Chinese Journal OF Rice Science ›› 2022, Vol. 36 ›› Issue (6): 623-638.DOI: 10.16819/j.1001-7216.2022.211102
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MA Jingjing, PAN Yanyan, YANG Sunyuyue, WANG Jiaqi, JIANG Donghua()
Received:
2021-11-02
Revised:
2022-04-05
Online:
2022-11-10
Published:
2022-11-10
Contact:
JIANG Donghua
通讯作者:
蒋冬花
基金资助:
MA Jingjing, PAN Yanyan, YANG Sunyuyue, WANG Jiaqi, JIANG Donghua. Control Effect of St-79 (Streptomyces thioluteus) on Rice Bacterial Blight and Its Growth-promoting Effect[J]. Chinese Journal OF Rice Science, 2022, 36(6): 623-638.
马静静, 潘妍妍, 杨孙玉悦, 王嘉琦, 蒋冬花. 硫藤黄链霉菌St-79对水稻白叶枯病的防效和促生作用[J]. 中国水稻科学, 2022, 36(6): 623-638.
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URL: http://www.ricesci.cn/EN/10.16819/j.1001-7216.2022.211102
处理组 Treatment group | 处理方法 Treatment method |
---|---|
空白对照组(CK1) Blank control group(CK1) | 无菌剪刀45°剪叶,并套袋保持接种处湿润24 h。下同 Cut leaves with sterile scissors, and bag them to keep the inoculation site moist for for 24 h. The same below. |
Xoo处理对照组(CK2) Xoo treatment control group(CK2) | 无菌剪刀蘸取Xoo菌液后剪叶 Cut leaves with scissors following a dip into Xoo bacterial liquid. |
处理组1(T1) Treatment group 1(T1) | 按照CK2的处理方法接种Xoo病菌;6 h后用喷壶将稀释4倍的目标菌株发酵滤液均匀地喷洒在伤口处(剂量:10 mL/株),共喷洒3次(1 h喷1次)。下同 Inoculate Xoo bacteria according to the treatment method of CK2; 6 h later, use a watering can to evenly spray the 4-fold dilution fermentation liquid of the target strain on the wound (dose: 10 mL/plant) for a total of 3 sprays (once every hour). The same below. |
处理组2(T2) Treatment group 2(T2) | 按照CK2的处理方法接种Xoo病菌;6 h后用喷壶将稀释5倍的目标菌株发酵滤液均匀地喷洒在伤口处(剂量:10 mL/株) Inoculate Xoo bacteria according to the treatment method of CK2; 6 h later, use a watering can to evenly spray the 5-fold dilution fermentation liquid of the target strain on the wound (dose: 10 mL/plant). |
处理组3(T3) Treatment group 3(T3) | 按照CK2的处理方法接种Xoo病菌;6 h后用喷壶将稀释6倍的目标菌株发酵滤液均匀地喷洒在伤口处(剂量:10 mL/株) Inoculate Xoo bacteria according to the treatment method of CK2; 6 h later, use a watering can to evenly spray the 6-fold dilution fermentation liquid of the target strain on the wound (dose: 10 mL/plant). |
处理组4(T4) Treatment group 4(T4) | 无菌剪刀剪叶后立即喷洒稀释4倍的目标菌株发酵滤液(剂量:10 mL/株);6 h后按照CK2的处理方法接种Xoo病菌 Cut leaves with sterile scissors and spray the 4-fold dilution fermentation liquid of the target strain on the wound (dose: 10 mL/plant); 6 h post-spraying, inoculate Xoo bacteria according to the CK2 treatment method. |
Table 1. Experimental design of control effect.
处理组 Treatment group | 处理方法 Treatment method |
---|---|
空白对照组(CK1) Blank control group(CK1) | 无菌剪刀45°剪叶,并套袋保持接种处湿润24 h。下同 Cut leaves with sterile scissors, and bag them to keep the inoculation site moist for for 24 h. The same below. |
Xoo处理对照组(CK2) Xoo treatment control group(CK2) | 无菌剪刀蘸取Xoo菌液后剪叶 Cut leaves with scissors following a dip into Xoo bacterial liquid. |
处理组1(T1) Treatment group 1(T1) | 按照CK2的处理方法接种Xoo病菌;6 h后用喷壶将稀释4倍的目标菌株发酵滤液均匀地喷洒在伤口处(剂量:10 mL/株),共喷洒3次(1 h喷1次)。下同 Inoculate Xoo bacteria according to the treatment method of CK2; 6 h later, use a watering can to evenly spray the 4-fold dilution fermentation liquid of the target strain on the wound (dose: 10 mL/plant) for a total of 3 sprays (once every hour). The same below. |
处理组2(T2) Treatment group 2(T2) | 按照CK2的处理方法接种Xoo病菌;6 h后用喷壶将稀释5倍的目标菌株发酵滤液均匀地喷洒在伤口处(剂量:10 mL/株) Inoculate Xoo bacteria according to the treatment method of CK2; 6 h later, use a watering can to evenly spray the 5-fold dilution fermentation liquid of the target strain on the wound (dose: 10 mL/plant). |
处理组3(T3) Treatment group 3(T3) | 按照CK2的处理方法接种Xoo病菌;6 h后用喷壶将稀释6倍的目标菌株发酵滤液均匀地喷洒在伤口处(剂量:10 mL/株) Inoculate Xoo bacteria according to the treatment method of CK2; 6 h later, use a watering can to evenly spray the 6-fold dilution fermentation liquid of the target strain on the wound (dose: 10 mL/plant). |
处理组4(T4) Treatment group 4(T4) | 无菌剪刀剪叶后立即喷洒稀释4倍的目标菌株发酵滤液(剂量:10 mL/株);6 h后按照CK2的处理方法接种Xoo病菌 Cut leaves with sterile scissors and spray the 4-fold dilution fermentation liquid of the target strain on the wound (dose: 10 mL/plant); 6 h post-spraying, inoculate Xoo bacteria according to the CK2 treatment method. |
Fig. 2. Inhibition zone of fermentation liquid of the strain St-79 against Xoo. A, Control with Kohl’s 1 liquid; B, Treatment with fermentation liquid of strain St-79.
Fig. 3. Colony and microscopic observation of strain St-79. A and B, Colonies of strain St-79; C and D, Aerial mycelium and chains of spores under an optical microscope.
试验项目 Test item | 试验结果 Reaction | 试验项目 Test items | 试验结果 Reaction |
---|---|---|---|
淀粉水解Hydrolysis of starch | ++ | 硫化氢产生Hydrogen sulfide production | +++ |
纤维素水解Hydrolysis of cellulose | ++ | 黑色素产生Melanin production | − |
明胶液化Gelatin liquefaction test | +++ | 过氧化氢酶Catalase | +++ |
MR实验Methyl Red test | − | 脲酶Urease | − |
V-P实验Voges-Proskauer test | − | 脂肪酶Lipase | + |
Table 2. Physiological and biochemical test results of strain St-79.
试验项目 Test item | 试验结果 Reaction | 试验项目 Test items | 试验结果 Reaction |
---|---|---|---|
淀粉水解Hydrolysis of starch | ++ | 硫化氢产生Hydrogen sulfide production | +++ |
纤维素水解Hydrolysis of cellulose | ++ | 黑色素产生Melanin production | − |
明胶液化Gelatin liquefaction test | +++ | 过氧化氢酶Catalase | +++ |
MR实验Methyl Red test | − | 脲酶Urease | − |
V-P实验Voges-Proskauer test | − | 脂肪酶Lipase | + |
碳源种类 Type of carbon source | 试验结果 Test result | 氮源种类 Type of nitrogen source | 试验结果 Test result |
---|---|---|---|
葡萄糖Glucose | +++ | 蛋白胨Peptone | +++ |
乳糖Lactose | ++ | KNO3 | ++ |
麦芽糖Maltose | ++ | (NH4)2SO4 | +++ |
木糖Xylose | + | 谷氨酸Glutamate | ++ |
蔗糖Sucrose | ++ | 甲硫氨酸Methionine | + |
鼠李糖Rhamnose | ++ | 赖氨酸Lysine | +++ |
肌醇Inositol | ++ | 亮氨酸Leucine | + |
阿拉伯糖L-arabinose | ++ | 组氨酸Histidine | ++ |
甘露醇mannitol | +++ |
Table 3. Test results of utilization of carbon source and nitrogen source of strain St-79.
碳源种类 Type of carbon source | 试验结果 Test result | 氮源种类 Type of nitrogen source | 试验结果 Test result |
---|---|---|---|
葡萄糖Glucose | +++ | 蛋白胨Peptone | +++ |
乳糖Lactose | ++ | KNO3 | ++ |
麦芽糖Maltose | ++ | (NH4)2SO4 | +++ |
木糖Xylose | + | 谷氨酸Glutamate | ++ |
蔗糖Sucrose | ++ | 甲硫氨酸Methionine | + |
鼠李糖Rhamnose | ++ | 赖氨酸Lysine | +++ |
肌醇Inositol | ++ | 亮氨酸Leucine | + |
阿拉伯糖L-arabinose | ++ | 组氨酸Histidine | ++ |
甘露醇mannitol | +++ |
Fig. 5. Antibacterial effect of fermentation liquid of strain St-79 against four kinds of plant pathogenic bacteria. A, X. oryzae pv. oryzicola; B, X. campestris pv. campestris; C, X. axonopodis pv. glycines; D, P. syringae pv. tomato.
植物病原细菌 Phytopathogenic bacteria | 抑菌圈直径 Inhibition zone diameter /mm |
---|---|
水稻细菌性条斑病菌X. oryzae pv. oryzicola | 34.85±0.49 b |
油菜黑腐病菌X. campestris pv. campestris | 33.95±0.35 b |
大豆细菌性斑疹病菌X. axonopodis pv. glycines | 32.25±0.70 b |
番茄细菌性叶斑病菌P. syringae pv. tomato | 19.95±0.21 a |
Table 4. Inhibition zone of fermentation liquid from strain St-79 against four kinds of plant pathogenic bacteria.
植物病原细菌 Phytopathogenic bacteria | 抑菌圈直径 Inhibition zone diameter /mm |
---|---|
水稻细菌性条斑病菌X. oryzae pv. oryzicola | 34.85±0.49 b |
油菜黑腐病菌X. campestris pv. campestris | 33.95±0.35 b |
大豆细菌性斑疹病菌X. axonopodis pv. glycines | 32.25±0.70 b |
番茄细菌性叶斑病菌P. syringae pv. tomato | 19.95±0.21 a |
菌株St-79乙酸乙酯粗提物浓度 Concentration of crude extracts of strain St-79/(μg·mL−1) | 粗提物作用24 h后培养液浊度 Turbidity of culture medium after treatment with crude extracts for 24 h | 平板上菌体生长情况 Growth of bacteria on the plate |
---|---|---|
0 | 浑浊Muddy | 有菌生长With Xoo |
2 | 浑浊Muddy | 有菌生长With Xoo |
4 | 浑浊Muddy | 有菌生长With Xoo |
8 | 澄清Clarification | 有菌生长With Xoo |
16 | 澄清Clarification | 有菌生长With Xoo |
32 | 澄清Clarification | 无菌生长Without Xoo |
64 | 澄清Clarification | 无菌生长Without Xoo |
128 | 澄清Clarification | 无菌生长Without Xoo |
256 | 澄清Clarification | 无菌生长Without Xoo |
Table 5. MIC and MBC of crude extracts of target strain against Xoo.
菌株St-79乙酸乙酯粗提物浓度 Concentration of crude extracts of strain St-79/(μg·mL−1) | 粗提物作用24 h后培养液浊度 Turbidity of culture medium after treatment with crude extracts for 24 h | 平板上菌体生长情况 Growth of bacteria on the plate |
---|---|---|
0 | 浑浊Muddy | 有菌生长With Xoo |
2 | 浑浊Muddy | 有菌生长With Xoo |
4 | 浑浊Muddy | 有菌生长With Xoo |
8 | 澄清Clarification | 有菌生长With Xoo |
16 | 澄清Clarification | 有菌生长With Xoo |
32 | 澄清Clarification | 无菌生长Without Xoo |
64 | 澄清Clarification | 无菌生长Without Xoo |
128 | 澄清Clarification | 无菌生长Without Xoo |
256 | 澄清Clarification | 无菌生长Without Xoo |
Fig. 7. Xoo cells treated with crude extracts at different concentrations for 4 h under a scanning electron microscopy. A, Control; B, 4 μg/mL; C, 8 μg/mL; D, 16 μg/mL; E, 32 μg/mL.
Fig. 9. Hydrophobicity assay of Xoo treated with different concentrations of crude extracts for 4 h. Different letters indicate significant difference at the P < 0.05 level.
处理 Treatment | 处理后天数Days after treatment/d | ||
---|---|---|---|
4 | 6 | 8 | |
发酵滤液原液Crude fermentation liquid | 0.00±0.00 a | 0.00±0.00 a | 0.00±0.00 a |
高氏1号液体培养基原液 Crude Gauze’s medium No. 1 | 0.00±0.00 a | 0.00±0.00 a | 0.00±0.00 a |
发酵滤液10倍稀释液1/10 fermentation liquid | 25.33±3.06 b | 63.33±1.15 b | 72.67±1.15 c |
高氏1号液体培养基10倍稀释液1/10 Gauze’s medium No. 1 | 23.00±3.00 b | 59.33±1.15 b | 66.00±2.00 b |
发酵滤液50倍稀释液1/50 fermentation liquid | 32.00±3.46 cd | 72.00±2.00 c | 78.00±2.00 d |
高氏1号液体培养基50倍稀释液1/50 Gauze’s medium No. 1 | 28.67±3.06 bc | 70.67±1.15 c | 76.67±3.06 d |
发酵滤液100倍稀释液1/100 fermentation liquid | 53.33±3.06 e | 91.33±2.31 f | 96.67±1.15 g |
高氏1号液体培养基100倍稀释液1/100 Gauze’s medium No. 1 | 29.33±1.15 bc | 79.33±1.15 de | 87.00±1.00 f |
发酵滤液500倍稀释液1/500 fermentation liquid | 39.33±1.15 d | 82.67±2.31 e | 84.67±1.15 ef |
高氏1号液体培养基500倍稀释液1/500 Gauze’s medium No. 1 | 30.67±3.06 bc | 76.00±2.00 cd | 80.67±1.15 de |
无菌水H2O | 30.00±2.00 bc | 80.67±1.15 de | 88.67±1.15 f |
Table 6. Effect of fermentation liquid of St-79 on rice seed germination percentage. %
处理 Treatment | 处理后天数Days after treatment/d | ||
---|---|---|---|
4 | 6 | 8 | |
发酵滤液原液Crude fermentation liquid | 0.00±0.00 a | 0.00±0.00 a | 0.00±0.00 a |
高氏1号液体培养基原液 Crude Gauze’s medium No. 1 | 0.00±0.00 a | 0.00±0.00 a | 0.00±0.00 a |
发酵滤液10倍稀释液1/10 fermentation liquid | 25.33±3.06 b | 63.33±1.15 b | 72.67±1.15 c |
高氏1号液体培养基10倍稀释液1/10 Gauze’s medium No. 1 | 23.00±3.00 b | 59.33±1.15 b | 66.00±2.00 b |
发酵滤液50倍稀释液1/50 fermentation liquid | 32.00±3.46 cd | 72.00±2.00 c | 78.00±2.00 d |
高氏1号液体培养基50倍稀释液1/50 Gauze’s medium No. 1 | 28.67±3.06 bc | 70.67±1.15 c | 76.67±3.06 d |
发酵滤液100倍稀释液1/100 fermentation liquid | 53.33±3.06 e | 91.33±2.31 f | 96.67±1.15 g |
高氏1号液体培养基100倍稀释液1/100 Gauze’s medium No. 1 | 29.33±1.15 bc | 79.33±1.15 de | 87.00±1.00 f |
发酵滤液500倍稀释液1/500 fermentation liquid | 39.33±1.15 d | 82.67±2.31 e | 84.67±1.15 ef |
高氏1号液体培养基500倍稀释液1/500 Gauze’s medium No. 1 | 30.67±3.06 bc | 76.00±2.00 cd | 80.67±1.15 de |
无菌水H2O | 30.00±2.00 bc | 80.67±1.15 de | 88.67±1.15 f |
Fig. 10. Germination of rice seeds soaked in fermentation liquid for 8 days. Different letters indicate significant differences at the P < 0.05 level.
Fig. 11. Effect of fermentation liquid of strain St-79 on rice seed germination percentage in soil. FL100, FL500 indicate 1/100 and 1/500 fermentation liquid of strain St-79 respectively; KN100, KN500 indicate 1/100 and 1/500 Gauze’s medium No. 1 respectively. The same below. Different letters indicate significant differences at the P < 0.05 level.
Fig. 13. Effects of fermentation liquid of strain St-79 on plant height, root length and stem diameter of rice seedlings. A, Plant height; B, Root length; C, Stem diameter. Different letters indicate significant differences at the P < 0.05 level.
Fig. 14. Effects of fermentation liquid of strain St-79 on wet weight and dry weight of rice seedlings. A, Effects of fermentation liquid of strain St-79 on fresh weight, shoot fresh weight and root fresh weight of rice seedlings; B, Effects of fermentation liquid of strain St-79 on dry weight, shoot dry weight and root dry weight of rice seedlings. Different letters indicate significant differences at the P < 0.05 level.
Fig. 15. Lesion length of rice varieties under different treatments. A, Yongyou 1540; B, Yongyou 15; C, Nipponbare. CK1, Blank control; CK2, Control with inoculating Xoo only; T1, Inoculation with Xoo before treatment with 4-fold dilution fermentation liquid; T2, Inoculation with Xoo before treatment with 5-fold dilution fermentation liquid; T3, Inoculation with Xoo before treatment with 6-fold dilution fermentation liquid; T4, Treatment with 4-fold dilution fermentation liquid before inoculation with Xoo, the same blow.
Fig. 16. Lesion inhibition rate of rice varieties with different treatments For a variety, common letters indicate no significant difference at the P < 0.05 level.
处理 Treatment | 病情指数Disease index | 相对防效Relative control effect/% | |||||
---|---|---|---|---|---|---|---|
甬优1540 Yongyou 1540 | 甬优15 Yongyou 15 | 日本晴 Nipponbare | 甬优1540 Yongyou 1540 | 甬优15 Yongyou 15 | 日本晴 Nipponbare | ||
CK1 | 0 | 0 | 0 | / | / | / | |
CK2 | 95.66±0.29 e | 95.93±0.52 e | 97.41±0.52 e | 0 | 0 | 0 | |
T1 | 16.93±0.42 b | 22.02±0.55 b | 18.16±1.06 b | 82.31±0.29 c | 77.05±0.70 c | 81.35±1.19 c | |
T2 | 23.13±1.05 c | 27.40±1.59 c | 23.19±0.84 c | 75.84±0.86 b | 71.23±1.53 b | 76.20±1.00 b | |
T3 | 30.93±0.26 d | 32.76±0.24 d | 27.98±0.24 d | 67.67±0.55 a | 65.85±0.07 a | 71.28±0.09 a | |
T4 | 12.23±1.58 a | 15.00±0.79 a | 14.07±0.52 a | 87.23±1.53 d | 84.38±0.74 d | 85.56±0.62 d |
Table 7. Disease index and relative control effect on three rice varieties under different treatments.
处理 Treatment | 病情指数Disease index | 相对防效Relative control effect/% | |||||
---|---|---|---|---|---|---|---|
甬优1540 Yongyou 1540 | 甬优15 Yongyou 15 | 日本晴 Nipponbare | 甬优1540 Yongyou 1540 | 甬优15 Yongyou 15 | 日本晴 Nipponbare | ||
CK1 | 0 | 0 | 0 | / | / | / | |
CK2 | 95.66±0.29 e | 95.93±0.52 e | 97.41±0.52 e | 0 | 0 | 0 | |
T1 | 16.93±0.42 b | 22.02±0.55 b | 18.16±1.06 b | 82.31±0.29 c | 77.05±0.70 c | 81.35±1.19 c | |
T2 | 23.13±1.05 c | 27.40±1.59 c | 23.19±0.84 c | 75.84±0.86 b | 71.23±1.53 b | 76.20±1.00 b | |
T3 | 30.93±0.26 d | 32.76±0.24 d | 27.98±0.24 d | 67.67±0.55 a | 65.85±0.07 a | 71.28±0.09 a | |
T4 | 12.23±1.58 a | 15.00±0.79 a | 14.07±0.52 a | 87.23±1.53 d | 84.38±0.74 d | 85.56±0.62 d |
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