Chinese Journal OF Rice Science ›› 2023, Vol. 37 ›› Issue (2): 200-212.DOI: 10.16819/j.1001-7216.2023.220802
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WANG Xuandong, YU Junjie, GAO Runjie, LAN Heting, JIANG Yingzi, QI Wenjie, SONG Zhen, JIANG Donghua(
)
Received:2022-08-04
Revised:2022-09-05
Online:2023-03-10
Published:2023-03-10
Contact:
JIANG Donghua
王炫栋, 余俊杰, 高润杰, 兰赫婷, 江樱姿, 齐文杰, 宋振, 蒋冬花()
通讯作者:
蒋冬花
基金资助:WANG Xuandong, YU Junjie, GAO Runjie, LAN Heting, JIANG Yingzi, QI Wenjie, SONG Zhen, JIANG Donghua. Streptomyces zaomyceticus Sz-11, a Potential Biocontrol Agent with the Functions of Preventing Plant Diseases and Promoting Plant Growth[J]. Chinese Journal OF Rice Science, 2023, 37(2): 200-212.
王炫栋, 余俊杰, 高润杰, 兰赫婷, 江樱姿, 齐文杰, 宋振, 蒋冬花. 一株兼具防病促生功能的沙阿霉素链霉菌Sz-11[J]. 中国水稻科学, 2023, 37(2): 200-212.
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URL: http://www.ricesci.cn/EN/10.16819/j.1001-7216.2023.220802
| 菌株 Strain | 侵染植物 Infected plant | 抗性 Resistance |
|---|---|---|
| Xanthomonas oryzae pv. oryzicola, Xooc, RS105 | 水稻Oryza sativa L. | Rif |
| Xanthomonas oryzae pv. oryzae, Xoo, P6 | 水稻Oryza sativa L. | / |
| Pseudomonas syringae pv. tomato, Pst, DC3000 | 番茄Lycopersicon esculentum Miller | / |
| Pseudomonas syringae pv. glycinea, Psg, NJAU | 大豆Glycine max (Linn.) Merr. | Cb |
| Xanthomonas axonopodis pv. phaseoli, Xap, XS2 | 菜豆Phaseolus vulgaris Linn. | Cb |
| Xanthomonas axonopodis pv. glycines, Xag, NEAU001 | 大豆Glycine max (Linn.) Merr. | Cb/Amp |
| Ralstonia solanacearum, FR2 | 烟草Nicotiana tabacum L. | / |
| Gaeumannomyces graminis, MG201 | 小麦Triticum aestivum L. | / |
| Fusarium oxysporum f. sp. momordicae, Fom, FM04 | 苦瓜Momordica charantia L. | / |
| Fusarium oxysporum f. sp. cucumerinum, Foc, FC27 | 黄瓜Cucumis sativus L. | / |
| Fusarium pseudograminearum, FP68 | 小麦Triticum aestivum L. | / |
| Penicillium expansum, MP001 | 苹果Malus pumila Mill. | / |
| Alternaria solani, AS24 | 番茄Lycopersicon esculentum Miller | / |
| Alternaria alternata, AL08 | 烟草Nicotiana tabacum L. | / |
| Alternaria tomato, AT01 | 番茄Lycopersicon esculentum Miller | / |
| Colletotrichum orbiculare, WT204 | 西瓜Citrullus lanatus (Thunb.) Matsum. et Nakai | / |
| Gibberella zeae, GZ69 | 小麦Triticum aestivum L. | / |
| Fusarium solani, FS023 | 杨树Populus L. | / |
Table 1. Strains subjected to antibacterial spectrum test.
| 菌株 Strain | 侵染植物 Infected plant | 抗性 Resistance |
|---|---|---|
| Xanthomonas oryzae pv. oryzicola, Xooc, RS105 | 水稻Oryza sativa L. | Rif |
| Xanthomonas oryzae pv. oryzae, Xoo, P6 | 水稻Oryza sativa L. | / |
| Pseudomonas syringae pv. tomato, Pst, DC3000 | 番茄Lycopersicon esculentum Miller | / |
| Pseudomonas syringae pv. glycinea, Psg, NJAU | 大豆Glycine max (Linn.) Merr. | Cb |
| Xanthomonas axonopodis pv. phaseoli, Xap, XS2 | 菜豆Phaseolus vulgaris Linn. | Cb |
| Xanthomonas axonopodis pv. glycines, Xag, NEAU001 | 大豆Glycine max (Linn.) Merr. | Cb/Amp |
| Ralstonia solanacearum, FR2 | 烟草Nicotiana tabacum L. | / |
| Gaeumannomyces graminis, MG201 | 小麦Triticum aestivum L. | / |
| Fusarium oxysporum f. sp. momordicae, Fom, FM04 | 苦瓜Momordica charantia L. | / |
| Fusarium oxysporum f. sp. cucumerinum, Foc, FC27 | 黄瓜Cucumis sativus L. | / |
| Fusarium pseudograminearum, FP68 | 小麦Triticum aestivum L. | / |
| Penicillium expansum, MP001 | 苹果Malus pumila Mill. | / |
| Alternaria solani, AS24 | 番茄Lycopersicon esculentum Miller | / |
| Alternaria alternata, AL08 | 烟草Nicotiana tabacum L. | / |
| Alternaria tomato, AT01 | 番茄Lycopersicon esculentum Miller | / |
| Colletotrichum orbiculare, WT204 | 西瓜Citrullus lanatus (Thunb.) Matsum. et Nakai | / |
| Gibberella zeae, GZ69 | 小麦Triticum aestivum L. | / |
| Fusarium solani, FS023 | 杨树Populus L. | / |
| 处理组 Treatment groups | 处理方法 Treatment methods |
|---|---|
| 无菌水阴性对照组（CK1） Sterile water group, negative control (CK1) | 无菌剪刀45°剪叶后，喷雾无菌水，并套袋保持剪叶处湿润（24 h） Cut with sterile scissors at 45°followed by sterile water spraying and 20-h bagging to keep them moist by bagging |
| Xooc菌液阳性对照组（CK2） Xooc positive control group (CK2) | 无菌剪刀45°剪叶后，喷雾接种Xooc水悬液，并套袋保持接种部位湿润（24 h） Cut leaves with sterile scissors at 45° followed by Xooc water suspension spraying and 24-h bagging to keep the inoculation site moist |
| 发酵液治疗处理组（T1） Fermentation liquid treatment group (T1) | 无菌剪刀45°剪叶后，喷雾接种Xooc水悬液，套袋保持接种部位湿润；接种3 h后，在叶片正反两面均匀喷洒目标菌株发酵液（剂量：100 µL/cm2），套袋保持接种部位湿润（24 h） Cut leaves with sterile scissors at 45° followed by Xooc water suspension spraying and bagging to keep the inoculation site moist. The fermentation liquid of the target strain was evenly sprayed on both sides of the leaves at a dosage of 100 µL/cm2 3hours after inoculation, and the inoculation site was hugged to keep it moist for 24 hours |
| 发酵液预防处理组（T2） Fermentation liquid prevention treatment group (T2) | 先用喷壶在叶片正反两面均匀喷洒目标菌株发酵液（剂量：100 µL/cm2），套袋保持接种部位湿润；3 h后，无菌剪刀45°剪叶，喷雾接种Xooc水悬液，套袋保持接种部位湿润（24 h） Firstly, the fermentation liquid of the target strain was evenly sprayed on both sides of the leaves with a watering can at the dosage of 100 µL/cm2 3 hours after inoculation, and the inoculation site was bagged to keep it moist. Sterile scissors were used to cut leaves at 45°3 h later, Xooc water suspension was sprayed, and the inoculation site was bagged to make it moist for 24 hours |
| 发酵滤液治疗处理组（T3） Fermentation filtrate treatment group (T3) | 处理方法同T1，但将目标菌株发酵液替换成过滤后不含菌体的发酵滤液 The treatment method was the same as T1, but the fermentation liquid of the target strain was replaced with a fermentation filtrate without bacteria after filtration |
| 发酵滤液预防处理组（T4） Fermentation filtrate prevention treatment group (T4) | 处理方法同T2，但将目标菌株发酵液替换成过滤后不含菌体的发酵滤液 The treatment method was the same as T2, but the fermentation liquid of the target strain was replaced with a fermentation filtrate without bacteria after filtration |
Table 2. Treatment methods of rice bacterial leaf streak control effect test.
| 处理组 Treatment groups | 处理方法 Treatment methods |
|---|---|
| 无菌水阴性对照组（CK1） Sterile water group, negative control (CK1) | 无菌剪刀45°剪叶后，喷雾无菌水，并套袋保持剪叶处湿润（24 h） Cut with sterile scissors at 45°followed by sterile water spraying and 20-h bagging to keep them moist by bagging |
| Xooc菌液阳性对照组（CK2） Xooc positive control group (CK2) | 无菌剪刀45°剪叶后，喷雾接种Xooc水悬液，并套袋保持接种部位湿润（24 h） Cut leaves with sterile scissors at 45° followed by Xooc water suspension spraying and 24-h bagging to keep the inoculation site moist |
| 发酵液治疗处理组（T1） Fermentation liquid treatment group (T1) | 无菌剪刀45°剪叶后，喷雾接种Xooc水悬液，套袋保持接种部位湿润；接种3 h后，在叶片正反两面均匀喷洒目标菌株发酵液（剂量：100 µL/cm2），套袋保持接种部位湿润（24 h） Cut leaves with sterile scissors at 45° followed by Xooc water suspension spraying and bagging to keep the inoculation site moist. The fermentation liquid of the target strain was evenly sprayed on both sides of the leaves at a dosage of 100 µL/cm2 3hours after inoculation, and the inoculation site was hugged to keep it moist for 24 hours |
| 发酵液预防处理组（T2） Fermentation liquid prevention treatment group (T2) | 先用喷壶在叶片正反两面均匀喷洒目标菌株发酵液（剂量：100 µL/cm2），套袋保持接种部位湿润；3 h后，无菌剪刀45°剪叶，喷雾接种Xooc水悬液，套袋保持接种部位湿润（24 h） Firstly, the fermentation liquid of the target strain was evenly sprayed on both sides of the leaves with a watering can at the dosage of 100 µL/cm2 3 hours after inoculation, and the inoculation site was bagged to keep it moist. Sterile scissors were used to cut leaves at 45°3 h later, Xooc water suspension was sprayed, and the inoculation site was bagged to make it moist for 24 hours |
| 发酵滤液治疗处理组（T3） Fermentation filtrate treatment group (T3) | 处理方法同T1，但将目标菌株发酵液替换成过滤后不含菌体的发酵滤液 The treatment method was the same as T1, but the fermentation liquid of the target strain was replaced with a fermentation filtrate without bacteria after filtration |
| 发酵滤液预防处理组（T4） Fermentation filtrate prevention treatment group (T4) | 处理方法同T2，但将目标菌株发酵液替换成过滤后不含菌体的发酵滤液 The treatment method was the same as T2, but the fermentation liquid of the target strain was replaced with a fermentation filtrate without bacteria after filtration |
Fig. 1. Antibacterial spectrum of strain Sz-11. Different lowercase letters represent significant differences between treatments(P<0.05). 1?Gaeumannomyces graminis, MG201; 2?Fusarium oxysporum f. sp. momordicae, Fom, FM04; 3?Penicillium expansum, MP001; 4?Alternaria solani, AS24; 5?Colletotrichum orbiculare, WT204; 6?Alternaria alternata, AL08; 7?Alternaria tomato, AT01; 8?Fusarium pseudograminearum, FP68; 9?Fusarium oxysporum f. sp. cucumerinum, Foc, FC27.
Fig. 2. Results of rice growth promoting experiment by soaking seed with Sz-11 fermentation liquid. Different lowercase letters represent significant differences between the treatments at P<0.05.
Fig. 3. Results of rice growth promoting experiment by irrigating with Sz-11 fermentation liquid.
Fig. 4. Results of SDS-PAGE protein electrophoresis(A) and DNA agarose gel electrophoresis (B) of Xooc treated with Sz-11 fermentation filtrate. M, Marker; N, Gause 1 medium treatment; 1, 1/2, 1/4, 1/8, Different concentrations of fermentation filtrate treatment.
Fig. 5. Effects of Sz-11 fermentation filtrate treatment on cell membrane permeability and surface hydrophobicity of Xooc. A, CLSM figure; B, Xooc intracellular β-galactosidase leakage; C, Changes of hydrophobicity on Xooc cell surface. Different lowercase letters represent significant differences between the treatments at P<0.05.
Fig. 6. SEM results of Xooc cell morphology treated with Sz-11 fermentation filtrate at different concentrations.
Fig. 7. Infrared absorption spectra of Xooc cell treated with different concentrations of Sz-11 fermentation filtrate. Control, Gause 1 medium treatment; T1, 1/2 fermentation filtrate; T2, 1/4 fermentation filtrate.
| 振动峰分配 Definition of the spectral assignment | 分类 Classification | 振动峰 Peak variation/(cm-1) | ||
|---|---|---|---|---|
| 对照Control | T1 | T2 | ||
| 碳水化合物中的C-O, C-C str, C-O-H, C-O-C伸缩振动峰 C-O, C-C str, C-O-H, C-O-C def of carbohydrates | 糖原和核酸 Glycogen and nucleic acids | 1063.47 | 1063.62 | 1060.50 |
| 磷酸二酯>PO2中P=O的不对称伸缩振动峰 P=O str (asym) of>PO2 phosphodiesters | 主要是核酸 Mainly nucleic acids | 1244.96 | 1239.16 | 1236.03 |
| COO中C=O对称伸缩振动峰 C=O str (sym) of COO | 氨基酸侧链，脂肪酸 Amino acid side chains, fatty acids | 1393.72 | 1393.87 | 1393.75 |
| 酰胺II中蛋白质N-H弯曲振动峰和C-N拉伸振动峰 Amide II (protein N-H bend, C-N stretch) | α螺旋 α helices | 1533.55 | 1539.65 | 1539.50 |
| 酰胺I的β-折叠结构Amide I of β-pleated sheet structures | β-折叠 β-pleated sheet | 1640.66 | 1634.86 | 1634.71 |
| >CH2中C-H不对称伸缩振动峰 C-H str (asym) of>CH2 | 主要是脂类Mainly lipids | 2931.90 | 2926.08 | 2925.95 |
| 羟基O-H伸缩振动峰O-H str of hydroxyl groups | 多糖和蛋白质Polysaccharides, proteins | 3288.93 | 3283.10 | 3274.05 |
Table 3. Changes of vibrational peaks of Xooc cell treated with different concentrations of Sz-11 fermentation filtrate.
| 振动峰分配 Definition of the spectral assignment | 分类 Classification | 振动峰 Peak variation/(cm-1) | ||
|---|---|---|---|---|
| 对照Control | T1 | T2 | ||
| 碳水化合物中的C-O, C-C str, C-O-H, C-O-C伸缩振动峰 C-O, C-C str, C-O-H, C-O-C def of carbohydrates | 糖原和核酸 Glycogen and nucleic acids | 1063.47 | 1063.62 | 1060.50 |
| 磷酸二酯>PO2中P=O的不对称伸缩振动峰 P=O str (asym) of>PO2 phosphodiesters | 主要是核酸 Mainly nucleic acids | 1244.96 | 1239.16 | 1236.03 |
| COO中C=O对称伸缩振动峰 C=O str (sym) of COO | 氨基酸侧链，脂肪酸 Amino acid side chains, fatty acids | 1393.72 | 1393.87 | 1393.75 |
| 酰胺II中蛋白质N-H弯曲振动峰和C-N拉伸振动峰 Amide II (protein N-H bend, C-N stretch) | α螺旋 α helices | 1533.55 | 1539.65 | 1539.50 |
| 酰胺I的β-折叠结构Amide I of β-pleated sheet structures | β-折叠 β-pleated sheet | 1640.66 | 1634.86 | 1634.71 |
| >CH2中C-H不对称伸缩振动峰 C-H str (asym) of>CH2 | 主要是脂类Mainly lipids | 2931.90 | 2926.08 | 2925.95 |
| 羟基O-H伸缩振动峰O-H str of hydroxyl groups | 多糖和蛋白质Polysaccharides, proteins | 3288.93 | 3283.10 | 3274.05 |
Fig. 8. Inhibition rate and control effect of four rice varieties. CK1, Sterile water negative control; CK2, Xooc positive control; T1, Fermentation liquid cure treatment; T2, Fermentation liquid prevention treatment; T3, Fermentation filtrate cure treatment; T4, Fermentation filtrate prevention treatment. Different lowercase letters indicate significant difference at P<0.05.
| 处理 Treatment | 病情指数 Disease index | 相对防效 Relative control effect / % | ||||||
|---|---|---|---|---|---|---|---|---|
| 湘两优900 Xiangliangyou 900 | 甬优15 Yongyou 15 | 嘉丰优2号 Jiafengyou 2 | 甬优1540 Yongyou 1540 | 湘两优900 Xiangliangyou 900 | 甬优15 Yongyou 15 | 嘉丰优2号 Jiafengyou 2 | 甬优1540 Yongyou 1540 | |
| CK1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| CK2 | 60.00 e | 64.00 e | 48.67 d | 54.00 e | 0 | 0 | 0 | 0 |
| T1 | 29.33 b | 39.33 c | 18.67 b | 24.00 b | 51.11 c | 38.54 b | 61.65 b | 55.56 b |
| T2 | 19.33 a | 10.00 a | 10.00 a | 10.00 a | 67.78 d | 84.38 d | 79.45 c | 81.48 d |
| T3 | 50.00 d | 46.00 d | 36.00 c | 32.67 d | 16.67 a | 28.13 a | 26.03 a | 39.51 a |
| T4 | 40.00 c | 22.00 b | 18.67 b | 23.33 c | 33.33 b | 65.63 c | 61.65 b | 56.79 c |
Table 4. Disease index and relative control effect of four rice varieties.
| 处理 Treatment | 病情指数 Disease index | 相对防效 Relative control effect / % | ||||||
|---|---|---|---|---|---|---|---|---|
| 湘两优900 Xiangliangyou 900 | 甬优15 Yongyou 15 | 嘉丰优2号 Jiafengyou 2 | 甬优1540 Yongyou 1540 | 湘两优900 Xiangliangyou 900 | 甬优15 Yongyou 15 | 嘉丰优2号 Jiafengyou 2 | 甬优1540 Yongyou 1540 | |
| CK1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| CK2 | 60.00 e | 64.00 e | 48.67 d | 54.00 e | 0 | 0 | 0 | 0 |
| T1 | 29.33 b | 39.33 c | 18.67 b | 24.00 b | 51.11 c | 38.54 b | 61.65 b | 55.56 b |
| T2 | 19.33 a | 10.00 a | 10.00 a | 10.00 a | 67.78 d | 84.38 d | 79.45 c | 81.48 d |
| T3 | 50.00 d | 46.00 d | 36.00 c | 32.67 d | 16.67 a | 28.13 a | 26.03 a | 39.51 a |
| T4 | 40.00 c | 22.00 b | 18.67 b | 23.33 c | 33.33 b | 65.63 c | 61.65 b | 56.79 c |
Fig. 9. Antagonistic mechanism model of Sz-11 fermentation liquid against Xooc.
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