分子植物育种
(
网络版
), 2016
年
,
第
14
卷
,
第
1087-1094
页
Fenzi Zhiwu Yuzhong (Online), 2016, Vol.14, 1087-1094
Copyright © 2016 BioPublisher 1087
研究报告
Research Report
解除独行菜种子低温萌发停滞的温度响应蛋白筛选及表达分析
李萍萍
1
,
曾卫军
1
,
周茜
1
,
李艳红
1
,
葛风伟
1
,
祝长青
1
,
赵君洁
1
,
卢函
1
,
赵惠新
1
,
赵和平
2
1
新疆师范大学生命科学学院，新疆特殊环境物种保护与调控生物学实验室
,
乌鲁木齐
, 830054
2
北京师范大学生命科学学院
,
抗性基因资源与分子发育北京市重点实验室
,
北京
, 100875
通讯作者
,
;
;
作者
分子植物育种
, 2016
年
,
第
14
卷
,
第
13
篇
doi: 10.5376/mpb.cn.2016.14.0013
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
)
：
李萍萍等
, 2016,
解除独行菜种子低温萌发停滞的温度响应蛋白筛选及表达分析
,
分子植物育种
(online), 14(13): 1087-1094 (doi:
10.5376/mpb.cn.2016.14.0013)
引用格式
(
英文
)
：
Li et al., 2016,
Screening and Expression Analysis of Temperature-responding Proteins that can Remove Low Temperature Stagnancy of Germination in
Lepidium
Seeds Fenzi Zhiwu Yuzhong (online) (Molecular Plant Breeding), 14(13): 1087-1094 (doi: 10.5376/mpb.cn.2016.14.0013)
摘 要
新疆北部早春短命植物独行菜
(
Lepidium apetalum Willd.
)
种子萌发对温度响应独特：低温层积可以促进种子的萌发
并显著增加整齐度，但在
4
℃条件下存在低温萌发停滞现象，而经
25
℃处理
45~55 min
及更长时间后，种子则可以在
4
℃条
件下萌发。这种短暂的高温诱导解除低温萌发停滞的机制尚不清楚。本研究对处于低温层积前、萌发停滞状态及
25
℃高温
处理解除低温萌发停滞后的种子进行蛋白质差异分析，筛选出解除独行菜种子低温萌发停滞的温度响应蛋白，采用定量
qRT-PCR
技术对相应基因的表达进行了验证。结果表明：低温层积前、后独行菜种子之间差异蛋白不显著；
25
℃高温处理
45 min
解除低温萌发停滞的种子与前二者种子蛋白差异显著。从中筛选出
37
个差异蛋白，其中
14
个上调，
23
个下调。选
择显著上调的
6
个蛋白进行
LC/MS-MS
鉴定，并在生物信息学分析基础上，进一步选择其中的
Cdc48E
、
Hsp17.6
和
Per12
进行
qRT-PCR
验证，结果表明：这三个蛋白的基因在解除低温萌发停滞的种子中表达量极显著增高；随
25
℃处理时间增加，
三个基因表达均呈上升趋势，且处理在
45~55 min
时上调表达达到最高，之后维持这一水平表达，这与经
25
℃处理
30~60 min
能够解除低温萌发停滞进行萌发的种子比率显著增加的特性相一致。以上结果说明
Cdc48E
、
Hsp17.6
和
Per12
可能与解除独
行菜种子低温萌发停滞有密切关系，此结果为植物温度信号响应的研究提供了新的线索。
关键词
独行菜
,
种子萌发
,
低温停滞
,
响应蛋白
Screening and Expression Analysis of Temperature-responding
Proteins that can Remove Low Temperature Stagnancy of
Germination in
Lepidium
Seeds
Li Pingping
1
, Zeng Weijun
1
, Zhou Xi
1
, Li Yanhong
1
, Ge Fengwei1 , Zhu Changqing
1
, Zhao Junjie
1
,
Lu Han
1
, Zhao Huixin
1
, Zhao Heping
2
1 Xinjiang Key Laboratory of Special Species Conservation and Regulatory Biology, College of Life Science, Xinjiang Normal University, Urumqi, 830054
2 Beijing Key Laboratory of Gene Resource and Molecular Development, College of Life Science, Beijing Normal University, Beijing, 100875
Corresponding authors,
;
Authors
Abstract
As a kind of ephemeral plant,
Lepidium apetalum
Willd. in northern XinJiang shows special responsible characteristics
to temperature during the germination: low temperature stratification could improve germination rate and homogeneity of
Lepidium
seeds. But it could not germinate and be at stagnation stage under 4
℃
, while this phenomenon could be removed by the treatment
with 25
℃
for 45 minutes or longer time. The mechanism why the treatment by higher temperature for a short time could remove
germination stagnation under low temperature is not clear yet. In this study the total proteins difference of seeds with three treatments
were analyzed by 2D, which the seeds were without stratification, stayed at germination stagnation stage under low temperature and
removed stagnation with 25
℃
treatment. Screened proteins responding to temperature which were related to remove germination
stagnation under low temperature, and the corresponding genes expression were confirmed by qRT-PCR. The result showed that
there was no obvious difference between the seeds without stratification and the seeds at germination stagnation stage by low
temperature. But the proteins expression was quite different between the seeds treated with 25
℃
for 45 minutes to remove the
germination stagnation and the other two treated samples. 37 distinct protein spots were found in the 2-DE map with 14 up-regulated
spots, 23 down-regulated spots. Among them, 6 up-regulated proteins of 14 spots were identified by LC-MS/MS, CDC48E, HSP17.6
