Effects of chemical substances on melanin formation of Rhizoctonia solani AG-1 IA
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摘要:目的
水稻纹枯病是由立枯丝核菌Rhizoctonia solaniAG-1 IA融合群引起的真菌病害，本研究旨在明确外界培养条件与水稻纹枯病菌黑色素形成的关系。
方法采用菌丝生长速率法和紫外分光光度法，测定化学肥料[500 μg/mL K2SO4、NaH2PO4、CO(NH2)2]、金属离子(5 μg/mL CuSO 4、FeSO4、ZnSO4)、抗氧化剂(5 μg/mL槲皮素、桑色素、抗坏血酸)和对照(300 μg/mL莨菪碱、50 μg/mL儿茶酚、水)对水稻纹枯病菌菌丝生长速率、菌核数量、菌核鲜质量和干质量以及黑色素含量的影响。
结果500 μg/mL K2SO4、NaH2PO4和CO(NH2)2，5 μg/mL CuSO4和ZnSO4，5 μg/mL槲皮素、桑色素和抗坏血酸以及50 μg/mL儿茶酚溶液对水稻纹枯病菌黑色素的形成均有促进作用。其中，500 μg/mL的CO(NH2)2处理下，水稻纹枯病菌产生黑色素最多，为113.2 mg；而300 μg/mL的莨菪碱处理下，水稻纹枯病菌产生黑色素最少，为37.4 mg。
结论水稻纹枯病菌黑色素的产生与菌丝生长速率和菌核发育没有必然的联系，即抑制菌丝生长和菌核发育的化学物质，不一定能够抑制黑色素的形成。本研究结果可为水稻纹枯病防控机理的研究提供依据。
Abstract:ObjectiveRice sheath blight, caused by Rhizoctonia solaniAG-1 IA, is an important fungal disease. The purpose of this study was to determine the relationship between external culture conditions and melanin formation of R. solani AG-1 IA.
MethodThe effects of four different treatment groups, i. e. chemical fertilizer group [500 μg/mL K2SO4, NaH2PO4, CO(NH2)2], metal ion compound group(5 μg/mL CuSO4, FeSO4, ZnSO4), antioxidant group (5 μg/mL quercetin, morin, vitamin C) and control group (300 μg/mL hyoscyamine, 50 μg/mL catechol, water), on the mycelial growth rate, sclerotia number, sclerotial fresh weight and dry weight as well as melanin content of R. solani AG-1 IA were determined by measuring mycelial growth rate and ultraviolet spectrophotometry.
ResultChemical fertilizer [500 μg/mL K2SO4, NaH2PO4, CO(NH2)2], metal ion compound (5 μg/mL CuSO4, ZnSO4), antioxidant(5 μg/mL quercetin, morin, vitamin C) and catechol(50 μg/mL) could promote the formation of melanin. Under the treatment of 500 μg/mL CO(NH2)2, the melanin content of R. solani AG-1 IA was the highest (113.2 mg), while under the treatment of 300 μg/mL hyoscyamine, the melanin content of R. solani AG-1 IA was the lowest (37.4 mg).
ConclusionMelanin formation is not necessarily related to mycelial growth and sclerotial development, i.e. the chemical substances, which have inhibitory effect on mycelial growth and sclerotial development, might have no inhibition on melanin formation. The results of this study provide a basis for understanding the mechanism for prevention and control of rice sheath blight.
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Keywords:
- Rhizoctonia solani AG-1 IA /
- mycelium /
- sclerotium /
- melanin /
- chemical substance
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图 1 不同化学物质处理的水稻纹枯病菌菌核形态及分布
Figure 1. Morphology and distribution of sclerotia of Rhizoctonia solani AG-1 IA treated by different chemicals
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图 2 不同化学物质对水稻纹枯病菌菌核数量的影响
相同图标的柱子上的不同小写字母表示差异显著(P<0.05，Duncan’s法)
Figure 2. Effects of different chemicals on sclerotia numbers of Rhizoctonia solaniAG-1 IA
Different lowercase letters on pillars of the same icon indicate significant differences (P<0.05，Duncan’s test)
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图 3 水稻纹枯病菌黑色素的紫外−可见光吸收光谱
Figure 3. Ultraviolet-visible light absorption spectra of the melanin of Rhizoctonia solani AG-1 IA
表 1 本研究使用的化学物质及浓度
Table 1 Chemicals and concentrations used in this study
组别
Group化学物质 Chemical 母液 Stock solution 组分 Component ρ/(μg·mL−1) 溶剂 Solvent ρ/(mg·mL−1) 金属离子
Metal ion compoundCuSO4 5 ddH2O 50 ZnSO4 5 ddH2O 50 FeSO4 5 ddH2O 50 抗氧化剂
Antioxidant槲皮素 Quercetin 5 无水乙醇 Absolute ethanol 5 桑色素 Morin 5 无水乙醇 Absolute ethanol 5 抗坏血酸 Ascorbic acid 5 无水乙醇 Absolute ethanol 5 化学肥料
Chemical fertilizerCO(NH2)2 500 ddH2O 50 NaH2PO4 500 ddH2O 50 K2SO4 500 ddH2O 50 对照
Control莨菪碱 Hyoscyamine 300 无水乙醇 Absolute ethanol 150 儿茶酚 Catechol 50 无水乙醇 Absolute ethanol 100 水 Water 
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表 2 化学物质对水稻纹枯病菌菌丝生长速率和菌核质量 (m) 的影响1)
Table 2 Effects of chemicals on mycelial growth rate and sclerotial weight (m) of Rhizoctonia solani AG-1 IA
组别
Group化学物质
Chemicalρ(化学物质)/(μg·mL−1)
Chemical contentd(菌落)/mm
Colony diameterm鲜/g
Fresh weightm干/g
Dry weight化学肥料
Chemical fertilizerK2SO4 500 45.66±1.26b 0.487±0.041a 0.154±0.008a NaH2PO4 500 40.89±0.87c 0.473±0.008a 0.139±0.004a CO(NH2)2 500 53.83±1.20a 0.499±0.027a 0.128±0.015a 金属离子
Metal ion compoundCuSO4 5 50.11±1.57b 0.175±0.003b 0.068±0.002b ZnSO4 5 47.52±1.13c 0.152±0.009b 0.057±0.004b FeSO4 5 58.13±0.82a 0.212±0.006a 0.081±0.004a 抗氧化剂
Antioxidant槲皮素 Quercetin 5 43.55±1.19b 0.347±0.021b 0.117±0.006b 桑色素 Morin 5 37.71±1.28c 0.509±0.021a 0.149±0.003a 抗坏血酸 Ascorbic acid 5 50.04±0.79a 0.414±0.009b 0.133±0.007b 对照
Control莨菪碱 Hyoscyamine 300 52.75±0.47a 0.434±0.009a 0.138±0.004a 儿茶酚 Catechol 50 23.40±0.72c 0.401±0.023a 0.132±0.007a 水 Water 49.23±0.62b 0.500±0.047a 0.148±0.011a 1)相同组别、同列数据后的不同小写字母表示差异显著(P<0.05，Duncan’s法)
1)Different lowercase letters in the same column of the same group indicate significant differences (P<0.05，Duncan’s test)
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表 3 化学物质对水稻纹枯病菌黑色素产生的影响
Table 3 Effects of chemicals on the melanin formation of Rhizoctonia solani AG-1 IA
组别
Group化学物质
Chemicalρ/(μg·mL−1) m(黑色素)/mg
Melanin mass化学肥料
Chemical fertilizerK2SO4 500 65.1 NaH2PO4 500 85.5 CO(NH2)2 500 113.2 金属离子
Metal ion compoundCuSO4 5 65.7 ZnSO4 5 59.5 FeSO4 5 44.3 抗氧化剂
Antioxidant槲皮素 Quercetin 5 75.7 桑色素 Morin 5 91.1 抗坏血酸 Ascorbic acid 5 69.5 对照
Control莨菪碱 Hyoscyamine 300 37.4 儿茶酚 Catechol 50 55.7 水 Water 0 46.2
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