球毛壳菌ND35菌肥肥效试验及其生防机制的研究

发布时间：2018-03-14 07:50

本文选题：球毛壳菌ND35菌株　切入点：微生物肥料　出处：《山东农业大学》2015年硕士论文　论文类型：学位论文

【摘要】：化肥的长期使用导致耕地肥力下降,环境污染,微生物肥料的研究与推广越来越受到重视。球毛壳菌(Chaetomium globosum)ND35菌株是分离自健康毛白杨体内的一株优势内生真菌,其培养性状优良,生防潜力良好,既能够促进植株的生长发育、提高作物产量,又能防治多种植物病害。因此可以将其制备成微生物肥料,加以推广利用。本文对毛壳菌生物菌肥的固体发酵技术进行了优化、在大田条件下进行了不同作物的肥效试验、并研究了毛壳菌生物菌肥对大田黄瓜根际土壤酶活性及微生物数量的影响、还研究了球毛壳菌ND35菌株发酵液粗提物对杨树腐烂病菌抑菌机理,试验结果如下:在前期ND35菌肥培养方法的基础上,对其加以完善,选择最优的培养方案制备毛壳菌生物菌肥。研究发现菌肥营养液中PDB的含量以及菌肥培养过程中的透气性对菌肥的产孢量的影响较大。营养液中PDB与水的配比设置了3:2、4:1、2:3、1:4、1:1五个梯度,透气性设置了袋口系绳和棉塞加套环两个处理。结果表明:袋口系绳按1:4的营养配比制作的菌肥,产孢量达3.8亿/g。将新制备的毛壳菌生物菌肥施用在大田作物甘薯、马铃薯、玉米、黄瓜上研究其促生作用,测定各作物的生长指标及产量。结果表明:甘薯上,ND35菌肥处理的最长蔓长比对照增长了6.19%、茎粗比对照提高了9.24%、单株产量较对照增产16.12%;马铃薯上,ND35菌肥处理较对照组增产25.49%;玉米上,ND35菌肥处理的小区产量显著提高,较对照提高了34.23%;黄瓜上,育苗时施用菌肥比移栽时施用增产效果明显,育苗移栽均施用菌肥应注意用量不可过多。以黄瓜为例,施用菌肥后测定黄瓜各生育期根际土壤中土壤酶活性及土壤微生物的数量变化。结果表明:土壤脲酶、蔗糖酶、过氧化氢酶活性在施用菌肥后均出现不同程度的升高,在结实期达到最高值,于拉秧期降低。施用菌肥后根际土壤中细菌和放线菌数量较对照显著升高,各处理细菌数量于苗期、开花期达到最高值,放线菌数量于开花期、结实期达到最高值;而真菌数量只在开花期出现迅速升高,之后又迅速下降,各菌肥处理的真菌数量低于对照组。球毛壳菌ND35菌株能够防治多种植物病害的发生,以杨树腐烂病菌为例,研究ND35菌株的次生代谢物对病原菌生理指标及代谢途径关键酶活性的影响。结果表明:经粗提物处理后,病原菌菌丝体丙二醛含量较对照显著升高,且始终高于对照组;病原菌可溶性蛋白质含量自处理开始即迅速下降,并始终低于对照组,48 h时处理组蛋白质含量仅为对照组的59.1%,差异显著;粗提物处理组电导率值迅速升高,后期趋于平稳,显著高于对照组,24 h时两组电导率差值最大,处理组高出对照组49.1%;糖酵解途径中的己糖激酶(HK)、丙酮酸激酶(PK)、乳酸脱氢酶(LDH),三羧酸循环途径中的琥珀酸脱氢酶(SDH)、苹果酸脱氢酶(MDH)及能量代谢中的总ATP酶六种酶活性经粗提物处理后均显著下降,明显低于对照组活性,说明病原菌的代谢途径遭到破坏,菌体不能进行正常的生命活动。
[Abstract]:The long-term use of chemical fertilizer leads to the decline of farmland fertility, environmental pollution, research and promotion of more and more attention. The microbial fertilizer c.globosum (Chaetomium globosum) ND35 strain was isolated from healthy Populus tomentosa in vivo one strains of endophytic fungi, the cultivation of good character, good biocontrol potential, which can promote plant growth the development, increase crop yield, but also can control a variety of plant diseases. It can be prepared into microbial fertilizer, to promote the use of technology. In this paper, the solid fermentation of Chaetomium biological fertilizer were optimized, the fertilizer experiment of different crops under field conditions, and studied the Chaetomium biological fertilizer on influence of the number of soil microbial and enzyme activities in rhizosphere of cucumber field, also studied the fermentation broth of strain ND35 of chaetomiumglobosum extracts the antibacterial mechanism of poplar rot pathogen, the results are as follows: in the early stage of ND35 bacteria Fertilizer cultivation method on the basis of perfecting the preparation scheme of Chaetomium biological fertilizer culture to choose the best. The study found that the fertilizer in the nutrient solution and the content of PDB in the process of culture, influence of air bacterial sporulation of fertilizer of larger. Water and PDB in nutrient solution ratio of 3:2,4:1,2:3,1:4,1:1 is set to five a gradient, breathable set bag and cotton plug and ring two tether processing. The results show that the bag tether made by nutritional ratio of 1:4 fertilizer, sporulation capacity of 380 million /g. biological bacteria fertilizer prepared from Chaetomium in field crops, sweet potato, potato, corn, cucumber on its growth promoting effect determination, growth and yield of various crops. The results showed that the sweet potato, the longest vine ND35 fertilizer longer than control increased 6.19%, stem diameter increased by 9.24%, the yield increased by 16.12%; potato, ND35 bacteria Fertilizer treatment compared with the control group increased by 25.49%; maize, ND35 fertilizer treatments significantly increased yield, increased by 34.23%; cucumber seedlings, microbial fertilizer compared with the yield increasing effect was obvious when transplanting, seedling transplanting all microbial fertilizer should pay attention to consumption is not too much. A yellow melon for example, changes in the number of soil enzyme and the activity of soil microorganism in the rhizosphere soil of Cucumber in different growth stages after determination of microbial fertilizer. The results showed that soil urease, invertase and catalase activity were increased significantly in bacterial manure, during the grain filling period reached the highest value, to reduce the number of uprooting. Bacteria and actinomycetes in the rhizosphere soil of control significantly higher than that of microbial fertilizer, the number of bacteria in each treatment at seedling stage, flowering stage reached the highest value, the number of actinomycetes in the flowering stage, fruiting period reached the highest value; while the number of fungi only in the flowering period is increased rapidly after The number of fungi decreased rapidly, the fertilizer treatment is lower than the control group. C.globosum strain ND35 can control many plant diseases occur in poplar canker for example, study the effect of secondary metabolites of strain ND35 on key enzymes of pathogen physiological indices and metabolic pathways. The results showed that the crude extract after treatment the content of MDA, the pathogen hypha body increased significantly compared with the control, and is always higher than that of control group; soluble protein content from beginning pathogenic bacteria decreased rapidly, and is always lower than the control group, 48 h treatment group was only a protein content of 59.1% in the control group, the difference was significant; the crude extract treatment group conductivity increases, late stabilized, significantly higher than the control group, two groups of difference in conductivity of a maximum of 24 h, treatment group higher than control group 49.1%; the glycolytic pathway of hexokinase (HK), pyruvate kinase (PK), lactate dehydrogenase (LDH) Three, the citric acid cycle pathway of succinate dehydrogenase (SDH), malate dehydrogenase (MDH) and energy metabolism in total ATP enzyme activities of six enzymes in the crude extract after treatment decreased significantly, significantly lower than the control group activity, illustrate the metabolic pathways of pathogenic bacteria were damaged, bacteria can not normal life activities.

【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S144;S476.1

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