Biology Reference
In-Depth Information
Chapter 38
Differential Proteome and Secretome Analysis During
Rice-Pathogen Interaction
Yiming Wang , Sang Gon Kim , Jingni Wu , Sun Tae Kim ,
and Kyu Young Kang
Abstract
Substantial evidences implicate that sample preparation and protein extraction in proteomic studies of
plant-pathogen interactions are critical to understand cross talk between host and pathogen. Therefore,
interest is growing in applying proteomics techniques to investigate simultaneously secreted proteins from
rice and pathogen. We have found, however, that most proteins of interest are low abundant so that proper
prefractionation or extraction of secreted proteins from extracellular space (ECS) in the rice leaf is required
to excavate relevant protein. This chapter describes the preparation of sample and extraction procedure to
enrich the proteins interested before separation by 2-DE or LC-MS/MS. This method signifi cantly
increases the sensitivity of proteomic comparisons.
Key words
Rice proteomics, Rice,
Magnaporthe oryzae
,
Xanthomonas oryzae
, 2-DE
1
Introduction
A number of rapid defense responses in plants are initiated after
pathogen attack and are activated rapidly hereafter [
1
,
2
]. In general,
an interaction is incompatible when the rice plant recognizes the
invading pathogen early enough and activates the host resistance
genes, resulting in a hypersensitive response (HR) and the triggering
of rapid and effective defense responses including oxidative burst,
the production of pathogenesis-related (PR) proteins, and phyto-
alexins [
3
]. In contrast, an interaction is compatible when the rice
plant responds too late to restrict ingress of the pathogen.
Upon interactions, the extracellular space (ECS) of the rice leaf
serves as a front line of battle fi eld of defense against the invading
pathogen. Both species secrete a diversity of functional compo-
nents into the ECS. It is likely that host arranges various molecular
events such as strengthening of the cell wall, and producing
antimicrobial activity by pathogenesis-related (PR) proteins,
