全 文 ：广 西 植 物 Guihaia 28(1)：100— 106 2008年 1月
Growth response and t
of Agrostis stolonifera to Cu2+
Zn2+，Cd2+or Pb2+stress
ZHAO Shu-Lan，DU0 Li_An
(College of Chemistry and Li Sciences，Tianjin Normal University，Tianjin 300387，China)
Abstract：By sand cuItural method，the experiment was conducted to investigate growth response and threshold con—
centration of Agrostis stoloni[era to Cuz+，ZnZ+，CdZ+ or pbz+stress．It was found that rate of seed germination
decreased with increasing supply of Cue+，Znz+，CdZ+or pb2+．Heavy metal supply at concentration≤100 mg／L
increased plant height of A．stoloni fera，whereas Cue+，Zn0+，Cd +or Pb0+supply higher than 100 mg／L decreased
plant height．Cue+ inhibited root growth significantly，and root length decreased with increasing Cu抖 supply．Com—
pared with the contro1．root length of A．stolonifera was lowered by 93．75 at Cu +concentration of 600 mg／L．
Aboveground biomass increased at Cue+，Znz+or pbz+concentration≤200 mg／L，but decreased with increasing
concentrations when their concentrations were higher than 200 mg／k Chlorophyl content increased at Cu抖 ，Znz+
concentration≤100 mg／L or CdB+，pbB+concentration≤200 mg／L，but tended to decrease at Cu抖 ，Zn +eoncen—
tration≥IOO mg／L or Cde+，pb2+concentration≥200 mg／L．Only Cd。+significantly lowered chlorophyl content
of A．stoloni[era by 43．55 at concentration of 600 mg／L．Synthetical effect of heavy metal on A．stolonifera
growth indicated that A．stoloni[era was most sensitive to Cue+ stress with low threshold，but the effect of Zn +
stress on A．stolonifera growth was the least．
Key words：heavy metal stress~Agrostis stolonifera；growth response~threshold concentration
CLC number：Q948．116 Docum ent code：A Article ID：1000—3142(2008)01—0100—07
Nowadays heavy metal contamination is prevalent
throughout the world，mainly coming from various hu—
man activities such as mining，smelting，manufacturing
processes of metals，vehicle emi ssions，and dumping of
industrial wastes(Charkhabl et a1．，2005；Aksoy et
a1．，2005；Lin et a1．，2005；Speir et a1．，2003)．On
this aspect，researchers have conducted extensive and
thorough work (Shu et a1．，2004；Chander et a1．，
2001；Chen et a1．，2003)． Heavy metals have a Iow
biodegradability and high persistence in the environ—
ment(Liao，1992)．Their high toxicity on biological
organisms，especially to human beings through food
chain has been drawn great attention in recent years．
However，heavy metal contami nation on crops and veg—
etables has received increasing attention(Athar et a1．，
2002；Du a1．，2003；Moreno et a1．，2002；Cui et a1．，
2003)，and phytoremediation of heavy metal—polluted
soil has become a hot issue in the world(Lasat 2002；
Tandy eta1．，2004；W alker eta1．，2004；Kayser eta1．，
2000)．Cu and Z are essential elements for plant
growth and development，but may be harmful at high
concentrations． Cd2+ and Pb + are not essentia1 ele—
ments in metabolic processes in plants，they can accu—
mulate to concentrations that are to·Xl’C or lethal to
plants．Cu ，Znz ，Cdz and Pb pollution is com—
mon in urban environment with rapid industrial devel一
收稿日期：2006-07-21修 回日期：2006—12-26
基金项目：天津市高等学校科技发展基金(20040716)；天津市科技发展计划项目(043100611)；国家自然科学基金(59878033)[Supported by the
Science and Technology Development Foundation for High Schools of T1a in(2OO4O716)；Tianjin Municipal Science and Technology Development
Program(043100611)；the National Natural Sc{ence Foundation of China(59878O33)f
作者简介：赵树兰(1968一)，天津市人，硕士，副教授，主要研究方向为污染生态。
通讯作者(Author for correspondence)
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赵树兰等：匍茎剪股颖对 Cu 、Zn 一、Cd抖与 Ph 胁迫的生长响应与阈限浓度 101
opment．W hat is needed to point out，in urban ecosys—
tem ，turfgrass plays an important role in alleviating
contamination and protecting environment(Chen et
a1．，1991)．But heavy metal toxicity on turfgrass has
not been paid much attention (Cui et a1．，2004)．
Agrostis stolonifera is one of the common turfgrasses
used for ornamental lawn and sports turf．It is planted
and applied in large areas，has great values of beautif—
ying environment and serving society (Chen et a1．，
1991)．The objective of the current study was to in—
vestigate growth responses and threshold concentra—
tions of A．stoloni to Cu。 ，Zn2 ，Cd and Pb
stress．It could provide a scientific basis for regulating
standard of heavy metal concentration in lawn irriga—
tion water in China and for turfgrass remediation of
heavy metal—polluted places．
1 Materials and methods
1．1 M aterials
Sand was used as turfgrass meUium． It was
sieved，rinsed with distilled water repeatedly，then ov—
en-dried at 105 ℃ for 1 h and then 80 ℃ for 8 h．
Agrostis stolonife L was chosen as plant materia1．
Heavy metal treatments were conducted by solving Cu—
S04·5H2O，ZnS04·7H2O，3CdS04·8H2O or Pb
(NO3)2．Cu ，Zn ，Cd。 and Pb抖 treatment con—
centrations were all 0，1O0，200，300，400，500 and 600
mg／L respectively(Zhao et a1．，2002a)．
1．2 M ethods
1．2．1 Turfgrass culture and heavy metal stress 70 g
sand me dium was loaded into culture dishes in diameter
of 9 cm and 1 00 seeds of A．stolonifera were even
sown in each dish．At the same time，above heavy met—
al solutions with equal volumes were added to each
dish respectively until the solution immersed the seeds，
with 0 mg／L used as the control(CK)．Each treatment
was replicated three times．After seed germination of
A．stoloni
．
，tufgrass culture was conducted in light
incubator for 15 d under the following con~tions：30℃
during the day and 25℃ at night，relative humidity of
5O ～7O ，light culture in 24 h．Watering was per—
formed with equal amount daily throughout the experi—
ment to maintain adequate medium moisture．
1．2．2 Index measurment Seed germination rate was
recorded daily until the seedling stage． 10 random
seedlings per dish were chosen to determ ine plant
height and their average was assumed as the plant
height of the dish．After the experiment was fulfiled，
plants of each dish were mowed，washed and then oven
dried at 105℃ for 1 h to kill cells。followed by 80℃
for approximately 8 h to constant weight．They were
weighed in dry weight to determine aboveground bio—
mass．The dried samples were extracted wi th 8O ac—
etone to determine chlorophyll content (Li，2000)．
O．O2 g dried plant leaf was precisely weighed and put
into mortar．A little quartz sand and calcium carbonate
was added． Then the compound was ground and ex—
tracted fully with 80 acetone．It was placed in dark—
ness for 3～ 5 mi n．then filtrated into 25 mL volumetric
flask and fixed volume with 80 acetone．The extract
was determined absorhance at wavelength 663 nm and
645 nm and chlorophyll content was calculated．The
roots of each dish were washed fairly free of sand．10
random plants were selected to determ ine root length，
and the longest fibrous root was assumed as the root
length of the plant．The root lengths of 1 0 plants were
averaged．
1．2．3 Data evaluation an d an alysis Synthetical effect
index(SEI)was applied to evaluate the synthetical
effect of Cu ，Zn ，Cd or Pb on A．stoloni Ⅱ
growth(Duo et a1．，1999)．The SEI can be calculated
by the folowing formula：SEI一(∑ +∑Br十∑Rr十
XCr)／4，where∑ is the sum of difference of relative
percentage of seed germination rate with Cu ．Zn2 ．
Cd2 or Pb。 each stress concentration and contro1．
EBr is the sum of difference of relative percentage of
aboveground biomass wi th Cu ，Zn2 ，Cd or Pb。
each stress concentration and control，∑Rr is the sum
of difference of relative percentage of root length with
Cu2 ，Z ，Cd2 or Pb each stress concentration
and control，and ECr is the sum of difference of relative
percentage of chlorophyll content with Cu抖 ，Zn抖 ，
Cd + or Pb + each stress concentration and control
(Duo eta1．，1999)．
Threshold concentration(TC)is an index to indi—
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1O2 广 西 植 物 28卷
cate the sensitivity of plants to pollutant stress(Zhao
et a1．，2002b)．In current study，as the representatives
of aboveground and underground growth indices，
aboveground biomass and root length were chosen as
growth indices to calculate threshold concentration to
each heavy metal stress． The synthetical threshold
concentration can be calculated by：TC=(Br30 4-Rr30)／
2，where Rr30 is the threshold concentration of each
heavy metal when aboveground biomass of A．stolonif-
era decreased 30 of the control，Rm is the threshold
concentration of each heavy metal when root length of
A．stolonifera decreased 30 of the control(Zhao et
n2．，2002b)．
Analysis of variance was performed on all the data
sets． The least significant difference was applied for
multiple comparison．Statistical analysis of the data was
perform ed using SPSS statistical package(SPSS 12．0)．
0 200 400 600 800
Heavy meta I concent rat i on(mg／L)
Fig．1 Effects of heavy metal stress on seed
germination of A．stoloni}em
2 Results
2．1 Effects of heavy metal on seed germination
The addition of Cu +，Zn +，Cd +or Pb +led to a
reduction in seed germination rate across the treat—
ments，and germination rate decreased as heavy metal
concentration increased (Fig．1)．Among four heavy
metals，Cd抖 inhibited seed germination of A．stolonif-
era significantly，espedally at high concentrations．At
Cd concentration of 600 mg／L，seed germination rate
decreased by 63．04 as compared with contro1．Sig—
nificant differences(P< 0．01)were found in seed ger—
mination rate among treatments of each heavy meta1．
For Cu stress，the result of further multiple compari—
son was 0 ，100 ，200似 ，300眦 ，400 ，500∞ ，600。
(mg／L)，among which different letters denoted signifi—
cant difference(P< 0．01)in seed germination rate be—
tween Cu2+ concentrations． For Zn2+，Cd + and Pb +
stress，the resuIts were 0 ，100 ，200AB，300A~c，400~c，
500 ，600 (mg／L)；0 ，100 ，200 ，300 ，400 ，500 ，
600c(mg／L)and 0 ，100 ，200眦 ，300K，400m，
500雎，600。(mg／L)respectively．
2．2 Effects of heavy metal on plant height
Except Pb stress，the differences in plant height
of A．stolonifera were statistically significant among
treatments of other three heavy metals(P< O．01，Fig．
2)． Cu +，Zn +，Cd + and Pb + supply at concentra—
tions≤ 100 mg／L promoted plant height growth，but
inhibited it at concentrations>100 mg／L．As com—
pared with control，plant height decreased by 4O．72 ，
31．22 ，48．87％ and 19．91 respectively at Cu抖，
Zn 。Cd and Pb concentration of 600 mg／L Sim—
ilar to seed germi nation，Cdz inhibited plant height
growth significantly at 600 mg／L．Analysis of variance
indicated the results of 0Bc，100AB，200ABc，3O0AB，
400AB，500 ，600 (mg／L)for Cu stress，0AB，100AB，
200 ，300AB，400AB，500AB，600 (mg／L)for Zn2 stress
and 0 ，100 ，200．~ ，3O0AB，400AB，500AB，600 (mg／L)
fOr Cd stress in which different letters showed signif—
icant difference (P< 0．0 1) in plant height between
stress concentrations of each heavy meta1．
4
0
0 200 400 600 800
Heavy meta I concent rat i on(mg／L)
Fig．2 Effects of heavy metal stress on
plant height of A．stolonifera
2．3 Effects ofheavy metal on aboveground biomass
Fig．3 showed the results of shoot biomass under
different stress concentrations of four heavy metals．
As compared with the control，Cu ，Zn2 ，Pb in一
∞ ∞ ∞ ∞ 加 0
co 一 仍c—EL∞龇 D∞∞疗
一E0一 ∞ c
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赵树兰等：匍茎剪股颖对 Cu 、zn抖、Cd。 与 Pb。 胁迫的生长响应与阈限浓度 103
creased aboveground biomass at concentrations≤200
mg／L，but aboveground biomass decreased with in—
creasing stress concentrations when concentration was
higher than 200 mg／L。However，for CA stress，the
maximum shoot biomass was observed at concentration
of 100 mg／L． Cu。+，Zn。+，Cd。+and Pb。+stress at
high concentration clearly reduced shoot biomass
． As
compared with the control，the shoot biomass of A．
stoloni
． r“decreased by 40．00 ，30．00 ，30。30％
and 20．20 respectively at Cu。 ，Zn。 ，Cd。 and
Pb stress concentration of 600 mg／L．After the
maximum biomass，significantly negative correlation
lied between biomass(Br)and stress concentration(C)
of each heavy meta1． The regressive equations for
Cu抖 ，Z ，Cd。 and Pb抖 stress were Brc 2 一O
． 162
— 0．0002 2+，r一 0．9948 ；Brz 2+ 一 0．210—
0．0002c， 2+，r= 0．9825一 ；B 。+ = 0．1653—
0．0002 dH ，r一 0．9955 and Breb¨ = 0。178—
0．0002Cpb2+，r= 0。9815一 ，respectively． Analysis of
variance showed that there was significant difference
(P< O．0 1)in biomass among treatments of each heavy
meta1．The results of multiple comparison were 0AB，
lOO ，200 ，300Aa，400Au，5OO ，600 (mg／L) for
Cu。 stress，OAB，1OOAB，200 ，300Au，4OOAB，5OOAB，600
(mg／L)for Znz stress，O ，100 ，2OO拙，3OO ，
4。O ，500~c，600 (mg／L)for Cd2 stress and 0Au，
1OO ，200 ，300Au，400A~，5OO ，600。(mg／L) for
Pb。 stress in which different letters denoted signifi—
cant difference(P< 0。O1)in aboveground biomass be—
tween stress concentrations of each heavy metal。
2．4 Effects of heavy metal on root length
Except Cd stress，there were significant differ—
ences in root length among treatments of other three
heavy metals(P< 0。01)。Zn ，Cd。 and Pb。 stress
showed the same tendency，namely，root length in—
creased first and then decreased，and t he maximum ap—
peared at concentration of 100 mg／L．However，Cu。
inhibited root growth of A。stolonife greatly．moreo—
ver，root length decreased significantly with increasing
Cu2 supply。 As compared with control，root length
decreased by 93。75 at Cu。 concentration of 600
mg／L(Fig．4)。After the maximum，negative correla—
tion was found between root length (Rr) and stress
concentration(C)of Zn。十，Cd。十and Pb。十．The regres—
site equations for Cu。十．Zn。J。．Cd。 and Pb。 stress
were Rrc 2十一0。85— 0。0016Cc 2+，r一 0．8940一 ；
Rrz 2+一 2。02— 0．002Cz H ，r一 0．8915一 ；R， 。+一
1．5193—0．0011( 2十，r= 0．9606一 ；Rrpb2+一 1．934
— 0．0018CPb2十，r一0．9088一 respectively．Analysis of
variance showed the following results：0 ，1 00 ，200 ，
3OOcD，400。，500。，600 (mg／L)for Cu。+ stress，0 ，
100 ，200 ，300 ，400 ，500 ，600 (mg／L)for Zn。
stress and 0 ，100 ，200 ，300 ，400 ，500 ，600 (mg／
L)for Pb stress in which different letters denoted
significant difference(P< 0．01) in root length be—
tween stress concentrations of each heavy meta1．
0 1 8
0．1 6
0 14
0．1 2
[
0．1
0 08
警 0．06
—
0 04
0．02
O
蚕
}
0 200
Heavy meta
400
COncent rat
600 8O0
on(mg／L)
Fig．3 Effects of heavy metal stress O11
aboveground biomass of A．stolonifern
2．5
2
1 5
1
0．5
0
0 200 400 600 800
Heavy meta J concent rat i on(mg／L)
Fig．4 Effects of heavy metal stress on root
length of A。stolonifern
2．5 Effects of heavy metal on chlorophyl content
Significant differences were found in chlorophyll
content among treatments of each heavy metal(P<
0。01，Fig。5)。Cu。 ，Zn。 ，Cd。 and Pb。 stress at low
concentrations increased chlorophyll content． But at
high concentrations chlorophyll content decreased，
probably because chlorophyll synthesis was hindered or
∞∞∞Eo—cI uc3oL >o cI( 一Eu—L{ ￡ 一 oo
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赵树兰等：匍茎剪股颖对 CuH、Zn 、Cd。 与Pb 胁迫的生长响应与阈限浓度 105
creased，the effects of four heavy metal ions on plant
height，root length，biomass and chlorophyll content
showed the sarne tendency，namely，heavy metals at
low concentrations(≤ 100 mg／L or 200 rag／L)had
positive effect，above indices appeared maxiinuin，but
after the rnaximum，t he3,decreased with the increase of
heavy metal concentrations．
From t he angle of applied value of turfgrass，root
growth and greenness are of vital importance because
root development would affect turfgrass growth and
greenness would directly affect ornamental properties
of turfgrass．The basic deleterious effect of Cu。。 on
growth is related to the root system． Faust et a1．
(2000) reported that rOOt development of Penncross
creeping bentgrass decreased as Cu。 concentration in—
creased．Similarly，in the present study，Cu。。 inhibited
root elongation of A．stoloniJera significantly，root
length decreased by 93．75 at Cu。。 concentration of
600 nag／[ when compared with contro1． So Cu。
would probably limit turf establishment of A．stoloni{一
era．Analysis on synthetical effect of heavy metals and
threshol d concentration showed the same results
． A．
st()kmi r_was FIOSI sensitive to Cu2 stress due to
marked inhibidon of root growth． However，A．stolo
niJk,,-a was tolerant to Zn with high concentrations，
which could be confirmed by other report(Palazzo et
a1．，2003)． So in practical application，irrigation of
turfgrass should strictIy controI concentration of CuH ．
1)L1c to high toxicity of Cd 。。and Pb ．their concentra—
tions should be regulated low，but concentration of
Zn纠 may be higher
．
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匍茎翦股颖对 Cu2+、Zn2+、Cd2+与 P1)2+
胁迫的生长响应与阈限浓度
赵树兰，多立安
(天津师范大学 化学与生命科学学院，天津 300387)
摘 要：采用砂培法，研究了匍茎翦股颖对Cuz+、Zn抖、Cd +与 Pb +胁迫的生长响应及阈限浓度，结果表明：种
子萌发率随着4种重金属浓度的增加而下降。对株高的影响是当重金属浓度小于100 mg／L时会促进株高生
长，高于 100 mg／L则产生抑制作用。Cu +显著抑制根系生长，并随浓度的增加抑制效应愈加显著；在Cu 浓
度为600 mg／L时匍茎翦股颖的根长比对照下降了93．75 。cu21卜、Zn2+、Pb +浓度小于200 mg／L时会促进地
上生物量的增加，但高于 200 mg／L时，地上生物量会随着 3种重金属的增加而减少。Cu 、Zn2+浓度小于 100
mg／L或 CdZ+、Pbz+浓度小于 200 mg／L会增加叶绿素的含量，高浓度会降低叶绿素的含量；Cd +在浓度为 600
mg／L时显著降低叶绿素含量，与对照相比，下降了43．55 。匍茎翦股颖生长的综合效应分析表明，匍茎翦股
颖对Cuz+胁迫最敏感，具有较低的阈限浓度，而Zn2+胁迫对匍茎翦股颖的生长影响最小，阈限浓度相对较高。
关键词：重金属胁迫；匍茎翦股颖；生长响应；阈限浓度
石 N e 石 { 暑 暑石 暑 暑石 暑石 暑 ＼! ＼! 0 暑 = 暑 ＼! 嘧 ：
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为进行物种多样性保护研究提供有益的旁证。为对
五列木种群适应特性以及该种群对于群落构建的生
态学意义等问题进一步展开研究提供有益帮助。为
五列木种群保护和管理提供依据。保护珍稀濒危植
物是维持生物多样性必不可少的措施之一，也是扩大
该种群的有效途径，通过研究五列木种群生态特征，
可为五列木种群保护和管理提供依据。由于五列木
种群Ⅲ级小树最多，发展潜力大，因此，五列木种群的
年龄结构呈发展型。人为干扰轻，种群发展趋势良
好，因此深入开展研究和保护是必要的。且重在保
护，尤其是保护其生境，使种群呈良性发展。怎样进
行保护呢?笔者认为除了对五列木的资源状况、生境
和群落特征进行研究外，还需进一步弄明白五列木种
群的濒危机制、濒危过程、该物种生存所需的条件等。
生科院O3级韦玉梅、叶又玮、苏立宁、陈秋霞、
徐作东，O2级的孟世勇、潘华勇、谢磊林、王亮同学
参加了取样工作。对以上同学特以致谢!
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