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草业学报 ›› 2023, Vol. 32 ›› Issue (4): 161-172.DOI: 10.11686/cyxb2022311

• 研究论文 • 上一篇    

欧美进口紫花苜蓿可培养种带细菌及其对动植物的致病性

张振粉1,2(), 黄荣1,2, 姚博1,2, 张旺东3, 杨成德4, 陈秀蓉4   

  1. 1.甘肃农业大学草业学院,甘肃 兰州 730070
    2.草业生态系统教育部重点实验室,中-美草地畜牧业可持续发展研究中心,甘肃 兰州 730070
    3.甘肃农业大学动物医学院,甘肃 兰州 730070
    4.甘肃农业大学植物保护学院,甘肃 兰州 730070
  • 收稿日期:2022-08-02 修回日期:2022-09-05 出版日期:2023-04-20 发布日期:2023-01-29
  • 通讯作者: 张振粉
  • 作者简介:张振粉(1984-),男,福建霞浦人,副教授,博士。 E-mail: zhangzf@gsau.edu.cn
  • 基金资助:
    国家自然科学基金(32060369);兰州市人才创新创业项目(2019-RC-116);2020年度陇原创新创业人才项目,甘肃农业大学青年导师基金项目(GAU-QDFC-2019-08);甘肃省杰出青年基金(20JR10RA562)

Culturable seed-borne bacteria of lucerne imported from Europe and North America and their pathogenicity to plants and animals

Zhen-fen ZHANG1,2(), Rong HUANG1,2, Bo YAO1,2, Wang-dong ZHANG3, Cheng-de YANG4, Xiu-rong CHEN4   

  1. 1.College of Pratacultural Science,Gansu Agricultural University,Lanzhou 730070,China
    2.Key Laboratory of Grassland Ecosystem,Ministry of Education,Sino-U. S. Center for Grassland Ecosystem Sustainability,Lanzhou 730070,China
    3.College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China
    4.College of Plant Protection,Gansu Agricultural University,Lanzhou 730070,China
  • Received:2022-08-02 Revised:2022-09-05 Online:2023-04-20 Published:2023-01-29
  • Contact: Zhen-fen ZHANG

摘要:

为了探究进口紫花苜蓿种带细菌的多样性及其对动植物的致病性,本研究从北美和欧洲共收集到紫花苜蓿种子样品34份,所有样品经室内研磨稀释分离培养,共获得39株种带细菌分离物,结合常规表型特征及16S rDNA鉴定方法确定它们的分类地位;并在室内采用菌悬液皿内发芽及盆栽接种法和腹腔注射法分别测定了21株代表细菌对供试紫花苜蓿和昆明小鼠的致病性。结果显示:1)39株细菌隶属3门15属,门分别为厚壁菌门、变形菌门和放线菌门,其中优势门为厚壁菌门;属地位的分别为芽孢杆菌属、假单胞菌属、马赛菌属、短芽孢杆菌属、欧文氏菌属、泛菌属、不动杆菌属、肠杆菌属、埃希氏肠杆菌属、假芽孢杆菌属、假节杆菌属、红球菌属、葡萄球菌属、土壤芽孢杆菌属和微杆菌属,其中优势属为芽孢杆菌属和假单胞菌属。2)红球菌属GCKH菌株仅对紫花苜蓿致病;不动杆菌属ZSR17、埃希氏肠杆菌属ZSR25和马赛菌属R1菌株仅对小鼠具有致病性;而欧文氏菌属ZF1和ZS3、泛菌属CQ10和ZS6菌株既可以引起紫花苜蓿致病,又可以引起小白鼠发病,是潜在的植物和动物跨界侵染共致病病原细菌。研究结果初步探明了欧美进口紫花苜蓿种带细菌的分类地位及其危害性,为综合防控这一类通过牧草种子携带的植物和动物独立致病细菌,以及植物和动物跨界侵染共致病病原细菌的侵入和传播奠定了理论依据。

关键词: 紫花苜蓿, 种带细菌, 多样性, 植物和动物跨界共患, 病原细菌

Abstract:

To explore the diversity of seed-borne bacteria of imported alfalfa (Medicago sativa) and their pathogenicity to animals and plants, 34 samples of lucerne seeds imported from North America and Europe were collected and analyzed. After Petri-dish isolation and culture, 39 seed-borne bacterial isolates were obtained. The taxonomic status of the bacterial isolates was determined through observations of phenotypic characteristics and 16S rDNA amplicon sequencing. Twenty-one representative strains with potential pathogenicity were selected for evaluation in a plant model (alfalfa, M. sativa) and an animal model (mouse, Mus musculus). Petri-dishes test and pot experiments were performed to study the impact of the bacterial isolates on alfalfa plants. The mouse infection model consisted of intraperitoneal injection of a bacterial suspension (109 CFU·mL-1) at a dose of 0.1 mL·10 g-1. We found that the alfalfa seeds harbored a diverse community of bacteria. These bacterial isolates were grouped in 39 morphospecies that corresponded to three phyla and 15 genera. Out of 39 isolates, 24 (61.54%) were Gram positive and 15 (38.46%) Gram negative. Among them, Firmicutes was the predominant phylum observed in all groups and Bacillus was the dominant genus. The results of pathogenicity tests showed that bacterial strain GCKH (Rhodococcus sp.) was only pathogenic to alfalfa while ZSR17 (Acinetobacter sp.), ZSR25 (Escherichia sp.), and R1 (Massilia sp.) were pathogenic to mice. Notably, ZF1 (Erwinia sp.), ZS3 (Erwinia sp.), CQ10 (Pantoea sp.), and ZS6 (Pantoea sp.) could cause disease in both alfalfa and mouse hosts, indicating that they are potential co-pathogenic bacteria capable of cross-kingdom infection. The results provide a preliminary overview of the taxonomic status of seed-borne bacteria carried by lucerne seeds imported from Europe and America and the potential harmfulness of these bacterial strains. These findings lay a theoretical foundation for the integrated management and prevention of the invasion and transmission of the plant-pathogenic, animal-pathogenic, or co-pathogenic bacteria on forage seeds that have the potential to cause infections in plants and animals.

Key words: lucerne, seed-borne bacteria, diversity, cross-kingdom co-pathogen of plants and animals, pathogenic bacteria