PR1) -SA induced...defensin (PDF1.2) - JA/ET induced