Identification of Root Rot Pathogens on Polygonatum cyrtonema in Fujian
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摘要:
目的 明确福建地区多花黄精根腐病的发病率和病原菌种类,并为该病害的防治提供理论依据。 方法 调查福建省3个多花黄精种植基地的根腐病发病率,并采集具有典型根腐病症状的植株和块根病样,分离纯化获得病原菌,利用形态学特征、分子生物学特征及致病性测定对其进行鉴定。 结果 福建省多花黄精种苗的根腐病平均发病率为10.50%,采收时根茎的根腐病平均发病率为17.65%。分离纯化获得98株菌株,结合菌株的形态学特征、特异性引物FOF1/FOR1和F8/R8、tef-1α 基因序列分析,分别鉴定为尖孢镰刀菌Fusarium oxysporum、腐皮镰刀菌F. solani和藤仓镰刀菌F. fujikuroi,三者的分离频率依次为75.51%、20.41%和4.08%。代表性菌株经回接根茎进行致病性检测发现其发病症状与田间表现一致,符合柯赫氏法则。 结论 尖孢镰刀菌、腐皮镰刀菌和藤仓镰刀菌是福建地区多花黄精根腐病的主要病原菌,首次报道藤仓镰刀菌可引起我国多花黄精根腐病。 Abstract:Objective Pathogens that caused the root rot disease on Polygonatum cyrtonema Hua in Fujian were identified. Methods Based on the survey of P. cyrtonema root rot disease in three planting areas in the province, infected plants and rhizomes were sampled to isolate the microbes. Suspected pathogens were examined morphologically and molecular biologically prior to a pathogenicity test for a confirmed identification. Results Approximately 10.50% the typical disease symptoms appeared on the seedlings, and 17.65% on the rhizomes at harvest. Based on the morphological characteristics, FOF1/FOR1 and F8/R8 PCR amplification of specific primers, and sequences of translation elongation factor-1α (tef-1α) gene, the pathogens that caused the infection were identified to include Fusarium oxysporum, F. solani,> and F. fujikuroi from 98 isolates. The frequency of isolating F. oxysporum from the collected samples was 75.51%, 20.41% on F. solani, and 4.08% on F. fujikuroi. Inoculating the isolated pathogens on healthy rhizomes reproduced the same symptoms as observed in the field and followed Koch’s rules. Conclusion F. oxysporum, F. solani, and F. fujikuroi were identified to be the pathogens of the root rot disease on P. cyrtonema in Fujian. F. fujikuroi was the first time to be associated with the outbreak in China. -
Key words:
- Polygonatum cyrtonema Hua /
- root rot disease /
- Fusarium oxysporum /
- Fusarium solani /
- Fusarium fujikuroi
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图 2 多花黄精根腐病病原菌的第I类菌株形态特征
A~B:菌落在PDA培养基上的形态;C:大型和小型分生孢子;D:厚垣孢子;E:产孢细胞。图3同。
Figure 2. Morphological characteristics for Class I pathogens causing root rot disease on P. cyrtonema
A–B: colony grown on potato dextrose agar; C: macroconidia and microconidia; D: chlamydospore; E: sporulation cells. Same for Fig. 3.
图 5 基于尖孢镰刀菌特异性引物FOF1/FOR1的代表性供试菌株的PCR扩增结果
M:DL2 000;1~20:分离菌株;21~23:分别为尖孢镰刀菌、腐皮镰刀菌和藤仓镰刀菌的阳性对照;24:无菌水阴性对照。图6同。
Figure 5. PCR amplification of DNA for representative tested pathogens using specific primer set FOF1/FOR1 of F. oxysporum
M: DL2 000; Lanes 1–20: tested strains; Lanes 21–23: positive control for F. oxysporum, F. solani, and F. fujikuroi, respectively; Lane 24: sterile water for negative control. Same for Fig. 6.
图 8 接种不同类型病原菌14d时多花黄精根茎的发病症状
A~B:尖孢镰刀菌FJAT-33155;C~D:腐皮镰刀菌FJAT-33165;E~F:藤仓镰刀菌FJAT-33320;G~H:尖孢镰刀菌西瓜专化型菌株FJAT-31362;I~J:无菌水对照。
Figure 8. Disease symptoms on P. cyrtonema rhizomes inoculated with isolated pathogens for 14 d
A–B: F. oxysporum FJAT-33155; C–D: F. solani FJAT-33165; E–F: F. fujikuroi FJAT-33320; G-–H: F. oxysporum f. sp. niveum FJAT-31362; I–J: sterile water (CK).
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