• ISSN 1008-505X
  • CN 11-3996/S
张嘉慧, 邢佳佳, 彭丽媛, 邬奇峰, 陈俊辉, 徐秋芳, 秦华. 丛枝菌根真菌提高感染青枯菌番茄根际土壤细菌群落多样性和稳定性及有益菌属相对丰度[J]. 植物营养与肥料学报, 2023, 29(1): 120-131. DOI: 10.11674/zwyf.2022260
引用本文: 张嘉慧, 邢佳佳, 彭丽媛, 邬奇峰, 陈俊辉, 徐秋芳, 秦华. 丛枝菌根真菌提高感染青枯菌番茄根际土壤细菌群落多样性和稳定性及有益菌属相对丰度[J]. 植物营养与肥料学报, 2023, 29(1): 120-131. DOI: 10.11674/zwyf.2022260
ZHANG Jia-hui, XING Jia-jia, PENG Li-yuan, WU Qi-feng, CHEN Jun-hui, XU Qiu-fang, QIN Hua. Arbuscular mycorrhizal fungi improves diversity and stability of bacterial community and abundance of beneficial bacteria genus in the rhizosphere of tomato infected with Ralstonia solanacearum[J]. Journal of Plant Nutrition and Fertilizers, 2023, 29(1): 120-131. DOI: 10.11674/zwyf.2022260
Citation: ZHANG Jia-hui, XING Jia-jia, PENG Li-yuan, WU Qi-feng, CHEN Jun-hui, XU Qiu-fang, QIN Hua. Arbuscular mycorrhizal fungi improves diversity and stability of bacterial community and abundance of beneficial bacteria genus in the rhizosphere of tomato infected with Ralstonia solanacearum[J]. Journal of Plant Nutrition and Fertilizers, 2023, 29(1): 120-131. DOI: 10.11674/zwyf.2022260

丛枝菌根真菌提高感染青枯菌番茄根际土壤细菌群落多样性和稳定性及有益菌属相对丰度

Arbuscular mycorrhizal fungi improves diversity and stability of bacterial community and abundance of beneficial bacteria genus in the rhizosphere of tomato infected with Ralstonia solanacearum

  • 摘要:
    目的 青枯病是由茄科雷尔氏菌 (Ralstonia solanacearum, 亦称青枯菌) 诱导产生的一种高温高湿型土传病害,土壤温度高、湿度大时易于青枯菌的繁殖进而引发青枯病。丛枝菌根真菌 (arbuscular mycorrhiza, AM) 可能通过调控根际微生物区系对病原体产生影响,我们研究了AM真菌对青枯菌入侵条件下土壤细菌群落的影响。
    方法 以番茄 (Solanum lycopersicum) 为试材进行盆栽试验,供试AM真菌为摩西管柄囊霉 (Funneliformis mosseae) M47V,供试病原菌为茄科雷尔氏菌QL-RS 1115 (GenBank:GU390462)。催芽5日的番茄种子,接种AM菌剂的为菌根苗,未接种AM真菌的为非菌根苗。在番茄幼苗生长30天时,一半菌根苗和非菌根苗接种青枯菌,另一半不接种青枯菌,共4个处理。在接种青枯菌后1天和14天,采集番茄样品,采用抖土方法采集根际土壤,利用实时荧光PCR分析番茄根际青枯菌数量,采用16S rRNA高通量测序探究土壤细菌群落多样性和结构稳定性。
    结果 在接种青枯菌初期 (1天),非菌根苗接种青枯菌 (TR–AMF) 和菌根苗接种青枯菌 (TR+AMF) 两组处理的根际土壤细菌群落结构发生明显改变,Chao1指数、Shannon指数和Simpson指数显著降低 (P<0.05),共现网络的节点数和连接数明显减少,模块化程度降低,共现网络简化表明细菌群落结构的稳定性降低。接种青枯菌14天后,不动杆菌属 (Acinetobacter)、鞘氨醇单胞菌属 (Sphingomonas)、溶杆菌属 (Lysobacter)、假单胞菌属 (Pseudomonas) 等有益细菌属在感染青枯菌的番茄根际富集,细菌共现网络的节点数和连接数增加,模块化程度提高,表明细菌群落稳定性得到恢复。与非菌根苗相比,菌根苗接种青枯菌 (TR+AMF) 和菌根苗未接种青枯菌 (TN+AMF) 两个处理番茄根际土壤中青枯菌丰度显著降低 (P<0.05)。AM真菌显著提高Chao1指数和Shannon指数 (P<0.05),提高了感染青枯菌番茄根际土壤中黄杆菌属(Flavobacterium)、黄色土源菌属 (Flavisolibacter)、噬胞菌属 (Cytophaga) 和苔藓杆菌属 (Bryobacter) 的相对丰度,同时增加了共现网络的节点数和连接数,并促进番茄根际细菌物种之间的良性互作,提高细菌网络的复杂程度。
    结论 感染青枯菌的番茄根际会富集不动杆菌属 (Acinetobacter)、鞘氨醇单胞菌属 (Sphingomonas)、溶杆菌属 (Lysobacter)、假单胞菌属 (Pseudomonas) 等有益菌属以提高其抗病性,恢复细菌多样性和群落稳定性。接种AM真菌可显著降低番茄根际土壤中青枯菌的丰度,特别是侵染青枯菌后提高番茄根际的黄杆菌属 (Flavobacterium)、黄色土源菌属 (Flavisolibacter) 、噬胞菌属 (Cytophaga) 和苔藓杆菌属 (Bryobacter)的相对丰度,进而抑制土壤中青枯菌的生长,并通过提高细菌的多样性和丰富度,促进番茄根际细菌物种之间的稳定共生和良性互作,从而提高细菌群落对青枯菌的抵抗能力。

     

    Abstract:
    Objectives Bacterial wilt is a soil-borne disease with high temperature and humidity, which is caused by Ralstonia solanacearum. Arbuscular mycorrhiza (AM) fungi have the potential to inhibit specific pathogens in soil by regulating the microbial community of the host rhizosphere. However, the effects of AM fungi on soil bacterial community infected by Ralstonia solanacearum is still unclear.
    Methods Pot culture method was used in the research, the test plant was tomato (Solanum lycopersicum), the AM fungus was F. mosseae M47V, and the pathogen was Ralstonia solanacearum QL-RS 1115 (GenBank: GU390462). Mycorrhizal seedlings were prepared by inoculating tomato seeds with F. mosseae M47V after 5 days of germination, and non-mycorrhizal seedlings without AM fungus inoculation. Half of the mycorrhizal and non-mycorrhizal tomato seedlings of 30-days-old were inoculated with pathogen, while the other half consisted of only AM treatments. At 1 and 14 days after inoculation, tomato plants and rhizosphere soil were sampled. Real-time PCR was used to analyze the number of bacterial species in tomato rhizosphere, and 16S rRNA high-throughput sequencing was used to explore the diversity and structural stability of soil bacterial community.
    Results On the 1st day of pathogen inoculation, the Chao1 index, Shannon index and Simpson index of bacterial community structure in rhizosphere soils under the two QL-RS 1115 inoculation treatments reduced significantly (P<0.05), the number of nodes and connections of co-occurrence network significantly decreased, and the degree of modularity decreased. The simplified co-occurrence network indicated that the stability of bacterial community structure decreased. On the 14th day of pathogen inoculation, the beneficial bacteria such as Acinetobacter, Sphingomonas, Lysobacter, and Pseudomonas were enriched in rhizosphere soil, the number of nodes and connections of bacterial co-occurrence network increased, and the degree of modularity increased, indicating that the stability of bacterial community was restored. Compared with non-mycorrhizal seedlings treatments, both the mycorrhizal seedlings with and without pathogen showed significantly lower abundance of pathogen in rhizosphere soil (P<0.05). AM fungi significantly increased the Chao1 index and Shannon index (P<0.05), and increased the relative abundance of Flavobacterium, Flavisolibacter, Cytophaga, and Bryobacter in the rhizosphere soil of tomato infected with pathogen. At the same time, the number of nodes and connections of the co-occurrence network were increased, and the benign interaction between bacterial species in tomato rhizosphere was promoted, and the complexity of the bacterial network was improved.
    Conclusions Tomato infected with Ralstonia solanacearum induces the enrichment of beneficial bacteria, including Acinetobacter, Sphingomonas, Lysobacter, and Pseudomonas in the rhizosphere to restore the microbial diversity and stability. Inoculation of AM fungi significantly decreases the relative abundance of Ralstonia solanacearum but increases that of Flavobacterium, and further improves the diversity and stability of bacterial community in the rhizosphere soil of tomato, so enhances the resistance to Ralstonia solanacearum.

     

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