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Received: 2008-08-07
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Tong-bao LIU, Jian-ping LU, Xiao-hong LIU, Hang MIN, Fu-cheng LIN. A simple and effective method for total RNA isolation of appressoria in Magnaporthe oryzae[J]. Journal of Zhejiang University Science B, 2008, 9(10): 811-817.
@article{title="A simple and effective method for total RNA isolation of appressoria in Magnaporthe oryzae",
author="Tong-bao LIU, Jian-ping LU, Xiao-hong LIU, Hang MIN, Fu-cheng LIN",
journal="Journal of Zhejiang University Science B",
volume="9",
number="10",
pages="811-817",
year="2008",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B0860011"
}
%0 Journal Article
%T A simple and effective method for total RNA isolation of appressoria in Magnaporthe oryzae
%A Tong-bao LIU
%A Jian-ping LU
%A Xiao-hong LIU
%A Hang MIN
%A Fu-cheng LIN
%J Journal of Zhejiang University SCIENCE B
%V 9
%N 10
%P 811-817
%@ 1673-1581
%D 2008
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B0860011
TY - JOUR
T1 - A simple and effective method for total RNA isolation of appressoria in Magnaporthe oryzae
A1 - Tong-bao LIU
A1 - Jian-ping LU
A1 - Xiao-hong LIU
A1 - Hang MIN
A1 - Fu-cheng LIN
J0 - Journal of Zhejiang University Science B
VL - 9
IS - 10
SP - 811
EP - 817
%@ 1673-1581
Y1 - 2008
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B0860011
Abstract: appressorium formation is an important event in establishing a successful interaction between the rice blast fungus, Magnaporthe oryzae, and its host plant, rice. An understanding of molecular events occurring in appressorium differentiation will give new strategies to control rice blast. A quick and reliable method to extract total RNA from appressorium is essential for studying gene expression during appressorium formation and its mechanism. We found that duplicate film is an efficient substratum for appressorium formation, even when inoculated with high density conidia. When inoculated with conidia at 1×106 ml−1, the percentages of conidium germination and appressorium formation were (97.98±0.67)% and (97.88±0.45)%, respectively. We applied Trizol before appressorium collection for total RNA isolation, and as much as 113.6 μg total RNA was isolated from the mature appressoria at 24 h after inoculation. Functional analysis of two genes, MNH6 and MgATG1, isolated from the cDNA subtractive library, revealed that the quantity of RNA was good enough to construct a cDNA (complementary DNA) library or a cDNA subtractive library. This method may be also applicable for the appressorium RNA isolation of other pathogenic fungi in which conidia differentiate into appressoria in the early stages of host infection.
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