To study the interaction between auxin and sugar signal transduction

pathway in model plant system Arabidopsis thaliana

Bhuwaneshwar S. Mishra
Ph.D. student (2nd year)

Supervisor: Dr. Ashverya Laxmi

Sugar signaling
Photosynthesis

SUGAR Energy
Biosynthesis

Storage

Structure
Osmotic regulation

Signaling molecules
Function of Glucose

•Seed and embryo development.

•Seed germination.

•Seedling development.

•Hypocotyl development.

•Nodule growth.

•Pollen development.

•Formation of adventitious root.

•Induction of senescence.
 The Glucose Sensor
Glucose Signaling Is AtHXK Dependent

6% Glucose

AtHXK Sense Antisense Control

6% Mannitol

Jang et al. 1997

Three glucose signal transduction pathways in plants

HXK sensor Activate or repressor Gene Expression

(CAB, ERA1, PC,
AtHXK1 dependent pathway PLD, RBCS)

Glucose HXK Unknown sensor Gene Expression

(PR1, PR5)
Glycolysis Dependent Pathway

Unknown sensor Gene Expression (AGPase, AS1, CHS,

CIN1, PAL1, PAL3)
AtHXK1 independent pathway

Xiao et al. 2000

Overview of interactions between glucose and
hormone signaling in Arabidopsis

Rolland et al, 2006

Auxin Signaling
 Auxin
The term auxin is applied to a group of weak organic acid in which the
acidic group is situated at the end of side chain; attached to an
unsaturated ring system (indole ring) and are responsible for a
diverse range of plant developmental physiological process.

Acetic acid
Indole ring side chain
Functions
Stimulates cell elongation.

Stimulates cell division.

Stimulates differentiation of phloem and xylem.

Stimulates root initiation on stem cuttings and lateral root

development.

Mediates the tropistic response of bending in response to gravity

and light.

The auxin supply from the apical bud suppresses growth of lateral
buds.

Delays leaf senescence.

Stimulates growth of flower parts.

 Synthesis

Synthesized in meristems, young leaves, fruits, roots and seeds

primarily.

All tissues can produce trace levels of IAA.

Synthesized from either tryptophan dependent or independent

pathways.
 Auxin Transport
Non polar transport

via phloem sap.

5-10cm/hr.
no direct requirement of energy.

Polar transport

5-10mm/hr.
polar auxin transport forms an auxin gradient
extending from shoot tip to root tip.
This longitudinal gradient affect proper
lateral root development, vascular
development, phyllotaxis, embryonic axis
development and tropism.
Polar transport requires energy and is gravity
independent.
A crucial feature of the polar transport model
is that auxin transport is mediated by several
transport protein.
Chemiosmotic Hypothesis for Polar Auxin Transport

Vanneste et al. 2009

 Transcription regulation of Auxin

Auxin inducible genes are regulated by members of 2 protein families

ARF (Auxin Response Factor) gene family Aux/IAA gene family

23 members 29 members
 Auxin receptor

The F-box protein TIR1 is an auxin receptor

Nihal Dharmasiri, Sunethra Dharmasiri, Mark Estelle
Nature Vol. 435|26 May 2005|

The Arabidopsis F-box protein TIR1 is an auxin receptor

Stefan Kepinski, Ottoline Leyser
Nature Vol. 435|26 May 2005|
 Auxin promotes SCF-TIR1 substrate binding by
acting as a “molecular glue rather than an allosteric
switch

Tan et al. 2007

Mechanism of auxin action

Paciorek et al. 2006

Sugar-Auxin signaling interaction
Auxin signaling mutants are glucose insensitive

Moore et al. 2003

A turanose-insensitive mutant suggests a role for WOX5 in
Auxin homeostasis in Arabidopsis thaliana

Turanose is a reducing disaccharide analog of sucrose not

metabolized by higher plants.

Turanose insensitive (tin), encodes for WOX5 gene, which

is responsible for auxin homeostasis and maintaining auxin
maxima in the root tip.

Gonzali et al. 2005

HLS1 functions in sugar and IAA responsive gene expression
in leaves

Ohto et al. 2006

Objectives

1. To study the developmental, morphological and physiological

parameters regulated by interaction of auxin and glucose.

2. To understand the molecular basis of sugar auxin interaction

globally using whole genomics approach.

3. Molecular and functional characterization of gene/genes

simultaneously regulated by auxin and glucose.
 Comparison of root growth and development of
Columbia seedlings grown in different concentrations of
glucose containing medium

Mishra et al. 2009

 Effect of glucose on IAA regulated genes
604
IAA>2 fold up or down

346 258
up down

191 (55%) gene also affected up 186 (72%) gene also affected up
or down by glucose or down by glucose

135(71%) genes 56(29%)genes 122(66%) genes 64(34%) genes

Up by glucose Down by glucose Down by glucose Up by glucose

62% genes are regulated by glucose

68% genes agonistically regulated
32% genes antagonistically regulated
 Presence of glucose can affect the extent of
up-regulation and down-regulation of IAA regulated genes

604
IAA>2 fold up or down

346 up 258 down

227(66%) genes IAA induction 153(59%) genes IAA suppression

up or down by glucose up or down by glucose

100(44%) 127(56%) 51(33%) down 102(67%) down

up by glucose down by glucose regulating up by glucose regulation down by glucose

89(89%) genes 50(39%) genes 51(100%) genes 57(56%) genes

Up by gluc. alone down by gluc. alone down by gluc. alone Up by gluc. alone

• Synergistically affected genes were mainly transcriptionally regulated by glucose

• Antagonistically affected genes were not transcriptionally regulated by glucose alone

GUS expression analysis of HS:AXR3NT::GUS
seedlings

Mishra et al. 2009

PIN2::GFP expression in root tip and basipetal transport

Mishra et al. 2009

Comparison of root growth and development of auxin related
mutants grown in different concentration
of glucose containing medium
0% 1% 3% 5% 1% 5%

Mishra et al. 2009

 Conclusion

Glucose increases root length, lateral roots, root hair and modulates gravitropic
responses.

Glucose antagonistically or synergistically regulates IAA regulated genes.

Glucose negatively regulates the degradation of Aux/IAA protein (repressor of

auxin signaling).

Glucose increases the accumulation of PIN2 protein on plasma membrane and

increases basipetal auxin polar transport.

Glucose auxin interaction is finally proved by the fact that glucose regulated
responses are defective in auxin perception and signaling mutants.
Thank you…