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1、石河子大学 硕士学位论文 生物有机肥和拮抗细菌防治棉花黄萎病的生物效应研究 姓名：王飞 申请学位级别：硕士 专业：植物营养学 指导教师：李俊华 20100601 生物有机肥和拮抗细菌防治棉花黄萎病的生物效应研究 I 摘 要 本研究通过温度控制试验、含水量控制试验、盆栽模拟试验及自然环境下的病圃微区试验，研 究施加生物有机肥、棉花黄萎病拮抗细菌及拮抗细菌与有机肥混施后对棉花生长发育、土壤酶活性、 土壤微生物量碳氮及土壤微生物种群多样性的影响。主要结论如下： (1) 温度控制试验结果表明： 棉花黄萎病菌在 25 时生长繁殖最为旺盛， 在 15 下仍可缓慢生 长，但在 35 下表现为下降趋势。在本试
2、验中，由真菌、细菌数量的变化趋势可知：拮抗细菌在三 种温度条件下均对棉花黄萎病菌有较强的拮抗作用，且拮抗作用随温度的升高表现出增大的趋势。 (2) 含水量控制试验表明： 随土壤含水量的增大， 拮抗细菌对棉花黄萎病菌的拮抗作用逐渐减弱， 这主要是由于棉花黄萎病菌比较适于在高湿的条件下生长繁殖。 (3) 盆栽模拟试验结果表明：单施两种拮抗细菌(T3、T4)可以弥补棉花黄萎病菌对棉花植株生长 的抑制作用，肥料单施或有机肥与拮抗细菌混施(T5、T6、T7、T8)则对棉花生物量增加具有显著促 进作用。处理 T5、T6、T7、T8 促进了棉花对全 N、全 P、全 K 的吸收，其养分吸收量是接棉花黄 萎病菌
3、处理养分吸收量的 4.73、4.95、4.14 倍。处理 T3、T4 在前期可促进碱性磷酸酶、脲酶、蔗糖 酶及多酚氧化酶活性，后期对土壤蔗糖酶活性也有显著的促进作用。肥料混施和两种拮抗细菌与有 机肥混施后，显著增大了土壤中碱性磷酸酶、蛋白酶、脲酶(中后期)活性，同时试验发现人为向土壤 中加入有益微生物种群或植物病原菌菌群都可能导致过氧化氢酶活性的增大。对照的微生物量碳和 量氮均低于其他处理， 接棉花黄萎病菌的处理(T2)显著增大了土壤微生物量碳和量氮， 前期处理 T3、 T4 对量碳和量氮的促进作用显著，而处理 T5、T6、T7、T8 在后期促进作用显著。通过 PCR- DGGE 技术对土壤真
4、菌及细菌的群落多样性进行分析可知：与对照相比，其他处理的细菌种群的 Shannon- wiener 指数、均匀度和 Simpson 指数均有所增加，种群丰富度以处理 T2、T3、T4 较大；处 理 T3、T5、T6、T7、T8 的真菌种群的 Shannon- Wiener 指数较对照均增大，但各处理的均匀度指数 和 Simpson 指数无较大变化。 (4) 微区试验结果：生物有机肥和有机肥在蕾期、花期和铃期都能显著提高土壤蔗糖酶、多酚氧 化酶、碱性磷酸酶、蛋白酶和脲酶活性，在蕾期、花期也能显著提高土壤过氧化氢酶活性。三种拮 抗菌在蕾期、花期也能显著提高多酚氧化酶、碱性磷酸酶、蛋白酶、过氧化氢酶
5、活性。各处理对棉 花黄萎病的防病效果由强到弱为 S37 生物有机肥 S44 S228 有机肥，而各处理对棉花产量的影 响由高到低依次为生物有机肥 S44 有机肥 S37 S228，其每平方米产量分别比对照产量增加了 163.34 g、143.56 g、101.33 g、90.34 g、81.49 g。 关键词：关键词：棉花黄萎病，拮抗细菌，土壤酶，微生物量，微生物多样性，PCR- DGGE 生物有机肥和拮抗细菌防治棉花黄萎病的生物效应研究 II Abstract The temperature controlling, water content controlling, pots and p
6、lots experiments were carried out to study on biological effect of controlling cotton virticillium wilt with applied antagonistic bacillus and bio- organic fertilizer. Biomass of cotton, enzyme activities of soil catalase, alkaline phosphatase, sucrase, protease, urease and polyphenol oxidase, and s
7、oil microorganism diversity were measured. (1) Result of temperature controlling experiment showed that the optimum temperature for verticillium dahiae kleb(V.D) growth was 25.The V.D could grow slowly in 15, but be inhibited growth in 35. The antagonism bacterium numbers were increased with the tem
8、perature increase. The antagonism bacterium had strongly antagonistic effects on the V.D in three temperature treatments. (2) Results of the water content controlling experiment showed that the antagonistic effect of bacterium on inhibiting V.D decreased with the increase of the water content in soi
9、l. Main reason was that the V.D were suitable to live in high humidity environment. (3) Results of the pots experiment showed that using two antagonism bacterium(T3, T4) could improve cotton growth, biomass of cotton were higher than V.D control treatment. The treatments of antagonism bacterium mixe
10、d with the organic fertilizer(T5、 T6、 T7、 T8)improved cotton to absorb N, P, K, the absorptions of nutrition were 4.73,4.95,4.14 times compared with V.D. Treatment T3,T4 could improve the activities of alkaline phosphatase, urease, sucrase, and polyphenol oxidase under the same fertilizer level in t
11、he prophase, and the activity of sucrase was significantly improved in the anaphase. The activities of alkaline phosphatase, protease and urease significantly increased, after using the mixture of antagonism bacterium, organic fertilizer and bio- organic fertilizer. While different influences on soi
12、l enzyme activity of antagonism bacterium had different effects on different periods of duration. The microorganism biomass carbon and biomass nitrogen of CK were lower than other treatments, and the T2s were notably increased. Promotion to microorganism biomass carbon and biomass nitrogen of T3, T4
13、 were notable in the prophase while the T5, T6, T7, T8s were notable in the anaphase. The analysis of fungal and bacterium diversity using the PCR- DGGE showed that the Shannon- wiener index, uniformity, and Simpson index of other treatments were all increased compared with CK. The population richne
14、ss of T2, T3, T4 were higher than others. The fungal Shannon- Wiener index of T3, T5, T6, T7, T8 were higher compared with CK. But the uniformity and Simpson of all treatments nearly had no changes. (4) Results of the plots experiment showed that the activities of alkaline phosphatase, sucrase, prot
15、ease, urease and polyphenol oxidase could be increase during the bud stage, flower season and boll stage after using the bio- organic fertilizer and organic fertilizer. The activities of soil catalase also increased in the bud stage and flower season. The resistances to V.D of all treatments from hi
16、gher to lower were S37, bio- organic fertilizer, S44, S228, organic fertilizer. The effects of cotton productions from higher to lower were bio- organic fertilizer, S44, organic fertilizer, S37, S228. The unit productions were increased 163.34 g、143.56 g、101.33 g、90.34 g、81.49 g compared with CK. Key words：cotton virticillium wilt; antagonistic bacillus; soil enzyme; microbial biomass; PCR- DGGE 生物有机肥和拮抗细菌防治棉花黄萎病的生物效应
