MPMI Proposal

Institute of Plant Biology, Master student, R98B42021 呂毓蘋

Topic
To define the mechanisms of stress imprint in plants

Background and rationale

A history of exposure to a range of different types of stress alters
subsequent plant responses. The process of priming or hardening
involves prior exposure to a biotic or an abiotic stress factor making a
plant more resistant to future exposure. This feature, in higher plants,
indicates some capacity for ‘‘memory’’. Sustained alterations in levels of
key signalling metabolites or transcription factors could provide an
explanation for how plant metabolism is altered by exposure to various
stresses. Alternatively epigenetic changes could play a role by enabling
long-term changes in gene expression. Exposure to a priming agent could
activate a gene or set of genes but instead of reverting to the
transcriptionally silent state once the stimulus is removed, an epigenetic
mark could perhaps be left, keeping the region in a permissive state,
facilitating quicker and more potent responses to subsequent attacks. In
the stress responses, defence genes show the ‘priming’ phenomenon;
they are able to respond faster and to a greater extent to subsequent
challenge. The promoters of many of these defence genes contain at
least one ’W-box’ that provides binding sites for WRKY transcription
factors. Genes encoding WRKY factors are themselves transcriptionally
induced by either pathogen infection or treatment with microbe-
associated molecular patterns, such as flagellin.
Chromatin structure is important for the regulation of gene
expression. Acetylation of lysines in the amino-terminal tails of histones
H3 and H4 has been associated with active genes. This modification
reduces the ionic interaction between positively charged lysine side
chains and the negatively charged DNA backbone. Moreover, lysine
acetylation provides docking sites for transcriptional coactivator
proteins. The strongest correlation between histone methylation and
gene activity is found for trimethylation of Lys 4 on histone H3
(H3K4me3) on promoters and coding sequences of active genes.
Although gene priming is a widespread phenomenon and has also
been described for the defence response in animals, little is known about
the mechanisms for it at the molecular level. Further studies needed to
establish the molecular mechanism by which plants store information on
stress exposure because biotic and abiotic stresses limit agricultural
production and stress responses often lead to down-regulation of yield
determining processes such as photosynthesis.

Hypothesis and aims

The existence of a stress imprint effect in plant responses to a variety of
biotic and abiotic stresses raises the question of how such effects occur
and what the underlying mechanism is. Previous study pointed out there
are molecular mechanisms responsible for priming are not well
understood but propose two potential mechanisms one involving
accumulation of signalling proteins and the other involving accumulation
of transcription factors. Here this review also propose an epigenetic
mechanism. The potential importance of transcriptional regulation of
gene expression has also been highlighted in plant responses to
dehydration and cold stresses. The hypothesis is that exposure to a
priming agent could activate a gene or set of genes but instead of
reverting to the transcriptionally silent state once the stimulus is
removed, an epigenetic mark (such as histone acetylation) could be left,
keeping the region in a permissive state perhaps facilitating quicker and
more potent responses to subsequent attacks. Thus I want to define
clearly the mechanisms of stress imprint in plants. If it is not in the
epigenetic level, maybe it is involved in the accumulation of proteins or
transcription factors.
Approaches
(1) To identify potential target genes of priming, test Arabidopsis genes
encoding WRKY transcription factors with abiotic stress.
(2) Treat with biotic stress after abiotic stress and qRT-PCR confirm the
gene expression.
(3) CHIP assay to know the histones modifications compare between the
primed and non-primed state.
(4) Use Salicylic acid(SA) accumulation deficient mutants npr1 and SA
constitutively expressed mutants sni1 to confirm the results observed
in wild type plants.

Expected results
(1) Get candidate genes

Stress
Psm

WRKYA WRKYB WRKYC

(2) Difference in modification patterns of histones

Psm
(3) Abiotic stress treat only modification patterns are different with
primed patterns.

Stress

WRKYA WRKYB WRKYC

(4) WRKY gene expression and histone modification in promoters of

mutants

Psm

WRKYA WRKYB WRKYC

Stress

WRKYA WRKYB WRKYC

Pitfall and alternative approaches
If the primed mechanism is not on the epigenetic level, than the
expected difference can’t be observed. Still, there is an alternative
mechanism possible for plant memory the stress. Therefore, I should try
to see is there any protein or transcription factors accumulated in
primed plants. First, analyzing PTI marker gene expression to know it still
can be induced after primed with abiotic stress. Second, use northern
and western blot to analyze transcript and protein level changes with
and without primed. If there are difference, it suggest that plants use the
mechanism of accumulating proteins and transcription factors to
memory the first abiotic stress, which can help plants stronger respond
to subsequent biotic challenge.

References
1. Bruce TJA, Matthes MC, Napier JA and Pickett JA. (2007) Stressful
“memories” of plants: evidence and possible mechanisms. Plant Sci
173: 603–608

2. Yi HS, Heil M, Adame-Álvarez RM, Ballhorn DJ, and Ryu CM. (2009)
Airborne Induction and Priming of Plant Defenses against a Bacterial
Pathogen. Plant physiol 151(4): 2152–2161

3. Jaskiewicz M, Conrath U and Peterhansel C. (2011) Chromatin

modification acts as amemory for systemic acquired resistance in the
plant stress response. EMBO reports 12: 50 - 55

4. Xia, Y., Gao, Q.M., Yu, K., Lapchyk, L., Navarre, D., Hildebrand, D.,
Kachroo, A., and Kachroo, P. (2009) An intact cuticle in distal tissues
is essential for the induction of systemic acquired resistance in plants.
Cell Host Microbe 5: 151–165