Molecular Plant Breeding2016, Vol.7, No.18, 1
-
8
1
Research Article Open Access
Screening and Expression Analysis of Temperature-responsing Proteins that Can
Remove Low Temperature Stagnancy of Seeds Germination of
Lepidium apetalum
Zhao Junjie
1
, Zhao Huixin
1
, Li Pingping
1
, ZengWeijun
1
, Li Yanhong
1
, Ge Fengwei
1,
, Zhu Changqing
1
, Lu Han
1
, Zhao Heping
2,
1 Xinjiang Key Laboratory of Special Species Conservation and Regulatory Biology, College of Life Science, Xinjiang Normal University, Urumqi, 830054, China
2 Beijing Key Laboratory of Gene Resource and Molecular Development, College of Life Science, Beijing Normal University, Beijing, 100875, China
Corresponding authors Email
Molecular Plant Breeding, 2016, Vol.7, No.18 do
Received: 29 Mar., 2016
Accepted: 03 May, 2016
Published: 12 May, 2016
Copyright© 2016
Zhao et al., This is an open access article published under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article
:
Zhao J.J., Zhao H.X., Li P.P., Zeng W.J., Li Y.H., Ge F.W., Zhu C.Q., Lu H., and Zhao H.P., 2016, Screening and Expression Analysis of
Temperature-responsing Proteins that Can Remove Low Temperature Stagnancy of Seeds Germination of
Lepidium
apetalum
, Molecular Plant Breeding,
7(18): 1-8 (doi
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
L.apetalum
seeds. But it could not germinate and be at stagnation stage under 4°C, while this phenomenon could be removed by the treatment
with 25°C 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 2-DE, which the seeds were without stratification, stayed at germination stagnation stage under low
temperature and removed stagnation with 25°C treatment. Screened proteins responsing 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°C 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
and
PER12
were further chosen to be confirmed expression analysis by qRT-PCR. The result showed that their relative expression
levels were significantly higher in seeds removed stagnation than in seeds before breaking stagnation. In addition, their expression
levels were obviously increased with the time prolonging treated by 25°C with the highest level with the dealt for 45-55min and then
maintainng this level of expression. The result showed that the genes expression levels were positively correlated to the germination
percentagetreated with 25°Cfrom 30-60 minutes. All the above suggested that Cdc48E, Hsp17.6, and Per12 induced by 25°C might
be related to the germination stagnation removing of the seeds of
L.apetalum
. This conclusion provides a new clue for the study on
the signal response to temperature in plants.
Keywords
Lepidium apetalum
Willd.; Seed Germination; Germination Stagnation by low temperature; Response protein
Introduction
Lepidium apetalum
Willd., an annual herbaceous ephemeralsis of
Cruciferae Lepidium
, its root, stem and leaf
have large parenchyma cells (Yu and Liu, 1997), and its vascular bundle possess a characteristic anatomy called
kranz anatomy with high light effect, belongs to C4 plants (Li, 2009).
L. apetalum
seeds are flat egg shaped and in
reddish brown, its rich in fat, protein, sugar, mustard glycosides, and have effect of cardiac, antiasthmatic, diuretic.
Similar to
Lepidium perfoliatum
seeds,
L. apetalum
seeds’ dry surface covered with a layer of mucus quality seed
coating, which main component is polysaccharide. In case of water, the seed coating emit outside like rays (Gu,
2008), and absorb moisture quickly from the surrounding soil for germination, but it barely work on seedling
growth (Yuan et al., 2006). The
L. apetalum
live in the desert of Northern Xinjiang, is pioneer plant in flora (Li,
2014), so it has important ecological significance. And the
L. apetalum
has the characteristics of typical ephemeral
plants, which can tolerant low temperature to germinate and grow in early spring. Zhao Huixin et al., (2012)
screened and acquired 18 genes of cold up-regulated expression genes from
L. apetalum
seedlings using
cDNA-AFLP technology, this work provides some clues for the study of the molecular basis of
L. apetalum
