山茶花腐病病原菌的分离鉴定及生物学特性

普梅英; 武自强; 张诗文; 李艳杰; 牛锦华; 付芳丽; 耿芳; 陈龙清; 王超

doi:10.19303/j.issn.1008-0384.2022.010.011

山茶花腐病病原菌的分离鉴定及生物学特性

doi: 10.19303/j.issn.1008-0384.2022.010.011

西南林业大学/国家林业和草原局西南风景园林工程技术研究中心/云南省功能性花卉资源及产业化技术工程研究中心/云南省森林灾害预警与控制实验室，云南　昆明　650224

基金项目: 国家重点研发计划项目（2019YFD100100005）；云南省教育厅科学研究基金项目（2021Y259、2021J0172）；西南山地森林保育与利用教育部重点实验室开放基金项目（KLESWFU-202005）；国家基础科学公共科学数据中心项目（ NBSDC-DB-03）

详细信息

作者简介:
普梅英（1997−），女，硕士研究生，研究方向：植物保护（E-mail：670868861@qq.com）

通讯作者:
王超（1980−），男，博士，研究方向：植物资源及利用（E-mail：47188127@qq.com）

中图分类号: S 432
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出版历程
- 收稿日期: 2022-06-23
- 修回日期: 2022-07-18
- 网络出版日期: 2022-10-21
- 刊出日期: 2022-10-30

Pathogen Identification and Biological Characterization of Camellia Petal Blight Disease

Southwest Research Center for Engineering Technology of Landscape Architecture, State Forestry and Grassland Administration/Yunnan Engineering Research Center for Functional Flower Resources and Industrialization/College of Landscape Architecture and Horticulture Sciences/Key Laboratory of Forest Disaster Warning and Control in Universities of Yunnan Province/Southwest Forestry University, Kunming, Yunnan　650224, China

摘要

摘要: 目的明确山茶花腐病的致病菌及其生物学特性，提升山茶花期品质和观赏价值。方法采用组织分离法对云南省昆明市昆明植物园山茶的花腐病进行病原菌分离，通过致病性测定、形态学和分子生物学鉴定确定病原菌，并进行生物学特性的研究。结果从病组织中分离得到1株菌株，将其回接于健康花瓣上，接种后划破的花瓣病症较为明显，花朵失色变黄褐，严重时能使花瓣干腐或湿腐，并且长菌丝体，与田间的症状一致，符合柯赫氏法则的致病性测定。PDA培养基上菌丝偏黄，培养后期长黑色的孢子，子囊孢子柠檬状，经合并多基因（ITS、LSU、EF1）系统发育分析，其与假螺卷毛壳菌（Chaetomium pseudocochliodes）聚集在同一分支。对假螺卷毛壳菌（C. pseudocochliodes）的生物学特性研究结果显示，该病原菌的最适培养温度为25 ℃，对光照条件没有特殊要求，最适培养基为LB和OMA培养基，以牛肉膏作为氮源和以果糖作为碳源的利用率最高，致死温度为55 ℃/10 min。结论假螺卷毛壳菌（C. pseudocochliodes）是山茶花腐病的病原菌，该病原菌的适宜温度为25 ℃，不受光照影响，培养基为LB和OMA培养基，以牛肉膏作为氮源和以果糖作为碳源的利用率最高，致死温度为55 ℃/10 min。
- 山茶 /
- 花腐病 /
- 假螺卷毛壳菌 /
- 生物学特性
Abstract: Objective The pathogen and its biological characteristics of camellia petal blight disease were investigated for improving the quality and protecting the ornamental value of the floral plant. Method A pathogen was isolated from of diseased Camellia japonica at the Botanical Garden in Kunming City by tissue separation method and confirmed by pathogenicity assay as well as morphological and molecular biological identifications. Biological characteristics of the isolate were studied. Result A suspected microbe was isolated from the diseased tissue and inoculated onto healthy camellia flower petals. With the appearance of yellowing and browning petals with dry or wet rots and fungal mycelial growth, the similar symptoms shown on the infected plants in the field, the isolate was studied further for a confirmed identification. On a PDA medium, the fungal colonies displayed yellow mycelia and produced black spores and lemon-like ascospores at late stage. The phylogenetic analysis of multiple genes of ITS, LSU, and EF1 clustered the isolate with Chaetomium pseudocochliodes clade. Subsequently, the optimal conditions for the growth of the isolated fungus were found to be at 25 ℃ on LB and OMA culture medium,without special requirement of light,beef paste as a nitrogen source and fructose as a carbon source have the highest utilization rates, and the lethality at 55 ℃ in 10 min. Conclusion C. pseudocochliodes was identified as the pathogen that caused floral petal blight disease on C. japonica. The pathogen grew optimally at 25 ℃ on LB and OMA culture medium ,without a specific light requirement, beef paste as a nitrogen source and fructose as a carbon source have the highest utilization rates, and it was killed by exposure to 55 ℃ for 10 min.
- Camellia japonica /
- petal blight disease /
- Chaetomium pseudocochliodes /
- biological characteristics

HTML全文

图 1 山茶花腐病田间和接种后症状及病原菌在PDA培养基上的形态学特征

a. 田间感病病花症状；b. 田间健康花；c、d. 室内离体接种后花腐症状；e. 菌落形态；f. 子囊孢子。

Figure 1. Symptoms of petal blight disease on field and inoculated C. japonica and morphology of pathogen on PDA

a: symptoms on field flower; b: healthy flower; c and d: symptoms on inoculated camellia flower;e: colony morphology; f: ascospores.

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图 2 山茶花腐病病原菌的基因电泳图

a、b、c分别为ITS、LSU、EF1区间电泳图，M为DL2 000的DNA Marker，1为SX-3，2为SX Ⅰ，3为YH Ⅰ。

Figure 2. Gene electrophoresis of camellia petal blight disease pathogen

a, b, and c: electrophoretic map of primer ITS, LSU and EF1 respectively; M: DNA marker of DL2 000; 1: SX-3; 2: SX Ⅰ; 3: YH Ⅰ.

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图 3 基于ITS、LSU和EF1的多基因系统发育进化树

Figure 3. Multigene phylogenetic tree based on ITS, LSU and EF1 gene sequences

下载: 全尺寸图片幻灯片

表 1 引物序列表

Table 1. Primer sequences

基因 Gene	引物 Primer	序列 Sequence	文献来源 Reference
ITS	ITS1	5′-TCCGTAGGTGAACCTGCGG-3′	Henson等^[17]
ITS	ITS4	5′-TCCTCCGCTTATTGATATG-3′	Henson等^[17]
LSU	NL1	5′-GCATATCAATAAGCGGAGGAAAAG -3′	刘彬等^[18]
LSU	NL4	5′-GGTCCGTGTTTCAAGACGG-3′	刘彬等^[18]
EF1	EF1-983F	5′-GCYCCYGGHCAYCGTGAYTTYAT-3′	谭秀梅等^[19]
EF1	EF1-2218R	5′-ATGACACCRACRGCRACRGTYTG-3′	谭秀梅等^[19]

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表 2 温度对菌株SX-3菌丝生长的影响

Table 2. Effect of temperature on mycelial growth of SX-3

温度 Temperature/℃	菌落直径 Colony diameter/cm	温度 Temperature/℃	菌落直径 Colony diameter/cm
10	3.967±0.0577 d	30	7.833±0.2887 b
15	5.500±0.5000 c	35	6.267±0.0577 c
20	7.733±1.1676 b	40	1.033±0.0577 e
25	8.833±0.2887 a
不同小写字母表示各处理菌落直径差异显著（P＜0.05），下同。
The different lowercase alphabet shows that the factor to the diameter of the colony has significantly difference (P＜0.05), The same applies below.

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表 3 光照对菌株SX-3菌丝生长的影响

Table 3. Effect of illumination on mycelial growth of SX-3

光照情况 Lighting condition	菌落直径 Colony diameter/ cm	光照情况 Lighting condition	菌落直径 Colony diameter/ cm
24 h光照 24 h lighting	8.800±0.1732 a	24 h黑暗 24 h darkness	8.600±0.6928 a
12 h交替 12 h alternation	8.500±0.0000 a	正常光照 Normal lighting	8.667±0.0577 a

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表 4 培养基对菌株SX-3菌丝生长的影响

Table 4. Effect of culture medium on mycelial growth of SX-3

培养基 Culture medium	菌落直径 colony diameter/ cm	培养基 Culture medium	菌落直径 Colony diameter/ cm
OMA	8.500±0.0000 b	PDA	8.167±0.2887 c
CMA	8.500±0.0000 b	LB	9.000±0.0000 a

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表 5 氮源对菌株SX-3菌丝生长的影响

Table 5. Effect of nitrogen sources on mycelial growth of SX-3

氮源 Nitrogen source	菌落直径 Colony diameter/cm	氮源 Nitrogen source	菌落直径 Colony diameter/cm
硝酸钠 Sodium nitrate	8.500±0.0000 a	硝酸钾 Potassium nitrate	8.333±0.2887 a
蛋白胨 Peptone	8.500±0.0000 a	甘氨酸 Glycine	8.667±0.2887 a
牛肉膏 Beef extract	8.500±0.0000 a	缺氮 Nitrogen deficiency	7.200±0.1000 c
酵母膏 Yeast cream	7.967±0.2517 b

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表 6 碳源对菌株SX-3菌丝生长的影响

Table 6. Effect of carbon sources on mycelial growth of SX-3

碳源 Carbon source	菌落直径 Colony diameter/cm	碳源 Carbon source	菌落直径 Colony diameter/cm
蔗糖 Sucrose	7.967±0.4509 b	阿拉伯树胶糖 Gum Arabic sugar	3.567±0.3055 c
葡萄糖 Glucose	8.433±0.0577 a	果糖 Fructose	8.500±0.0000 a
木糖 Xylose	8.467±0.0577 a	缺碳 Carbon deficiency	7.500±0.0000 b
麦芽糖 Maltose	8.333±0.1155 ab

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表 7 菌株SX-3致死温度测定

Table 7. Lethal temperature of SX-3

致死温度 Lethal temperature/ ℃	菌落直径 Colony diameter/cm	致死温度 Lethal temperature/ ℃	菌落直径 Colony diameter/cm
35	7.633±0.3215 b	55	0.000±0.0000 d
40	8.367±0.1528 a	60	0.000±0.0000 d
45	7.133±0.9074 b	65	0.000±0.0000 d
50	2.467±0.1528 c

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参考文献(41)

[1]	卓丽环, 陈龙清. 园林树木学[M]. 北京: 中国农业出版社, 2004.
[2]	李峻红, 陆俊, 单蓓, 等. 保鲜剂对山茶花切花保鲜效果的研究 [J]. 农村实用技术, 2021(9):59−60. LI J H, LU J, SHAN B, et al. Study on the effect of preservatives on fresh-keeping of cut camellia flowers [J]. Rural Practical Technology, 2021(9): 59−60.(in Chinese)
[3]	WANG Y, ZHUANG H, SHEN Y, et al. The Dataset of Camellia Cultivars Names in the World[J]. Biodiversity Data Journal, 2021, 9(12). DOI: 10.3897/BDJ.9.e61646
[4]	杨英. 山茶花常见病虫害及其防治对策 [J]. 农业科技与信息, 2016, 13(10):101−101,103. doi: 10.3969/j.issn.1003-6997.2016.10.070 YANG Y. Common diseases and insect pests of Camellia and their control countermeasures [J]. Agricultural Technology and Information, 2016, 13(10): 101−101,103.(in Chinese) doi: 10.3969/j.issn.1003-6997.2016.10.070
[5]	张齐发, 胡一民(摄). 中国十大名花之山茶花常见主要病虫害的识别与防治(上) [J]. 花木盆景(上半月), 2006(11):24−26. ZHANG Q F, HU Y M. Identification and Control of Common Main Diseases and Insects of Camellia, the Top Ten Famous Flowers in China (Part 1) [J]. Bonsai (Flowers and Horticulture), 2006(11): 24−26.(in Chinese)
[6]	刘国信. 山茶花常见病虫害防治 [J]. 植物医生, 2009(1):28−29. doi: 10.3969/j.issn.1007-1067.2009.01.018 LIU G X. Prevention and control of camellia pests and diseases [J]. Plant Doctor, 2009(1): 28−29.(in Chinese) doi: 10.3969/j.issn.1007-1067.2009.01.018
[7]	罗会生. 山茶病虫害防治 [J]. 中国花卉园艺, 2016(4):32−35. doi: 10.3969/j.issn.1009-8496.2016.04.013 LUO H S. Prevention and control of camellia pests and diseases [J]. China Flower and Horticulture, 2016(4): 32−35.(in Chinese) doi: 10.3969/j.issn.1009-8496.2016.04.013
[8]	王建伟, 钱显明, 张炳欣. 浙江省山茶花病害 [J]. 植物病理学报, 1990(2):89−92. WANG J W, QIAN X M, ZHANG B X. The diseases of Camellia in Zhejiang Province [J]. Acta Phytopathologica Sinica, 1990(2): 89−92.(in Chinese)
[9]	何美仙. 山茶花主要病害的发生及防治 [J]. 内蒙古林业科技, 2005(1):51−52. doi: 10.3969/j.issn.1007-4066.2005.01.020 HE M X. Occurrence and control of main disease for Camellia japomica [J]. Inner Mongolia Forestry Science & Technology, 2005(1): 51−52.(in Chinese) doi: 10.3969/j.issn.1007-4066.2005.01.020
[10]	RAABE R D, MCCAIN A H, PAULUS A O. Diseases and pests[M]//Feathers D L, Brown M H. The camellia its history culture genetics and a look into its future development. Columbia South Carolina, USA: Southern Californian Camellia Society Inc, 1978: 279－285.
[11]	RON VAN TOOR. Development of Biocontrol Methods for Camellia Flower Blight Caused by Ciborinia camelliae [J]. American Camellia Yearbook, 2003: 29−33.
[12]	KONDRATEV N, DENTON-GILES M, BRADSHAW R E, et al. Camellia plant resistance and susceptibility to petal blight disease are defined by the timing of defense responses [J]. Mol Plant Microbe Interact, 2020, 33(7): 982−995. doi: 10.1094/MPMI-10-19-0304-R
[13]	陈秀虹, 伍建榕. 观赏植物病害诊断与治理[M]. 北京: 中国建筑工业出版社, 2009.
[14]	方中达. 植病研究方法[M]. 3版. 北京: 中国农业出版社, 1998.
[15]	任纬恒. 高山杜鹃病害的病原菌分离鉴定与防治基础研究[D]. 贵阳: 贵州师范大学, 2019. REN W H. Basic research on isolation, identification and control of pathogen of Rhododendron lapponicum disease [D]. Gui yang: Guizhou Normal University, 2019. (in Chinese)
[16]	朱轶慧, 刘玉军, 毕飞虎, 等. 红叶石楠叶斑病病原鉴定及其生物学特性研究 [J]. 植物保护, 2021(5):210−215, 235. ZHU Y H, LIU Y J, BI F H, et al. Identification and biological characteristics of the pathogen causing leaf spot disease on Photinia × fraseri [J]. Plant Protection, 2021(5): 210−215, 235.(in Chinese)
[17]	HENSON J M, FRENCH R. The polymerase chain reaction and plant disease diagnosis [J]. Annual Review of Phytopathology, 1993, 31: 81−109. doi: 10.1146/annurev.py.31.090193.000501
[18]	刘彬, 张祥林, 王羽中, 等. 向日葵白锈病菌巢式PCR检测方法的研究 [J]. 新疆农业科学, 2011(5):859−863. doi: 10.6048/j.issn.1001-4330.2011.05.013 LIU B, ZHANG X L, WANG Y Z, et al. Study on the nest-PCR to detect the Albugo tragopogi (persoon) schruter var helianthi novotlenova [J]. Xinjiang Agricultural Sciences, 2011(5): 859−863.(in Chinese) doi: 10.6048/j.issn.1001-4330.2011.05.013
[19]	谭秀梅, 阿地力·沙塔尔, 朴春根, 等. 无柄灵芝遗传多样性的SRAP、ITS、TEF1-α和LSU分析 [J]. 微生物学通报, 2016, 43(12):2667−2677. TAN X M, ADIL S, PIAO C G. Genetic diversity of Ganoderma resinaceum by SRAP, ITS, TEF1-α and LSU markers [J]. Microbiology China, 2016, 43(12): 2667−2677.(in Chinese)
[20]	李雪玲, 谢美华, 杨海艳, 等. 高山榕叶枯病病原菌分离鉴定和生物学特性的初步研究 [J]. 西南农业学报, 2015(6):2558−2562. LI X L, XIE M H, YANG H Y, et al. Pathogen identification and biological characterstics of leaf blight of Ficus altissima [J]. Southwest China Journal of Agricultural Sciences, 2015(6): 2558−2562.(in Chinese)
[21]	黎肇家, 李汶晋, 毛跃彦, 等. 栾树枝枯病病原菌鉴定及其生物学特性研究 [J]. 西北农林科技大学学报(自然科学版), 2020(9):53−63. LI Z J, LI W J, MAO Y Y, et al. Identification and biological characteristics of branch withered disease causing pathogen to Koelreuteria paniculata Laxm [J]. Journal of Northwest A & F University (Natural Science Edition), 2020(9): 53−63.(in Chinese)
[22]	张凡, 冯婷, 韩正敏, 等. 落羽杉赤枯病病原菌的分离鉴定及生物学特性分析 [J]. 植物保护学报, 2018(4):900−907. ZHANG F, FENG T, HAN Z M, et al. Biological characteristics and molecular identification the pathogen of Taxodium red blight [J]. Journal of Plant Protection, 2018(4): 900−907.(in Chinese)
[23]	彭成彬, 王泽榕, 阮俊峰, 等. 柳叶蜡梅真菌病害分离鉴定及生物学特性 [J]. 福建农业学报, 2021(4):457−463. PENG C B, WANG Z R, RUAN J F, et al. Identification and biology of leaf spot pathogen on Chimonanthus salicifolius [J]. Fujian Journal of Agricultural Sciences, 2021(4): 457−463.(in Chinese)
[24]	姚锦爱, 黄鹏, 兰成忠, 等. 福建省铁皮石斛茎腐病病原菌鉴定及其生物学特性 [J]. 福建农业学报, 2021(12):1457−1463. YAO J A, HUANG P, LAN C Z, et al. Identification and biological characteristics of stem rot pathogen of Dendrobium officinale [J]. Fujian Journal of Agricultural Sciences, 2021(12): 1457−1463.(in Chinese)
[25]	李润根, 卢其能, 何咪, 等. 百合新病原菌假短孢弯孢生物学特性及其对杀菌剂的敏感性 [J]. 植物保护, 2020(6):41−46. LI R G, LU Q N, HE M, et al. The biological characteristics of Curvularia pseudobrachyspora, a new causal agent of lily leaf spot, and its sensitivity to fungicides [J]. Plant Protection, 2020(6): 41−46.(in Chinese)
[26]	杨迪, 杜婵娟, 叶云峰, 等. 广西香蕉煤污病病原菌的分离鉴定及其生物学特性 [J]. 植物病理学报, 2021(4):507−514. YANG D, DU C J, YE Y F, et al. Isolation, identification and characterization of the pathogen causing banana sooty blotch in Guangxi [J]. Acta Phytopathologica Sinica, 2021(4): 507−514.(in Chinese)
[27]	张建强, 吴康莉, 张晓梦, 等. 芹菜叶斑病病原菌的分离鉴定、生物学特性及其生防菌筛选 [J]. 西北农业学报, 2021(7):1089−1099. ZHANG J Q, WU K L, ZHANG X M, et al. Isolation, identification, biological characteristics and biocontrol bacteria screening of celery leaf spot pathogen [J]. Acta Agriculturae Boreali-Occidentalis Sinica, 2021(7): 1089−1099.(in Chinese)
[28]	WANG X W, LOMBARD L, GROENEWALD J Z, et al. Phylogenetic reassessment of the Chaetomium globosum species complex [J]. Persoonia, 2016, 36: 83−133. doi: 10.3767/003158516X689657
[29]	廖宏娟, 张志斌, 江玉梅, 等. 球毛壳菌对植物病原真菌和根结线虫的生物防治潜力 [J]. 天然产物研究与开发, 2022(6):1076−1089. doi: 10.16333/j.1001-6880.2022.6.021 LIAO H J, ZHANG Z B, JIANG Y M, et al. Biocontrol potential of Chaetomium globosum against plant pathogenic fungi and root-knot Nematodes: A review [J]. Natural Product Research and Development, 2022(6): 1076−1089.(in Chinese) doi: 10.16333/j.1001-6880.2022.6.021
[30]	谭悠久. 毛壳菌科(Chaetomiaceae)分类及分子系统发育研究[D]. 杨凌: 西北农林科技大学, 2005. TAN Y J. Taxonomy and molecular phylogeny of Chaetomiaceae [D]. Yangling: Northwest A&F University, 2005. (in Chinese )
[31]	TVEIT M. Control of a seed-borne disease by Chaetomium cochlioides Pall., under natural conditions [J]. Nature, 1953, 172(4366): 39. doi: 10.1038/172039a0
[32]	陈利军, 史洪中, 陈月华. 油菜内生球毛壳菌抑菌作用初步测定 [J]. 河南农业科学, 2005(7):54−56. doi: 10.3969/j.issn.1004-3268.2005.07.018 CHEN L J, SHI H Z, CHEN Y H. Preliminary study on the anti-fungal action of Chaetomium globosum from endophytic fungi of rapeseed [J]. Journal of Henan Agricultural Sciences, 2005(7): 54−56.(in Chinese) doi: 10.3969/j.issn.1004-3268.2005.07.018
[33]	兰楠, 祁高富, 喻子牛, 等. 油菜内生真菌的分离鉴定及抑菌作用 [J]. 华中农业大学学报, 2011(3):270−275. LAN N, QI G F, YU Z N, et al. Isolation, identification and anti-fungal action of endophytic fungi of rapeseed [J]. Journal of Huazhong Agricultural University, 2011(3): 270−275.(in Chinese)
[34]	赵林艳. 球毛壳菌对三七根腐病镰刀菌的生防潜力[D]. 昆明: 云南大学, 2018. ZHAO L Y. Biocontrol potential of Chaetomium globosum against Fusarium spp. —the pathogenic fungi of Panax notoginseng root rot diseases[D]. Kunming: Yunnan University, 2018. (in Chinese )
[35]	莫维弟, 卯婷婷. 辣椒枯萎病土壤拮抗真菌的筛选与鉴定 [J]. 贵州农业科学, 2019(8):66−70. doi: 10.3969/j.issn.1001-3601.2019.08.015 MO W D, MAO T T. Screening and identification of antagonistic fungi in soil against Fusarium oxysporum in pepper [J]. Guizhou Agricultural Sciences, 2019(8): 66−70.(in Chinese) doi: 10.3969/j.issn.1001-3601.2019.08.015
[36]	周春元, 朴向民, 闫梅霞, 等. 人参黑斑病菌生防内生真菌的分离筛选、鉴定及抑菌特性 [J]. 中国中药杂志, 2019(2):274−277. ZHOU C Y, PIAO X M, YAN M X, et al. Isolation, screening and identification of endophytic fungi and detection of its antifungal effects against Alternaria panax [J]. China Journal of Chinese Materia Medica, 2019(2): 274−277.(in Chinese)
[37]	马婉琴, 蔡苏, 钱永生, 等. 球毛壳菌引起杭白菊叶枯病的首次报道 [J]. 杭州师范大学学报(自然科学版), 2015(4):390−393. MA W Q, CAI S, QIAN Y S, et al. First report of leaf blight disease of Chrysanthemum morifolium caused by Chaetomium globosum [J]. Journal of Hangzhou Normal University (Natural Science Edition), 2015(4): 390−393.(in Chinese)
[38]	钱永生, 蔡苏, 吴剑丙, 等. 浙江省“浙八味”道地药材真菌病害调查及病原鉴定 [J]. 湖北农业科学, 2015(19):4740−4745,4748. QIAN Y S, CAI S, WU J B, et al. Investigation and identification of the pathogenic fungi in trueborn “Zhe-ba-Wei”Medicinal herbs in Zhejiang Province [J]. Hubei Agricultural Sciences, 2015(19): 4740−4745,4748.(in Chinese)
[39]	杨丽芬, 朱心, 周春香, 等. 石榴叶斑病球毛壳菌生长习性及杀菌剂筛选 [J]. 安徽农业大学学报, 2018(1):195−200. YANG L F, ZHU X, ZHOU C X, et al. Growth characteristics of mycelia of Chaetomium globusum causing pomegranate leaf spot disease and fungicides screening [J]. Journal of Anhui Agricultural University, 2018(1): 195−200.(in Chinese)
[40]	刘南南, 王桂清. 红果臭椿苗期病害致病菌的分离与鉴定 [J]. 北方园艺, 2019(17):89−94. doi: 10.11937/bfyy.20184580 LIU N N, WANG G Q. Isolation and identification of pathogenic fungi from red fruit Ailanthus altissima seedling disease [J]. Northern Horticulture, 2019(17): 89−94.(in Chinese) doi: 10.11937/bfyy.20184580
[41]	蒋桂芝, 田海, 朱国渊, 等. 橡胶籽苗黑根病病原鉴定及生物学特性研究 [J]. 植物保护, 2020(1):223−228. JIANG G Z, TIAN H, ZHU G Y, et al. Pathogen identification and biological characteristics of black root disease in rubber seedlings [J]. Plant Protection, 2020(1): 223−228.(in Chinese)