Identification of the pathogens causing root rot on Polygonatum cyrtonema Hua 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 多花黄精种苗的根腐病平均发病率约为10.50%，采收时根茎的根腐病平均发病率为17.65%。分离纯化获得到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，且符合柯赫氏法则。 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，其中藤仓镰刀菌可引起我国多花黄精根腐病为首次报道。 Abstract: :Objective The purpose of this paper is to clarify the root rot incidence rate of Polygonatum cyrtonema Hua in Fujian area and identify the causing pathogens, which will provide a theoretical basis for the control of root rot disease. Methods The incidence of root rot were investigate in the three planting areas of P. cyrtonema in Fujian province. The roots and rhizomes with typical root rot symptoms were collected from the fields. The pathogens were isolated and identified with morphology, molecular biology and pathogenicity test. Results The average incidence of root rot was about 10.50% in the seedlings phase and 17.65% in the rhizomes during harvesting phase. A total of 98 isolates were obtained from the diseased samples. The pathogens were finally identified as Fusarium oxysporum, F. solani and F. fujikuroi based on morphological characteristics, FOF1/FOR1 and F8/R8 PCR amplification of specific primer, and sequence analysis of translation elongation factor-1α (tef-1α) gene. The separation frequencies of the tree pathogens were 75.51%, 20.41% and 4.08%, respectively. Inoculating the pathogens on the healthy rhizomes reproduced the same symptoms previously observed in the field. Pathogenicities of the tree pathogens were confirmed by fulfilling the Koch’s postulates. Conclusion It was confirmed that F. oxysporum, F. solani and F. fujikuroi were the pathogens of root rot of P. cyrtonema in Fujian province. This is the first report of F. fujikuroi causing root rot of P. cyrtonema in China. -
Key words:
- Polygonatum cyrtonema Hua /
- Root rot /
- Fusarium oxysporum /
- Fusarium solani /
- Fusarium fujikuroi
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图 1 多花黄精的根腐病田间症状
A-D：种苗；E：生长期植株；F-I根茎；J-L：根茎切面。
Figure 1. Disease symptoms of root rot on Polygonatum cyrtonema in the field
A-D: the seedling plans; E: the growing stage plants; F-I: rhizomes; J-L: longitudinal section of rhizomes.
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。
Figure 2. Morphological characteristics for the class I of the pathogen causing root rot of Polygonatum cyrtonema
A-B: Colony grown on potato dextrose agar; C: macroconidia and microconidia; D: chlamydospore; E: sporulation cells.
图 3 多花黄精根腐病病原菌的第II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。
Figure 3. Morphological characteristics for the class II of the pathogen causing root rot of Polygonatum cyrtonema
A-B: Colony grown on potato dextrose agar; C: macroconidia and microconidia; D: chlamydospore; E: sporulation cells.
图 4 多花黄精根腐病病原菌的第III类菌株形态特征
A-B：菌落在PDA培养基上的形态；C：大型分生孢子；D：小型分生孢子；E：产孢细胞。
Figure 4. Morphological characteristics for the class III of the pathogen causing root rot of Polygonatum cyrtonema
A-B: Colony grown on potato dextrose agar; C: macroconidia; D: microconidia; E: sporulation cells.
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。
Figure 5. PCR amplification of DNA for the representative tested pathogen strains using specific primer set FOF1/FOR1 of Fusarium oxysporum
M: DL2 000; Lane 1-20: the tested strains; Lane 21-23: positive control for F. oxysporum，F. solani and F. fujikuroi; Lane 24: negative control for sterile water.
图 6 基于腐皮镰刀菌特异性引物F8/R8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。
Figure 6. PCR amplification of DNA for the representative tested pathogen strains using specific primer set F8/R8 of Fusarium solani
M: DL2 000; Lane 1-20：the tested strains; Lane 21-23: positive control for F. oxysporum，F. solani and F. fujikuroi; Lane 24: negative control for sterile water.
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. The disease symptoms on the rhizome of Polygonatum cyrtonema inoculated with the pathogens for 14d
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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