Identification,Biological Characteristics and Fungicide Sensitivity of the Pathogen Causing Brown Spot Disease on Forsythia

doi:10.16420/j.issn.0513-353x.2021-0335

Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (8): 1805-1814.doi: 10.16420/j.issn.0513-353x.2021-0335

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Identification,Biological Characteristics and Fungicide Sensitivity of the Pathogen Causing Brown Spot Disease on Forsythia

LIU Peng¹^,², LI Qin¹^,², ZHANG Weirui¹^,², HE Sheqi², ZHANG Suping², MA Xiaoxu², YUAN Wangjun¹^,²^,^*()

1. Institute of Traditional Chinese Medicine,Henan University,Kaifeng,Henan 475004,China
2. School of Pharmacy,Henan University,Kaifeng,Henan 475004,China

Received:2022-03-08 Revised:2022-05-30 Online:2022-08-25 Published:2022-09-05
Contact: YUAN Wangjun E-mail:10200068@vip.henu.edu.cn

Abstract

Abstract:

In order to clarify the species and biological characteristics of the pathogen of the brown spot of Forsythia suspensa（Thunb.）Vahl,the pathogen was isolated by tissue separation method and was tested for its pathogenicity following Koch’s postulation. Combing morphological feature with molecular identification of the ITS,LSU and SSU sequences,the pathogen was identified as Alternaria alternata. Growth response of the pathogen to different medium,light treatments,temperature,pH,carbon sources,and nitrogen sources were also studied. The toxicities of eight fungicides against the pathogen were tested using mycelial growth rate method. The results showed that the optimal conditions for mycelial growth were 25-30 ℃ at pH 7.0 with xylitol and mannitol as the carbon sources and peptone as the nitrogen source. The optimal sporulation conditions were 25 ℃ at pH 7.0 with xylitol as the carbon source and urea as nitrogen source. The optimal conditions for conidia germination were 15-35 ℃ at pH 5-9 with xylitol as the carbon source and beef extract powder as the nitro gen source. Among eight fungicides tested in toxicity assays,Cyprodinil（EC₅₀= 0.032 mg · L^-1）and Difenoconazole（EC₅₀= 0.054 mg · L^-1） had stronger inhibitory effect on A. alternata.

Key words: Forsythia suspensa, brown spot, Alternaria alternata, biological characteristics, pathogen, identification, fungicide screening.

CLC Number:

S685.24

LIU Peng, LI Qin, ZHANG Weirui, HE Sheqi, ZHANG Suping, MA Xiaoxu, YUAN Wangjun. Identification,Biological Characteristics and Fungicide Sensitivity of the Pathogen Causing Brown Spot Disease on Forsythia[J]. Acta Horticulturae Sinica, 2022, 49(8): 1805-1814.

Figures/Tables 7

References 23

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碳源 Carbon source	萌发率 Germination rate	氮源 Nitrogen source	萌发率 Germination rate
木糖醇 Xylitol	78.01 ± 5.25 a	氯化铵 Ammonium chloride	0 c
葡萄糖 Glucose	40.14 ± 4.28 d	草酸铵 Ammonium oxalate monohydrate	0 c
蔗糖 Sucrose	74.83 ± 5.38 ab	甘氨酸 Glycine	69.23 ± 6.51 a
山梨醇 Sorbitol	74.15 ± 5.59 ab	尿素 Urea	62.77 ± 3.41 a
乳糖 Lactose	56.40 ± 7.44 c	牛肉浸粉 Beef Powder	66.35 ± 5.94 a
甘露醇 Mannitol	74.14 ± 2.28 ab	硝酸钠 Sodium nitrate	62.44 ± 7.42 a
果糖 Fructose	68.19 ± 5.10 b	蛋白胨 Peptone bacteriological	43.80 ± 3.30 b
对照 Control	67.29 ± 4.38 b	对照 Control	67.29 ± 4.38 a

碳源 Carbon source	萌发率 Germination rate	氮源 Nitrogen source	萌发率 Germination rate
木糖醇 Xylitol	78.01 ± 5.25 a	氯化铵 Ammonium chloride	0 c
葡萄糖 Glucose	40.14 ± 4.28 d	草酸铵 Ammonium oxalate monohydrate	0 c
蔗糖 Sucrose	74.83 ± 5.38 ab	甘氨酸 Glycine	69.23 ± 6.51 a
山梨醇 Sorbitol	74.15 ± 5.59 ab	尿素 Urea	62.77 ± 3.41 a
乳糖 Lactose	56.40 ± 7.44 c	牛肉浸粉 Beef Powder	66.35 ± 5.94 a
甘露醇 Mannitol	74.14 ± 2.28 ab	硝酸钠 Sodium nitrate	62.44 ± 7.42 a
果糖 Fructose	68.19 ± 5.10 b	蛋白胨 Peptone bacteriological	43.80 ± 3.30 b
对照 Control	67.29 ± 4.38 b	对照 Control	67.29 ± 4.38 a

温度/℃ Tempearture	萌发率 Germination rate	pH	萌发率 Germination rate	光照条件 Photoperiod	萌发率 Germination rate
5	0 c	4	64.29 ± 4.15 b	黑暗 Dark	63.23 ± 5.60 a
10	22.45 ± 9.36 b	5	76.36 ± 7.01 a	半光 Half light	64.16 ± 2.21 a
15	62.22 ± 9.26 a	6	72.16 ± 4.36 a	全光 Full light	71.08 ± 6.57 a
20	61.32 ± 6.29 a	7	75.03 ± 1.25 a
25	67.29 ± 4.38 a	8	76.69 ± 8.03 a
30	55.48 ± 11.72 a	9	76.24 ± 4.09 a
35	58.47 ± 4.48 a	10	65.21 ± 2.63 b

温度/℃ Tempearture	萌发率 Germination rate	pH	萌发率 Germination rate	光照条件 Photoperiod	萌发率 Germination rate
5	0 c	4	64.29 ± 4.15 b	黑暗 Dark	63.23 ± 5.60 a
10	22.45 ± 9.36 b	5	76.36 ± 7.01 a	半光 Half light	64.16 ± 2.21 a
15	62.22 ± 9.26 a	6	72.16 ± 4.36 a	全光 Full light	71.08 ± 6.57 a
20	61.32 ± 6.29 a	7	75.03 ± 1.25 a
25	67.29 ± 4.38 a	8	76.69 ± 8.03 a
30	55.48 ± 11.72 a	9	76.24 ± 4.09 a
35	58.47 ± 4.48 a	10	65.21 ± 2.63 b

药剂 Fungicide	毒力回归方程 Regression equation	决定系数 R² Coefficient of determination	抑制中浓度/（mg · L^-1） EC₅₀
嘧菌环胺 Cyprodinil	y = 0.415x + 6.8651	0.9614	0.032
苯醚甲环唑 Difenoconazole	y = 0.2601x + 6.11	0.9513	0.054
吡唑醚菌酯 Pyraclostrobine	y = 0.2014x + 5.4204	0.9138	8.2
百菌清 Chlorothalonil	y = 0.2481x + 5.279	0.9435	75.1
春雷霉素 Kasugamycin	y = 1.8876x + 6.0587	0.9941	274.9
甲基硫菌灵 Thiophanate-methyl	y = 1.2704x + 4.1356	0.9629	4 790.9
枯草芽孢杆菌 Bacillus subtilis	y = -0.7751x + 6.0136	0.9499	20 309.6
嘧菌酯 Azoxystrobin	y = 0.4271x + 3.5136	0.9428	3 021 450

Identification,Biological Characteristics and Fungicide Sensitivity of the Pathogen Causing Brown Spot Disease on Forsythia

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