虽然不依赖培养的方法为植物相关微生物群的组成、多样性和功能潜力提供了有价值的见解，但依赖培养的方法对于研究单个物种的作用及其在群落中的相互作用的简化方法是必不可少的

A minimal cross-kingdom SynCom promotes plant growth and suppresses wheat crown rot via coordinated rhizosphere microbiome remodeling

摘要

（1）Wheat crown rot (WCR) caused by Fusarium pseudograminearum threatens wheat productivity, and sustainable control strategies are urgently needed. 由假禾谷镰孢菌引起的小麦冠腐病(WCR)威胁着小麦的生产力，迫切需要可持续的控制策略。

（2）We constructed a minimal cross-kingdom synthetic community (SynCom) consisting of Trichoderma harzianum T19 and Bacillus rugosus PM16, and evaluated its effects on wheat growth and WCR suppression. Rhizosphere microbiome assembly (full-length 16S rRNA/ITS sequencing) and metabolomic shifts were assessed to elucidate mechanisms. 我们构建了由哈茨木霉T19和皱纹芽孢杆菌PM16组成的最小跨王国合成群落(SynCom ),并评价了其对小麦生长和WCR抑制的影响。根际微生物群组装(全长16S rRNA/ITS测序)和代谢组学变化被评估以阐明机制。

（3）The SynCom significantly suppressed WCR and promoted wheat growth under pathogen pressure, improving biomass, chlorophyll content, and yield-related traits. SynCom显著抑制WCR并促进小麦在病原体压力下的生长，改善生物量、叶绿素含量和产量相关性状。  

（4）SynCom inoculation remodeled the rhizosphere microbiome by enriching beneficial taxa (e.g., Mortierella) and reducing pathogen-associated fungi, and it enhanced rhizosphere enzyme activities and nutrient availability. SynCom接种通过富集有益类群(如被孢霉)和减少病原相关真菌重塑了根际微生物群，并增强了根际酶活性和养分有效性。  

（5）Metabolomics revealed accumulation of growth-promoting and defense-related metabolites, supporting coordinated microbiome–metabolome regulation. 代谢组学揭示了促进生长和防御相关代谢物的积累，支持协调的微生物组-代谢组调节。

（6） A minimal cross-kingdom SynCom can establish a disease-suppressive and growth-promoting rhizosphere through coordinated restructuring of microbial communities and metabolites, highlighting its potential as an eco-friendly strategy for WCR management. 一个最小的跨王国SynCom可以通过微生物群落和代谢物的协调重组建立一个抑制病害和促进生长的根际，突出了其作为WCR管理的生态友好策略的潜力。

（7）结果

（8）Selection and identification of core strains in the minimal SynCom 最小同步体中核心菌株的筛选和鉴定

Fig 1 拮抗真菌T19与促生细菌PM16的鉴定及互作机制分析

（A）T19的菌落形态及分生孢子显微结构 （B）PM16的菌落形态及芽孢染色 （C）T19与PM16的相容性测定 （D）T19的系统发育树 （E）PM16的系统发育树 （F）由T19和PM16构建的跨界简易合成菌群TB在小麦中的作用机制模型

（9）Effects of the minimal SynCom TB on wheat growth and disease resistance 微量同步结核对小麦生长和抗病性的影响

Disease severity index 疾病严重指数 Fp为接种病原菌

（10） Regulation of antioxidant and soil enzyme activities by TB 结核杆菌对抗氧化剂和土壤酶活性的调节

（11） TB improves rhizosphere soil physicochemical properties and nutrient availability TB改善根际土壤理化性质和养分有效性

（12） SynComs significantly altered the microbial diversity and community composition in the rhizosphere soil SynComs显著改变了根际土壤的微生物多样性和群落组成

（13） SynComs altered the structure of the microbial community in the rhizosphere soil as well as the composition of core species SynComs改变了根际土壤中微生物群落的结构以及核心物种的组成

TB组(SynCom生长促进组)显示出更强的微生物多样性，特别是有益微生物如被孢霉和Sonoraphyctis的增加。这表明SynComs的引入有效地促进了植物生长并改善了根际土壤的微生物结构。

（14） Microbial co-occurrence network analysis 微生物共现网络分析

（15） TB treatment altered and enhanced the composition of rhizosphere soil metabolites, leading to the accumulation of growth-promoting and stress-responsive compounds TB处理改变和增强了根际土壤代谢物的组成，导致生长促进和胁迫响应化合物的积累

（16） Integrative analysis reveals nutrient–enzyme–microbiome pathways underpinning TB-induced disease resistance and yield benefits 综合分析揭示了营养-酶-微生物群途径，这些途径支撑着结核诱导的疾病抗性和产量效益

（17）方法

  将木霉菌株T19的分生孢子加入到羧甲基纤维素液体培养基中，并在28℃下培养5天。然后用无菌水将孢子悬浮液调节至最终浓度为106孢子/mL。菌株PM16在37℃的营养肉汤中培养8小时，将细胞密度调节至OD600 = 0.1，然后再悬浮于无菌水中。将等体积(v/v)的T19孢子悬浮液和PM16细菌悬浮液混合，以构建两个成员的最小跨王国合成群落(SynCom，TB)

We constructed a minimal cross-kingdom synthetic community (SynCom) consisting of Trichoderma harzianum T19 and Bacillus rugosus PM16, and evaluated its effects on wheat growth and WCR suppression. Rhizosphere microbiome assembly (full-length 16S rRNA/ITS sequencing) and metabolomic shifts were assessed to elucidate mechanisms.