Phosphorus uptake by

Ectomycorrhiza and
Endomycorhhiza
To:
Dr. Abdul Nasir
Khalid
1
 Contents
 Introduction
 Phosphorus in the rhizosphere
 Limitations in Phosphorus uptake
 Sources of P in the soil
 Mobilization of P in the rhizosphere
 Uptake of P from soil by extraredical mycelium
 Fungal metabolism
 Translocation of P
 Transfer of P from fungus to plant
 Conclusion
2
 Introduction
 Phosphorus(P) is the second most important plant nutrient
after nitrogen that is critical for plant growth and
productivity.

 It contributes up to about 0.2% dry mass.

 P is a component of nucleic acids, phospholipids, and ATP

and is involved in controlling enzyme reactions and the
regulation of the metabolic pathways.

3
 Phosphorus in the Rhizosphere
 P may be present in relatively large amounts in rhizosphere.
But Plants can only take up P as free phosphate ions, H 2PO4−
and HPO4 2−.

 poorly available because of the very low solubility of

phosphates of iron, aluminium, and calcium. Due to its
 low solubility
 low mobility in soil
 high fixation by soil matrix

 P can be rapidly depleted in the rhizosphere by root uptake and

it is considered as one of the most limiting macronutrients for
plant growth (Rakshit, Bhadoria 2009). 4
 5
http://nysgolfbmp.cals.cornell.edu/phosphorus/
Limitations in P uptake from soil
 The limiting steps in the uptake of P by plants from soil
are:

(1) diffusion of phosphate ions from soil solution to plant

roots
(2) decrease the concentration of phosphate at the root Surface
(3) the release of phosphate ions from the soil particles.
But
“Infection of plant roots with a mycorrhizal
fungus alter any of these rate-limiting steps and
increase phosphate uptake by plants”
6
http://journal.frontiersin.org/article/10.3389/fpls.2014.00337/full

7
 Role of Mycorrhiza in Phosphorus
Transport
 Plants evolved a range of strategies that increase Pi uptake
capacity or availability of Pi in soil.
 Most widespread strategy worldwide is Mycorhiza symbiosis.

 Mycorrhizal fungi are ecologically significant because they form

relationships in and on the roots of a host plant in a symbiotic
association so that mycorrhizas, not roots, are the chief organs of
nutrient uptake by plants.

 Mycorrhizas form a critical link between the aboveground plant

and the soil by influencing plant nutrient cycling and soil structure

8
http://journal.frontiersin.org/article/10.3389/fpls.2014.00337/full

9
Continue..

 Mycorrhizal plants deplete phosphate to lower levels and to a

much larger soil volume per unit root length than non
mycorrhizal plants.

 Influx of P in roots colonized by AM fungi can be three to

five times that in non mycorrhizal roots (Smith, Read 1997).

 Jakobsen (1986) reported a two to three times increase in

influx of P to roots of pea and clover plants, respectively, due
to the effect of mycorrhiza.

10
 Nutrient uptake Pathways in Arbuscular
Mycorrhizal or Ectomycorrhizal Roots
 Mycorrhizal plants can take up nutrients from the soil via two
pathways:

1: the ‘Plant pathway’

that involves the direct uptake of nutrients from the soil by the
root epidermis and its root hairs

2: the ‘Mycorrhizal pathway’

that involves the uptake of nutrients via the ERM of the fungus
and the transport to the Hartig net in ECM interactions or to the
IRM in AM interactions, and the uptake by the plant from the
interfacial apoplast
11
12
 Difference in nutrient uptake pathways
in ECM and AM roots
 AM roots do not form a fungal sheath and use both
pathways for nutrient uptake. So, in the AM
symbiosis both uptake pathways act additively.

 ECM roots are characterized by a more or less dense

fungal sheath that surrounds the mycorrhizal root
completely. So, these roots usually rely on the
mycorrhizal pathway for nutrient uptake.

13
Comparison of nutrient uptake mechanisms
in ECM and AM interactions

14
http://www.cell.com/trends/plant-science/fulltext/S1360-1385(14)00168-X
15
Mechanism of Phosphorus uptake
 Mechanism of P uptake relies on the:

Sources of P in the soil

Mobilization of P in the rhizosphere
Uptake of P from soil by extraredical mycelium
Fungal metabolism
Translocation of P
Transfer of P from fungus to plant (Fungal plant
interface)
16
1
Sources and mobilization of P in the Soil
 P compounds present in soils are broadly categorized into:
1. Inorganic P
2. Organic P
 Relative distribution of these two forms in soils depends on
various factors include:
 type of vegetation
 fertilizer history
 microbial activity
 cultivation and soil type

17
 1. Organic source of P in the Soil
 A large proportion of organic P is found as Po compounds.
 Most Po is in the form of phosphate esters (C-O- P bonds)
such as phosphate monoesters (e.g., sugar–phosphates) and
phosphate diesters (nucleic acids and phospholipids).

 Inositol phosphate, phospholipids and nucleic acid are the

predominant organic phosphate compounds.

 Organic P occurs:
 as soluble P in soil solution
 as insoluble P adsorbed onto soil particles
 as a component of soil organic matter.

18
 Phytate
 Phytate (myo-inositol hexa kisphosphate) is
the potential source of P for plants, because
it is found in many ecosystems including
forest ecosystems.

 It is the principal storage form of

phosphorus in many plant tissues.

 Preserved specially in seeds and hydrolyzed

during germination by intracellular plant
phytases to supply Pi to young seedlings.

Structure of Phytate
 How ever, if the seeds do not germinate,
their phytate content will fill the pool of soil
phytate. 19
 P mobilization

 Organic P is immobilized and is generally mineralized to

inorganic P before it becomes plant-available in the soil.

 Mineralization is caused either by hydrolysis or by enzymatic

dephosphorylation.

 Phytases are the enzymes which are produced by mycorrhizal

fungi and plants (in less amount) that help in the mobilization
of organic phosphates.

20
Continue..
 Recent studies relies on the exploitation of the interactions
between plants, ECM fungi, bacteria, and their grazers within
the rhizosphere combined with the capacity of bacteria to
degrade phytate.

 Bacteria inhabiting the plant rhizosphere are able to mineralize

phytate and increase the available P pool.
 However, bacteria are more competitive than plants and
ectomycorrhizal fungi to take up P released by phytase. So, a
large fraction of P released from phytate is immobilized and
locked in the soil bacterial biomass.

 Here Bacterial grazers (nematodes) significantly improve plant

P nutrition through re-mineralization of the microbial P pool into
21
soil.
P mobilization by mycorrhiza and bacteria
22
 2. Inorganic source of P in the Soil
 Inorganic P (Pi) can occur in soil solution, which
constitutes only a small proportion of total P
(<1%) but its is the most immediate P source used
when require by plant.

 Most of the Pi (Orthophosphate) is adsorbed onto

the soil surfaces or precipitated as Fe and Al
phosphates in acid soils and as Ca and Mg
phosphates in alkaline and calcareous soils.

23
Continue..

 Pi is readily available in soil at 6.5 pH.

 Rock phosphates and Apatite are also inorganic P

source for soil P used by some ECM fungi.

 Mycorrhizal fungi release citrate, oxalate or

malonate (organic acids) and acid phosphatases
which release P from its inorganic source and
mobilize it.

24
https://www.researchgate.net/figure/7547191_fig4_Figure-3-Plant-and-microbial-
25
mechanisms-to-increase-phosphorus-P-availability-in-the
2
Uptake of P from the Soil
 Mycorrhizal fungi involve the:
1. Exploration of large soil volumes by the ERM in which
orthophosphate (Pi) is scavenged and delivered to plant
cortical cells, by passing the plant pathway for P uptake.

2. Small hyphal diameter allows the fungus to penetrate into

small soil cores in search for P, and higher P influx rates per
surface unit.

3. Store P in form of poly P that allows the fungus to keep the

internal Pi concentration relatively low and allows an
efficient transfer of P from the ERM to the IRM.
26
 Fungal uptake Systems
 Fungi have two uptake systems for P:

(1) a high affinity system that works against an

electrochemical potential gradient, which takes up Pi
from the soil via proton co transport and

(2) a low affinity system which facilitates the

diffusion of Pi across the fungal plasma membrane.

27
Continue..

 AM and ECM fungi express high affinity P

transporters in the ERM that are involved in the P
uptake from the soil.

 The expression of these transporters is regulated in

response to the externally available P concentration,
and to the P demand of the fungus.

 Under Pi starvation the transcript levels generally

increase.
28
3
Fungal Metabolism
Once Orthophosphate (Pi) absorbed from the soil
solution into the fungal cytoplasm by the ERM, it can:

1. replenish the cytoplasmic, metabolically active Pi

pool
2. be incorporated into phospholipids, RNA-, DNA-
and protein-phosphates
3. can be transferred into a storage pool of short- or
long-chained polyphosphates (Poly P).

• . 29
 Poly P molecules
 Inorganic Poly P are linear polymers in which Pi
(orthophosphate) residues are linked by energy-rich
phospho-anhydride bonds.

 Two types of Poly P can be distinguished in mycorrhizal

fungi: short chain Poly P and long chained Poly P.

 The average length of short chained Poly P in AM fungi

has been estimated as 11-20 Pi and of long chained Poly
P as 190 to 300 Pi residues.
30
Fungal P Metabolism
31

https://www.nature.com/articles/ncomms1046
 Function of Poly P
 P is transferred mainly as Poly P from the ERM to the root.

 The chain lengths of Poly P in the ERM are longer than in the
IRM, suggesting that Poly P are primarily formed in the ERM
and re-mobilized in the IRM.

 Poly P is synthesized by a wide range of microorganisms and

fungi and the most likely function is as a storage molecule,
buffering the cytoplasmic orthophosphate concentration within
physiologically acceptable limits.

32
 Balance of Poly P
 PolyP are poly anions and their negative charge is balanced by
cations.

 The cations K+ and Mg2+ are mainly involved in charge

balance but polyP can also serve as trap for toxic cations such
as heavy metals.

 The basic aminoacid Arg+ can also be involved in the charge

balance of polyP and it has been suggested that in ECM fungi
polyP can also store significant amounts of N.

33
4
Translocation of P

 In ECM and AM fungi a motile tubular vacuole system has

been identified that allows the poly P transport through the
hyphae separately from the cytoplasmic compartment and
enables the fungus to fine-tune its local cytoplasmic Pi
concentration.

 Long-chain poly P are mainly involved in long term storage of

P, whereas short-chain poly P are correlated to the P
translocation in the symbiosis.

34
 Translocation of P

35

https://www.researchgate.net/figure/236188851_fig5_Fig-5-Phosphate-transportome-
during-mycorrhizal-interactions-and-P-transfer-mechanisms
Continue..

 ECM fungal vacuoles hold substantial amounts of P mainly in

the form of polyphosphates and responsible for short distance
P translocation at the mm to cm scale. However, long-distance
translocation of Poly P is observed in AM hyphae.

 In ECM fungi, the tubules are found in extraradical hyphae

and in the fungal sheath.

 A similar tubular vacuole system in AM fungi has been

observed in extraradical and intercellular hyphae of the AM
fungus Gigaspora margarita.

36
5
Transfer of P from fungus to plant
(Fungal plant interface)
 Transfer of P from fungus to plant involves membrane
transport steps at living interfaces, which are comprised of the
membranes of both symbionts and an apoplastic region
between them.

 Nutrient transfer models generally involve:

(1) the passive efflux of phosphate and carbohydrates through the
fungal and plant plasma membranes into the interfacial
apoplasm and
(2) the active absorption of nutrients by both symbionts driven
by an H+- ATPase.

37
 ECM interface
 Bücking and Heyser(2001) showed that Pi accumulated
rapidly in the ECM sheath and was slowly translocated off the
Hartig net to the cortical cells.
 Because there is no direct symplastic continuity between the
ECM fungus and the roots, Pi has to move into the interfacial
apoplast before it can be absorbed by the plant.

AM interface
 Early workers assumed that degeneration of arbuscules was
the mechanism that made fungal P available to the plants.
 it is now assumed that the site of transfer is the mature
arbuscular interface and intracelluar coils in AM
mycorrhizas.
38
 ECM interface AM interface

39
 Phosphate transporters involve in
transfer process
 Phosphate transporters are involved in transfer of Pi from
fungus to plant cell.
 Contrary to AM symbiosis, little is still known about plant
transporters responsible for Pi acquisition in the Hartig net of
roots with ECM fungi.

 Loth-Peredaetal.(2011) showed that Populus colonization by

both AM and ECM fungi led to the up-regulation of two Pht1
transporters, PtPT9, and PtPT12.

40
41
Continue..

 Families of transporter proteins that facilitate P

transfer through plant and fungal membranes are
reported in AM Fungi.

 These transporters have been identified in the

Solanum tuberosum, S. lycopersicon, Medicago
truncatula, Oryza sativa, Triticum aestivum, and
Zea mays.

42
Arbuscular mycorrhizal root

43
http://www.plantphysiol.org/content/156/3/1050/F1.expansion.html
 Conclusion

Mycorrhizal plant roots form

relationships with fungi and increase the
uptake of phosphorus than non
mycorrhizal plants

44
 References
 Finzi, A.C.; Abramoff, R.Z.; Spiller, K.S.; Brzostek, E.R.; Darby, B.A.; Kramer, M.A.;
Phillips, R.P. Rhizosphere processes are quantitatively important components of terrestrial
carbon and nutrient cycles. Glob. Chang. Biol. 2015, 21, 2082–2094.
 Gachomo E, Allen JW, Pfeffer PE, Govindarajulu M, Douds DD, Jin HR, Nagahashi G,
Lammers PJ, Shachar-Hill Y, Bücking H. Germinating Spores of Glomus intraradices can use
Internal and Exogenous Nitrogen Sources for de novo Biosynthesis of Amino Acids. New
Phytologist 2009;184(2): 399-411.
 George, E.; Häussler, K.U.; Vetterlein, D.; Gorgus, E.; Marschner, H. Water and nutrient
translocation by hyphae of Glomus mosseae. Can. J. Bot. 1992, 70, 2130–2137.
 Duponnois R. Responses of Pinus halepensis Growth, Soil Microbial Catabolic Functions and
Phosphate-Solubilizing Bacteria after Rock Phosphate Amendment and Ectomycorrhizal
Inoculation. Plant and Soil 2009;320(1-2): 169-79.
 Paszkowski U, Kroken U, Roux C, Briggs SP. Rice Phosphate Transporters Include an
Evolutionarily Divergent Gene Specifically Activated in Arbuscular Mycorrhizal Symbiosis.
Proceedings of the National Academy of Sciences, U.S.A. 2002;99(20): 13324- 9.
 Persson, B.L., J.O. Lagerstedt, J.R. Pratt, J. Pattison-Granberg, K. Lundh, S. Shokrollahzadeh
and F. Lundh. 2003. Regulation of phosphate acquisition in Saccharomyces cerevisiae. Curr.
Genet. 43:225–244.

45
 Harrison MJ, Dewbre GR, Liu J. A Phosphate Transporter from Medicago
truncatula Involved in the Acquisition of Phosphate Released by Arbuscular
Mycorrhizal Fungi. The Plant Cell 2002;14: 2413-29.
 Harrison MJ, van Buuren ML. A Phosphate Transporter from the Mycorrhizal
Fungus Glomus versiforme. Nature 1995;378: 627-9.
 Hawkins HJ, Johansen A, George E. Uptake and Transport of Organic and
Inorganic Nitrogen by Arbuscular Mycorrhizal Fungi. Plant and Soil 2000;226:
275-85.
 Hawkins, H.J.; George, E. Reduced 15N-nitrogen transport through arbuscular
mycorrhizal hyphae to Triticum aestivum L. supplied with ammonium vs. nitrate
nutrition. Ann. Bot. 2001, 87, 303–311.
 Preuss CP, Huang CY, Gilliham M, Tyerman SD. Channel-like Characteristics of
the Low-Affinity Barley Phosphate Transporter PHT1;6 when Expressed in
Xenopus Oocytes. Plant Physiology 2010;152(3): 1431-41.
 Raghothama K.G. 1999. Phosphate acquisition. Annu. Rev. Plant Physiol. Mol.
Biol. 50: 665–693.
 Rakshit A., Bhadoria P.B.S., Mittra B.N. 2002. Nutrient use efficiency for bumper
harvest. Yojana, Dehli, May 2002, 12–15.
 Richardson A.E., Lynch J.P., Ryan P.R., Delhaize E., Smith F.A. 2011. Plant and
microbial strategies to improve the phosphorus efficiency of agriculture. Plant Soil
349: 121–156. 46
Thankyou
47