Basic HTML Version
分子植物育种
(
网络版
), 2012
年
,
第
10
卷
,
第
1179
-
1184
页
Fenzi Zhiwu Yuzhong (Online), 2012, Vol.10, 1179
-
1184
http://mpb.5th.sophiapublisher.com
1179
研究报告
A Letter
氮饥饿诱导对稻瘟菌分泌蛋白组影响的研究
孔垂思
1,2
,
苏源
1,3
,
余萍
1
,
杨静
1
,
刘林
1
,
朱有勇
1
,
李成云
1
1
云南农业大学教育部生物多样性与病虫害控制重点实验室
,
昆明
, 650201;
2
云南省农业科学院农业环境资源研究所
,
昆明
, 650205;
3
昆明学院生命科学与技术系
,
昆明
, 650214
通讯作者
:
li.chengyun@gmail.com
作者
分子植物育种
, 2012
年
,
第
10
卷
,
第
25
篇
doi: 10.5376/mpb.cn.2012.10.0025
收稿日期：
2012
年
05
月
09
日
接受日期：
2012
年
05
月
27
日
发表日期：
2012
年
06
月
04
日
这是一篇采用
Creative Commons Attribution License
进行授权的开放取阅论文。只要对本原作有恰当的引用
,
版权所有人允许并同意第三方无条
件的使用与传播。
引用格式
(
中文
)
：
孔垂思等
, 2012,
氮饥饿诱导对稻瘟菌分泌蛋白组影响的研究
,
分子植物育种
(online) Vol.10 No.25 pp.1179-1184 (doi: 10.5376/mpb. cn.2012.10.0025)
引用格式
(
英文
)
：
Kong et al., 2012, Proteomic Analysis of Nitrogen Starvation Induced Secretome from
Magnaporthe oryzae
, Fenzi Zhiwu Yuzhong (online) (Molecular
Plant Breeding) Vol.10 No.25 pp.1179-1184 (doi: 10.5376/mpb.cn.2012.10.0025)
摘
要
氮是生物必不可少的营养元素，病原菌在与寄主的互作中对氮元素的竞争也就显得尤为重要。利用蛋白质双向凝
胶电泳技术分析在含氮培养条件下稻瘟病菌蛋白质谱的变化，可了解该病菌氮营养代谢的特征，分析不同组分蛋白质的毒性，
可了解这些蛋白在与寄主互作中氮营养争夺的特点，从营养争夺的角度分析稻瘟病菌与水稻互作的机制。本实验首先利用液
相等电聚焦电泳分离稻瘟病菌
Y98
-
16
菌株在氮饥饿诱导下的分泌蛋白，并在水稻感病品种丽江新团黑谷
(LTH)
的叶片上进
行致伤接种。结果显示，等电点
4.2~6.9
范围内的分泌蛋白中对
LTH
叶片有较强的毒性。然后利用
pI4
-
7
分离范围的胶条对
全氮培养与氮饥饿诱导下稻瘟病菌分泌蛋白进行比较蛋白质组学的研究。结果表明
Y98
-
16
在全氮培养条件下平均检测到
238
个蛋白点，在氮饥饿诱导下平均检测到
173
个蛋白点，其中表达量有显著变化的蛋白点有
51
个
(
上调
22
个
,
下调
29
个
)
，
利用串联质谱鉴定了其中
26
个蛋白质。对这些蛋白质功能的分析表明，这些蛋白功能涉及代谢、催化、应激、发育、防御
/
抗胁迫、细胞分裂等过程。
关键词
稻瘟菌
;
分泌蛋白
;
双向电泳
;
氮饥饿
Proteomic Analysis of Nitrogen Starvation Induced Secretome from
Magna
-
porthe oryzae
Kong Chuisi
1,2
, Su Yuan
1,3
, Yu Ping
1
, Yang Jing
1
, Liu Ling
1
, Zhu Youyong
1
, Li Chengyun
1
1. Key Laboratory of Ministry of Education for Agricultural Biodiversity and Plant Disease Control, Yunnan Agricultural University, Kunming, 650201, P.R.
China;
2. Institute of Agricultural Environment and Resources, Yunnan Academy of Agricultural Sciences, Kunming, 650205, P.R. China;
3. Department of Life Science and Technology, Kunming University, Kunming, 650214, P.R. China
Corresponding author, li.chengyun@gmail.com;
Authors
Abstract
Nitrogen is necessary nutrition for organisms, and it is very important for pathogen and host interactions. The
characteristics of nitrogen metabolism in
M. oryzae
which grows in medium containing nitrogen could be detected through 2
-
DE by
using MS analysis, and then the toxicities of different protein fractions could be analyzed, as well as their features in nitrogen
nutrition competition between pathogen and host, thereby the interaction mechanism between pathogen and host could be analyzed
from nutrition competition. The total secreted proteins extracted from isolate Y98
-
16 were separated into 20 fractions by using
Rotofor (Bio Rad product) preparative isoelectric focusing apparatus. The virulences of all fractions were tested on LTH, a
susceptible rice cultivar. The results show that the pI span of the strongest virulence fraction is from pI4.2 to pI6.9. Then, the
comparative proteome analysis of the secreted proteins from Y98-16 at nitrogen starvation induction and nitrogen culture was carried
out by using two-dimensional gel electrophoresis with pI4
-
7 IPG strips. 173 and 238 spots were detected on the 2
-
DE maps of the
secreted proteins at nitrogen starvation induction and at nitrogen culture respectively. Among them, 51 protein spots have significant
differences (up regulation 22 and down regulation 29), and moreover 26 of these spots were identified through MALDI TOF/TOF.
According to function analysis, these differential proteins are involved in molecular functions including metabolism, catalysis, stress,
development and cell division and so on.
Keywords
Magnaporthe oryzae
; Secreted protein; Two-dimensional gel electrophoresis; Nitrogen starvation