全 文 ：广 西 植 物 Guihaia 24(3)：259— 262 2004年 5月
黑杨水培中营养液抑菌处理研究
周立刚，宋卫堂，谢光辉，王勇军，刘西莉，李健强
(中国农业大学农学与生物技术学院，北京 100094)
摘 要：水培条件下进行快速繁殖是大量生产黑杨种苗的途径之一，营养液的抑茵处理是水培的一个重要方
面。该研究进行了二硫氰甲烷、中生菌素、敌磺钠、链霉紊、青霉紊等五种杀细菌剂和多茵灵、福美双、三唑酮
等三种杀真菌剂的抑茵试验，结果表明，杀细菌剂对微生物生长的抑制率要高于杀真菌剂，二硫氰甲烷和中生
菌素能较好地抑制营养液中微生物的生长，浓度分别为 20 mg／L和 15 mg／L时，抑制率分别为 75．6O 和
78．94 。杀真菌剂和杀细菌剂复配对营养液中微生物生长的抑制表现出较好的加和作用，其中以中生菌素
(15 mg／L)与福美双(5 mg／L)复配的效果最好，抑制率为 87．48 。
关键词：黑杨；营养液；杀细菌剂；杀真菌剂；抑茵处理
中图分类号：Q945 文献标识码：A 文章编号：1000—3142(2004)03—0259—04
● J■ - l - l J J n · · · Antlm icro }ia treatm ent 0tnutrient
solution in hydroponic cultivation
of Populus nigra
ZHOU Li—gang，SONG Wei—tang，XIE Guang—hui，
WANG Yong-jun，LIU Xi—li，LI Jian-qiang
(Colege of Agronomy and Biotechnology，China Agricultural tiniversitv。Beiinz 100094。China)
Abstract；Fast micro—propagation by hydroponic cultivation is one of the methods in large scale production of
Populus nigra seedlings．Antimicrobial treatment of nutrient solution is an important aspect in hydroponic
cultivation．Inhibition experiments on microbial growth were carried out by using five bactericides which were
dlisothiocyanatomethane，zhongshengmycin，fenaminosulf，streptomycin，penicillin，and three fungicides which
were carbendazim，thiram and triadimefon．The results showed that the inhibition effects of bactericides a—
galnst microorganisms in the nutrient solution were better than those of the fungicides．Of bactericides。both
diisothiocyanatomethane and zhongshengmycin at their concentrations of 20 mg／L and 15 mg／L strongly inhib—
ited the growth of the microorganisms which the corresponding inhibitory rate were 75．60％ and 78．94％sep—
arately．It was found that microbial growth were inhibited to a great extent by the combination of one bacteri—
cide with one fungicide，and an additional effects of the agrochemicals were also observed．The inhibition rate
of zhongshengmycin at 1 5 mg／L in combination with thiram at 5 mg／L Was 87．48 which was the highest in—
hibition rate of al combination effects．
Key words：Populus nigra L．；nutrient solution；bactericide；fungicide~antimicrobial treatment
Received date：2003·。07·。17 Accepted date：2003·。09’-24
基金项目：北京市科委工厂化高效栽培技术研究项目(HO2O72003O63O)；河北省科技计划项目(O322O169D)。
作者简介：周立刚(1965一)，男，湖南长沙人，博士，主要从事植物生物技术、植物与其它生物相互作用的研究。E-mail：[gzhou@
cau．edu．cn
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260 广 西 植 物 24卷
1 Intr0ducti0n
Poplar(P0p Z spp．)plants，which belongs to
family Saliaceae，have many characters such as fast
growth，drought resistance，easily hybrid and asex—
ual cuttage propagation． They are suitable for
short—rotation，intensive cultivation．Poplar plants
are of considerable ecological value and capable of
producing utilizable timber(Ceulemans et a1．，
1999)． There are more than 6O poplar species
widely distributed in many parts of China(Delectis
Florae Rerpublicae Popuparis Sinicae，1984)．Black
Poplar(P．nigra L．)was introduced from Europe
to China in 1 980s，and its farming has been estab—
lished as this species has many good characters o—
ver other poplar species．It has become one of the
first forest tree species to be cultivated in most
parts of China(Shi et a1．。2000)．
Poplar asexual cuttage method has been used
widely instead of traditional seed propagation(Ken—
ny et a1．，1990)．As poplar seedlings are in great
demand，tube micropropagation and hydroponic
propagation which need fl small amounts of plant
materials have also been studied(Jafari et a1．，
1995)．In the process of hydroponic propagation，
nutrient solution can be polluted easily by microor—
ganisms．This would lead to fast replacement of
nutrient solution，cast increasing，stem segment
rot，and seedling disease occurring．In order to in—
hibit microbial growth in hydroponic system，some
methods such as ultraviolet，ozone，surface activa—
tor，filtration，adsorption，osmotic pressure treat—
ments have been studied (Zhang，1999)．To the
best of our knowledge，there is still no published
report on antimicrobial treatment of nutrient solu—
tion with agrochemicals in hydroponic cultivation
of P．nigra．In this study，appropriate antimicrobi—
al agrochemicals were screened．The objective of
our work was to lay a foundation for the hydropo—
nic propagation of black poplar in large scale，and
also to provide some evidences for other plants hy—
droponic propagation in order to prevent microor—
ganism pollution．
2 Materials and Methods
2．1 M aterials
The materials of Black Poplar(P．nigra L．)
No．1 10 was provided by Dadongliu Seedling Nurs—
ery of Beij ing Forestry Bureau． The stems of
healthy male poplar were selected and cut into seg—
ments as long as five to six centimeters．One bud
was kept in morphological upper part on each seg—
ment．The morphological lower part would be kept
in nutrient solution．All the segments were washed
with tap water for 10 min。and then kept in 1 of
fresh sodium hypochlorite solution for 1 0 min，then
washed with tap water for 5 min．The sterilized seg—
ments would be utilized for further experiments．
2．2 Culture condition
The poplar stem segments were germinated
and grown in fl greenhouse with 12 h of light and
12 h of dark intervals。at 30~35℃ in daytime and
2O～ 25℃ at night．Relative humidity was kept in
6O％～8O％．The light density was about 3 000 l】【．
2．3 Plant nutrient solution
The compositions of Black Poplar nutrient so—
lution was as follows．In one liter of nutrient solu—
tion，there were 500 mg of Ca(NO3)2，125 mg of
MgSO‘·7H2O，125 mg of KH2PO‘，60 mg of
KC1。1。43 mg of H3BO3。0．11 mg of ZnC12，0．04
mg of CuSO‘·5H2O，0．01 mg of H2MoO‘，O．O4
mg of MnCl2，2．68 mg of Na,EDTA，2 mg of Fe—
SO4·7H2O，2 mg of gibberellin，0．2 mg of a—naph—
thalene acetic acid。1．5 mg of benzyl adenine．The
pH value of nutrient solution was adjusted to 5．8．
2．4 Agrochemicals and their inhibition on microor-
ganisms
All the agrochemicals were kindly supplied by
the Seed Coating Chemical Center of China Agri—
cultural University． Agrochemicals included five
bactericides which were diisothiocyanatomethone，
zhongshengmycin，fenaminosulf，streptomycin and
penicillin，and three fungicides which were carben-
dazim。thiram and triadimefon． When the poplar
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3期 周立刚等：黑杨水培中营养液抑菌处理研究 261
segments were cultured in nutrient solution，the
agrochemicals were added in at the same time．Ten
replicates for each test nutrient solution were set
up．Aher three weeks co—culture，the percentage
growth inhibition was measured by following the
absorbance at 655 nm and was defined as 100一times
the ratio of the change in absorbance of the average
growth in the control(△C)minus the change in
absorbance in the test nutrient solution(△T)，the
difference then divided by△C；i．e．[(△C一△T)]／
△CX 100(Marcus et a1．，1997)．
Table 1 Inhibition of agrochemica[s on microbial
growth in nutrient solution of P．nigra )
A gr oc
。
h
。
e-
C oncen-
t
I nhib i
t
-
e 蕊t e
There were both bacteria and fungi in the nutrient solution of P
nigra}Inhibition rate of the control was 0．
3 Results
3．1 Inhibition of agrochemicals on microbial growth
in nutrient solution
It was found that all eight agrochemicals had
no inhibitory effects on black poplar stem segments
growth with the tested concentration in nutrient
solution．Their inhibitory effects on microorgan—
isms were shown in table 1．Appropriate concen—
tration for agrochemicals to inhibit microbial
growth were as follows：diisothiocyanatomethone
20 mg／L(inhibition rate 75．60 0A)，zhongshengmy—
cin 15 mg／L(inhibition rate 78．94 )，fenamin—
osulf 35 mg／L(inhibition rate 2O．30％)，strepto—
mycin 50 mg／L(inhibition rate 75．52％)，penicilin
50 mg／L(inhibition rate 72．49 )，carbendazim 50
mg／L(inhibition rate 10．53 )，thiram 5 mg／L(in—
hibition rate 6．70 )and triadimefon 50 mg／L(inhibi—
tion rate 9．11 )．Of five bactericides，diisot~ocyanat-
omethone was the most effective one．Effects of bacte—
fiddes were better than those of fungicides．
3．2 Inhibition of agrochemical cornbination on mi-
crobial growth in nutrient solution
In order to get fl better inhibitory result，five
bactericides with their appropriate concentration sepa—
rately combined with three fungicides at their different
concentrations．Inhibition effects of chemical complex
on microbial growth of nutrient solution of black pop—
lfir was shown in fi 1．Results showed that combina—
tion of one fungicide with a bactericide have an ad~ng
efects．Of them，15 mg／L of zhongshengmydn in
combination with 5 mg／L of thlram(Fig．1：B)，which
inhibitory efect was 87．48％，had fl good effect than
any other combinations．
4 Discussion
The poplar stem segments(or called stem aN—
plants)were easily infected by microorganisms in—
eluding pathogenic and non—pathogenic ones in in
vitro culture．Otherwise，microorganisms also easi—
ly grew in nutrient solution with an open culture
system．They can contaminate whole culture sys-
tern in fl short time．So it is very necessary to pre—
vent poplar seedlings in nutrient solution from mi-
croorganism contamination． M ain microorganisms
in nutrient solution were bacteria，and fungi were
also found．Separation，purification and identifica—
tion of the microorganisms in poplar nutrient solu—
tion have not been finished．In this paper，we just
reported the results of some bactericides and fungi—
tides to inhibit microorganism growth in nutrient
solution．Our results showed that appropriate ag—
rochemicals with their combination in nutrient so—
lution were very effective to inhibit microorganisms
growth，and these agrochemicals have been proved
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262 广 西 植 物 24卷
to have no inhibitory effects on black poplar seed—
ling growing．Our results of the agrochemicals on
poplar seedling growing are j ust primary，further
investigation is needed． Otherwise，all the agro—
chemicals which we tested were low toxic to envi—
ronment as well as to human．Combina【tion of bac—
l00
80
60
40
100
80
60
40
20
0
0 0．5 5 50
Concentration
Concentration(mg／L)
lOO
(mg／L)
1OO
害 8O
是 6O
上
C
一 40
tericides and fungicides at their appropriate concen—
trations i．e．1 5 mg／L of zhongshengmycin in com—
bination with 5 mg／L of thiram，had fl good inhibi—
tory effect on microorganism growth in nutrient
solution．The nutrient solution could be replaced
every 60 days with agrochemical treatment instead
80
40
0 0．5 5 50 1OO
Concentration(mg／L)
O O．5 5 5O
lOO
Concentration
lOO
(mg／L)
0 0．5 5 50 lO0
Concentration(mg／L)
Fig．1 Inhibition of agrochemical complex on microbial growth in nutrient solution of P．nigra
A was 20 mg／L of disothiocyanatomethane combination with three fungicides separatelyI B was 15 mg／L of zhongshengmycin combination
with three fungicides separately‘C was 35 mg／L of fenaminosulf combination with three fungicides separately‘D was 50 mg／L of strepto—
mycin combination with three fungicides separately)E was 50 mg／L of penicilin combination with three fungicides separately．X-eoordi—
hate was the concentration of earbendazim．thiram and triadimefon IY-coordinate was the inhibition rate of microbial growth 口was the
carbendazim complex I 口 was the thiram complex I 图 was the triadimefon complex．
of previous 15 days without agrochemicals treat—
ment(data not shown)．This study will be benefit
to hydroponics cultivation of black poplar as well
as to other plant hydroponics culture．It will lay fl
foundation to decrease culture cost，provide healthy
seedlings for poplar cultivation in future．
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