Identification and Etiology of a New Dangerous Neopestalotiopsis Leaf Fall Disease of Rubber Tree (Hevea brasiliensis) in China

doi:10.3969/j.issn.1000-2561.2020.08.016

Abstract

Abstract:

Based on the current outbreak and epidemic situation of a novel Neopestalotiopsis leaf fall disease of rubber trees (Hevea brasiliensis) in major rubber planting countries abroad and its great impact on natural rubber industry economy, the authors conducted a preliminary inspection on 13 farms and rubber plantations in main rubber producing areas in Hainan, Yunnan and Guangdong provinces, and found suspected disease samples in Baoxian Farm, Zhizhong Town, Ledong County, Hainan. In this study, the pathogens of the suspected disease samples were isolated, purified and the pathogenicity was determined. Through morphological and molecular phylogenetic analysis, the pathogen was identified as Neopestalotiopsis aotearoa Maharachch., K.D. Hyde & Crous. The basic biological characteristics showed that the suitable temperature for the mycelial growth and spore germination was 28℃, the lethal temperature of spores was 50℃. The optimum pH value was 6.5, the optimum carbon sources were sucrose and D-glucose, the optimum nitrogen sources were potassium nitrate and soybean peptone, and the optimum medium was PDA, which could produce spores after eight days. The pathogen and its basic biological characteristics were preliminarily identified, which would provide theoretical basis for the reserve of related technical measures for the prevention and control of the disease in advance.

Key words: Hevea brasiliensis, Neopestalotiopsis leaf fall disease, pathogen identification, biological characteristics

CLC Number:

LI Boxun,LIU Xianbao,SHI Tao,CAI Jimiao,ZHENG Xiaolan,FENG Yanli,XU Chunhua,HUANG Guixiu. Identification and Etiology of a New Dangerous Neopestalotiopsis Leaf Fall Disease of Rubber Tree (Hevea brasiliensis) in China[J]. Chinese Journal of Tropical Crops, 2020, 41(8): 1616-1624.

Figures/Tables 12

References 20

[1]	李达, 张绍文. 天然橡胶产业发展政策分析: 红线与红利[J]. 林业经济问题, 2020,40(2):208-215.
[2]	周雪敏, 刘先宝, 李博勋, 等. 橡胶树新发茎杆溃疡病病原鉴定及其生物学特性测定[J]. 热带作物学报, 2016,37(4):758-765.
[3]	Li B X, Shi T, Liu X B, et al. First report of rubber tree stem rot caused by Fusarium oxysporum in China[J]. Plant Disease, 2014,98(7):1008-1008. DOI URL PMID
[4]	Cai Z Y, Liu Y X, Li G H, et al. First report of Alternaria alternata causing black spot of rubber tree in China[J]. Plant Disease, 2014,98(7):1011 URL PMID
[5]	Nyaka Ngobisa A I C, Zainal Abidin M A, Wong M Y, et al. Neofusicoccum ribis associated with leaf blight on rubber (Hevea brasiliensis) in Peninsular Malaysia[J]. Plant Pathology Journal, 2013,29(1):10-16. DOI URL PMID
[6]	Tajuddin Ismail. Pestalotiopsis outbreak in Bumi Rambang plantation in Palembang: Report of meeting on collaborative research on Pestalotiopsis[Z]. Indonesia and Malaysia, 2019.
[7]	International Rubber Research Institute(IRRI). Identification, bioecology and management to control Pestalotiopsis leaf fall diseases in rubber[Z]. Indonesia, 2019.
[8]	Tajuddin Ismail. Recent outbreak of Pestalotiopsis leaf diseases in South East Asia: Meeting of experts on Pestalotiopsis leaf disease[Z]. Surat Thani Province,Thailand, 2020.
[9]	方中达. 植病研究方法[M]. 3版. 北京: 中国农业出版社, 1998: 63.
[10]	Pornsuriya C, Chairin T, Thaochan N, et al. Identification and characterization of Neopestalotiopsis fungi associated with a novel leaf fall disease of rubber trees (Hevea brasi-liensis) in Thailand Journal of Phytopathology. 2020[2020-07-25].https://www.researchgate.net/publication/341512722_Identification_and_characterization_of_Neopestalotiopsis_fungi_associated_with_a_novel_leaf_fall_ disease_of_rubber_trees_Hevea_brasiliensis_in_Thailand.
[11]	Georgen Agrios. 植物病理学[M].沈崇尧, 译. 北京: 中国农业大学出版社, 2009.
[12]	李博勋, 刘先宝, 林春花, 等. 国内橡胶树主要种质对棒孢霉落叶病的抗性评价[J]. 植物保护, 2014,40(5):86-92.
[13]	时涛, 李博勋, 郑肖兰, 等. 橡胶树新发拟盘多毛孢叶斑病的危害与防控对策[J]. 热带农业科学, 2020,40(1):1-6.
[14]	Maharachchikumbura S S N, Hyde K D, Groenewald J Z, et al Pestalotiopsis revisited[J]. Studies in Mycology, 2014,79(11):121-186.
[15]	Adam Malik Ahmad. Outbreak of Pestalotiopsis leaf fall in Malaysia: Meeting of experts on Pestalotiopsis leaf disease[Z]. Surat Thani Province, Thailand, 2020.
[16]	Maharachchikumbura S S N, Liang-Dong G, Ekachai C, et al. A destructive new disease of Syzygium samarangense in Thailand caused by the new species Pestalotiopsis samarangensis[J]. Tropical Plant Pathology, 2013,38(3):227-235.
[17]	宋玉. 拟盘多毛孢属(Pestalotiopsis)真菌系统学研究及条形码基因的评价[D]. 贵阳: 贵州大学, 2015.
[18]	Steyaert R L. Contribution a l'etude monographique de Pestalotia de Not et Monochaetia Sacc. (Truncatella gen. nov. et Pestalotiopsis gen. nov.)[J]. Bulletin du Jardin Botanique de 16tat Bruxelles, 1949,19:285-358.
[19]	韦继光, 徐同, 郭良栋, 等. 根据形态学和分子系统学特征界定拟盘多毛孢属的种[J]. 广西农业生物科学, 2005(4):304-313.
[20]	Febbiyanti T R. Outbreak of Pestaloiopsis in Indonesia: IRRDB meeting of the experts on new leaf disease[Z]. Kuala Lumpur, 2019.

类群 Species	菌株编号 Stains	采集时间 Date	地理来源 Geographical origin	寄主 Host/substrate	ITS登录号 GeneBank	TEF登录号 GeneBank	TUB登录号 GeneBank
Neopestalotiopsis aotearoa	HNPeHNLD2001	2020	中国海南省乐东县志仲镇	Hevea brasiliensis	MT764947	MT800516	MT796262
N. aotearoa	HNPeHNLD2002	2020	中国海南省乐东县志仲镇	Hevea brasiliensis	MT764948	MT800517	MT796263
N. aotearoa	CBS 367.54^*	1954	新西兰	Canvas	KM199369	KM199526	KM199454
N. australis	CBS 114159^*	1999	澳大利亚新南威尔士州	Telopea sp.	KM199348	KM199432	KM199537
N. clavispora	CBS 447.73	1949	斯里兰卡	Decaying wood	KM199374	KM199539	KM199443
N. cubana	CBS 600.96^*	1996	古巴	Leaf litter	KM199347	KM199521	KM199438
N. ellipsospora	CBS 115113	2012	中国香港	Ardisia crenata	KM199343	KM199544	KM199450
N. eucalypticola	CBS 264.37^*	2012		Eucalyputus globulus	KM199376	KM199551	KM199431
N. formicarum	CBS 362.72^*	1971	加纳	Dead formicidae（ant）	KM199358	KM199517	KM199455
N. formicarum	PSU-R-L02	2019	泰国	Hevea brasiliensis	LC521861	LC521869	LC521875
N. honoluluana	CBS 111535	1998	美国夏威夷	Telopea sp.	KM199363	KM199546	KM199461
N. javaensis	CBS 257.31^*	2011	印度尼西亚	Cocos nucifera	KM199357	KM199543	KM199437
N. mesopotamica	CBS 336.86^*	1969	伊拉克	Pinus brutia	KM199362	KM199555	KM199441
N. piceana	CBS 394.48^*	1998	英国	Picea sp.	KM199368	KM199527	KM199453
N. protearum	CBS 114178^*	1998	津巴布韦	Leucospermum cumeiforme cv.	JN712498	KM199542	KM199463
N. rosae	CBS 101057^*	1998	新西兰	Rosa sp.	KM199359	KM199429	KM199523
N. saprophytica	CBS 115452	2001	中国香港	Litsea rotundifolia	KM199345	KM199433	KM199538
N. surinamensis	CBS 450.74^*	1974	苏里南	Soil under Elaeis fuineensis	KM199351	KM199518	KM199465
N. zimbabwana	CBS 111495^*	1998	津巴布韦	Leucospermum cumeiforme cv.	JX556231	KM199545	KM199456
Pseudopestalotiopsis cocos	CBS 272.29^*		印度尼西亚	Cocos nucifera	KM199378	KM199553	KM199467
Ps. indica	CBS 459.78^*		印度	Hibiscus rosa-sinensis	KM199381	KM199560	KM199470

类群 Species	菌株编号 Stains	采集时间 Date	地理来源 Geographical origin	寄主 Host/substrate	ITS登录号 GeneBank	TEF登录号 GeneBank	TUB登录号 GeneBank
Neopestalotiopsis aotearoa	HNPeHNLD2001	2020	中国海南省乐东县志仲镇	Hevea brasiliensis	MT764947	MT800516	MT796262
N. aotearoa	HNPeHNLD2002	2020	中国海南省乐东县志仲镇	Hevea brasiliensis	MT764948	MT800517	MT796263
N. aotearoa	CBS 367.54^*	1954	新西兰	Canvas	KM199369	KM199526	KM199454
N. australis	CBS 114159^*	1999	澳大利亚新南威尔士州	Telopea sp.	KM199348	KM199432	KM199537
N. clavispora	CBS 447.73	1949	斯里兰卡	Decaying wood	KM199374	KM199539	KM199443
N. cubana	CBS 600.96^*	1996	古巴	Leaf litter	KM199347	KM199521	KM199438
N. ellipsospora	CBS 115113	2012	中国香港	Ardisia crenata	KM199343	KM199544	KM199450
N. eucalypticola	CBS 264.37^*	2012		Eucalyputus globulus	KM199376	KM199551	KM199431
N. formicarum	CBS 362.72^*	1971	加纳	Dead formicidae（ant）	KM199358	KM199517	KM199455
N. formicarum	PSU-R-L02	2019	泰国	Hevea brasiliensis	LC521861	LC521869	LC521875
N. honoluluana	CBS 111535	1998	美国夏威夷	Telopea sp.	KM199363	KM199546	KM199461
N. javaensis	CBS 257.31^*	2011	印度尼西亚	Cocos nucifera	KM199357	KM199543	KM199437
N. mesopotamica	CBS 336.86^*	1969	伊拉克	Pinus brutia	KM199362	KM199555	KM199441
N. piceana	CBS 394.48^*	1998	英国	Picea sp.	KM199368	KM199527	KM199453
N. protearum	CBS 114178^*	1998	津巴布韦	Leucospermum cumeiforme cv.	JN712498	KM199542	KM199463
N. rosae	CBS 101057^*	1998	新西兰	Rosa sp.	KM199359	KM199429	KM199523
N. saprophytica	CBS 115452	2001	中国香港	Litsea rotundifolia	KM199345	KM199433	KM199538
N. surinamensis	CBS 450.74^*	1974	苏里南	Soil under Elaeis fuineensis	KM199351	KM199518	KM199465
N. zimbabwana	CBS 111495^*	1998	津巴布韦	Leucospermum cumeiforme cv.	JX556231	KM199545	KM199456
Pseudopestalotiopsis cocos	CBS 272.29^*		印度尼西亚	Cocos nucifera	KM199378	KM199553	KM199467
Ps. indica	CBS 459.78^*		印度	Hibiscus rosa-sinensis	KM199381	KM199560	KM199470

基因 Gene	引物 Primer	序列 Sequence (5°-3°)	退火温度 Annealing temperature/℃	文献来源 Reference
ITS	ITS1	TCCGTAGGTGAACCTGCGG	53	Pornsuriya等^[10]
ITS	ITS4	TCCTCCGCTTATTGATATGC	53
EF-1α	EF1-728F	CATCGAGAAGTTCGAGAAGG	53
EF-1α	EF1-986R	TACTTGAAGGAACCCTTACC	53
TUB	T1	AACATGCGTGAGATTGTAAGT	56
TUB	Bt2b	ACCCTCAGTGTAGTGACCCTTGGC	56

基因 Gene	引物 Primer	序列 Sequence (5°-3°)	退火温度 Annealing temperature/℃	文献来源 Reference
ITS	ITS1	TCCGTAGGTGAACCTGCGG	53	Pornsuriya等^[10]
ITS	ITS4	TCCTCCGCTTATTGATATGC	53
EF-1α	EF1-728F	CATCGAGAAGTTCGAGAAGG	53
EF-1α	EF1-986R	TACTTGAAGGAACCCTTACC	53
TUB	T1	AACATGCGTGAGATTGTAAGT	56
TUB	Bt2b	ACCCTCAGTGTAGTGACCCTTGGC	56

形态特征 Morphological characteristic	N. aotearoa	HNPeHNLD2001	HNPeHNLD2002
PDA上的培养性状	菌丝纯白色、稠密,菌落圆形、平铺、边缘整齐	菌落呈圆形、最初边缘成波浪状;菌丝纯白色、生长旺盛,一层一层的向外延伸,菌落背面略微黄色	菌落呈圆形、边缘整齐无波浪状;菌丝纯白色、生长旺盛菌落背面略微黄色
孢子颜色	深棕色至黑色	深棕色	深棕色
细胞数量/个	5	5	5
孢子长/μm	19.5~29.0	18.35~27.12	17.38~28.34
孢子宽/μm	6.0~9.0	4.11~7.03	4.21~6.94
有色细胞长/μm	13.0~18.5	12.53~18.97	11.93~18.67
附属丝数量/根	2~3	2~3	2~3
附属丝长/μm	3.0~13.0	14.7~25.3	12.4~24.3
基部附属丝数量/根	1	1	1
基部附属丝长/μm	1.5~4.0	3.13~7.13	2.97~6.59
参考	Maharachchikumbura等^[14]	本研究	本研究