Isolation and identification of an anthracnose pathogen on Vicia sativa

doi:10.11686/cyxb2019299

Abstract

Abstract: Anthracnose in Vicia sativa is an important fungal disease affecting leaves and stems. This research combined morphological and multiple gene phylogenetic analyses, using gene sequences of an internal transcribed spacer region, actin, chitin synthase 1 and glyceraldehyde-3-phosphate dehydrogenase to identify an anthracnose pathogen (WQ) isolated from leaves and stems of vetch plants (cv. Lanjian No.2) in Xiahe County, Gansu Province. Koch’s postulates were applied to identify pathogenicity of the strains. It was found that colonies of the fungus on potato dextrose agar (PDA) were flat, with surface dark gray-olivaceous to olivaceous-black, white margin, covered by short floccose whitish aerial mycelium, reverse gray-olivaceous to iron-gray. Conidia were hyaline, unicellular, (17.5-25.0) μm×(3.75-5.00) μm, and slightly falcate. Multiple-gene phylogenetic analysis indicated that the isolate WQ and a strain of CBS 128.57 were clustered into one clade, with a confidence level of 70%. The strains (WQ) could infect V. sativa leaves through wounds. It was confirmed that the pathogen was Colletotrichum spinaciae. This is the first report of C. spinaciae on V. sativa in China. A host range test showed that C. spinaciae exhibited high pathogenicity to Onobrychis viciifolia and Avena sativa, and moderate pathogenicity to four V. sativa cultivars, and low pathogenicity to Medicago sativa and Trifolium repens. However, Colletotrichum spinaciae had no pathogenicity to Lolium multiflorum.

Key words: Vicia sativa, anthracnose, Colletotrichum spinaciae, pathogen identification, host range

WANG Qiong, DUAN Ting-yu, NAN Zhi-biao. Isolation and identification of an anthracnose pathogen on Vicia sativa[J]. Acta Prataculturae Sinica, 2020, 29(6): 127-136.

References

[1] Wang Y L, Zheng M N, Han Z S. Growth dynamics and harvest period of Vicia sativa. Chinese Agricultural Science Bulletin, 2016, 32(9): 71-76.
王雁丽, 郑敏娜, 韩志顺. 春箭筈豌豆生长动态及收割期的研究. 中国农学通报, 2016, 32(9): 71-76.
[2] Liu Y, Wang Y R. Effect of temperature and soil moisture on seedling growth of Vicia sativa. Pratacultural Science, 2014, 31(7): 1302-1309.
刘勇, 王彦荣. 温度和水分对箭筈豌豆幼苗生长的影响. 草业科学, 2014, 31(7): 1302-1309.
[3] Sayar M S. Path coefficient and correlation analysis between forage yield and its affecting components in common vetch (Vicia sativa L.). Legume Research, 2014, 37(5): 445-452.
[4] Mao K, Zhou S R, Wang S M, et al. Study on the dynamics of biomass and interspecific competition of mixture communities of common vetch with Italian ryegrass. Acta Agrestia Sinica, 1997, 5(1): 8-14.
毛凯, 周寿荣, 王四敏, 等. 箭筈豌豆混播黑麦草生物量和种间竞争的研究. 草地学报, 1997, 5(1): 8-14.
[5] Mikiĉ A, Mihailoviĉ V, Ĉupina B, et al. Forage yield components and classification of common vetch (Vicia sativa L.) cultivars of diverse geographic origin. Grass & Forage Science, 2014, 69(2): 315-322.
[6] Tao X L, Ma L C, Nie B, et al. The draft and characterization of the complete chloroplast genome of Vicia sativa cv. Lanjian No.3. Pratacultural Science, 2017, 34(2): 321-330.
陶晓丽, 马利超, 聂斌, 等. “兰箭3号”春箭筈豌豆叶绿体全基因组草图及特征分析. 草业科学, 2017, 34(2): 321-330.
[7] Huang Y F, Gao X L, Nan Z B, et al. Potential value of the common vetch (Vicia sativa L.) as an animal feedstuff: A review. Journal of Animal Physiology and Animal Nutrition, 2017, 101(5): 807-823.
[8] Akmakci S, Aclikgöz E. Components of seed and straw yield in common vetch (Vicia sativa L.). Plant Breeding, 2010, 113(1): 71-74.
[9] Flrlnclog Lu H K, Erbekta? E, DogRuyol L, et al. Phenotypic variation of autumn and spring-sown vetch (Vicia sativa ssp.) populations in central turkey. Spanish Journal of Agricultural Research, 2009, 7(3): 596-606.
[10] Erol A, Kaplan M. Oats (Avena sativa)-common vetch (Vicia sativa) mixtures grown on a low-input basis for a sustainable agriculture. Tropical Grasslands, 2009, 43(1): 191-196.
[11] Li J F. Cultivation techniques and benefit analysis of common vetch in Gulang County. Gansu Animal and Veterinary Sciences, 2017, 47(5): 109-110.
李金芳. 箭筈豌豆在古浪县的栽培技术及效益分析. 甘肃畜牧兽医, 2017, 47(5): 109-110.
[12] Gao X L. Responses of four Vicia sativa cultivars to phosphorus and potassium fertilizers. Lanzhou: Lanzhou University, 2018.
高小莉. 四个春箭筈豌豆品种对磷钾肥的响应. 兰州: 兰州大学, 2018.
[13] Nan Z B. Establishing sustainable management system for diseases of pasture crops in China. Acta Prataculturae Sinica, 2000, 9(2): 1-9.
南志标. 建立中国的牧草病害可持续管理体系. 草业学报, 2000, 9(2): 1-9.
[14] Xu S, Li Y Z. Research advances on fungal diseases of Vicia sativa. Acta Prataculturae Sinica, 2016, 25(7): 203-214.
徐杉, 李彦忠. 箭筈豌豆真菌病害研究进展. 草业学报, 2016, 25(7): 203-214.
[15] Dearness J. New and noteworthy fungi. IV. Mycologia, 1926, 18(5): 236-255.
[16] Horn N L. A comparative study of two species of Colletotrichum on vetch. Phytopathology, 1952, 42: 670-674.
[17] Weimer J L. A new species of Colletotrichum on vetch. Phytopathology, 1945, 35(12): 977-990.
[18] Sawada K. Descriptive catalogue of Taiwan (Formosan) fungi. Part XI. Special Publication College Agriculture Taiwan University, 1959.
[19] Liu R, Hou T J. Preliminary list of fungal diseases of leguminous forge in Northern China. Chinese Journal of Grassland, 1984, (1): 56-60, 80.
刘若, 侯天爵. 我国北方豆科牧草真菌病害初步名录. 中国草地学报, 1984, (1): 56-60, 80.
[20] Xu S, Li Y Z. First report of common vetch anthracnose caused by Colletotrichum lentis in China. Plant Disease, 2015, 99(12): 1859.
[21] Xu S, Christensen M J, Li Y Z. Pathogenicity and characterization of Colletotrichum lentis: A causal agent of anthracnose in common vetch (Vicia sativa). European Journal of Plant Pathology, 2017, 149(3): 1-13.
[22] Li Y Z, Nan Z B. The methods of diagnose, investigation and loss evaluation for diseases. Nanjing: Jiangsu Fenghuang Science and Technology Press, 2015.
李彦忠, 南志标. 牧草病害诊断调查与损失评定方法. 南京: 江苏凤凰科学技术出版社, 2015.
[23] Ma R, Xu G, Zheng F, et al. Identification of the pathogen causing anthracnose on Curcuma wenyujin in Hainan, China. Plant Protection, 2018, 44(4): 81-86.
马瑞, 徐刚, 郑樊, 等. 海南省温郁金炭疽病的病原鉴定. 植物保护, 2018, 44(4): 81-86.
[24] Carbone I, Kohn L M. A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia, 1999, 91(30): 553-556.
[25] Guerber J C, Liu B, Correll J C, et al. Characterization of diversity in Colletotrichum acutatum sensu lato by sequence analysis of two gene introns, mtdna and intron rflps, and mating compatibility. Mycologia, 2003, 95(5): 872-895.
[26] Damm U, Woudenberg J, Cannon P, et al. Colletotrichum species with curved conidia from herbaceous hosts. Fungal Diversity, 2009, 39: 45-87.
[27] Kurt ?, Uysal A, Akgül D. First report of anthracnose caused by Colletotrichum spinaciae on spinach in the Mediterranean region of Turkey. Plant Disease, 2016, 100(1): 219.
[28] Yang J W, Zhao Z L, Zhang G Q, et al. Identification and biological characterization of anthrax bacterial in xianlajiao chili pepper in Shanxi province. Acta Agriculturae Boreali-occidentalis Sinica, 2017, 26(11): 1695-1705.
杨佳文, 赵尊练, 张管曲, 等. 陕西线辣椒炭疽病原菌的鉴定及生物学特性研究. 西北农业学报, 2017, 26(11): 1695-1705.
[29] Uysal A, Kurt ?. Influence of inoculum density, temperature, wetness duration, and leaf age on infection and development of spinach anthracnose caused by the fungal pathogen Colletotrichum spinaciae. European Journal of Plant Pathology, 2017, 149(4): 1041-1052.
[30] Correll J C, Morelock T E, Black M C, et al. Economically important diseases of spinach. Plant Disease, 1994, 78(7): 653-660.
[31] Gourley C. The pathogenicity of Colletotrichum dematium to table beets and other hosts. Canadian Journal of Plant Science, 1966, 46(5): 531-536.
[32] Cerkauskas R, Nauta R, Mcdonald M. First report of spinach anthracnose in ontario. Plant Disease, 1991, 75(1): 101.
[33] Washington W, Irvine G, Aldaoud R, et al. First record of anthracnose of spinach caused by Colletotrichum dematium in Australia. Australasian Plant Pathology, 2006, 35(1): 89-91.
[34] Zhang J P. Research on poplar leaf veins spot pathogen and occurrence regularity. Urumqi: Xinjiang Agricultural University, 2016.
张金萍. 杨树叶纹斑病病原菌及发生规律的研究. 乌鲁木齐: 新疆农业大学, 2016.
[35] Li C, Jiang J X, Leng J H, et al. Isolation and identification of pathogenic fungi causing fruit rot of kiwifruit in Fengxin County. Acta Agriculturae Universitatis Jiangxiensis, 2012, 34(2): 259-263.
李诚, 蒋军喜, 冷建华, 等. 奉新县猕猴桃果实腐烂病病原菌分离鉴定. 江西农业大学学报, 2012, 34(2): 259-263.
[36] Shang J. Study on the infection cycle and field epidemic of Lonsdalea quercina subsp. populi on Populus xeuramericana. Beijing: Beijing Forestry University, 2014.
商靖. 欧美杨细菌性溃疡病菌侵染循环及病害流行研究. 北京: 北京林业大学, 2014.
[37] Xiong C W, Ruan C J, Huang H, et al. Identification and homology analysis of pathogens causing anthracnose of Camellia oleifera growing in Yuping. Molecular Plant Breeding, 2019, 17(1): 169-174.
熊朝伟, 阮成江, 黄河, 等. 玉屏油茶炭疽病病原菌鉴定及同源性分析. 分子植物育种, 2019, 17(1): 169-174.
[38] Smith B, Black L. Morphological, cultural, and pathogenic variation among Colletotrichum species isolated from strawberry. Plant Disease, 1990, 74(1): 69-76.
[39] Wang Q H, Liu X H, Fan K, et al. Identification and biological characteristics of pathogen from Colletotrichum gloeosporioides. Journal of Shandong Agricultural University (Natural Science Edition), 2016, 47(1): 9-14.
王清海, 刘幸红, 范昆, 等. 核桃炭疽病病原菌鉴定及生物学特性. 山东农业大学学报(自然科学版), 2016, 47(1): 9-14.
[40] Jing R, Zhao F J, Liu Y F, et al. Isolation and identification of potato Verticillium wilt pathogen in Ningxia and its host range. Acta Phytopathologica Sinica, 2019, 49(1): 11-19.
景瑞, 赵方杰, 刘一凡, 等. 宁夏马铃薯黄萎病病原菌分离鉴定及寄主范围测定. 植物病理学报, 2019, 49(1): 11-19.