Maize Genetics Cooperation Newsletter vol 84 2010
Please Note: Notes submitted to the Maize Genetics
Cooperation Newsletter may be cited only with consent of authors.
PIACENZA, Italy
Institute of Agronomy, Genetics and
Field Crops, Universit� Cattolica del Sacro Cuore (UCSC)
Gene expression analysis in maize ears and silks after Fusarium verticillioides infection
Lanubile, A; Pasini, L; Marocco, A.
The genus Fusarium includes numerous plant
pathogens that cause destructive diseases on some of the world�s most
agriculturally important plant species, including maize. In particular F. verticillioides is one of the most
economically important Fusarium
species that causes root, stalk and ear rots, blights and wilts, and can
contaminate kernels with a family of closely related mycotoxins known as
fumonisins.
Plant defenses consist of physical
barriers such as cell wall and its modifications as well as chemical desense
mechanisms that are induced in response to external stimuli. In addition plant
defense involves expression of pathogenesis-related (PR) proteins (Kitajima and
Sato, 1999 J Biochem 125:1-8). PR proteins are grouped into 17 indipendent
families and antimicrobial properties have been described for some of them. The
PR-2 proteins display β-1,3-glucanase activity, whereas the PR-3 proteins
(as well as PR-4, PR-8 and PR-11 proteins) show endochitinase activity (Campo
et al., 2004 Proteomics 4:383-396).
In maize few detailed studies are
presently available on its response to F.
verticillioides infection and although differences in susceptibility to
this pathogen attack have been described, resistant mazie varieties have not
yet been developed (Bluhm and Woloshuk, 2005 Mol. Plant-Microbe Interactions
18:1333-1339). Objective of the research was the identification of genes
expressed in kernels and silks of maize tolerant and susceptible lines, during F. verticillioides infection, using the
microarray technology.
The tolerant line CO441 and the
susceptible line CO354 were used. Their ears were infected with a
fumonisin-producing strain of F.
verticillioides, with the pin-bar technique and harvested 48 hours after
infection and also from uninfected ears. RNAs extracted, reverse transcribed
and labelled with fluorophor dyes, were hybridized on the array slides. Venn
Diagrams showed that only 60 genes were differentially expressed in CO441 e
CO354 48h after infection. Most of genic sequences were peculiar to both the
lines. Similar functional categories of genes were involved in the response to
infection in both tolerant and susceptible lines, such as: defense response
proteins, oxidative burst-associated enzymes, enzymes involved in sugar
metabolism and proteins involved in aminoacid synthesis, folding and
stabilization. The most significant differentially expressed genes were
validated in RealTime-PCR.
The main PR genes identified in the
array experiments were also tested in silks, infected and harvested 12, 24, 48
and 72h after infection, and from uninfected samples using RealTime-PCR.
To monitorate the presence and the
activity of F.verticillioides through
absolute quantification in seeds and silks, the costitutive gene for the
β-tubulin and the target gene FUM21,
a transcriptional regulator of fumonisins metabolic pathway, of Fusarium verticillioides were also
analyzed. The results showed a higher copy number for both the genes in the
susceptible line CO354.