Abstract: To screen for efficient degrading functional microorganisms and construct a compound microbial agent suitable for the resource utilization of Polygonum cuspidatum residues，so as to solve the problems including difficult degradation of Polygonum cuspidatum residues in the natural environment and the resulting environmental pollution and resource waste，compost pile，soil from Polygonum cuspidatum planting fields，decomposed straw，decomposed Polygonum cuspidatum，and the laboratory strain collection were used as the sources for strain screening. Functional microorganisms with high cellulase and protease production were directionally screened by clear⁃zone tests，filter⁃paper disintegration tests，and residue degradation tests，and the enzyme activities of the strains were determined.The strains were identified by molecular biology techniques，and antagonism tests were conducted to evaluate the compatibility among the strains. Solid microbial agents were prepared from the selected strains by solid⁃state fermentation. The formulation of the microbial agent was optimized by the U₆*（66）uniform design method，and the maturation effect was evaluated based on composting trials，with the maximum composting temperature，duration of the thermophilic phase，seed germination index，and T value（final C/N ratio divided by initial C/N ratio） used as the evaluation indicators to determine the optimal formulation of the compound microbial agent. The strain screening results showed that three fungal strains（X1，X2，and TR） with high cellulase production，capable of efficiently degrading Polygonum cuspidatum residues and ten bacterial strains （D9，D11，D12，F5.1，F5.2，F5.3，F7.3，JD7.1，Z7.1，and BS）with high protease production were obtained. Among them，Trichoderma reesei TR and Bacillus subtilis BS were previously obtained and preserved in the laboratory strain collection.Molecular identification showed that X1 was Aspergillus niger，X2 was Trichoderma longibrachiatum，D9，F7.3，and JD7.1 were Bacillus amyloliquefaciens，D11 was Bacillus velezensis，F5.1，D12，and Z7.1 were Bacillus subtilis，and F5.2 and F5.3 were Priestia megaterium. Based on the enzyme activities and identification results，six superior strains （X1，X2，TR，D9，D11，and F5. 1） with strong functional complementarity and high enzyme activities were selected.The antagonism test results showed no obvious antagonism among the six strains，which could coexist well and meet the compatibility requirements of the compound microbial agent. The composting results showed that the optimal formulation of the compound microbial agent was TR∶X1∶X2∶D9∶D11∶F5. 1=5∶3∶1∶6∶4∶2. This formulation was superior to the other treatment groups in effectively increasing the peak composting temperature，prolonging the thermophilic phase，improving the seed germination index，promoting organic matter degradation，and decreasing the T value of the compost. The organic matter content，total nutrient content，moisture content，pH value，and seed germination index of the mature compost were 41.38%，6.95%，29.95%，8.12，and 93.35%，respectively. In conclusion，multiple efficient degrading strains for Polygonum cuspidatum residues were successfully screened，and a compound microbial agent with good compatibility and excellent degradation performance was constructed，with the optimal formulation of TR∶X1∶X2∶D9∶D11∶F5. 1=5∶3∶1∶6∶4∶2.

Key words: Polygonum cuspidatum residue, Compound microbial agent, Degradation, Compost maturity effect, Resource utilization, Composting

摘要： 筛选高效降解功能微生物、研制适合虎杖药渣资源化利用的复合微生物菌剂，以解决虎杖药渣在自然环境中难以降解及由此造成的环境污染和资源浪费等问题。以堆肥、虎杖种植田土壤、腐烂秸秆、腐烂虎杖及实验室菌种库为菌株筛选来源，通过透明圈试验、滤纸崩解试验和药渣降解试验定向筛选高产纤维素酶、蛋白酶的功能微生物，并测定菌株酶活力，采用分子生物学技术鉴定菌株种类，通过拮抗试验评估菌株间相容性，对优选菌株进行固态发酵制备固体菌剂。采用U₆^*（6⁶）均匀设计法对菌剂配比进行优化，基于堆肥试验，评价腐熟效果，以堆肥最高温度、嗜热期时长、种子发芽指数和T值（终点碳氮比/初始碳氮比）等指标为评价依据，筛选复合菌剂最优配比。菌株筛选结果显示，筛选得到3株可高效降解虎杖药渣、高产纤维素酶的真菌菌株（X1、X2和TR）和10株高产蛋白酶的细菌菌株（D9、D11、D12、F5.1、F5.2、F5.3、F7.3、JD7.1、Z7.1和BS）。其中，里氏木霉（Trichoderma reesei）TR和枯草芽孢杆菌（Bacillus subtilis）BS为前期获得并保藏于实验室菌种库的菌株。分子生物学鉴定结果表明，X1为黑曲霉（Aspergillus niger），X2为长枝木霉（Trichoderma longibrachiatum），D9、F7.3和JD7.1为解淀粉芽孢杆菌（Bacillus amyloliquefaciens），D11为贝莱斯芽孢杆菌（Bacillus velezensis），F5.1、D12和Z7.1为枯草芽孢杆菌（Bacillus subtilis），F5.2和F5.3为巨大普里斯特氏菌（Priestia megaterium）。综合酶活力与菌株鉴定结果，从中选出X1、X2、TR、D9、D11和F5.1共6株功能互补性强、酶活力高的优势菌株。拮抗试验结果表明，6株菌株间无明显拮抗作用，可以良好共生共存，满足复合菌剂配伍要求。堆肥试验结果表明，复合菌剂最优配比为TR∶X1∶X2∶D9∶D11∶F5.1=5∶3∶1∶6∶4∶2。该配比菌剂在有效提升堆肥峰值温度、延长嗜热期、提高种子发芽指数、促进有机物降解、降低堆肥T值等方面优于其他处理组，堆肥成品的有机质含量、总养分含量、含水率、pH值、种子发芽指数分别为41.38%、6.95%、29.95%、8.12、93.35%。综上，筛选获得多株虎杖药渣高效降解功能菌株，构建出相容性良好、降解效果优良的复合微生物菌剂，其最优配比为TR∶X1∶X2∶D9∶D11∶F5.1=5∶3∶1∶6∶4∶2。

关键词: 虎杖药渣, 复合微生物菌剂, 降解, 腐熟效果, 资源化利用, 堆肥