全 文 ：十字花科根肿病菌在不同温
度、pH 值条件下致病性差异
研究
裴卫华 1，李向东 1，杨佩文 1，曹继芬 1，毕云青 1，
杨子林 2，芮文 3，马桂明 4，林兴华 2，周丽凤 1，杨
明英 1* （1.云南省农业科学院农业环境资源
研究所，云南昆明 650205；2.临沧市植保植检
站，云南临沧；3.丽江市农产品质量安全检测
中心，云南丽江 674100；4.丽江市植保植检站，
云南丽江 674100）
摘 要 [目的] 探讨十字花科白菜根肿病菌
在不同温度、pH 值条件下致病性。 [方法]采用
根肿病菌和土壤混合法， 在休眠孢子浓度 1×
108（个/g 土）条件下，设置 7 个温度及 6 个土
壤 pH 值梯度，进行根肿病菌对白菜致病性测
定，90 d 后拔根调查根肿病菌侵染发病情况。
[结果]在 pH 值 6 时，不同温度梯度间白菜根
肿病发病株率存在较大差异，其发病株率从高
到低的温度为 30 ℃＞25 ℃＞20 ℃＞35 ℃＞15 ℃＞
10 ℃＞5 ℃。 5 ℃和 10 ℃条件下，其根肿病菌
侵染力弱， 不发病或发病株率较低；15～30 ℃
条件下随温度升高发病率增加，但在 35 ℃条
件下，发病株率降低。 温度 25 ℃条件下，pH值
为 4、5、6、7、8、9的白菜发病株率分别为 80.36%、
100%、65%、10.77%、3.23%、0.0%。 pH 值 4 发
病率虽高，但白菜生长受抑制，生长弱，存活率
低。 pH 值 7 和 8 时，发病率较低，pH 值 9 时没
有发病，在 pH 值 5 和 6 时，植株生长良好，发
病率也较高。 [结论]该研究结果表明根肿病菌
侵染发病与温度、土壤 pH 值关系密切。
关键词 根肿病菌；温度；pH 值；致病性
基金项目 云南省科技计划（2014RA061）；云
南省现代农业油菜产业技术体系。
作者简介 裴卫华(1982-），男，山西平遥人，
助理研究员， 主要从事植物病害应用技术研
究，E-mail：peiwh@qq.com。* 通讯作者，E-mail：
ymying18＠aliyun.com。
收稿日期 2014-09-30
修回日期 2014-12-05
Study on Pathogenicity Difference of
Plasmodiophora brassicae Under Different
Temperature and pH Conditions
Weihua PEI1, Xiangdong LI1, Peiwen YANG1, Jifen CAO1, Yunqing BI1, Zilin YANG2, Wen RUI3,
Guiming MA4, Xinghua LIN2, Lifeng ZHOU1, Mingying YANG1*
1. Institute of Agricultural Environment and Resources, Yunnan Academy of Agricultural Sciences, Kunming 650205, China;
2. Lincang Plant Protection Station, Lincang, 677000, China;
3. Supervision and Testing Center for Agricultural Product Quality of Lijiang City, Lijiang 674100, China;
4. Lijiang Plant Protection Station, Lijiang 674100, China
Supported by Science and Technology Project of Yunnan Province (2014RA061);
Special Fund for Modern Agriculture Research System for Rape of Yunnan Province.
*Corresponding author. E-mail: ymying18＠aliyun.com
Received: September 30, 2014 Accepted: December 5, 2014A
Agricultural Science & Technology, 2015, 16(1): 112-115
Copyright訫 2015, Information Institute of HAAS. All rights reserved Plant Protection
C lubroot on cruciferous plants isa worldwide disease causedby Plasmodiophora brassicae
Woron, which is also one of the seri-
ous soil-borne diseases. The disease
occurred in North America, South
America, Europe, South Africa, East
Asia and other places in the world. It
was first found in Taiwan as early as in
1936, and then it also occurred Hei-
longjiang, Liaoning, Shandong, Xin-
jiang, Gansu, Shaanxi, Jiangsu, Zhe-
jiang, Fujian, Jiangxi, Hubei, Hunan,
Guangdong, Guangxi, Sichuan, Yun-
nan, Tibet and other provinces and au-
tonomous regions. The losses in the
areas attacked by P. brassicae are
generally about 20%, even up to 70%-
80%, or 100% in some areas. So it is a
threat to the development of the veg-
etable industry[1-3].
In recent decades, much progress
has been made in the occurrence,
prevalence, forecasting and detection
of pathogens and pathogenic differen-
tiation of clubroot with the applications
of molecular biology in plant patholo-
gy. Yang et al. [4] carried out rapid de-
tection on clubroot of cabbage by
PCR, and found that the germination
of resting spores was a prerequisite for
pathogenic infection. The study of Guo
et al. [5] revealed that the germination
rate of resting spores was affected by
the environment temperature and pH,
and significantly inhibited by light. So,
temperature and soil pH are two im-
portant factors affecting the germina-
tion of spores in vitro. In this study,
the cabbage plants were grown in
Abstract [Objective] This study was conducted to investigate the pathogenicity of
Plasmodiophora brassicae on cabbage grown under different temperature and soil
pH conditions. [Method] The pathogenicity of P. brassicae were tested at seven dif-
ferent temperatures and at six different soil pH values with the resting spore con-
centration of 1×108 (spores/g) in the soil. The plant survival rate and incidence rate
of clubroot were investigated after 90 d. [Result] The incidence rate of clubroot on
cabbage among the different temperature sets varied in a descending order as fol-
lows: 30 ℃＞25 ℃＞20 ℃＞35 ℃＞15 ℃＞10 ℃＞5 ℃ at soil pH value of 6, indicating
that the pathogenicity of P. brassicae was weak at 5 and 10 ℃. The incidence rate
increased with soil temperature increasing from 15 to 30 ℃ , but decreased at 35
℃. The incidence rates of clubroot were 80.36%, 100%, 65%, 10.77%, 3.23% and
0% at soil pH 4, 5, 6, 7, 8 and 9 at 25 ℃ , respectively. The growth of cabbage
was inhibited and the survival rate was reduced at pH 4.The incidence rates of
clubroot were low at pH value of 7 and 8, and was 0% at pH 9. The Chinese
cabbage grew better at pH value of 5 and 6, but had high incidence rates of club-
root. [Conclusion] The results revealed that the incidence rate of clubroot on cab-
bage was closely related to the temperature and soil pH.
Key words Plasmodiophora brassicae; Temperature; pH value; Pathogenicity
DOI:10.16175/j.cnki.1009-4229.2015.01.030
Agricultural Science & Technology2015
Table 1 The grading standards of clubroot (at seedling stage)
Grade Symbol
Grade 0 Root system grow normally, without any obvious swelling
Grade 1 The taproot become swollen, and the diameter of swollen part is below 4 mm
Grade 3 The taproot becomes swollen, and the diameter of swollen part is 4-7 mm
Grade 5 Both the taproot and lateral roots become swollen, and the diameter of swollenpart is 7-10 mm
Grade 7 Both the taproot and lateral roots become swollen, and the diameter of swollenpart is above 10 mm
the soil mixed with resting spores of
Plasmodiophora brassicae at 1 ×108
(spores/g soil) in pot experiment, at
seven different temperature sets and
six soil pH values to investigate the
correlation of pathogenicity of P. bras-
sicae with temperature and soil pH,
with an attempt to provide theoretical
basis for the prevention and control of
disease in field by adjusting planting
pattern and soil pH.
Materials and Methods
Materials
The P. brassicae was collected
from fresh cabbage samples infected
by clubroot, and its pathogenicity was
tested on Chinese cabbage cultivar
83-1.
Preparation of soil sample
The Chinese cabbage samples
infected by clubroot were soaked in
water for 10 d. The spore concentra-
tion in the mixture of vermiculite and
disinfected humus at a ratio of 1∶1 was
adjusted to 1×108 spores/g soil, before
the soil sample was covered by film for
10 d, during which the soil sample was
flipped once every 5 d, so that the
pathogens and the soil could fully
mixed.
Cabbage cultivation under different
temperature and soil pH conditions
Cabbage cultivation under different
temperature conditions The cab-
bage seeds were sown in the soil
samples prepared in above steps in
pots of 30 cm in diameter and 20 cm in
height, and cultivated in a light incuba-
tor at 25 ℃ till germinating. Then, the
pots were placed into different light in-
cubators and cultivated at 5, 10, 15,
20, 25, 30 and 35 ℃ . Pathogen-free
soil was used in control group. Three
pots were prepared for each tempera-
ture treatment, and 12 plants were re-
mained in each pot.
Cabbage cultivation under different
soil pH condition The soil samples
were placed in pots of 30 cm in diam-
eter and 20 cm in height, and their pH
was respectively adjusted to 4, 5, 6, 7,
8 and 9 before cabbage seeds were
sown. Pathogen-free soil was used in
control group. Three pots were pre-
pared for each soil pH treatment, and
12 plants were remained in each pot.
Then, the pots were cultivated in a
light incubator at 25℃.
Other management measures The
plants were watered with sterile water
regularly, and no fungicides were ap-
plied during the entire experimental
stage.
Measurement items and methods
The plant survival rate in every
treatment was calculated 90 d after
seed sowing, while the cabbages were
uprooted for clubroot grading, accord-
ing to DB53/T 442-2012 Technologi-
cal Process for Identification of Rape
Resistance to Clubroot (Table 1).
Results and Analysis
Pathogenicity of P. brassicae on
Chinese cabbage at different tem-
perature sets
The results showed that the aver-
age survival rate was 55.0% , 75.2% ,
80.6%, 90.0%, 95.2%, 82.0%and45.3%;
and the average plant height was 5.5,
8.6, 9.4, 15.6, 12.2, 9.2 and 6.0 cm for
cabbage grown at 5, 10, 15, 20, 25, 30
and 35 ℃ , all indicating that the cab-
bage plant grew better at 20 -25 ℃
than at other temperature conditions
(Fig.1).
The incidence rate of clubroot on
cabbage grown at 5, 10, 15, 20, 25, 30
and 35℃was 0%, 3.8%, 15.2%, 61.8%,
89.6%, 90.2% and 53.6%, with the dis-
ease index of 0, 0.8, 3.5, 6.5, 15.6,
12.0 and 4.2, indicating that the inci-
dence of clubroot on cabbage plants
was higher at 25-30 ℃ than at other
temperature conditions, namely P.
brassicae had stronger pathogenicity
at 25-30℃ (Fig.2).
Pathogenicity of P. brassicae on
Chinese cabbage at different soil
pH values
As shown in Fig.3, the average
survival rate was 25.6%, 76.4%, 85.5%,
90.6%, 78.9%, 58.7 % and the aver-
age plant height was 6.5, 8.8, 16.5,
Fig.1 The effects of clubroot infection on Chinese cabbage growth under different temper-
ature conditions
Fig.2 The incidence of clubroot under different temperature conditions
113
Agricultural Science & Technology 2015
17.8, 15.2 and 13.6 cm at soil pH 4, 5,
6, 7, 8 and 9, respectively, suggesting
that the cabbage plant grew better
when the soil pH was 6, 7 and 8.
The incidence rate of clubroot on
cabbage grown in soil with pH value of
4, 5, 6, 7, 8 and 9 was 80.4%, 100%,
65%, 10.8%, 3.2% and 0.0%, with the
disease index of 6.8, 15.9, 14.2, 3.5,
1.5 and 0.0. The incidence rate of clu-
broot was higher, but the cabbage
growth was inhibited in the soil with pH
4. The incidence rate of clubroot was
lower at soil pH 7 and 8, and was zero
at pH 9. When the soil pH was 5 and 6,
the cabbage plant grew well, and the
incidence rate of clubroot was high,
suggesting that soil pH 5-6 was most
suitable for the survival and reproduc-
tion of P. brassicae. So, the occur-
rence of clubroot in field could be re-
duced through increasing the applica-
tion of organic fertilizers, plant ash and
alkaline fertilizers to adjust the soil pH.
Discussion
Clubroot pathogen on cruciferous
plant is an obligate parasite, and it has
strong vitality as it can survive for 7 to
9 years. Many factors can affect the
incidence of clubroot infection, such as
soil moisture, amount of pathogen
source, susceptible host, especially
soil temperature and pH. The study of
Yang et al. [6] revealed that the inci-
dence rate of clubroot was lower in soil
with water content of 20% , 30% and
80%, but higher in soil with water con-
tent of 70%-80% . By measuring the
vertical distribution of resting spores in
soil infected by clubroot, Yang et al. [6]
found that the amount of resting spore
of clubroot pathogen in soil of 10 cm
deep was higher than in surface soil,
both higher than in soil of 20 cm deep.
The study of Yang et al.[7] showed that,
if the infected tissue of cabbage was
soaked in water or exposed to sunlight
for 5, 10 and 15 d before inoculation,
the incidence rates of clubroot on
cabbage were 55.2% , 79.2% , 89.6%
and 85.7%, 100%, 100%, respectively;
and the incidence rate was 84.9% for
the cabbage inoculated with fresh in-
fected root. The study of Lv [8] showed
that resting spores of clubroot
pathogen could germinate at tempera-
ture from 6 to 30℃, and 21-22℃ was
the most suitable soil temperature.
Guo et al.[5] found that the germination
rate of the resting spores of clubroot
pathogen at different temperature in a
descending order was 24 ℃> 20 ℃>
28℃>16℃>32℃> 8℃> 36℃> 4℃>
40 ℃ . All the above studies revealed
the effects of temperature and pH on
the germination of resting spores of
clubroot pathogen in vitro. The present
study simulated the natural field condi-
tions to investigate the effects of tem-
perature and pH on the pathogenicity
of clubroot pathogen, and to find out
the main factors affecting clubroot
occurrence, which could provide a
scientific basis for developing mea-
sure to prevent and control clubroot.
The results showed that the incidence
rate of clubroot on cabbage increased
with soil temperature increasing from
15 to 30 ℃ but decreased at 35 ℃ .
The pathogenicity of P. brassicae was
weak at 5 and 10 ℃. So, soil tempera-
ture from 25 to 30 ℃ was most suit-
able for the propagation of clubroot
pathogen. In summary, 20-30 ℃ was
the most suitable temperature for the
occurrence of clubroot infection in soil
and the germination of resting spores
in vitro. Besides, clubroot could occur
at temperature from 15 to 30 ℃ ,
which covered the major vegetable
producing areas in Yunnan, so club-
root occured widely here. The occur-
rence of clubroot was less when the
temperature was below 15 ℃ or
above 30 ℃[9-10].
Our results also showed that
among the soil pH values of 4, 5, 6, 7,
8 and 9, the incidence rate of clubroot
was higher at pH 4, but the growth of
cabbage was inhibited and its survival
rate was reduced. The incidence rates
of clubroot were low at pH value of
7 and 8, and was 0% at pH 9. The
Chinese cabbage grew better at pH
value of 5 and 6, but had high inci-
dence rates of clubroot. Thus, the soil
pH at 5 -6 was most suitable for the
occurrence and infection of clubroot on
cruciferous plants. However, the soil
pH in most vegetable growing areas in
Yunnan Province was 5-6, so, club-
root widely occurred in these areas
and even very severely in same of the
areas. So, the application of alkaline
fertilizers can reduce the occurrence of
clubroot[11].
Sustained and comprehensive
measures are necessary for the pre-
vention and control of clubroot on cru-
ciferous plants. Suitable measures
should be conducted according to the
climatic and soil situation, such as ro-
tation of different crops, introduction or
screening of disease-resistant (toler-
Fig.3 The effects of clubroot infection on Chinese cabbage growth under different soil pH
conditions
Fig.4 The incidence of clubroot under different soil pH value conditions
114
Agricultural Science & Technology2015
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(Continued from page 69)
Responsible editor: Qingqing YIN Responsible proofreader: Xiaoyan WU
able for the growth of Chinese kale.
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