非洲哈茨木霉与贝莱斯芽胞杆菌联用对藜麦茎基腐病防效及根际微生态的影响

doi:10.13560/j.cnki.biotech.bull.1985.2025-1271

摘要/Abstract

摘要：

目的探究非洲哈茨木霉Trichoderma afroharzianum LMNS-M9可湿性粉剂与贝莱斯芽胞杆菌Bacillus velezensis YPTJ-1可湿性粉剂复配施用对藜麦茎基腐病的防治效果及其对根际微生物群落结构的调控作用。方法采用盆栽试验比较单一菌剂与复配菌剂对藜麦茎基腐病的防治效果；进一步通过大田试验评估不同浓度复配菌剂的防效，并结合高通量测序分析各处理对藜麦根际微生物群落的影响。结果复配菌剂对藜麦茎基腐病的盆栽防效达73.55%，显著优于2种单菌剂处理；田间试验中，复配菌剂稀释300倍时防效最佳，为67.92%，其防治效果在盆栽与田间条件下均与化学农药咯菌腈相当。2种菌剂联用显著提高了土壤中细菌和真菌的OTU数目及Alpha多样性指数，增加了放线菌门Actinomycetota、绿弯菌门Chloroflexota等有益菌群的相对丰度，并降低了致病菌的相对丰度。同时，复配处理增强了藜麦根际微生物网络结构的稳定性和复杂度。群落功能预测结果显示，复配处理下细菌氨基酸转运与代谢功能表达量与藜麦发病率呈负相关；真菌群落中共生型与内生菌等功能类群相对丰度显著上升，植物病原类群显著降低。结论复配菌剂可通过优化根际微生物群落结构有效抑制藜麦茎基腐病发生，为该病的绿色防控提供了理论依据与新途径。

关键词: 茄病镰孢菌, 非洲哈茨木霉, 贝莱斯芽胞杆菌, 微生物多样性

Abstract:

Objective This study evaluated the control efficacy of a wettable powder formulation containing Trichoderma afroharzianum LMNS-M9 combined with another formulation containing Bacillus velezensis YPTJ-1 against quinoa basal stem rot and its regulatory effects on the rhizosphere microbial community structure. Method We conducted pot experiments to compare the control efficacy of the single and combined microbial agents, and we evaluated the efficacy of the combined agent at different concentrations in field experiments. We then used high-throughput sequencing to analyze the effects of the different treatments on the quinoa rhizosphere microbial communities. Result The control efficacy of the combined microbial agent against quinoa basal stem rot in the pot experiment was 73.55%, which was significantly higher than that of either single agent. In the field experiment, the best control efficacy (67.92%) was achieved when the combined microbial agent was diluted 300-fold. The efficacy of this combined treatment was comparable to that of the chemical pesticide fludioxonil in both pot and field conditions. The combination of the two agents significantly increased the number of OTUs and the alpha diversity indices of both bacterial and fungal communities in the soil. It also significantly increased the relative abundance of beneficial taxa, such as Actinomycetota and Chloroflexota, while reducing the abundance of pathogenic fungi. Furthermore, the combined treatment enhanced the stability and complexity of the microbial network structure in the quinoa rhizosphere. Functional prediction of the microbial communities revealed that the expression of bacterial functions related to amino acid transport and metabolism under the combined treatment was negatively correlated with the incidence of quinoa disease. Within the fungal community, the relative abundance of functional groups such as symbiotrophic and endophytic fungi increased significantly, whereas plant pathogenic groups decreased significantly. Conclusion The combined microbial agent can effectively inhibit the occurrence of quinoa basal stem rot by optimizing the rhizosphere microbial community structure, providing a theoretical basis and a new strategy for the green control of this disease.

Key words: Fusarium solani, Trichoderma afroharzianum, Bacillus velezensis, microbial diversity

张文静, 马馨蕊, 徐紫璐, 吕红, 秦楠, 殷辉, 赵晓军, 任璐. 非洲哈茨木霉与贝莱斯芽胞杆菌联用对藜麦茎基腐病防效及根际微生态的影响[J]. 生物技术通报, 2026, 42(9): 327-341.

ZHANG Wen-jing, MA Xin-rui, XU Zi-lu, LYU Hong, QIN Nan, YIN Hui, ZHAO Xiao-jun, REN Lu. Effects of Combined Application of Trichoderma afroharzianum and Bacillus velezensis on Quinoa Basal Stem Rot and Rhizosphere Microecology[J]. Biotechnology Bulletin, 2026, 42(9): 327-341.

图/表 12

表1 不同处理对藜麦茎基腐病的田间防治效果

Table 1 Field control efficacy of different treatments against quinoa basal stem rot

处理 Treatment	发病率 Incidence （%）	防治效果 Control effect （%）
T1	27.92±1.10b	57.86±1.66c
T2	24.17±0.42c	63.52±0.63b
T3	21.25±0.72cd	67.92±1.09ab
T4	30.42±0.83b	54.09±1.26c
T5	19.17±1.82d	71.07±2.74a
T6	66.25±1.44a	-

图1 不同处理下藜麦茎基腐病的发病情况T1：YPTJ-1 WP 500倍稀释，T2：LMNS-M9 WP 500倍稀释，T3：复配菌剂WP 500倍稀释（YPTJ-1 WP：LMNS-M9 WP的比例为7∶3），T4：50%咯菌腈WP 5 000倍稀释（阳性化学药剂对照），T5：仅接种F. solani孢子液（阴性对照），T6：仅浇灌清水（空白对照），下同

Fig. 1 Disease occurrence of quinoa basal stem rot under different treatmentsT1: YPTJ-1 WP 500-fold dilution, T2: LMNS-M9 WP 500-fold dilution. T3: Mixed microbial agent WP 500-fold dilution (The ratio of YPTJ-1 WP to LMNS-M9 WP is 7∶3). T4: 50% fludioxonil WP 5 000-fold dilution (positive chemical control). T5: Only F. solani spore suspension (negative control). T6: Only watering water (blank control). The same below

图2 不同处理对藜麦茎基腐病的盆栽防效不同小写字母表示差异显著（P<0.05）。下同

Fig. 2 Pot culture control efficacy of different treatments on quinoa basal stem rotDifferent lowercase letters indicate significant difference (P<0.05), the same below

图3 不同处理下藜麦根际细菌（A）和真菌（B）的主坐标分析（PCoA）

Fig. 3 Principal coordinate analysis (PCoA) of bacteria (A) and fungi (B) in the quinoa rhizosphere under different treatments

图4 不同处理下藜麦根际土壤细菌（A）和真菌（B）在门水平上的相对丰度Other为相对丰度低于1%的类群，下同

Fig. 4 Relative abundance of bacteria (A) and fungi (B) in the rhizosphere soil of quinoa under different treatments at the phylum levelMicrobial groups with a relative abundance of less than 1% are classified as other, the same below

图5 不同处理下藜麦根际土壤细菌（A）和真菌（B）在属水平上的相对丰度

Fig. 5 Relative abundance of bacteria (A) and fungi (B) in the rhizosphere soil of quinoa under different treatments at the genus level

图6 藜麦根际土壤微生物群落差异物种进化分支图图中由内至外辐射的圆圈表示由属至门的分类级别，不同颜色节点表示在对应组别中显著富集，淡黄色节点表示对组间差异无显著影响的微生物类群

Fig. 6 Evolutionary branch diagram of differentially abundant microbial species in the quinoa rhizosphere soilThe circles radiated from the inside out in the figures represent the classification levels from genus to phylum. Different color nodes indicate significant enrichment in the corresponding groups, and light yellow nodes indicate microbial groups that have no significant effect on the differences between groups

图7 藜麦根际土壤中细菌（A）和真菌（B）的LEfSe分析

Fig. 7 LEfSe analysis of bacteria (A) and fungi (B) in quinoa rhizosphere soil

图8 不同处理下的土壤细菌共现网络分析CK1：空白对照，只浇灌清水；CK2：阴性对照，只接种茄病镰孢菌；D：菌剂对照，枯草芽胞杆菌处理；E：复配菌剂处理（稀释300倍），下同

Fig. 8 Co-occurrence network analysis of soil bacteria under different treatmentsCK1: blank control, water only; CK2: negative control, inoculated with F.solani only; D: microbial agent control, B. subtilis treatment; E: compound microbial agent treatment (diluted 300-fold). The same below

图9 不同处理下的土壤真菌共现网络分析

Fig. 9 Co-occurrence network analysis of soil fungi under different treatments

表2 不同处理下土壤细菌和真菌群落共现网络拓扑性质

Table 2 Topological properties of the co-occurrence network of soil bacterial and fungal communities under different treatments

微生物 Microorganism	处理 Treatment	节点数 Node number	边数 Edge number	平均度 Average degree	平均聚类系数 Average cluster coefficient	平均路径长度 Average path distance	模块化 Modularity
细菌 Bacteria	CK1	100	616	12.32	0.572	3.404	0.558
	CK2	100	1 579	31.58	0.768	2.183	0.384
	D	100	2 464	49.28	0.765	1.843	0.260
	E	100	1 923	38.46	0.764	1.914	0.375
真菌 Fungi	CK1	100	365	7.300	0.516	3.565	1.665
	CK2	97	407	8.392	0.521	3.499	0.710
	D	93	335	7.204	0.572	4.078	1.689
	E	97	430	8.866	0.589	3.790	0.624

图10 不同处理组的藜麦根际土壤细菌（A）和真菌（B、C）功能预测

Fig. 10 Functional prediction of bacteria (A) and fungi (B, C) in quinoa rhizosphere soil of different treatment groups

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