Identification of Pathogen and Biocontrol Agents on Southern Blight of Pseudostellaria heterophylla
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摘要:
目的 太子参白绢病是一种可造成严重产量损失的土传病害。为有效防控病害，对其病原菌种类进行分离鉴定，并筛选有效生防菌。 方法 从具有典型白绢病症状的太子参植株和块根病样中，分离纯化获得病原菌；利用形态学、分子生物学和致病性测定进行病原菌鉴定；取代表性菌株用平板对峙法进行生防菌的筛选。 结果 病原菌的菌落呈圆形，菌丝为白色绢状辐射生长、有明显隔膜和分枝，后期有不同颜色的菌核形成；病原菌经ITS、LSU、TEF-1α等基因序列分析，确定为罗耳阿太菌Athelia rolfsii；致病测定发病症状与田间一致，符合柯赫氏法则。筛选获得对该病原菌具有良好抑制效果的生防菌贝莱斯芽胞杆菌FJAT-17931和暹罗芽胞杆菌FJAT-52595，抑菌率分别为73.23%和71.16%。 结论 鉴定出太子参白绢病的病原菌为A. rolfsii，筛选获得两株对该病原菌具有良好防效的生防菌，为太子参白绢病的防治提供依据。 Abstract:Objective Pathogen and potential biocontrol agents of the southern blight, a major soil-borne disease, of Pseudostellaria heterophylla were identified. Method P. heterophylla plant parts and tubers showing the typical southern blight symptoms were collected for the pathogen isolation. The isolate was examined by morphological, molecular biological, and pathogenicity tests for taxonomic confirmation. Microbes with an antibacterial property against the isolate were screened using the plate confrontation culture method to evaluate the potential of their being used as a biocontrol agent on the blight. Result On culture plate, the round colonies of the suspected pathogen had white, silky, spreading mycelia with septa and ramifications that, at later stage, formed sclerotia in different colors. The isolate was further identified as Athelia rolfsii by ITS, LSU, and TEF-1α gene sequence analyses. The disease symptoms on the plants inoculated with the isolate were identical to those observed in the field and in accordance with Koch's rule. On the biocontrol efficacy evaluation, Bacillus velezensis FJAT-17931 and B. siamensis FJAT-52595 displayed high inhibitory effect against A. rolfsii with the rates of 72.23 % and 71.16%, respectively. Conclusion The pathogen of the southern blight on P. heterophylla was identified as A. rolfsii. Two Bacillus strains were found to be potentially applicable as biocontrol agents on the disease that can cause serious yield loss on the crop. -
Key words:
- Pseudostellaria heterophylla /
- southern blight /
- Athelia rolfsii /
- identification /
- biocontrol agent
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图 1 太子参白绢病田间发病症状
注：A，植株症状；B～D，块根症状。
Figure 1. Disease symptoms of southern blight on P. heterophylla in the field
Note: A, disease symptoms on aboveground plant parts; B-D, disease symptoms on tuberous roots.
图 2 太子参白绢病病原菌的形态特征
注：A～C，菌落在PDA培养基上的形态；D，菌丝形态。
Figure 2. Morphology of southern blight pathogen
Note: A-C, Colonies grown on PDA medium; D, Hyphae.
图 3 病原菌系统发育分析
注：A～C分别为基于rDNA-ITS、rDNA-LSU、TEF-1α序列构建的系统发育树。
Figure 3. Phylogenetic trees of pathogen
Note: A-C, phylogenetic trees constructed based on rDNA-ITS, rDNA-LSU, and TEF-1α.
图 4 接种太子参白绢病病原菌后太子参植株的发病症状
注：A，接种病原菌6 d 的根际周围；B，接种病原菌6 d 的植株根茎部；C，接种病原菌6 d的地上部植株。
Figure 4. Disease symptoms on P. heterophylla plants artificially inoculated with isolated pathogen
Note: A, rhizosphere at 6 days post inoculation (dpi) with pathogen; B, plant rhizome at 6 dpi with pathogen; C, aboveground plant parts at 6 dpi with pathogen.
图 5 菌株FJAT-32609和FJAT-31609接种太子参块根后，不同时间的发病症状
注：A～B，块根接种FJAT-32609第7天；C～D，块根接种FJAT-32609第8～9天；E～F，块根接种FJAT-32610第8～9天；G～H，块根接种FJAT-32609 10天后；I～J，无菌水对照处理。
Figure 5. Disease symptoms on tubers of P. heterophylla plants artificially inoculated with isolated pathogen
Note: A-B, tubers at 7 dpi with FJAT-32609; C-D, tubers at 8–9 dpi with FJAT-32609; E-F, tubers at 8–9 dpi with FJAT-32610; G-H, tubers at 10 dpi with FJAT-32609; I-J, control soaked in sterile water.
表 1 不同芽胞杆菌对太子参白绢病病原菌的抑菌率
Table 1. Inhibition rates of Bacillus spp. on A. rolfsii
菌株 Strain 抑菌率 Inhibitory rate/% 贝莱斯芽胞杆菌FJAT-17931（B. velezensis） 73.23±1.38 a 暹罗芽胞杆菌FJAT-52595（B. siamensis） 71.16±0.65 ab 暹罗芽胞杆菌FJAT-52594（B. siamensis） 69.96±1.46 ab 贝莱斯芽胞杆菌FJAT-16022（B. velezensis） 67.96±2.04 ab 贝莱斯芽胞杆菌FJAT-47051（B. velezensis） 64.60±0.05 b 地衣芽胞杆菌FJAT-4（B. licheniformis） 32.03±5.03 c 注：同列数据后不同小写字母表示处理间差异显著（P＜0.05）。
Note: Data with different lowercase letters on same column indicate significant differences at P ＜ 0.05.
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[1] 江维克, 周涛. 太子参产业发展现状及其建议 [J]. 中国中药杂志, 2016, 41(13):2377−2380.JIANG W K, ZHOU T. Pesudostellariae Radix industry status and development countermeasures [J]. China Journal of Chinese Materia Medica, 2016, 41(13): 2377−2380.(in Chinese) [2] 肖荣凤, 陈燕萍, 陈梅春, 等. 太子参根腐病病原菌的鉴定及防治药剂筛选 [J]. 植物保护学报, 2020, 47(6):1333−1342.XIAO R F, CHEN Y P, CHEN M C, et al. Pathogen identification of root rot of Pseudostellaria heterophylla plant and fungicide screening for its efficient control [J]. Journal of Plant Protection, 2020, 47(6): 1333−1342.(in Chinese) [3] 曾桂萍, 赵致, 李忠. 贵州太子参白绢病发生及防治 [J]. 耕作与栽培, 2020(6):37−38,41.ZENG G P, ZHAO Z, LI Z. The occurrence and prevention of Pseudostellaria heterophylla southern blight in Guizhou [J]. Tillage and Cultivation, 2020(6): 37−38,41.(in Chinese) [4] 桑维钧, 熊继文, 宋宝安, 等. 贵州太子参主要真菌病害的调查与防治 [J]. 安徽农业科学, 2006, 34(14):3314−3316,3318. doi: 10.3969/j.issn.0517-6611.2006.14.083SANG W J, XIONG J W, SONG B A, et al. Investigation of main fungus diseases of ginseng in Guizhou Province [J]. Journal of Anhui Agricultural Sciences, 2006, 34(14): 3314−3316,3318.(in Chinese) doi: 10.3969/j.issn.0517-6611.2006.14.083 [5] 周丹, 程搏幸, 丁波. 太子参真菌病害和防治研究进展 [J]. 中国植保导刊, 2020, 40(9):23−27,32. doi: 10.3969/j.issn.1672-6820.2020.09.004ZHOU D, CHENG B X, DING B. Research progress on fungal diseases of Pseudostellaria heterophylla and their control [J]. China Plant Protection, 2020, 40(9): 23−27,32.(in Chinese) doi: 10.3969/j.issn.1672-6820.2020.09.004 [6] 陈桂华. 太子参白绢病药剂防治效果试验 [J]. 内蒙古农业科技, 2010, 38(2):54−54,57.CHEN G H. Control effect of Pseudostellaria heterophylla southern blight with fungicides [J]. Inner Mongolia Agricultural Science and Technology, 2010, 38(2): 54−54,57.(in Chinese) [7] 姜星, 祝前, 刘德江. 太子参白绢病田间药效试验初报[J]. 农业与技术, 2017, 37(18): 23.JIANG X, ZHU Q, LIU D J. Preliminary report on field efficacy test of white blight of Pseudostellaria heterophylla. Agriculture & Technology, 2017, 37(18): 23. (in Chinese) [8] 陈凤毛. 真菌ITS区序列结构及其应用 [J]. 林业科技开发, 2007, 21(2):5−7. doi: 10.3969/j.issn.1000-8101.2007.02.002CHEN F M. The sequence of ITS region and its application of fungus [J]. China Forestry Science and Technology, 2007, 21(2): 5−7.(in Chinese) doi: 10.3969/j.issn.1000-8101.2007.02.002 [9] HARLTON C E. Genetic diversity in Sclerotium (Athelia) rolfsii and related species [J]. Phytopathology, 1996, 85(10): 1269. [10] 肖仲久, 李小霞, 蒋选利. 贵州省白术白绢病菌遗传多样性评价 [J]. 热带作物学报, 2012, 33(9):1686−1689. doi: 10.3969/j.issn.1000-2561.2012.09.029XIAO Z J, LI X X, JIANG X L. Genetic diversity analysis of Sclerotium rolfsii Sacc. in rhizoma of Atractylodes macrocephala [J]. Chinese Journal of Tropical Crops, 2012, 33(9): 1686−1689.(in Chinese) doi: 10.3969/j.issn.1000-2561.2012.09.029 [11] 郭艳春, 姚嘉瑜, 张镕斌, 等. 中国黄麻炭疽病病原菌的分离鉴定及系统发育分析 [J]. 作物学报, 2021(8):1−17.GUO Y C, YAO J Y, ZHANG R B, et al. Identification and phylogenetic analysis of the pathogen of jute anthracnose in China [J]. Acta Agronomica Sinica, 2021(8): 1−17.(in Chinese) [12] ALPER I, FRENETTE M, LABRIE S. Ribosomal DNA polymorphisms in the yeast Geotrichum candidum [J]. Fungal Biology, 2011, 115(12): 1259−1269. doi: 10.1016/j.funbio.2011.09.002 [13] CHEN Q, LI J, MIAO Y, et al. First report of southern blight on Chrysanthemum morifolium caused by Sclerotium rolfsii in china [J]. Plant Disease, 2019, 104(2): 310. [14] 陈婧, 桑维钧. 道真玄参白绢病病原鉴定及杀菌剂毒力测定 [J]. 湖北农业科学, 2015, 54(20):5020−5021,5024.CHEN J, SANG W J. Identification and fungicide toxicity determination of the Sclerotium rolfsii Sacc. on Scrophularia ningpoensis Hemsl. in Daozhen [J]. Hubei Agricultural Sciences, 2015, 54(20): 5020−5021,5024.(in Chinese) [15] 张佳星, 徐颖菲, 徐艳芳, 等. 白术两种主要土传病害的分离、鉴定及杀菌剂的室内活性筛选 [J]. 植物保护, 2017, 43(6):177−181,186. doi: 10.3969/j.issn.0529-1542.2017.06.031ZHANG J X, XU Y F, XU Y F, et al. Isolation and identification of two main soil-borne pathogens on Atractylodes macrocephala and screening of fungicides in vitro [J]. Plant Protection, 2017, 43(6): 177−181,186.(in Chinese) doi: 10.3969/j.issn.0529-1542.2017.06.031 [16] 王国荣, 沈伟东, 孙超, 等. 铁皮石斛茎基部2种主要病害病原菌的分离与鉴定 [J]. 植物保护, 2017, 43(1):168−172. doi: 10.3969/j.issn.0529-1542.2017.01.030WANG G R, SHEN W D, SUN C, et al. Isolation and identification of two main basal stem pathogens on Dendrobium officinale [J]. Plant Protection, 2017, 43(1): 168−172.(in Chinese) doi: 10.3969/j.issn.0529-1542.2017.01.030 [17] 崔月贞, 郝兴顺, 魏芳勤, 等. 秦巴山区魔芋白绢病病原菌的鉴定及生物学特性的测定 [J]. 江西农业学报, 2020, 32(4):103−107.CUI Y Z, HAO X S, WEI F Q, et al. Identification and biological characteristics of Hypochnus centrifugus in Qinba Mountains area [J]. Acta Agriculturae Jiangxi, 2020, 32(4): 103−107.(in Chinese) [18] 金苹, 高晓余. 白绢病的研究 [J]. 农业灾害研究, 2011, 1(1):14−22.JIN P, GAO X Y. Research and overview of southern blight [J]. Journal of Agricultural Catastrophology, 2011, 1(1): 14−22.(in Chinese) [19] NIE Q, ZHENG T, LIU L, et al. First report of southern blight on Pinellia ternata caused by Athelia rolfsii in china [J]. Plant Disease, 2020, 104(5): 11. [20] KAMIL D, BAHADUR A, DEBNATH P, et al. First report of leaf sheath rot of Areca nut (Arecacatechu) caused by Athelia rolfsii from tripura, india [J]. Plant Disease, 2020, 104(11): 1−2. [21] SEO Y, PARK M J, BACK C G, et al. First report of stem rot caused by Athelia rolfsii on Zamioculcas zamiifolia in korea [J]. Plant Disease, 2019, 103(11): 1−4. [22] LI J, CHEN Q, WANG M, et al. First report of southern blight on cynanchum stauntonii caused by Athelia rolfsii in china [J]. Plant Disease, 2020, 104(6): 1−2. [23] XU Z H, HARRINGTON T C, GLEASON M L, et al. Phylogenetic placement of plant pathogenic sclerotium species among teleomorph genera [J]. Mycologia, 2010, 102(2): 337−346. doi: 10.3852/08-189 [24] 刘思睿. 贵州省太子参主要根部病害病原鉴定及防治[D]. 贵阳: 贵州大学, 2020.LIU S R. Identification and control of main root diseases of Pseudostellariae in Guizhou province[D]. Guiyang: Guizhou University, 2020. (in Chinese) [25] 张金凤, 成波, 曹延丽, 等. 花生白绢病生防菌筛选及防控作用研究 [J]. 花生学报, 2019, 48(3):65−70.ZHANG J F, CHENG B, CAO Y L, et al. Screening and controlling effect of biocontrol bacteria against peanut white blight [J]. Journal of Peanut Science, 2019, 48(3): 65−70.(in Chinese) [26] 张献强, 黎起秦, 甘启范, 等. 防治茉莉白绢病的药剂筛选研究 [J]. 安徽农业科学, 2010, 38(10):5152−5154,5228. doi: 10.3969/j.issn.0517-6611.2010.10.077ZHANG X Q, LI Q Q, GAN Q F, et al. Screening fungicides for controlling jasmine Sclerotium rolfsii Blight [J]. Journal of Anhui Agricultural Sciences, 2010, 38(10): 5152−5154,5228.(in Chinese) doi: 10.3969/j.issn.0517-6611.2010.10.077 -
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