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Changes of Membrane Stability in PotassiumStressed Plants

植物在缺钾胁迫中膜稳定性的变化



全 文 :植物在缺钾胁迫中膜稳定性的变化∗
王丹丹1ꎬ2ꎬ 郑国伟1ꎬ 李唯奇1ꎬ3∗∗
(1 中国科学院昆明植物研究所中国西南野生生物种质资源库ꎬ 云南 昆明  650201ꎻ 2 中国科学院大学ꎬ
北京  100039ꎻ 3 红河学院生物系ꎬ 云南 蒙自  661100)
摘要: 植物维持膜的功能是其抵御胁迫的关键问题ꎬ 而维持膜功能必须要保持膜的稳定性和合适的流动
性ꎮ 我们前期的研究发现植物主要是通过积累叶片膜脂和保持根部膜脂基本不变来适应长期缺钾ꎮ 在本研
究中ꎬ 以拟南芥和其具有耐受缺钾胁迫特性的近缘种须弥芥为对象ꎬ 研究了与膜的流动性密切相关的双键
指数 (double bond indexꎬ DBI) 的变化ꎬ 发现长期缺钾条件下ꎬ 两种植物叶片中总的 DBI保持不变ꎬ 根部
总的 DBI略有降低ꎮ 同时研究了与膜稳定性密切相关的溶血磷脂的含量和 DGDG / MGDG以及 PC / PE这两
个比值的变化ꎬ 发现长期缺钾后拟南芥和须弥芥叶片中溶血磷脂的总量呈上升趋势ꎬ 根部溶血磷脂总量基
本保持不变ꎻ 无论在对照还是缺钾条件下ꎬ 拟南芥溶血磷脂的总含量要高于须弥芥ꎮ 须弥芥叶片具有更高
的 DGDG / MGDG值ꎬ 根部具有更高的 PC / PE值ꎬ 说明长期缺钾条件下须弥芥膜的稳定性可能更好ꎮ 这可
能是须弥芥耐缺钾的原因之一ꎮ
关键词: 须弥芥ꎻ 拟南芥ꎻ 耐缺钾ꎻ 膜流动性ꎻ 膜稳定性
中图分类号: Q 945            文献标识码: A              文章编号: 2095-0845(2014)05-595-08
Changes of Membrane Stability in Potassium ̄Stressed Plants
WANG Dan ̄Dan1ꎬ2ꎬ ZHENG Guo ̄Wei1ꎬ LI Wei ̄Qi1ꎬ3∗∗
(1 The Germplasm Bank of Wild Speciesꎬ Kunming Institute of Botanyꎬ Chinese Academy of Sciencesꎬ
Kunming 650201ꎬ Chinaꎻ 2 University of Chinese Academy of Sciencesꎬ Beijing 100039ꎬ Chinaꎻ
3 Biology Departmentꎬ Honghe Universityꎬ Mengzi 661100ꎬ China)
Abstract: The maintenance of membrane function is critical to the ability of plants to resist environmental stressesꎻ
specificallyꎬ the stability and appropriate fluidity of membranes are crucial to their normal function. We previously
demonstrated that plants adapt to long ̄term potassium (K+) deficiency by accumulation of membrane lipids in leaves
and maintenance of the lipid composition in roots. In this studyꎬ which involved Arabidopsis thaliana and its K+  ̄defi ̄
ciency ̄tolerant relative Crucihimalaya himalaicaꎬ we first calculated the double ̄bond index (DBI) as an indicator of
membrane fluidity. After exposure to long ̄term K+  ̄deficiency stressꎬ the DBI of the total lipids in leaves of
A􀆰 thaliana and C􀆰 himalaica showed no significant changesꎬ whereas the DBI of the total lipids in the roots of these
species showed slight increases. Changes in lysophospholipids (lysoPLs) levelsꎬ and digalactosyldiacylglycerol / mo ̄
nogalactosyldiacylglycerol ( DGDG / MGDG) and phosphatidylcholine / phosphatidylethanolamine ( PC / PE) ratiosꎬ
all of which strongly reflect membrane stabilityꎬ were also studied in K+  ̄stressed A􀆰 thaliana and C􀆰 himalaica. After
long ̄term K+ deficiencyꎬ total lysoPLs levels increased in A􀆰 thaliana and C􀆰 himalaica leavesꎬ but showed no signif ̄
icant changes in roots. DGDG / MGDG and PC / PE ratios were higher in C􀆰 himalaica leaves and roots than in those of
A􀆰 thaliana. These results indicate that C􀆰 himalaica exhibits superior membrane stability compared with A􀆰 thaliana.
This may explain its superior growth and tolerance under K+  ̄deficient conditions.
植 物 分 类 与 资 源 学 报  2014ꎬ 36 (5): 595~602
Plant Diversity and Resources                                    DOI: 10.7677 / ynzwyj201413106

∗∗
Funding: The National Natural Science Foundation of China (NSFC 30670474ꎬ 30870571 and 31070262)
Author for correspondenceꎻ E ̄mail: weiqili@mail􀆰 kib􀆰 ac􀆰 cn
Received date: 2013-05-05ꎬ Accepted date: 2013-07-15
作者简介: 王丹丹 (1983-) 女ꎬ 助理研究员ꎬ 主要从事植物逆境分子生理学研究ꎮ E ̄mail: wangdandan@mail􀆰 kib􀆰 ac􀆰 cn
Key words: Crucihimalaya himalaicaꎻ Arabidopsis thalianaꎻ Tolerance of K+ deficiencyꎻ Membrane fluidityꎻ Mem ̄
brane stability
Abbreviations: digalactosyldiacylglycerolꎬ DGDGꎻ monogalactosyldiacylglycerolꎬ MGDGꎻ phosphatidylglycerolꎬ
PGꎻ phosphatidylcholineꎬ PCꎻ phosphatidylethanolamineꎬ PEꎻ phosphatidylinositolꎬ PIꎻ phosphatidylserineꎬ PSꎻ
phosphatidic acidꎬ PAꎻ double ̄bond indexꎬ DBIꎻ lysophospholipidꎬ lysoPLꎻ lysophosphatidylglycerolꎬ lysoPGꎻ ly ̄
sophosphatidylcholineꎬ lysoPCꎻ lysophosphatidylethanolamineꎬ lysoPEꎻ electrospray ionization tandem mass spec ̄
trometryꎬ ESI ̄MS / MS
  In our previous studyꎬ we found that Crucihima ̄
laya himalaicaꎬ a relative of Arabidopsis thalianaꎬ is
tolerant to K+ ̄deficient conditions ( Wang et al.ꎬ
2014). Compared with A􀆰 thalianaꎬ the higher ratios
of root / shoot dry weight and shoot K+ / root K+ in
C􀆰 himalaica might account for its superior growth and
lower levels of chlorosis under K+ ̄deficient condi ̄
tions. Detailed analysis of the lipid composition in
A􀆰 thaliana and C􀆰 himalaica indicated that plants
adapt to long ̄term K+ deficiency by the accumulation
of membrane lipids in leaves and the maintenance of
the lipid composition in roots (Wang et al.ꎬ 2014).
In addition to adjustment of the lipid compositionꎬ
the double ̄bond index (DBI) and level of lysophos ̄
pholipids (lysoPLs) in plants also change under ad ̄
verse conditions. It is well known that the degree of
unsaturation of glycerolipidsꎬ which is reflected by
DBIꎬ affects membrane fluidity. DBI is the average
number of double bonds in the two fatty acid chains
of a glycerolipid molecular species. DBI is increased
or decreased in order to enhance or reduce membrane
fluidity as an adaptation to low ̄temperature or high ̄
temperature stressꎬ respectively.
LysoPLsꎬ which include lysophosphatidylglycer ̄
ol (lysoPG)ꎬ lysophosphatidylcholine ( lysoPC) and
lysophosphatidylethanolamine ( lysoPE)ꎬ are minor
constituents of membrane lipidsꎬ but the levels of
lysoPLs are very sensitive to environmental stimuli.
Previous studies indicated that mechanical damage
(Lee et al.ꎬ 1997)ꎬ low ̄temperature stress ( Li et
al.ꎬ 2008ꎻ Welti et al.ꎬ 2002ꎻ Zhang et al.ꎬ 2013)
and abscisic acid ̄promoted senescence (Jia and Liꎬ
2013) induced the accumulation of lysoPLs in plants.
An increase in lysoPLs was also observed in drought ̄
and salt ̄stressed A􀆰 thaliana and Thellungiella halo ̄
phila plants (our unpublished data).
Different lipids have different effects on the
properties of membranes. It has been reported that
digalactosyldiacylglycerol ( DGDG) and phosphati ̄
dylcholine ( PC) are bilayer ̄preferring lipidsꎻ in
contrastꎬ monogalactosyldiacylglycerol (MGDG) and
phosphatidylethanolamine (PE) tend to form a hex ̄
agonal II phase ( Cullis et al.ꎬ 1986ꎻ Webb and
Greenꎬ 1991)ꎬ which has a strong propensity to re ̄
sult in membrane leakage. Many studies have shown
positive correlations between stress tolerance and
higher ratios of DGDG / MGDG and PC / PE (Chen et
al.ꎬ 2006ꎻ Quartacci et al.ꎬ 1997ꎻ Suss and Yor ̄
danovꎬ 1986ꎻ Toumi et al.ꎬ 2008ꎻ Zhang et al.ꎬ
2012). Howeverꎬ it remains unclear how DBIꎬ lyso ̄
PLsꎬ and the ratios of DGDG / MGDG and PC / PE
change with long ̄term K+ deficiency.
In this studyꎬ electrospray ionization tandem
mass spectrometry ( ESI ̄MS / MS) was employed to
determine the changes in the levels of lysoPLs in K+  ̄
stressed A􀆰 thaliana and C􀆰 himalaica. DBI was also
used as an indicator of membrane fluidity. DGDG /
MGDG and PC / PE ratios were calculated to estimate
the membrane stability of shoots and rootsꎬ respec ̄
tively. The results obtained indicate thatꎬ under K+  ̄
deficient conditionsꎬ changes of the DBI of total lip ̄
ids in the leaves of A􀆰 thaliana and C􀆰 himalaica dif ̄
fered from those of roots. The extent of the change in
the levels of lysoPLs in K+  ̄stressed A􀆰 thaliana and
C􀆰 himalaica leaves was larger than that in roots. In
additionꎬ the ratios of DGDG / MGDG and PC / PE
were greater in C􀆰 himalaica than in A􀆰 thaliana un ̄
der both control and K+  ̄deficient conditions.
695                                  植 物 分 类 与 资 源 学 报                            第 36卷
1  Methods and materials
1􀆰 1  Plant material
Seed ̄germinated plantlets of A􀆰 thaliana ( Co ̄
lumbia ecotype) and C􀆰 himalaica were used throug ̄
hout this study. Seeds of C􀆰 himalaica were collected
from the alpine cold desert soil of Baima Snow
Mountainꎬ which is in De Qinꎬ Yunnan.
1􀆰 2  Growth conditions and K+ ̄deficient treatment
Seeds of A􀆰 thaliana and C􀆰 himalaica were
planted on Murashige and Skoog medium and germi ̄
nated in a growth chamber at 22 ℃ with 12 h light /
12 h darkness photoperiod with a light intensity of
120 μmol m-2s-1 . For further growthꎬ plantlets were
transferred to a hydroponic system (Tocquin et al.ꎬ
2003 ). To assess the effects of K+ ̄deficiencyꎬ
A􀆰 thaliana and C􀆰 himalaica plants with rosettes of
the same diameter (30 ̄day ̄old A􀆰 thaliana plants and
35 ̄day ̄old C􀆰 himalaica plants) were transferred to a
modified hydroponic system with different potassium
levels: K+ sufficiency (5􀆰 1 mmol􀅰L-1 KNO3)ꎬ mild
K+ limitation (0􀆰 51 mmol􀅰L-1 KNO3)ꎬ severe K

limitation (0􀆰 051 mmol􀅰L-1 KNO3)ꎬ and K
+ depri ̄
vation (0 mmol􀅰L-1 KNO3). The nitrogen level was
balanced using NH4NO3 and the growth medium was
changed every six days.
1􀆰 3  Lipid extraction and ESI ̄MS / MS analysis
Lipid extractionꎬ ESI ̄MS / MS analysisꎬ and
lipid quantification were performed as described pre ̄
viously with minor changes (Welti et al.ꎬ 2002).
Leaves and roots of the plants were collected sepa ̄
rately and immediately transferred to isopropanol
with 0􀆰 1% butylated hydroxytoluene at 75 ℃ to in ̄
hibit lipolytic activities. They were then extracted
several times using chloroform / methanol (2 ∶ 1) with
0􀆰 1% butylated hydroxytoluene. The extracts were
gathered andꎬ when the extract solvent appeared
whiteꎬ the extraction procedure was terminated.
1􀆰 4  Data analysis
The Q ̄test was performed to remove discordant
data. The remaining data were then subjected to one ̄
way factorial ANOVA using SPSS 16􀆰 0. Differences
between means were tested by Fisher’s least signifi ̄
cant difference ( LSD) method. Double ̄bond index
(DBI) was calculated using the following formula: DBI
=[∑(N × mol% lipid)] / 100ꎬ where N is the total
number of double bonds in the two fatty acid chains of
each glycerolipid molecule (Zheng et al.ꎬ 2011).
2  Results and discussion
2􀆰 1  The patterns of DBI change differed between
plastidic and extraplastidic lipids in A􀆰 thaliana
leaves
In A􀆰 thaliana leavesꎬ the DBI of DGDG and
MGDG were higher than those of any other lipid
classꎬ whereas the DBI of PI and PS were the lowest
(Table 1). The DBI of MGDG was 5􀆰 88ꎬ whereas
that of PS was only 2􀆰 74. Under K+  ̄deficient condi ̄
tionsꎬ changes of DBI of total lipids showed no sig ̄
nificant difference compared with the control. Under
K+  ̄deficient conditionsꎬ DBI changes in most lipid
classes showed no significant difference compared
with the control. The largest change in DBI occurred
for PAꎬ which showed a 6􀆰 84% increase under 0
mmol􀅰L-1 K+ condition. Under K+  ̄deficient condi ̄
tionsꎬ patterns of DBI change differed between plas ̄
tidic and extraplastidic lipids. After long ̄term K+ de ̄
ficiencyꎬ the DBI of plastidic lipids showed a de ̄
creaseꎬ but that of extraplastidic lipids increased
(Table 1). The extent of DBI change of extraplastid ̄
ic lipids was larger than that of plastidic lipids. For
exampleꎬ the DBI of DGDG decreased 1􀆰 32%ꎬ
1􀆰 32% and 0􀆰 38% under conditions of 0􀆰 51ꎬ 0􀆰 051
and 0 mmol􀅰L-1 K+ꎬ respectivelyꎬ compared with
the control. In contrastꎬ the DBI of PC showed in ̄
creases of 2􀆰 30%ꎬ 3􀆰 57% and 3􀆰 57% under the
same K+ conditions (Table 1).
2􀆰 2   The extent of DBI change in A􀆰 thaliana
roots was larger than that in leaves
Under K+  ̄sufficient conditionꎬ the DBI of total
lipids in A. thaliana root was 3􀆰 45 (Table 2)ꎻ this
was much lower than that in leafꎬ which was
5􀆰 35. In additionꎬ the DBI of all lipid classes in
A􀆰 thaliana roots were lower than those in its leaves.
This phenomenon also occurred under K+  ̄deficient
7955期            WANG Dan ̄Dan et al.: Changes of Membrane Stability in Potassium ̄Stressed Plants             
conditionsꎬ which indicated that the membrane fluid ̄
ity in A􀆰 thaliana leaves was superior to that in roots.
Under control conditionsꎬ the DBI of MGDG was
5􀆰 69ꎬ which was much higher than those of other
lipids. The lowest DBI was exhibited for PGꎬ which
was only 2􀆰 11. Howeverꎬ in contrast to the case for
leavesꎬ there were no clear trends between K+  ̄defi ̄
ciency stress and changes in DBI in roots. Insteadꎬ
complex and varied patterns were displayedꎬ and the
changes were larger than those in leaves. The DBI of
total lipids in A􀆰 thaliana roots showed a 1􀆰 16% de ̄
creaseꎬ and 0􀆰 29% and 4􀆰 06% increases under
conditions of 0􀆰 51ꎬ 0􀆰 051ꎬ and 0 mmol􀅰L-1 K+
(Table 2)ꎬ respectivelyꎬ compared with the control.
Howeverꎬ there was a 0􀆰 75% decreaseꎬ a 0􀆰 56%
increaseꎬ and a 0􀆰 19% decrease in A􀆰 thaliana leav ̄
es under the same K+ conditions ( Table 1). The
changes of DBI in extraplastidic lipids were larger
than those in plastidic lipids.
2􀆰 3  The patterns of DBI change in C􀆰 himalaica
was similar to that in A􀆰 thaliana
In C􀆰 himalaica leavesꎬ the DBI of MGDG was
5􀆰 70ꎬ which was much higher than those of other
lipids. PG had the lowest DBIꎬ which was only 2􀆰 41
(Table 1). Under control conditionꎬ the DBI of each
lipid class and total lipids in C􀆰 himalaica leaves
were lower than those of A􀆰 thalianaꎬ a phenomenon
that also occurred in K+  ̄deficient C􀆰 himalaica and
A􀆰 thaliana leaves. This indicates that the membrane
fluidity in C􀆰 himalaica leaves was lower than that in
A􀆰 thaliana. Under K+  ̄deficient conditionsꎬ changes
in the DBI of total lipids in C􀆰 himalaica leaves
showed no significant difference compared with the
control. The DBI of extraplastidic lipids showed an
increase after the imposition of K+  ̄deficiency. For
plastidic lipidsꎬ the DBI of DGDG and PG increased
after exposure to both 0􀆰 051 and 0 mmol􀅰L-1 K+ .
Meanwhileꎬ the DBI of MGDG showed a 0􀆰 35%
Table 1  DBI of membrane lipids in A􀆰 thaliana and C􀆰 himalaica leaves following exposure to different levels of K+ . Values in the same
row with different letters differ significantly at P<0􀆰 05 (Fisher’s least significant difference) . An asterisk indicates that the value
differs significantly from that of A􀆰 thaliana under the same conditions (P<0􀆰 05)
Lipid
class Plant species
Double ̄bond index
Control 
0􀆰 51 
/ mmol􀅰L-1  
0􀆰 051 
/ mmol􀅰L-1  
0 
/ mmol􀅰L-1  
Relative change / %
0􀆰 51
/ mmol􀅰L-1
0􀆰 051
/ mmol􀅰L-1

/ mmol􀅰L-1
DGDG A􀆰 thaliana 5􀆰 29±0􀆰 03
a 5􀆰 22±0􀆰 05b 5􀆰 22±0􀆰 04b 5􀆰 27±0􀆰 07ab -1􀆰 32 -1􀆰 32 -0􀆰 38
C􀆰 himalaica 5􀆰 20±0􀆰 03a∗ 5􀆰 20±0􀆰 02a 5􀆰 16±0􀆰 04a∗ 5􀆰 17±0􀆰 01a∗ 0  -0􀆰 77 -0􀆰 58
MGDG A􀆰 thaliana 5􀆰 88±0􀆰 01
a 5􀆰 86±0􀆰 02ab 5􀆰 86±0􀆰 00ab 5􀆰 86±0􀆰 01b -0􀆰 34 -0􀆰 34 -0􀆰 34
C􀆰 himalaica 5􀆰 70±0􀆰 02ab∗ 5􀆰 72±0􀆰 01a∗ 5􀆰 69±0􀆰 02b∗ 5􀆰 70±0􀆰 02ab∗ 0􀆰 35 -0􀆰 17 0 
PG A􀆰 thaliana 3􀆰 20±0􀆰 04
a 3􀆰 25±0􀆰 14a 3􀆰 26±0􀆰 08a 3􀆰 20±0􀆰 06a 1􀆰 56 1􀆰 87 0 
C􀆰 himalaica 2􀆰 41±0􀆰 05ab∗ 2􀆰 43±0􀆰 05a∗ 2􀆰 35±0􀆰 02ab∗ 2􀆰 34±0􀆰 02b∗ 0􀆰 83 -2􀆰 49 -2􀆰 90
PC A􀆰 thaliana 3􀆰 92±0􀆰 07
b 4􀆰 01±0􀆰 10a 4􀆰 06±0􀆰 04a 4􀆰 06±0􀆰 05a 2􀆰 30 3􀆰 57 3􀆰 57
C􀆰 himalaica 3􀆰 59±0􀆰 02c∗ 3􀆰 79±0􀆰 04b∗ 3􀆰 81±0􀆰 05b∗ 3􀆰 90±0􀆰 02a∗ 5􀆰 57 6􀆰 13 8􀆰 63
PE A􀆰 thaliana 3􀆰 42±0􀆰 04
b 3􀆰 41±0􀆰 01b 3􀆰 53±0􀆰 05a 3􀆰 57±0􀆰 03a -0􀆰 29 3􀆰 22 4􀆰 39
C􀆰 himalaica 3􀆰 27±0􀆰 02c∗ 3􀆰 32±0􀆰 04b∗ 3􀆰 40±0􀆰 02a∗ 3􀆰 44±0􀆰 02a∗ 1􀆰 53 3􀆰 98 5􀆰 20
PI A􀆰 thaliana 2􀆰 79±0􀆰 01
b 2􀆰 74±0􀆰 04c 2􀆰 80±0􀆰 03ab 2􀆰 82±0􀆰 04a -1􀆰 79 0􀆰 36 1􀆰 07
C􀆰 himalaica 2􀆰 59±0􀆰 02c∗ 2􀆰 65±0􀆰 01b∗ 2􀆰 68±0􀆰 02ab∗ 2􀆰 71±0􀆰 01a∗ 2􀆰 32 3􀆰 47 4􀆰 63
PS A􀆰 thaliana 2􀆰 74±0􀆰 03
b 2􀆰 71±0􀆰 04b 2􀆰 72±0􀆰 02b 2􀆰 80±0􀆰 03a -1􀆰 09 -0􀆰 73 2􀆰 19
C􀆰 himalaica 2􀆰 58±0􀆰 02c∗ 2􀆰 62±0􀆰 02b∗ 2􀆰 66±0􀆰 03a∗ 2􀆰 69±0􀆰 01a∗ 1􀆰 55 3􀆰 10 4􀆰 26
PA A􀆰 thaliana 3􀆰 51±0􀆰 04
b 3􀆰 46±0􀆰 11b 3􀆰 56±0􀆰 07b 3􀆰 75±0􀆰 12a -1􀆰 42 1􀆰 42 6􀆰 84
C􀆰 himalaica 3􀆰 24±0􀆰 05b∗ 3􀆰 40±0􀆰 27ab 3􀆰 23±0􀆰 20b∗ 3􀆰 46±0􀆰 13a∗ 4􀆰 94 -0􀆰 31 6􀆰 79
Total A􀆰 thaliana 5􀆰 35±0􀆰 03a 5􀆰 31±0􀆰 09a 5􀆰 38±0􀆰 03a 5􀆰 34±0􀆰 20a -0􀆰 75 0􀆰 56 -0􀆰 19
lipids C􀆰 himalaica 5􀆰 17±0􀆰 04a∗ 5􀆰 25±0􀆰 07a 5􀆰 20±0􀆰 04a∗ 5􀆰 20±0􀆰 02a∗ 1􀆰 55 0􀆰 58 0􀆰 58
895                                  植 物 分 类 与 资 源 学 报                            第 36卷
Table 2  DBI of membrane lipids in A􀆰 thaliana and C􀆰 himalaica roots following exposure to different levels of K+ . Values in the same
row with different letters differ significantly at P<0􀆰 05 (Fisher’s least significant difference) . An asterisk indicates that the value
differs significantly from that of A􀆰 thaliana under the same conditions (P<0􀆰 05)
Lipid
class Plant species
Double ̄bond index
Control 
0􀆰 51 
/ mmol􀅰L-1  
0􀆰 051 
/ mmol􀅰L-1  
0 
/ mmol􀅰L-1  
Relative change / %
0􀆰 51
/ mmol􀅰L-1
0􀆰 051
/ mmol􀅰L-1

/ mmol􀅰L-1
DGDG A􀆰 thaliana 4􀆰 98±0􀆰 07
b 5􀆰 04±0􀆰 11ab 5􀆰 04±0􀆰 07ab 5􀆰 17±0􀆰 12a 1􀆰 20 1􀆰 20 3􀆰 81
C􀆰 himalaica 4􀆰 67±0􀆰 14a∗ 4􀆰 69±0􀆰 17a∗ 4􀆰 72±0􀆰 09a∗ 4􀆰 71±0􀆰 10a∗ 0􀆰 43 1􀆰 07 0􀆰 86
MGDG A􀆰 thaliana 5􀆰 69±0􀆰 06
a 5􀆰 47±0􀆰 44a 5􀆰 68±0􀆰 10a 5􀆰 73±0􀆰 05a -3􀆰 87 -0􀆰 18 0􀆰 70
C􀆰 himalaica 5􀆰 56±0􀆰 04a 5􀆰 37±0􀆰 30a 5􀆰 47±0􀆰 21a 5􀆰 46±0􀆰 25a -3􀆰 42 -1􀆰 62 -1􀆰 80
PG A􀆰 thaliana 2􀆰 11±0􀆰 04
b 2􀆰 20±0􀆰 04a 2􀆰 10±0􀆰 03b 2􀆰 18±0􀆰 05a 4􀆰 26 -0􀆰 47 3􀆰 32
C􀆰 himalaica 2􀆰 15±0􀆰 04ab 2􀆰 14±0􀆰 04ab∗ 2􀆰 12±0􀆰 05b 2􀆰 17±0􀆰 05a -0􀆰 46 -1􀆰 39 0􀆰 93
PC A􀆰 thaliana 3􀆰 50±0􀆰 05
ab 3􀆰 43±0􀆰 03c 3􀆰 44±0􀆰 06bc 3􀆰 52±0􀆰 02a -2􀆰 00 -1􀆰 71 0􀆰 57
C􀆰 himalaica 3􀆰 55±0􀆰 06b 3􀆰 56±0􀆰 03b∗ 3􀆰 68±0􀆰 04a∗ 3􀆰 70±0􀆰 11a∗ 0􀆰 28 3􀆰 66 4􀆰 22
PE A􀆰 thaliana 2􀆰 88±0􀆰 01
c 3􀆰 02±0􀆰 03b 3􀆰 07±0􀆰 02a 3􀆰 10±0􀆰 02a 4􀆰 86 6􀆰 60 7􀆰 64
C􀆰 himalaica 3􀆰 05±0􀆰 03c∗ 3􀆰 11±0􀆰 03b∗ 3􀆰 14±0􀆰 04ab∗ 3􀆰 17±0􀆰 05a∗ 1􀆰 97 2􀆰 95 3􀆰 93
PI A􀆰 thaliana 2􀆰 50±0􀆰 02
a 2􀆰 45±0􀆰 02b 2􀆰 46±0􀆰 02b 2􀆰 47±0􀆰 02ab -2􀆰 00 -1􀆰 60 -1􀆰 20
C􀆰 himalaica 2􀆰 54±0􀆰 04a 2􀆰 49±0􀆰 02b 2􀆰 55±0􀆰 02a∗ 2􀆰 55±0􀆰 06a∗ -1􀆰 97 0􀆰 39 0􀆰 39
PS A􀆰 thaliana 2􀆰 55±0􀆰 02
a 2􀆰 50±0􀆰 02b 2􀆰 56±0􀆰 04a 2􀆰 60±0􀆰 03a -1􀆰 96 0􀆰 39 1􀆰 96
C􀆰 himalaica 2􀆰 66±0􀆰 03b∗ 2􀆰 64±0􀆰 03b∗ 2􀆰 74±0􀆰 04a∗ 2􀆰 73±0􀆰 06a∗ -0􀆰 75 3􀆰 01 2􀆰 63
PA A􀆰 thaliana 3􀆰 29±0􀆰 03
a 3􀆰 27±0􀆰 06a 3􀆰 32±0􀆰 03a 3􀆰 32±0􀆰 08a -0􀆰 61 0􀆰 91 0􀆰 91
C􀆰 himalaica 3􀆰 36±0􀆰 05a 3􀆰 37±0􀆰 05a∗ 3􀆰 41±0􀆰 05a∗ 3􀆰 39±0􀆰 07a 0􀆰 30 1􀆰 49 0􀆰 89
Total A􀆰 thaliana 3􀆰 45±0􀆰 04b 3􀆰 41±0􀆰 13b 3􀆰 46±0􀆰 04b 3􀆰 59±0􀆰 03a -1􀆰 16 0􀆰 29 4􀆰 06
lipids C􀆰 himalaica 3􀆰 49±0􀆰 03bc 3􀆰 44±0􀆰 06b 3􀆰 61±0􀆰 09a∗ 3􀆰 58±0􀆰 14ab -1􀆰 43 3􀆰 44 2􀆰 58
increase and a 0􀆰 17% decrease under conditions of
0􀆰 51 and 0􀆰 051 mmol􀅰L-1 K+ꎬ respectivelyꎬ com ̄
pared with the control. The changes of DBI in extra ̄
plastidic lipids were larger than those in plastidic
lipids. The DBI of PC showed the largest changesꎬ
namelyꎬ 5􀆰 57%ꎬ 6􀆰 13% and 8􀆰 63% increases after
exposure to 0􀆰 51ꎬ 0􀆰 051 and 0 mmol􀅰L-1 K+ꎬ re ̄
spectivelyꎬ compared with the control.
Under K+  ̄sufficient conditionꎬ the DBI of total
lipids in C􀆰 himalaica root was 3􀆰 49 ( Table 2)ꎬ
which was much lower than that in leavesꎬ at 5􀆰 17.
Furthermoreꎬ the DBI of almost all lipid classes in
C􀆰 himalaica roots were lower than those in C􀆰 hima ̄
laica leavesꎬ except for PS and PA. This phenome ̄
non was also observed in plants subjected to K+ defi ̄
ciencyꎬ which indicates that the membrane fluidity
in C􀆰 himalaica leaves was superior to that in its
roots. This was similar to the case for A􀆰 thaliana. In
C􀆰 himalaica rootsꎬ lower DBI in PG and PI were
observed under both K+  ̄sufficient and K+  ̄deficient
conditionsꎬ whereas the DBI of DGDG and MGDG
were much higher than those of other lipids (Table
2). In contrast to leavesꎬ the DBI of most lipid clas ̄
ses and total lipids in roots were higher in C􀆰 himal ̄
aica than in A􀆰 thalianaꎬ with the exceptions of DG ̄
DG and MGDG. This suggests that membrane fluidity
in C􀆰 himalaica roots was higher than that in
A􀆰 thaliana. Under K+  ̄limited conditionsꎬ the chan ̄
ges in the DBI of extraplastidic lipids were larger
than those of plastidic lipidsꎬ and the DBI of PC and
PE showed greater changes than those of other lipids.
2􀆰 4  Total lysoPLs increased in K+ ̄stressed A􀆰 tha ̄
liana and C􀆰 himalaica leaves
LysoPLs are minor phospholipids of A􀆰 thalianaꎬ
but they are very sensitive to stresses such as ex ̄
treme temperature and drought. Under K+  ̄deficient
conditionsꎬ no tendency for change in each lysoPL
class in the leaves and roots of C􀆰 himalaica and
9955期            WANG Dan ̄Dan et al.: Changes of Membrane Stability in Potassium ̄Stressed Plants             
A􀆰 thaliana was observed (Table 3). We further cal ̄
culated the total amount of lysoPLs and found thatꎬ
under K+  ̄deficient conditionsꎬ it increased markedly
in C􀆰 himalaica and A􀆰 thaliana leaves (Table 4).
A. thaliana leaves showed 17􀆰 98%ꎬ 29􀆰 21%ꎬ and
60􀆰 67% increases under conditions of mild K+ limi ̄
tationꎬ severe K+ limitationꎬ and K+ deprivationꎬ re ̄
spectivelyꎬ compared with the control. In C􀆰 himala ̄
ica leavesꎬ when compared with the controlꎬ total ly ̄
soPLs showed 30􀆰 00% and 31􀆰 11% increases under
conditions of 0􀆰 051 and 0 mmol􀅰L-1 K+ . In con ̄
trastꎬ no significant changes in this variable were ob ̄
served in A􀆰 thaliana and C􀆰 himalaica roots.
The levels of total lysoPLs in K+  ̄stressed
A􀆰 thaliana leaves and roots were higher than those
in C􀆰 himalaica. At 4 ℃ꎬ lysoPCꎬ lysoPEꎬ and ly ̄
soPG in A􀆰 thaliana leaves were 0􀆰 09ꎬ 0􀆰 06ꎬ and
0􀆰 07 nmol􀅰mg-1ꎬ whereas when subjected to freez ̄
ing stress (-8 ℃)ꎬ they were 0􀆰 71ꎬ 0􀆰 8ꎬ and 0􀆰 12
nmol􀅰mg-1ꎬ respectively ( Li et al.ꎬ 2008). This
indicated that the level of lysoPL was positively re ̄
lated to the intensity of stress. The higher levels of
total lysoPLs in A􀆰 thaliana under K+  ̄deficient con ̄
ditions suggest that this species is far more sensitive
to K+ deficiency than C􀆰 himalaica.
2􀆰 5  The membrane stability of C􀆰 himalaica was
superior to that of A􀆰 thaliana under K+ ̄deficient
conditions
The ratios of DGDG / MGDG and PC / PE are
thought to be positively related to membrane stabili ̄
ty. Thereforeꎬ in this studyꎬ we used these ratios as
indicators of membrane stability in K+  ̄stressed
A􀆰 thaliana and C􀆰 himalaica. The ratio of DGDG to
MGDG can be used to estimate membrane stability in
leaves because they are the major constituents of
plastidic membranes in leaves. In the case of PC and
PEꎬ these are extraplastidic lipidsꎬ which account
for considerable proportions of root membrane lipids.
Thusꎬ the stability of root membranes and extraplas ̄
tidic membranes in leaves can be estimated using the
PC / PE ratio. K+ deficiency led to a slight decrease
in the DGDG / MGDG ratio in A􀆰 thaliana and C􀆰 him ̄
alaica leaves (Fig􀆰 1A)ꎬ but this ratio showed in ̄
creases in K+  ̄stressed A􀆰 thaliana and C􀆰 himalaica
roots. DGDG / MGDG ratios in C􀆰 himalaica leaves
were significantly higher than in A􀆰 thaliana under
both control and K+  ̄deficient conditions. Further ̄
moreꎬ C􀆰 himalaica roots also had a higher ratio of
DGDG / MGDG than those of A􀆰 thaliana under the
four different K+ conditions.
Fig􀆰 1  Changes in DGDG / MGDG and PC / PE ratios in A􀆰 thaliana and C􀆰 himalaica under K+  ̄deficient conditions. Data are means ± SD
(n= 4 or 5) . Bars with different letters differ significantly at P<0􀆰 05 (Fisher’s least significant difference) . Blank barsꎬ A􀆰 thalianaꎻ
dark gray barsꎬ C􀆰 himalaica. A. Changes in DGDG / MGDG following exposure to different K+ conditionsꎻ
B. Changes in PC / PE following exposure to different K+ conditions
006                                  植 物 分 类 与 资 源 学 报                            第 36卷
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1065期            WANG Dan ̄Dan et al.: Changes of Membrane Stability in Potassium ̄Stressed Plants             
    Under K+  ̄deficient conditionsꎬ a tendency for
an increase in the PC / PE ratio was observed for
both leaves and roots of both A􀆰 thaliana and
C􀆰 himalaica ( Fig􀆰 1B ). Under control and 0􀆰 51
mmol􀅰L-1 K+conditionsꎬ the PC / PE ratio in C􀆰 him ̄
alaica leaves was significantly higher than that in
A􀆰 thaliana. Howeverꎬ when grown under conditions
of 0􀆰 051 and 0 mmol􀅰L-1 K+ꎬ A􀆰 thaliana showed a
higher ratio of PC / PE than in the presence of 0􀆰 51
mmol􀅰L-1 K+ . Under the 0􀆰 51 mmol􀅰L-1 K+ condi ̄
tionꎬ the PC / PE ratio in A􀆰 thaliana root was higher
than that in C􀆰 himalaicaꎬ but under 5􀆰 1ꎬ 0􀆰 051
and 0 mmol􀅰L-1 K+ conditionsꎬ C􀆰 himalaica had
higher PC / PE ratios than A􀆰 thaliana.
In summaryꎬ the obtained results suggest thatꎬ
under K+ ̄deficient conditionsꎬ the DBI of most lipid
classes in A􀆰 thaliana and C􀆰 himalaica showed no
significant change when compared with that of the
control. The DBI of extraplastidic lipids showed an
increase under K+ ̄deficient conditionsꎬ whereas that
of plastidic lipids decreased with the decreasing avail ̄
ability of K+ . The membrane fluidity in A􀆰 thaliana
and C􀆰 himalaica leaves was higher than that in
roots. C􀆰 himalaica roots exhibited greater membrane
fluidity than A􀆰 thalianaꎻ in contrastꎬ the membrane
fluidity in A􀆰 thaliana leaves was superior to that in
C􀆰 himalaica. The total amount of lysoPLs was higher
in K+ ̄stressed A􀆰 thaliana than in C􀆰 himalaicaꎬ and
the ratios of DGDG / MGDG and PC / PE were higher
in C􀆰 himalaica leaves and roots than in A􀆰 thaliana.
These findings indicate that the membrane stability of
C􀆰 himalaica leaves and roots is superior to that of
A􀆰 thaliana under K+ ̄deficient conditions.
Acknowledgments: The authors thank Mary Roth (Kansas
Lipidomics Research Center) for lipid analysis.
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