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Pink Pigmented Facultative

Methylotrophs (PPFMs)
N. Karthikeyan

Compounds used for methylotrophic
growth include:
Trimethylamine (multicarbon but without

Obligate methylotrophs cannot grow on multicarbon
compounds (include most methanotrophs)
Facultative methylotrophs also grow on multicarbon
compounds (eg. Methylobacterium extorquens = Pseudomonas AM1
Restricted facultative methylotrophs grow on C1 or C2
compounds (eg. Hyphomicrobium)
There is a huge number of different types of methylotroph. Very
few are typical bacteria that had been described previously.
Although their oxidative metabolism is basically similar, there
are many different carbon assimilation pathways.
Most are Gram-negative. The few Gram positive methylotrophs
(Bacillus sp) have different oxidation enzymes


Sohngen. Bacillus methanicus growing on methane


Peel and Quayle. Pseudomonas AM1 (cited only 6 papers) A pink

facultative methyltroph

Anthony and Zatman. Description of Methanol dehydrogenase in

Pseudomonas M27 (=AM1)


Hirsch and Conti. Hyphomicrobium. A stalked facultative methylotroph


Whittenbury (with Wilkinson). 100 new methanotrophs (2 Types)


Ribbons Early work on methane oxidation

1970- Ogata, Sahm, Hazeu, van Dijken, Harder. Methylotrophic yeast.


Dalton Definitive work on methane oxidation


Anthony book. Cited about 1200 methylotroph references

C1 carbon assimilation
Methylotrophs assimilate C1 carbon
source via either ribulosemonophosphate pathway or the
serine pathway

Serine Pathway

Plant Probiotics
Probiotics are: Live
microorganisms which
when administered in
adequate amounts
confer a health benefit
on the host.
It should come as no
surprise that we
humans arent the only
organisms that benefit
from relationships with
the right kind of

Methylobacterium is a genus of bacteria (also

called PPFM bacteria) that seems to be associated
ubiquitously with plants.
These bacteria are found in relatively large
numbers on all kinds of plants and in seeds.
Given the amount of plant biomass on Earth,
they must be among the most abundant organisms
on the planet.
If you eat lunch today at the salad bar, you will
no doubt consume some of them. Go swimming in
lake water or digging in your vegetable garden and

Seeds cured of their PPFMs no longer germinate

well or

develop normally, but reinoculating

the cured seeds

with a population of the

bacteria restores germination

and growth.

The surprising conclusion to this story is that

sometimes poorly stored or aged seeds fail to
germinate because

their bacteria have died,

not because they are

themselves dead. This

discovery led to another patent


and a second award.

(Prof. Mark Holland, Salisbury

Members of the Methylobacterium genus are
classified as
-proteobacteria and include 51 reported species
This genus is composed of Gram-negative bacteria
that are generally with pink pigmentation due to
carotenoid synthesis rod shaped, strictly aerobic
and able to grow using compounds containing only
one carbon (C1), such as methanol and
Thus, these bacteria are denoted pink-pigmented
facultative methylotrophs (PPFMs).

The main characteristic of this group is the ability to
oxidize methanol using the methanol dehydrogenase
enzyme (MDH)
The mxaF gene encodes the large subunit of this
enzyme, which is key in methylotrophic metabolism and
is used to study that group of bacteria
Members of the Methylobacterium genus occupy
different habitats due to their great phenotypic plasticity,
including soil, water, leaf surfaces, nodules, grains, and
They can also be opportunistic pathogens in human

Growing in meristematic tissue, they can reach
populations of 104 to 106 colony-forming units (CFU) per
gram of plant tissue
In addition, they can form biofilms and use
methylotrophic metabolism as an adaptive advantage
during plant host colonization

The association between Methylobacterium spp. and host
plants varies from

strong or symbiotic to weak or epiphytic

and to intermediate or


During interactions with plants, M. nodulans and M.

Radiotolerans have been

reported to be involved in nitrogen

fixation and nodule formation,

while other Methylobacterium species may be related to

production or interacting with plant

pathogens , promoting plant growth

photosynthetic activity

and inducing higher

Dourado et al

Dourado et al

The aerial parts of living plants including leaves, stems,
buds, flowers and

fruit provide a habitat for microbes

called as Phyllosphere
The term phyllosphere coined by Ruinen (1961)



e (Flowers)



Under the microscope, aerial plant leaves resemble

eerie landscapes, with deep gorges, tall peaks and
gaping pits that riddle the waxy surface.Add to this
scenery a climate that features temperature highs of 50
C [122 F] or more, exposure to harmful ultraviolet
rays, erratic periods of drought and limited access to
nutrients, and one gets the picture that this is a hostile
environment. Still, many bacteria, fungi, yeast and
other microorganisms dwell in great abundance in this

Major bacterial genera detected in

phyllosphere samples ofArabidopsis thaliana,
soybean, clover and rice

PPFM- Plant Interactions

How ppfm gets methanol from plant leaves
1.Demethylation of the cell wall pectin by pectin
2.Intermediate reactions in tetrahydrofolate pathway
3.Pectin methyltransferases
4.Degradation of lignin in secondary cell walls.
5.Also produced during the growth of cells and the
formation of intercellular space

PPFM- Plant Interactions

How ppfm improves growth of plants
1.By the production of
1. Auxins
2. Cytokinins
3. Vitamin B12
4. Exopolysaccharides
5. osmoprotectants
2.Phosphate solubilisation
3.Siderophore production
4.Nitrogen fixation
5.ACC deaminase activity
6.Inhibition of plant pathogens

PPFM- Plant Interactions

PPFM- Plant Interactions

Nitrogen fixation in legumes
Methylobacterium nodulans nodulates several legumes including
Crotalaria podocarpa
Phosphate solubilisation
Methylobacterium oryzae- shown to have all enzymes like
phosphatase, phytase, Phosphonatase
Siderophore production
Are low molecular mass compounds with a high affinity for iron that
are produced by bacteria to solubilize iron to promote efficient its
efficient uptake
Methylobacterium extorquens and M.populi shown to have genes for
the production of siderophores
ACC deaminase activity
M. Oryzae showed acc deaminase activity

Effects of the PPFM bacteria on soybean germination

Heating dry seeds before germination lowers their
PPFM population


Effect of PPFM on male

sterility of plants




Beneficial effect on plant

Fasten seed germination and seedling growth

Accelerate vegetative growth
Increase leaf area index and chlorophyll content
Earliness in flowering, fruit set and maturation
Improves fruit quality, color and seed weight
Yield increase by 10%
Mitigate drought
Method of Application

Seed treatment Imbibe seed in 1.0 % volume for 5-10min (depending on seed)
Foliar Spray of 1% PPFM
Spray during morning or evening
Recommended for all crops
Spray at critical stage of crop growth (or) 30days interval
Precaution : Do not mix with pesticide / fungicide

Drought Mitigation Program in Rice Delta Region

of TamilNadu

PPFM 1000ml / acre

Foliar application

Thank you