Alternaria spp. Associated with Potato Foliar Diseases in China

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1 Alternaria spp. Associated with Potato Foliar Diseases in China 引起马铃薯叶部病害的链格孢菌种类鉴定 Prof. Xuehong Wu ( Department of Plant Pathology, China Agricultural University 吴学宏教授 / 博士生导师 ( 手机 : ; 电子邮件 :wuxuehong@cau.edu.cn) 中国农业大学植物病理学系

2 Main Contents ( 主要内容 ) 1. Introduction ( 研究背景 ) 2. Technical Route ( 研究内容 ) 3. Results ( 研究结果 ) 4. Conclusions and Discussions ( 结论和讨论 )

3 1.1 Importance of Potato Crop 1.1 马铃薯的重要性 Potato ranks as the fourth most important staple food crop after rice, wheat, and maize. 马铃薯是仅次于水稻 小麦 玉米之后的第四大粮食作物 In China, culture areas of potato is about 5 million hectares (25% of the world). 我国种植面积 500 万公顷 (25%)

4 World potato production (Metric tones by count) China :74,799,084 China has the largest growing area and consumption of potato in the world.

5 1.2 Potato Foliar Diseases 1.2 马铃薯叶部病害 1) Potato early blight (In 1892, USA), caused by Alternaria solani 1892 年美国首先报道茄病链格孢菌引起马铃薯早疫病 2) Potato brown spot (In 1984, Israel), caused by A. alternata 1984 年以色列报道交链格孢菌引起马铃薯褐斑病 3) Potato leaf spot (In 2010, Iran), caused by A. alternata, A.tenuissima, A. dumosa, A. solani, A. arborescens,a. infectoria and A. longipes 年伊朗报道交链格孢菌等 7 种引起马铃薯叶斑病

6 Early blight (1892, USA), Brown spot (1984, Israel), Leaf spot (2010, Iran) 早疫病 (1892, 美国 ) 褐斑病 (1984, 以色列 ) 叶斑病 (2010, 伊朗 )

7 1.3 Alternaria spp. associated with potato foliar diseases 1.3 引起马铃薯叶部病害的链格孢菌种类 Worldwide Alternaria solani, A. alternata, A. interrupta, A. grandis, A. tenuissima, A. dumosa, A. arborescens, A. infectoria. China (Hebei, Heilongjiang) Alternaria solani, A. alternata

8 We want to know An accurate understanding of the pathogen composition is crucial for efficient disease management. 引起病害的病原菌种类的准确鉴定才能进行有效地防治 Whether the other six species already existed or will appear later in the potato growing areas of China? How about their spatial distribution? 另外六种国际上已报道的链格孢菌在中国是否存在, 它们的分布情况如何?

9 2. Technical route 2. 研究内容 Sampling of diseased potato leaves with symptoms 采集发病的病害叶片 Alternaria fungi isolation, purification, and preserve 链格孢菌的分离 纯化和保存 Alternaria species identification (morphology, molecular biology) 采用形态学和分子生物学方法进行种类鉴定 Pathogenicity on potato (spore suspension inoculation) 致病性试验 ( 采用孢子悬浮液的方法 )

10 A total of 511 Alternaria isolates were obtained from 193 locations in 16 provinces, autonomous regions, and municipalities of China. 中国 16 个省市自治区采样分离得到 511 株链格孢菌 黑龙江 吉林 新疆 西藏 青海 辽宁 内蒙古 河北 宁夏 山西 山东 甘肃 陕西 河南 江苏 安徽 四川 重庆 湖北 浙江 湖南 江西 贵州 福建 云南台湾广东广西 海南

11 a c e b d f Fig.1 Leaf sample collection (a, b) ; Pathogen isolation (c, d) and purification (e, f)

12 2.1 Morphological traits observation Fig. 2 Colony and microscopic morphologies of Alternaria species a-c:colonies of A. tenuissima,a. alternata,a. solani ;d-f:conidia of A. tenuissima,a. alternata,a. solani ;g-h:sporulation pattern of A. tenuissima and A. alternata

13 Table 1 Morphological traits of Alternaria species Species Colony Sporulation Conidiophore Conidia Shape Size/ µm Shape Size/ µm Septum A.t loosely cottony, greyishgreen to olive brown unbranched conidial chains with one or two lateral branches occasionally short, arising singly typically ovoid to obclavate transverse septa:1-6 longitudinal septa:0-2 A.a densely turfy dark gray to black brown, with numerous secondary and occasionally tertiary chains single or fasciculat e, straight or bent obpyriform to ellipsoid transversal septa: 3-8 longitudinal septa: 0-3 A.s dense dark gray to black Not observed simple conidiophores bearing dark, multiseptate conidia with one or two beak transverse septa: 9-11 longitudinal septa: 1-2

14 2.2 Molecular identification

15 A. solani A. tenuissima / A. alternata Phylogenetic tree constructed based on the sequences of the ITS regions of rdna of 55 A. tenuissima, 23 A. alternata, 9 A. solani isolates, as well as 15 reference sequences retrieved from GenBank.

16 A. solani A. alternata A. tenuissima Phylogenetic tree constructed based on the partial coding sequences of histone 3 gene of 55 A. tenuissima isolates, 23 A. alternata isolates, 9 A. solani isolates, as well as 9 reference sequences retrieved from GenBank

17 Table 2 Alternaria isolates obtained from the diseased leaf samples Geographic origins Numbers of locations Numbers of Alternaria isolates A. tenuissima A. alternata A. solani Inner Mongolia Autonomous Region (I M) (27) 13 (10) 3 (3) Hebei province (HB) (19) 10 (2) 16 (16) Beijing Municipality (BJ) 1 3 (1) 1 (1) 0 Shandong Province (SD) (15) 18 (10) 0 Shanxi Province (SX) 1 1 (1) 0 0 Shaanxi Province (SAX) 1 3 (1) 1 (1) 0 Ningxia Hui Autonomous Region (NX) (18) 3 (3) 8 (8) Gansu Province (GS) (5) 21 (4) 3 (3) Heilongjiang Province (HLJ) (9) 5 (4) 0 Jilin Province (JL) 2 3 (2) 0 0 Liaoning Province (LN) 1 3 (1) 0 0 Chongqing Municipality (CQ) 4 10 (4) 7 (2) 0 Yunnan Province (YN) 3 8 (3) 6 (2) 0 Guizhou Province (GZ) 3 6 (2) 2 (1) 0 Sichuan Province (SC) 1 4 (1) 0 0 Xinjiang Uygur Autonomous Region (XJ) 2 8 (1) 8 (2) 0 Total (110) 95 (42) 30 (30) Ratio % 18.6% 5.9%

18 Alternaria 链格孢菌 Disease incidence 发病率 (%) Disease index 病情指数 A. tenuissima (30.9±6.7) (44.98±5.53) A. alternata (34.3±4.8) (46.61±6.26) A. solani (44.2±1.9) (57.75±1.31) A. tenuissima 细极链格孢 A. alternata 交链格孢 A. solani 茄病链格孢

19 4. Conclusions and discussions 4. 结论和讨论 4.1 A. tenuissima was the most prevalent (75.5%), and widely distributed in all the sixteen provinces, autonomous regions, and municipalities, followed by A. alternata (18.6%), and A. solani (5.9%). This is the first report of A. tenuissima causing potato foliar diseases in China. 细极链格孢菌为优势种群, 其次为交链格孢和茄病链格孢 这是国内首次系统研究报道 4.2 The other five Alternaria species including A. dumosa, A. arborescens, A. infectoria, A. grandis, and A. interrupta reported to cause potato foliar diseases in other countries, were not detected in the present study. 国际上已报道的另外 5 种链格孢菌未见报道

20 4.3 The ITS sequences of rdna were able to differentiate A. solani isolates from small-spored Alternaria isolates; histone 3 gene were successfully used in differentiating the small-spored Alternaria isolates that were hard to classify by morphological traits or ITS sequence. ITS 区可以用于区分大孢子链格孢与小孢子链格孢菌, 组蛋白基因还可以区分不同的小孢子链格孢菌 4.4 Since A. tenuissina is predominant in the isolates obtained, we therefore propose that A. tenuissima should be treated as the major target in decision of management strategies. 细极链格孢菌应作为生产中防治的重点对象

21 Publications related to Alternaria Causing potato diseases 1) Wu X. H (Corresponding Author) First Report of Alternaria Blight of Potato Caused by Alternaria tenuissima in China, Plant Disease. 97(9): ) Wu X. H (Corresponding Author) Characterization of Alternaria species associated with potato foliar diseases in China, Plant Pathology, 2015,64(2):425-43

22 Publications related to Rhizoctonia causing potato diseases 1)Wu X H (corresponding author). Characterization of a new anastomosis group (AG-W) of binucleate Rhizoctonia, causal agent for potato stem canker. Plant Disease, 2015,99(12): )Wu X H (corresponding author). Anastomosis group and pathogenicity of Rhizoctonia solani associated with stem canker and black scurf of potato in China. European Journal of Plant Pathology, 2015,143(1): )Wu X H (corresponding author). Potato stem canker caused by binucleate Rhizoctonia AG-G in China. Journal of General Plant Pathology, 2015,81(4): )Wu X H (corresponding author). Anastomosis groups and pathogenicity of binucleate Rhizoctonia isolates associated with stem canker of potato in China. European Journal of Plant Pathology, 2014, 139(3):

23 Thanks for your attention! Prof. Xuehong Wu Department of Plant Pathology, China Agricultural University, Beijing

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