Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (2): 427-436.doi: 10.16420/j.issn.0513-353x.2020-0929
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SHI Zhaorong1, ZHANG Guangrong2, SUN Shujun2, JIN Mengjun1, CUI Lingxiao1, LIU Yanchao1, WEI Lijuan1, MEI Dahai1, YANG Chengde1,*()
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
CLC Number:
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.
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URL: https://www.ahs.ac.cn/EN/10.16420/j.issn.0513-353x.2020-0929
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 |
Table 1 Primer information
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 |
Fig. 4 Phylogenetic tree of pathogens based on ITS,CAL and gpd sequences The GenBank accession numbers for nucleotide sequence data are shown in the brackets;Scale is the genetic distance of evolutionary branches.
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 |
Table 2 The effect of carbon source on mycelium growth,spore production and 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 | 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/(× 106 CFU · 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 |
Table 3 The effect of nitrogen source on mycelium growth,spore production and germination
氮源 Nitrogen source | 菌落直径/ cm Colony diameter | 产孢量/(× 106 CFU · 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 |
pH | 菌落直径/cm Colony diameter | 产孢量/(× 106 CFU · 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 |
Table 4 The effect of pH on mycelium growth,spore production and germination
pH | 菌落直径/cm Colony diameter | 产孢量/(× 106 CFU · 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/(× 106 CFU · 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 |
Table 5 The effect of temperature on mycelium growth,spore production and germination
温度/ ℃ Temperature | 菌落直径/ cm Colony diameter | 产孢量/(× 106 CFU · 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/(×106 CFU · 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 |
Table 6 The effect of light on mycelium growth,spore production and germination
光照 Light | 菌落直径/ cm Colony diameter | 产孢量/(×106 CFU · 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/(× 106 CFU · 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 |
Table 7 The effect of media on mycelium growth,spore production and germination
培养基 Media | 菌落直径/ cm Colony diameter | 产孢量/(× 106 CFU · 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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