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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