全 文 ：Variation of the Mineral Elements in the Barks of
Pinus armandii Infected by Cronartium ribicola
YANG Fa-zhong1，YANG Bin1* ，YANG De-qiang2，ZHU Hai-feng1，HE Mei-jun1
1． Southwest Forestry College，Yunnan Province，Kunming 650224，China;2． College of Computer Science and Information Technology，Yun-
nan Normal University，Kunming 650092，China
Abstract ［Objective］The difference of mineral element content in healthy bark of P． armandii and bark affected by C． ribicola were compared，
which would provide theoretical foundation for exploring new ways to control the disease by artificial water and fertilizer． ［Method］ Inductively Cou-
pled Plasma Optical Emission Spectrometer(ICP-OES)was adopted to measure the variation of mineral elements content in bark after P． armandii
was affected by C． ribicola． ［Result］45 mineral elements in the healthy bark and 46 mineral elements in affected bark caused by C． ribicola were
detected． These mineral elements contained 14 kinds of mineral elements which were necessary for plant growth and 31 kinds of other mineral ele-
ments (32 kinds after affected disease)． The total content of mineral elements in affected bark of P． armandii decreased for 15． 77%，in which nec-
essary elements for plant growth decreased for 15． 54% and other elements decreased for 17． 07%． The content of Ca decreased the most，de-
crease rate was about 75%． The order of content was also changed，they were Na ＞Ca ＞B ＞K ＞Si ＞Mg ＞S ＞P in healthy bark，and Na ＞B ＞K ＞
Ca ＞Mg ＞P ＞Si ＞S in affected bark． ［Conclusion］The results laid foundation for controlling C． ribicola through application of mineral elements．
Key words P． armandii;C． ribicola;Mineral element;ICP-OES
Received:December 18，2009 Accepted:February 13，2010
Supported by The 10th Five-Years Key Programs for Science and
Technology Development of Yunnan Province (2003NG12)
* Corresponding author． E-mail:yangbin48053@yahoo． com． cn
The blister rust of five-needled pines is one of the three
diseases in world forest caused by Cronartium ribicola J． C．
Fisch．［1 －2］． In China，the fungus mainly infected Pinus ko-
raiensis Sieb． et Zucc． and Pinus armandii Franch，which had
largely occurred in forest area of P． armandii in Shaanxi，Si-
chuan［3 －4］ and Yunnan in recent years． The mechanism and
control method of the disease had been studied extensively at
home and abroad，but spread and extension of the disease still
hadn＇t been controlled effectively．
Mineral elements are the necessary substances for normal
growth and development of plant． On the one hand，it can be
used as components of plant tissues which directly participates
in metabolism;on the other hand，it also has effect on growth
pattern，morpha，anatomical characteristic and biochemical
characteristic of plants，which can increase or decrease the re-
sistance against plant diseases and affect plant growth and pro-
duction［5］． Some researches have shown［6］ that plant diseases
can affect the absorption of mineral elements，water-fertilizer
management can reduce the occurrence of disease to inhibit
the development of disease． The study measured the content
of mineral elements in healthy and Cronartium ribicola-infected
barks of P． armandii，in order to deeply understand the impact
mechanism of the disease to P． armandii and provide theoreti-
cal basis for exploring new ways to control the disease by artifi-
cial water and fertilizer．
1 Materials and Methods
1．1 Materials，reagents and equipment
1． 1． 1 Sources of material． Healthy bark of P． armandii and
diseased bark which were infected by C． ribicola were collected
from Dongchuan national 222 forest farms in Kunming of Yun-
nan Province．
1． 1． 2 Reagents． HNO3，HCl，HF，H2O2 were all analytical
reagents，and their solutions were prepared with ultrapure wa-
ter． Reservior solution including 1． 0 mg/ml K，Ca，Mg，Fe，P，
B，Mn，Zn，etc． were provided by chemical laboratory of
Southwest Forestry College (purchased from the National Iron
and Steel Testing Center，Iron and Steel Research Institute) ，
medium was 5% HNO3 solution．
1． 1． 3 Equipment． Inductively Coupled Plasma Optical Emis-
sion Spectrometer(ICP-OES)was produced by VARIAN com-
pany in United States． Model:VISTA-MPX CCD Simultaneous
ICP-OES． Range of wavelength:175 － 785 nm full coverage，
completely without breakpoints． Focal length was 0． 4 m，
groove of echelle grating was 97． 4 lines /mm;RF generator fre-
quency:40．68 MHz;plasma output power was 700 －1 700 W;
plasma cooling air was 0 －22．5 L/min which was adjustable．
1． 2 Methods
1． 2． 1 Sample processing． According to reference［7 － 8］，
the sample was washed with deionized water，placed in drying
oven (50 ℃)and dried，accurately weighed for 0． 500 0 g(first
soaked with 5% HNO3 for 10 h，pulverized，and sieved through
120 mesh) ，placed in high-pressure digestion tank (soaked
with 5% HNO3 for 10 h before being used)added concentrated
HNO3 for 5 ml，and after 10 h，added 1 ml concentrated HNO3，
1 ml H2O2 and 0． 5 ml HF，digested for 5 h after regulating di-
gestion temperature for 180 ℃． Sealing devices were removed，
and the solution was slowly heated to dry． 2 ml concentrated
Plant Diseases and Pests 2010，1(1):14 －17
HClO4 were added into residue，and the solution were slowly
heated to emit white fume to get rid of excess fluorin． 2 ml con-
centrated HNO3 were added after slight cooling，and the solu-
tion were slowly heated to dissolve all the residue，while the so-
lution became yellow or colorless． After cooling，the solution
was set the volume to 50 ml for test． The blank experiment was
conducted with the same method．
1． 2． 2 Measurement of mineral element content． Inductively
Coupled Plasma Optical Emission Spectrometer(ICP-OES)
were adopted to measure the content of inorganic element．
Working condition:RF power:1． 0 kW;plasma gas flow
quantum:15． 0 L/min;auxiliary gas flow quantum:1． 50 L/min;
nebulizer pressure:200．00 kPa;peristaltic pump speed 15 r /min;
reading time:10．000 s，settling time:10 s．
Selection of element analysis line:consulted equipment
operating software line database，chose the wavelength of ana-
lyte element with less interference within spectral line and mod-
erate sensitivity． Every mineral element was repeatedly meas-
ured for three times，the average value was taken as the con-
tent of mineral elements．
1． 2． 3 Accuracy of measurement． Series of standard solvent
were sampled for three times，average intensity versus concen-
tration was used to chart． The results indicated that the con-
centration range that all elements showed good linear relation-
ship were as follows:B 0 －100．00 mg/L，Ca 0 －100．00 mg/L，
Fe 0 －10．00 mg/L，K 0 －100．00 mg/L，Mg 0 －100．00 mg/L，
Mn 0 －10．00 mg/L，Zn 0 －10． 00 mg/L，P 0 －100． 00 mg/L．
Linear regression equation and correlation coefficient R were
shown in Table 1．
Table 1 Standard curve regression equation and correlation coef-
ficient of different elements
Elements Regression equation Correlation coefficient R
B y =3 999．41x + 4 438．1 1．000 000
Ca y =3 221．95x + 2 620．87 0．999 935
Fe y =18 293．6x + 1 433．2 0．999 994
K y =139 620x + 327 697 0．999 102
Mg y =1 289．23x + 281．922 0．999 999
Mn y =109 786x + 1 251．4 0．999 998
Zn y =16 997．2x + 1 038．39 0．999 966
P y =785．074x + 115．021 0．999 997
2 Results and Analysis
2．1 Changes of total mineral element content in the bark
of infected P． armandii ICP-OES were adopted to detect
mineral elements． The results showed that there were 45 kinds
of mineral elements detected from healthy bark of P． armandii
and 46 kinds of mineral elements detected from the bark of
P． armandii which had affected disease． There was no signifi-
cant difference between the kinds of mineral elements in healthy
bark and infected bark． These mineral elements contained 14
kinds of mineral elements which were necessary for plant
growth and 32 kinds of other mineral elements．
Fig． 1 showed that the content of mineral elements in infec-
ted bark of P． armandii decreased，the total content decreased
for 15． 77%，in which necessary elements for plant growth in
affected bark decreased for 15． 54% and other elements de-
creased for 17． 07% compared with healthy bark．
Fig． 1 The total content of mineral elements in healthy and
affected bark of P． armandii
2．2 Changes of content of necessary mineral element for
plant growth in the bark of infected P． armandii Fig． 2
showed that 14 kinds of necessary mineral element were all de-
tected from healthy bark and infected bark of P． armandii by In-
ductively Coupled Plasma Optical Emission Spectrometer． The
order of content in healthy bark were Na ＞Ca ＞B ＞K ＞Si ＞Mg
＞S ＞P ＞Fe ＞ Zn ＞Mn ＞ Cu ＞ Mo ＞ Ni，while the order of con-
tent in infected bark were Na ＞ B ＞ K ＞ Ca ＞ Mg ＞ P ＞ Si ＞ S ＞
Fe ＞ Zn ＞ Mn ＞ Cu ＞ Mo ＞ Ni． Most necessary elements for
plant growth decreased when P． armandii had been infected，in
Fig． 2 Analysis of necessary mineral elements in healthy and affected bark of P． armandii
which the content of Ca decreased the most，decrease rate was about 75%;the content of K，Mg，P increased，increase
51YANG Fa-zhong et al． Variation of the Mineral Elements in the Barks of Pinus armandii Infected by Cronartium ribicola
rate were 30%，20%，79%，respectively． In addition，the con-
tent of three necessary elements，Cu，Mo，Ni，were all less
than 0． 1 mg/g (dry weight)．
2． 3 Content analysis of other mineral elements in healthy
and infected barks of P． armandii Table 2 showed that in
addition to necessary mineral elements for plant growth，31 other
mineral elements were also detected from healthy bark of
P． armandii，32 other mineral elements were detected from in-
fected bark of P． armandii． Bi was detected in healthy bark，
while it didnt emerge in infected bark;Pt and Tl were detected
in infected bark，while they didn＇t emerge in healthy bark． Te，
Se，Rb in bark were substantially increased after P． armandii
were infected by C． ribicola，the content of other 12 mineral el-
ements including Zr，Cr，Sb，W，As，Ce，La，Nd，Sn，Au，
Li，Pb were significantly decreased．
Table 2 Analysis of other mineral elements in healthy and affect-
ed bark of P． armandii
Mineral
elements
Content ∥mg/g
Healthy bark Affected barks Increasing percent∥%
Ag 0．001 7 0． 001 4 －17． 65
Bi 0． 000 5 － －
Cr 0． 008 5 0． 005 3 －37． 67
Gd 0． 000 5 0． 000 4 －20
Pd 0． 000 9 0． 000 7 －22． 22
Te 0． 000 7 0． 055 9 798 5
V 0． 000 9 0． 000 9 0
Al 9． 659 9 7． 992 0 －17． 27
Cd 0． 001 3 0． 000 9 －30． 77
Pr 0． 002 5 0． 001 4 －44
Sb 0． 013 5 0． 010 8 －20
Sm 0．000 7 0． 000 5 －28． 57
W 0．003 1 0． 002 6 －17． 13
Y 0． 001 8 0． 001 9 4． 45
As 0． 011 9 0． 010 5 －11． 76
Ce 0． 013 6 0． 005 4 －60． 29
La 0． 002 9 0． 002 1 －27． 59
Nd 0． 004 3 0． 002 8 －34． 88
Pt － 0． 000 5 －
Sc 0． 000 2 0． 000 2 0
Sn 0． 004 8 0． 001 0 －79． 17
Tl － 0． 000 4 －
Yb 0． 000 2 0． 000 2 0
Au 0． 004 2 0． 001 5 －64． 29
Eu 0． 000 1 0． 000 1 0
Li 0． 006 6 0． 005 8 －12． 12
Pb 0． 020 2 0． 006 1 －69． 8
Rb 0． 014 8 0． 029 8 101
Se 0． 000 6 0． 002 3 283
Sr 0． 028 3 0． 014 3 －49． 47
Zr 0． 049 6 0． 020 8 －55． 66
Ti 0． 070 7 0． 055 9 －20． 93
Co 0． 000 3 0． 000 3 0
3 Discussion
As we all know that absorption and imbalance supply of
mineral will lead to physiological diseases of plants，infection of
plant pathogen will affect the normal physiology and metabolism of
plants，including absorption and transportation of mineral ele-
ments［6］．
The results showed that after P． armandii was infected by
C． ribicola，the total content of mineral elements and necessary
elements in bark assumed obvious downtrend，especially the
content of element Ca which had close relationship with plant
disease were significantly decreased，the decrease rate was up
to about 75%． Calcium formed calcium polygalacturonate a-
mong the cells，which increased the stability of cell wall． The
content of Ca decreased，which would increase the permeabili-
ty of cell． Therefore，small molecule compounds (sugars，ami-
no acids，etc．)would be leakage，which would provide nutri-
tion for reproduction and infection of mycelium［9］． Some re-
searchers had reported that Si and B had close relationship with
plant diseases，the content of the two elements decreased after
the plant was infected［10］． This experiment confirmed that the
content of these two elements in bark had different degrees of
reduction after P． armandii was affected by C． ribicola． In addi-
tion，trace necessary elements including Fe，Mn，Cu，Zn，etc．
were components of many kinds of enzyme，when P． armandii
was affected by C． ribicola，lack of these elements in bark of
P． armandii could change the activity of enzyme，causing dis-
order of metabolism and physiology． Because many studies
had confirmed that some mineral elements in infected plants
were decreased，application of fertilizer or nutritional liquid were
tested，which could help reducing the severity of disease． For
example，application of silicon fertilizer and nutritional solution
or fertilizer with B would be benefit for resisting Erysiphe spp
and Uromyces spp［11 －12］． When P． armandii was affected by
C． ribicola，the content of mineral elements decreased，wheth-
er addition of mineral elements such as Ca，Si，B，etc． could
inhibit the occurrence and development of pathogenic was
worth for further study．
Author had studied the changes of secondary metabolites
in healthy bark and infected bark after P． armandii was affected
by C． ribicola using HPLC，and found that chemical composi-
tion and content of healthy bark and infected bark had signifi-
cant difference，indicating that the disease had significant influ-
ence on the metabolism of P． armandii［13］． These test results
showed C． ribicola could affect the absorption of mineral ele-
ments，which further confirmed that the disease had significant
influence on the metabolism of P． armandii from the perspec-
tive of mineral nutrition．
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茶藨生柱锈菌侵染后华山松树皮矿质元素的变化
杨发忠1，杨 斌1* ，杨德强2，朱海峰1，何美军1
(1．西南林学院，云南昆明 650224;2．云南师范大学计算机科学与信息技术学院，云南昆明 650092)
摘要 ［目的］比较华山松健康树皮、感疱锈病树皮中矿质元素含量的差异，为深入了解该病害对华山松的影响机理、探索水肥控制该病害的新
途径提供理论依据。［方法］采用电感耦合等离子体发射光谱法(ICP-OES)测定华山松感染疱锈病(Cronartium ribicola)后树皮矿质元素含量的
变化。［结果］从华山松健康树皮中检测出矿质元素 45种，从感染疱锈病树皮中检测出矿质元素 46种。这些矿质元素中植物生长必需矿质元
素均为 14种，其他矿质元素 31种(感病后为 32种)。华山松感病后矿质元素总量降低了 15． 77%，其中，植物生长必需元素降低了 15． 54%，其
他元素含量降低了 17． 07%，其中 Ca含量减少最多，减幅约达 75%。含量大小顺序也发生了变化，健康树皮中为 Na ＞ Ca ＞ B ＞ K ＞ Si ＞ Mg ＞ S ＞
P，而在感病树皮中为 Na ＞ B ＞ K ＞ Ca ＞ Mg ＞ P ＞ Si ＞ S。［结论］该研究结果为施用矿质元素控制疱锈病病害奠定了基础。
关键词 华山松;茶藨生柱锈菌;矿质元素;ICP-OES
基金项目 云南省“十五”科技攻关项目(2003NG12)。
作者简介 杨发忠(1974 －)，男，云南楚雄人，硕士，讲师，从事化学生态学方面的研究。
收稿日期 2009-12-18 修回日期
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2010-02-13
(Continued from page 13)
［4］FENG ZM，WANG XE，WAN KJ． Phytoplasma on witche＇s broom of
Paulownia:a review［J］． Journal of Xinyang Agricultural College，
2007(4):126 －128． (in Chinese)．
［5］Chuomacuo． Occurrence regularity and preventive measure of Pau-
lownia witches broom［J］． Modern Agricultural Science and Technol-
ogy，2008(11):156． (in Chinese)．
泡桐丛枝病的鉴别与防治
李栋海 (廊坊职业技术学院，河北廊坊 065000)
摘要 简述了泡桐丛枝病病原及特征、发生规律、为害症状，并总结了该病的防治方法。
关键词 泡桐;丛枝病;病原;发生现状;防治对策
作者简介 李栋海(1961 － )，男，河北三河人，高级讲师，从事植物保护研究。
收稿日期 2009-08-25 修回日期 2010-01-21
71YANG Fa-zhong et al． Variation of the Mineral Elements in the Barks of Pinus armandii Infected by Cronartium ribicola