全 文 ：第 51 卷 第 8 期
2 0 1 5 年 8 月
林 业 科 学
SCIENTIA SILVAE SINICAE
Vol. 51，No. 8
Aug．，2 0 1 5
doi:10．11707 / j．1001-7488．20150816
Ｒeceived date:2014 － 09 － 24;Ｒevised date:2015 － 04 － 01．
Foundation project:National Natural Science Foundation (U1204324) ;Ｒesearch Fund of Henan University of Science and Technology
(2011QN42)．
* Corresponding author:Zhou Zhou．
一种引起虎皮兰叶斑病的刺孢壳属真菌的生物学特性*
李永丽 周 洲 娄金凤 胡雅静 时苗均
(河南科技大学林学院 洛阳 471003)
摘 要: 【目的】研究一种引起虎皮兰叶斑病的刺孢壳属真菌 HPL06 的生物学特性，为了解其潜在危害及
制定科学的防治方案提供依据。【方法】将 HPL06 于多种光照(自然光、12 h光照 /12 h黑暗、24 h光照、24 h
黑暗)、温度(5，10，15，20，25，30，35，40 ℃)、pH(4，5，6，7，8，9，10，11)、氮源(天冬酰胺、氯化铵、硝酸钾、
硫酸铵、谷氨酸)和碳源(蔗糖、果糖、乳糖、麦芽糖、葡萄糖)条件下进行培养，采用十字交叉法测量菌落直
径，分析不同环境因子对病原菌生长的影响;用 HPL06 菌饼和孢子悬浮液进行离体叶片接种，检测病原菌对
23 科 62 种植物的致病性。【结果】HPL06 在 4 种光照条件下均能生长，光照有利于菌丝生长，24 h 光照对
菌丝生长最有利;在极端温度(5，10，40 ℃)条件下病原菌不能生长，其他温度下均能生长，菌丝生长的最适
温度为 25 ～ 30 ℃;病原菌能在 pH 为 4 ～ 11 的条件下生长，最适 pH 为 4;5 种供试碳源中，病原菌均可正常生
长，葡萄糖对菌丝生长最有利;5 种供试氮源中，天冬酰胺对病原菌的生长最有利，其菌丝在 PDA 培养基中生
长最快。HPL06 对 20 科 46 种植物具有致病性，其中菌饼接种发病的植物为 20 科 46 种，孢子悬浮液接种发
病的植物为 12 科 30 种。【结论】刺孢壳属真菌 HPL06 对环境因子适应性范围较广，仅在低于 10 ℃或高于
40 ℃时不能生长;而且寄主范围广，在被检测的 23 科 62 种植物中，对 74. 19%的植物均具有致病性，在自然
界中可能还存在更多的寄主植物。
关键词: 虎皮兰;刺孢壳属;生物学特性
中图分类号:S718. 81 文献标识码:A 文章编号:1001 － 7488(2015)08 － 0121 － 06
Biological Characteristics of Chaetomella sp．(Sphaeropsidaceae) ，
A Novel Causal Agent of Sansevieria Leaf Spot Disease
Li Yongli Zhou Zhou Lou Jinfeng Hu Yajing Shi Miaojun
(College of Forestry，Henan University of Science and Technology Luoyang 471003)
Abstract: 【Objective】This study aims at evaluating the effects of environmental factors and susceptibility of
various plant species to pathogens causing Sansevieria leaf spot disease for developing the control methods and
strategies． 【Method】We investigated the biological characteristics of the HPL06 isolate of Chaetomella sp． that
causes leaf spot on Sansevieria trifasciata Prain cv． ‘Hanii’ in China． The fungus was incubated in different
conditions (i． e． light environments， temperature，pH，nitrogen and carbon sources)． Colony diameters were
measured using the CM method． The optimal environmental factors for the fungal growth were analyzed． A host
spectrum of the isolate was also investigated on 62 plant species covering 23 families using a detached leaf inoculation
technique． 【Ｒesult】Ｒesults showed that the optimal illumination and temperature for hyphal growth were 24 h of
fluorescent light and 25 － 30 ℃，respectively． The pathogen could grow at pH 4 － 11，but the optimal pH was 4 ．
The optimal carbon and nitrogen sources were glucose and asparagine，respectively，and the hyphae exhibited optimal
growth on PDA plates． A total of 46 species belonging to 20 families were susceptible to Chaetomella sp． HPL06 ．
【Conclusion】The isolate could adapt to various environments． The fungus could not grow below 10 ℃，or above
40 ℃ ． Chaetomella sp． HPL06 can cause diseases in many plants． A number of plants were found to be susceptible
to Chaetomella sp． with 74 ． 19% of the total number of the tested 62 plant species． A large number of plants
susceptible to Chaetomella sp． HPL06 may exist in nature． In order to avoid even greater economic losses caused by
林 业 科 学 51 卷
the fungus，further field studies on the occurrence of the new Sansevieria leaf spot disease are needed to establish a
science-based disease management strategy．
Key words: Sansevieria trifasciata;Chaetomella ;biological characteristics
1 Introduction
Sansevieria trifasciata is native to west Africa． It is
widely grown as a potted ornamental plant in China
because it improves indoor air quality by passively
absorbing toxins，such as nitrogen oxides，formaldehyde
(Wolverton et al．，1989) ，and ozone (Papinchaki et al．，
2009)． S． trifasciata leaf spots are caused by Fusarium
moniliforme (Jones，1940) ，Pythium spinosum (Takeuchi
et al．，2002) ，and Colletotrichum sansevieriae (Nakamura
et al．，2006;Aldaoud et al．，2011)．
Chaetomella (Sphaeropsidaceae ) is a fungus
distributed worldwide that can infect numerous plant
species，such as Pennisetum sp． (Sydow et al．，1916) ，
Cycas revoluta， Ficus bengalensis， Bambusa sp．
(Ｒamchandre et al．，1965) ，Ｒosa rugosa (Margorie，
1930) ，sterile oak (DiCosmo et al．，1980) ，Cuphea spp．
(Singh et al．，1999) ，and Eucalyptus grandis (Pérez-
Vera et al．，2005)． In addition，Chaetomella raphigera is
an endophytic fungus that can produce anticancer drugs
(Gangadevi et al．，2009)． In China，a new leaf spot
disease has been reported to affect S． trifasciata and
constrain its growth (Li et al．，2013)． We identified the
pathogen as Chaetomella sp． using morphological and
molecular methods (Li et al．，2013)． The new leaf spot
disease should be biologically characterized to establish a
control method． This study aimed to control the disease by
evaluating the effects of environmental factors and
susceptibility of various plant species to pathogen causing
S． trifasciata leaf spot disease．
2 Materials and methods
2. 1 Pathogen
A fungal culture (HPL06)was isolated from diseased
leaf samples collected from S． trifasciata in Luoyang City，
Henan Province，in 2011． The isolate was incubated on
potato dextrose agar (PDA:200 g of potato，20 g of
glucose，and 15 g of agar in 1 000 mL of water;pH 7. 0)
plates．
2. 2 Biological characterization
To determine the effect of temperature on fungal
growth，we placed 5 mm mycelial discs in the middle of a
Petri dish containing PDA． These discs were removed from
five-day-old colonies cultured on PDA plates． The Petri
dish was then incubated at temperatures ranging from 5 ℃
to 40 ℃ at 5 ℃ intervals for 3 d (triplicates per
temperature)． Colony diameters were determined using
the cross-measurement (CM ) method． Triplicate
measurements were averaged to obtain colony diameters．
To investigate the effect of pH on fungal growth，we
adjusted the pH of PDA to various levels (pH 4 － 11)at
intervals of 1 unit by adding an appropriate amount of
1 mol·L －1 HCl or NaOH solution． We obtained 5 mm
mycelial discs from five-day-old colonies cultured on PDA
plates． These discs were inoculated onto the center of new
PDA plates with different pH levels (triplicates per pH
level)． Colony diameters were measured three times using
the CM method and then averaged．
To evaluate the effect of light environment on
fungal growth，we transferred 5 mm mycelial discs onto
the center of new PDA plates． These discs were
removed from a five-day-old colony cultured on PDA
plates． The Petri dish was placed in a chamber with an
illuminating lamp at 28 ℃ at a distance of 20 cm
between the lamp and dish． Four light environments
were used to assess the influence of cultural conditions
on fungal growth． The conditions of these light
environments were as follows:1) alternating cycles of
12 h of fluorescent light (50 － 60 klx·m － 2 light
intensity)/12 h darkness， 2) natural light， 3)
continuous irradiation with fluorescent light (50 － 60
klx·m － 2 light intensity) ，and 4)complete darkness．
Each treatment was performed in triplicate． The average
of three replicates was used to determine colony
diameters using the CM method．
To assess the effect of nitrogen and carbon sources on
fungal growth，we inoculated 5 mm mycelial discs onto the
centers of different media． These discs were obtained from
five-day-old colonies cultured on PDA plates． In PDA，
dextrose was replaced by sucrose，fructose，maltose，and
lactose as carbon sources． Mycelial discs were placed on
PDA medium as contrast． The total carbon content of these
carbon sources was equal． In Ｒichard medium (10 g of
KNO3，5 g of KH2PO4，2. 5 g of MgSO4，0. 02 g of
221
第 8 期 李永丽等:一种引起虎皮兰叶斑病的刺孢壳属真菌的生物学特性
FeCl3，30 g of sucrose，and 15 g of agar in 1 000 mL of
water) ，potassium nitrate was replaced by ammonium
sulfate， glutamic acid， asparagine， and ammonium
chloride as nitrogen sources． Mycelial discs were placed
on Ｒichard and PDA media as contrast． The total nitrogen
content of these nitrogen sources was equal． Each
treatment was performed in triplicate． The average of
triplicate measurements was used to determine colony
diameters using the CM method． The isolates were
incubated at 28 ℃ for 3 d under different pH values，light
environment，and nitrogen sources，and at 28 ℃ for 5 d
under different carbon sources．
2. 3 Host range investigation
The pathogenicity of the isolate was tested among 62
plant species covering 23 families． All test plants were
collected from Luoyang． All fully developed leaves from
the test plants collected from Luoyang were sterilized with
75% ethanol， and washed three times with sterilized
distilled water． Fresh wounds were performed with a
sterile needle on healthy leaves． Two methods were used
for inoculation． 1)A 5 mm mycelial disc cut from the
margin of a five-day-old colony cultured on a PDA plate
was placed on each pin-wounded leaf to ensure that the
mycelium was in contact with the wound． Non-colonized
PDA discs were placed on pin-wounded leaves as the
control treatment． Eight leaves of one plant were
inoculated． 2)The isolate was grown on PDA for 5 d and
then suspended in sterile distilled water to produce a
suspension with a final concentration of 1 × 105 spores·
mL －1 ． The isolate was spray-inoculated with a spore
suspension on three leaves of one plant． Sterile distilled
water was used as a negative control． The inoculated
leaves were stored in a moist chamber and examined daily
for symptom development．
3 Ｒesults
3. 1 Biological characteristics
The temperature for vegetative growth of the isolate
ranged from 15 ℃ to 35 ℃，and the optimal temperature
was 25 － 30 ℃ (Tab. 1)． The pH for vegetative growth of
the isolate ranged from 4 to 11，and the optimal pH was 4
(Tab. 2 ) The fungus could grow in four light
environments，but the colony diameters of the isolate
subjected to continuously irradiated fluorescent light were
larger than those under the other light conditions
(Tab. 3)． Chaetomella sp． grew well on media containing
five carbon sources，and dextrose was the optimal carbon
source (Tab. 4)． Moreover，Chaetomella sp． grew well on
media comprising potassium nitrate，ammonium sulfate，
glutamic acid，asparagine，and ammonium chloride as
nitrogen sources (Tab. 5)．
3. 2 Host range
Among the 62 plant species tested from 23 families，
46 species covering 20 families were found to be
susceptible to Chaetomella sp． via inoculation with
mycelial discs (Fig． 1 and Tab. 3)． A total of 30 species
covering 12 families were also determined to be susceptible
to Chaetomella sp． via inoculation with conidial
suspension (Tab. 6)．
Tab. 1 Effect of temperature on mycelium growth
Temperature /℃ Colony diameter /mm
5 5. 0 f
10 5. 0 f
15 12. 1 e
20 32. 4 c
25 38. 6 b
30 43. 2 a
35 27. 3 d
40 5. 0 f
Tab. 2 Effect of pH on mycelium growth
pH Colony diameter /mm
4 81． 1 a
5 77． 1 b
6 56． 2 c
7 32． 2 e
8 36． 8 d
9 27． 0 f
10 25． 2 f
11 13． 0 g
Tab. 3 Effect of light on mycelium growth
Light environment Colony diameter /mm
12 h of fluorescent light /12 h of darkness 35． 2 b
Natural light 34． 6 b
24 h of fluorescent light 39． 7 a
24 h of darkness 33． 2 b
Tab. 4 Effect of carbon sources on vegetative growth
Carbon source Colony diameter /mm
Sucrose 56． 0 d
Lactose 58． 4 cd
Glucose 81． 1 a
Maltose 64． 9 bc
Fructose 74． 1 ab
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林 业 科 学 51 卷
Tab. 5 Effect of nitrogen sources on vegetative growth①
Nitrogensource Colony diameter /mm
Asparagine 12． 9 b
Ammonium chloride 8． 8 d
Potassiumnitrate 8． 3 de
Ammonium sulfate 10． 7 c
Glutamic acid 7． 1 d
Control (PDA) 32． 2 a
①Values are means of triplicate measurements． Means followed by
similar letters are not significantly different．
Fig． 1 Symptoms caused by Chaetomella sp． in selected inoculated plants
a. Prunus serrulata;b. Eriobotrya japonica;c. Nandina domestica;d.
Syringa oblata;e. Osmanthus fragrans;f. Distylium chinense．
4 Discussions
Some species possess an ethnopharmacological
background;in particular，S． trifasciata grown in south
Africa and tropical America is used for the treatment of
inflammatory conditions，and marketed as a crude drug in
the market to treat victims of snakebite (Morton，1981)．
S． trifasciata is widely grown as a potted ornamental plant
in China because it can absorb toxins (Cao et al．，2009)．
It also demonstrates strong ability to adapt to the
environment． The new leaf spot disease caused by
Chaetomella sp． affects the health and ornamental value of
S． trifasciata．
Chaetomella sp． widely exists in nature，and can
cause diseases in many plants． The host range experiment
demonstrated that the isolate of Chaetomella sp． from S．
trifasciata could infect plants from 23 families (Tab. 6)．
The number of newly discovered plants susceptible to
Chaetomella sp． comprised 74. 19% of the total number of
plants tested (Tab. 6)． We speculated that a large
number of plants susceptible to Chaetomella sp． may exist
in nature． Such plants may be considered as potential
hosts of the pathogen． Chaetomella sp． may infect
potential hosts if these plants are commercially cultivated
on a large scale under conditions conducive to disease
development． On the other hand，plants not susceptible to
Chaetomella sp． might be due to growth suppression of the
pathogen by unknown antifungal substance． The non-host
plants might be considered as promising sources for
screening of botanical fungicide ( s ) against
Chaetomella sp．
The effects of illumination，temperature，pH，carbon
source levels，and nitrogen source levels on the vegetative
growth of Chaetomella sp． were also characterized
(Tab. 1 － 5)． The isolate could exist in various conditions
and adapt to various environments． Further field studies
on the occurrence of the new Sansevieria leaf spot disease
are needed to establish a science-based disease
management strategy． Such studies are important because
Chaetomella sp． is a new causal organism of Sansevieria
leaf spots and the host range of the pathogen has
increased．
Tab. 6 Susceptibility of test plants to Chaetomella sp． inoculation①
No． Family Scientific name
Days required for visible symptoms (method of inoculation)/d
Treated with conidial suspension Treated with mycelial disc
1 Ｒosaceae Photinia frasery 6 － 7 6 － 7
2 Ｒosa chinensis 10 － 11 3 － 4
3 Eriobotrya japonica 8 － 9 5 － 6
4 Kerria japonica NT NT
5 Prunus serrulata 12 － 13 6 － 7
6 Prunus cerasifera 3 － 7 7 － 10
7 Spiraea salicifolia 11 － 12 4 － 5
8 Leguminosae Cercis chinensis 5 － 6 5 － 6
9 Wisteria sinensis 6 － 7 5 － 6
10 Dolichos lablab 8 － 9 5 － 6
11 Trifolium repens 8 － 9 3 － 4
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第 8 期 李永丽等:一种引起虎皮兰叶斑病的刺孢壳属真菌的生物学特性
Continued
Serial number Family Scientific name
Days required for visible symptoms (method of inoculation)/d
Treated with conidial suspension Treated with mycelial disc
12 Medicago falcata 6 － 7 3 － 4
13 Berberidaceae Nandina domestica 6 － 7 3 － 4
14 Berberis thunbergii 6 － 7 3 － 4
15 Berberis wangii 9 － 10 3 － 4
16 Mahonia fortunei 11 － 17 4 － 5
17 Asteraceae Dendranthema grandiflorum NT NT
18 Dahlia pinnata NT NT
19 Tagetes erecta NT NT
20 Helianthus annuus NT NT
21 Magnoliaceae Magnolia grandiflora NT 3 － 4
22 Oleaceae Osmanthus fragrans 7 － 10 3 － 7
23 Ligustrun lucidum 3 － 7 3 － 7
24 Ligustrun quihoui 3 － 7 3 － 7
25 Syringa oblata 3 － 4 2 － 3
26 Jasminum nudiflorum 3 － 4 2 － 3
27 Liliaceae Ophiopogon japonicus NT 3 － 7
28 Chlorophytum comosum NT NT
29 Lilium casablanca 7 － 9 5 － 6
30 Solanaceae Capsicum annuum NT NT
31 Lycopersicon esculentum NT NT
32 Capsicum frutescens NT NT
33 Nicotiana tabacum NT NT
34 Lycium barbarum NT NT
35 Salicaceae Salix babylonica 7 － 8 4 － 5
36 Meliaceae Melia azedarach 3 － 4 2 － 3
37 Toona sinensis 3 － 4 2 － 3
38 Hamamelidaceae Distylium chinense 7 － 10 7 － 8
39 Buxaceae Buxus sinica NT 10 － 12
40 Buxus bodinieri NT 11 － 12
41 Bignoniaceae Ｒadermachera sinica NT 8 － 10
42 Gramineae Zea mays NT 8 － 10
43 Phyllostachys heterocycla NT 8 － 10
44 Papilionaceae Ｒobinia pseudoacacia 8 － 10 3 － 7
45 Sophora japonica 3 － 7 3 － 7
46 Zephyranthes grandiflora NT NT
47 Amaryllidaceae Agave sisalana NT 11 － 13
48 Dracaena marginata NT NT
49 Narcissus tazetta var． chinensis NT 3 － 7
50 Clivia miniata NT 3 － 7
51 Dracaena sanderiana NT 3 － 7
52 Lamiaceae Salvia splendens 8 － 10 3 － 7
53 Lavandula angustifolia 7 － 10 3 － 7
54 Mentha haplocalyx NT 3 － 7
55 Ｒanunculaceae Paeonia suffruticosa 2 － 3 2 － 3
56 Paeonia lactiflora NT NT
57 Calycanthaceae Chimonanthus praecox NT 3 － 7
58 Pittosporaceae Pittosporum tobira NT 9 － 11
59 Araceae Philodendron congo NT 2 － 3
60 Anthurium andraeanum NT NT
61 Lauraceae Cinnamomum camphora NT 10 － 12
62 Oxalidaceae Oxalis corniculata NT NT
①NT denotes plants without visible symptomsat 13 d after inoculation．
521
林 业 科 学 51 卷
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