Isolation and Identification of Melon Black Spot in Gansu Province

doi:10.16420/j.issn.0513-353x.2020-0929

Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (2): 427-436.doi: 10.16420/j.issn.0513-353x.2020-0929

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Isolation and Identification of Melon Black Spot in Gansu Province

SHI Zhaorong¹, ZHANG Guangrong², SUN Shujun², JIN Mengjun¹, CUI Lingxiao¹, LIU Yanchao¹, WEI Lijuan¹, MEI Dahai¹, YANG Chengde¹^,^*()

¹College of Plant Protection,Gansu Agricultural University,Biocontrol Engineering Laboratory of Crop Diseases and Pests of Gansu Province,Lanzhou 730070,China
²The Agri-Tech Extension Service Center of Baiyin City,Baiyin,Gansu 730900,China

Received:2021-04-21 Revised:2021-08-27 Online:2022-02-25 Published:2022-02-28
Contact: YANG Chengde E-mail:yangcd@gsau.edu.cn

Abstract

Abstract:

This experiment was conducted to isolate,identify and determine the biological characteristics of black spot pathogen on melon in Baiyin City,Gansu Province. The results showed that the colony of isolate 98BYT2-1 was green-brown,round,conidia solitary or clustered,inverted rod-shaped or oval,with a vertical and horizontal septum. The size of conidia was（21.94-52.10）μm ×（7.01-13.93）μm,combining ITS,CAL and gpd gene sequence analysis showed that the strain 98BYT2-1 had 100% homology with Alternaria tenuissima and they were clustered together on the phylogenetic tree. Together,the isolate was identified as A. tenuissima. Starch,lactose and glucose were the most appropriate carbon sources for mycelia growth,spore formation and spore germination,respectively. Yeast extract and beef extract were the most suitable nitrogen sources for mycelia growth,but peptone was the most suitable source of nitrogen for sporulation and spore germination. In addition,PDA facilitated the mycelia growth of pathogen,and PCA was suitable for sporulation and spore germination. The optimum pH for mycelia growth and spore germination was 7 and the value for sporulation was 6. The optimum temperature for the growth and spore germination of mycelia was 30 ℃,and the sporulation was 15 ℃. The dark held up the growth of mycelium but benefited to sporulation and spore germination. The results obtained will provide a basis for the diagnosis and comprehensive control of melon black spot in Gansu Province.

Key words: muskmelon alternaria black spot, Alternaria tenuissima, identification, biological characteristics, determination

CLC Number:

S652

SHI Zhaorong, ZHANG Guangrong, SUN Shujun, JIN Mengjun, CUI Lingxiao, LIU Yanchao, WEI Lijuan, MEI Dahai, YANG Chengde. Isolation and Identification of Melon Black Spot in Gansu Province[J]. Acta Horticulturae Sinica, 2022, 49(2): 427-436.

Figures/Tables 11

References 26

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基因 Gene	引物序列（5′-3′） Primer sequence
ITS	ITS1：TCCGTAGGTGAACAYCCTGCG G	ITS4：TCCTCCGCTTATTGATATGC
CAL	CALDR1：CTTCTGCATCATCAYCTGGACG	CALDF1：AGCAAGTCTCCGAGTTCAAGG
gpd	gpd-1：CAACGGCTTCGGTCGCATTG	gpd-2：GCCAAGCAGTTGGTTGTGC

基因 Gene	引物序列（5′-3′） Primer sequence
ITS	ITS1：TCCGTAGGTGAACAYCCTGCG G	ITS4：TCCTCCGCTTATTGATATGC
CAL	CALDR1：CTTCTGCATCATCAYCTGGACG	CALDF1：AGCAAGTCTCCGAGTTCAAGG
gpd	gpd-1：CAACGGCTTCGGTCGCATTG	gpd-2：GCCAAGCAGTTGGTTGTGC

碳源 Carbon source	菌落直径/cm Colony diameter	产孢量/（× 10⁶ CFU · mL^-1） Sporulation	孢子萌发率/ % Spore germination	碳源 Carbon source	菌落直径/ cm Colony diameter	产孢量/（× 10⁶ CFU · mL^-1） Sporulation	孢子萌发率/ % Spore germination
木糖 Xylose	2.47 ± 0.02 e	13.17 ± 2.46 e	52.00 ± 3.06 de	葡萄糖 Glucose	2.20 ± 0.05 f	21.67 ± 0.44 d	90.33 ± 1.45 a
甘露醇 Mannite	2.53 ± 0.04 e	23.50 ± 0.76 d	69.33 ± 2.03 c	乳糖 Lactrose	4.13 ± 0.22 c	61.50 ± 0.50 a	82.33 ± 2.03 b
肌醇 Inosite	2.83 ± 0.07 d	24.67 ± 1.01 d	53.00 ± 4.93 de	淀粉 Starch	5.47 ± 0.03 a	56.50 ± 1.52 a	49.00 ± 1.52 c
麦芽糖 Maltose	4.43 ± 0.01 b	50.00 ± 0.76 b	58.67 ± 1.33 d	蔗糖 Sucrose	3.03 ± 0.01 d	43.33 ± 3.94 c	47.00 ± 0.58 e

碳源 Carbon source	菌落直径/cm Colony diameter	产孢量/（× 10⁶ CFU · mL^-1） Sporulation	孢子萌发率/ % Spore germination	碳源 Carbon source	菌落直径/ cm Colony diameter	产孢量/（× 10⁶ CFU · mL^-1） Sporulation	孢子萌发率/ % Spore germination
木糖 Xylose	2.47 ± 0.02 e	13.17 ± 2.46 e	52.00 ± 3.06 de	葡萄糖 Glucose	2.20 ± 0.05 f	21.67 ± 0.44 d	90.33 ± 1.45 a
甘露醇 Mannite	2.53 ± 0.04 e	23.50 ± 0.76 d	69.33 ± 2.03 c	乳糖 Lactrose	4.13 ± 0.22 c	61.50 ± 0.50 a	82.33 ± 2.03 b
肌醇 Inosite	2.83 ± 0.07 d	24.67 ± 1.01 d	53.00 ± 4.93 de	淀粉 Starch	5.47 ± 0.03 a	56.50 ± 1.52 a	49.00 ± 1.52 c
麦芽糖 Maltose	4.43 ± 0.01 b	50.00 ± 0.76 b	58.67 ± 1.33 d	蔗糖 Sucrose	3.03 ± 0.01 d	43.33 ± 3.94 c	47.00 ± 0.58 e

氮源 Nitrogen source	菌落直径/ cm Colony diameter	产孢量/（× 10⁶CFU · mL^-1） Sporulation	孢子萌发率/ % Spore germination
氯化铵 Ammonium Chloride	1.28 ± 0.06 f	6.67 ± 0.33 e	32.67 ± 1.76 d
蛋白胨 Peptone	5.77 ± 0.05 b	47.50 ± 2.36 a	92.33 ± 0.89 a
尿素 Carbamide	3.23 ± 0.13 d	37.83 ± 2.05 bc	56.33 ± 4.67 b
硫酸铵 Ammonium Sulfate	1.83 ± 0.12 e	0.13 ± 0.02 f	32.67 ± 1.76 d
硝酸钾 Potassium Nitrate	3.68 ± 0.11 c	38.17 ± 1.59 bc	52.67 ± 5.46 bc
酵母膏 Yeast extrac	6.08 ± 0.02 a	26.00 ± 0.76 d	90.33 ± 2.33 a
牛肉膏 Beef extract	6.17 ± 0.17 a	33.50 ± 0.00 c	51.33 ± 2.19 bc
硝酸钠 Sodium Nitrate	3.03 ± 0.01 d	43.33 ± 3.94 ab	47.00 ± 0.58 c

Isolation and Identification of Melon Black Spot in Gansu Province

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pH	菌落直径/cm Colony diameter	产孢量/（× 10⁶CFU · mL^-1） Sporulation	孢子萌发率/% Spore germination
4	3.80 ± 0.03 e	33.33 ± 0.73 d	69.67 ± 1.33 cd
5	5.58 ± 0.03 d	38.50 ± 0.87 cd	79.33 ± 2.40 b
6	5.67 ± 0.04 d	64.83 ± 1.20 a	79.67 ± 2.96 b
7	6.87 ± 0.04 a	54.33 ± 2.59 b	91.00 ± 1.73 a
8	6.50 ± 0.03 b	53.17 ± 3.03 b	72.67 ± 1.76 c
9	6.43 ± 0.09 b	41.33 ± 1.09 c	63.33 ± 2.33 d
10	6.21 ± 0.04 c	35.83 ± 3.91 cd	56.33 ± 2.02 e

温度/ ℃ Temperature	菌落直径/ cm Colony diameter	产孢量/（× 10⁶CFU · mL^-1） Sporulation	孢子萌发率/% Spore germination
5	0.58 ± 0.01 f	7.50 ± 0.29 e	29.33 ± 1.76 d
10	0.58 ± 0.02 f	3.33 ± 0.17 ef	16.33 ± 1.76 e
15	0.84 ± 0.07 e	88.17 ± 2.20 a	17.33 ± 0.33 e
20	2.93 ± 0.02 d	77.83 ± 1.97 b	45.00 ± 1.53 c
25	6.00 ± 0.09 b	41.00 ± 1.32 d	69.00 ± 1.73 b
30	6.36 ± 0.05 a	45.67 ± 1.01 c	90.33 ± 2.02 a
35	5.60 ± 0.03 c	74.33 ± 2.59 b	36.00 ± 5.03 d
40	0.70 ± 0.02 f	0 f	19.67 ± 2.02 e

光照 Light	菌落直径/ cm Colony diameter	产孢量/（×10⁶CFU · mL^-1） Sporulation	孢子萌发率/ % Spore germination
全光照 All light	6.50 ± 0.08 a	1.83 ± 0.17 c	21.00 ± 2.52 c
光照交替 Dark/ Light	6.50 ± 0.08 a	47.33 ± 1.76 b	80.00 ± 1.53 b
全黑暗 All dark	5.88 ± 0.02 b	139.83 ±1.48 a	91.67 ± 0.09 a

培养基 Media	菌落直径/ cm Colony diameter	产孢量/（× 10⁶CFU · mL^-1） Sporulation	孢子萌发率/% Spore germination
WA	2.92 ± 0.03 f	2.67 ± 1.67 g	80.67 ± 3.93 b
PDA	6.00 ± 0.09 a	41.00 ± 1.32 d	73.67 ± 1.45 bc
PSA	5.75 ± 0.02 b	48.67 ± 1.01 c	76.00 ± 5.03 b
MEA	5.31 ± 0.01 c	60.83 ± 0.60 b	65.00 ± 2.65 c
CA	5.02 ± 0.01 d	33.00 ± 1.53 e	91.00 ± 1.00 a
OMA	4.58 ± 0.09 e	14.83 ± 1.20 f	80.00 ± 3.61 b
PCA	4.92 ± 0.03 d	73.83 ± 2.24 a	91.67 ± 1.45 a

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