Article(id=1259888473876644240, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1259888457367806489, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20250921, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1765382400000, receivedDateStr=2025-12-11, revisedDate=null, revisedDateStr=null, acceptedDate=1770048000000, acceptedDateStr=2026-02-03, onlineDate=1778310419767, onlineDateStr=2026-05-09, pubDate=1777824000000, pubDateStr=2026-05-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1778310419767, onlineIssueDateStr=2026-05-09, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1778310419767, creator=13701087609, updateTime=1778310419767, updator=13701087609, issue=Issue{id=1259888457367806489, tenantId=1146029695717560320, journalId=1192105938417971205, year='2026', volume='66', issue='5', pageStart='2031', pageEnd='2556', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=0, articleOrder=1, issueType=-1, specialIssue=null, createTime=1778310415832, creator=13701087609, updateTime=1778320153326, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1259929299465921482, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1259888457367806489, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1259929299465921483, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1259888457367806489, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=2208, endPage=2225, ext={EN=ArticleExt(id=1259888476326117800, articleId=1259888473876644240, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Screening of plant growth-promoting Actinomycetes strains and evaluation of plant growth-promoting effect of the synthetic microbial consortium in a maize-soybean rotation system, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

Objective To explore plant growth promoting rhizobacteria (PGPR) resources from the rhizosphere soil of maize in a maize-soybean rotation system and elucidate their roles in promoting the growth of maize and soybean, thus providing a theoretical basis and practical support at the microbial level for the sustainable development of agriculture. Methods Actinomycetes strains were isolated from the rhizosphere soil of maize via the dilution plating method with Gauze’s Synthetic Medium No. 1. The strains capable of secreting protease, producing siderophores, and fixing nitrogen were selected out. The isolated strains were identified by means of morphological observation and 16S rRNA gene sequence analysis. After optimization of the fermentation conditions and tests of stress tolerance, a synthetic microbial consortium (SMC) was prepared. Its growth-promoting effects on maize and soybean were evaluated through seed germination tests and pot experiments. Results A total of 105 Actinomycetes strains were isolated, five of which simultaneously exhibited the abilities of siderophore production, protease secretion, and nitrogen fixation. These strains were identified as Arthrobacter pokkalii (JM-18, JM-21), A. oryzae (JM-24), A. ginsengisoli (JM-47), and A. pascens (JM-48). They were mixed in equal proportions to form a SMC. Growth promotion assays showed that the SMC significantly improved maize seed germination and maize plant growth in pots. Specifically, the SMC increased the root length and shoot length in the seed germination assay by 120.22% and 20.94%, respectively, and it also significantly increased the plant height, root length, fresh weight, and dry weight of maize plants in pots. Moreover, the SMC markedly promoted soybean development, increasing soybean shoot length by 42.08% during seed germination. For potted soybean plants, the SMC increased the plant height, root length, fresh weight, and dry weight by 39.40%, 93.31%, 161.14%, and 163.57%, respectively. Conclusion We successfully identified five Actinomycetes strains capable of secreting protease, producing siderophores, and fixing nitrogen. The SMC constructed from these strains significantly enhances the growth of both maize and soybean. This study offers promising microbial resources for the development of efficient and environmentally friendly biofertilizers.

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E-mail: XING Jihong, ;
DONG Jingao,
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These authors contributed equally to this work.

, authorsList=Mengke QIAO, Liming WANG, Tingfeng LI, Kang ZHANG, Hongzhe CAO, Jihong XING, Jingao DONG), CN=ArticleExt(id=1259888497759011519, articleId=1259888473876644240, tenantId=1146029695717560320, journalId=1192105938417971205, language=CN, title=玉米-大豆轮作模式下根际促生放线菌的筛选及合成菌群的促生效果, columnId=1192149544164012138, journalTitle=微生物学报, columnName=研究报告, runingTitle=null, highlight=null, articleAbstract=

目的 挖掘玉米-大豆轮作模式下玉米根际土壤中的植物根际促生菌(plant growth promoting rhizobacteria, PGPR)资源,明确其在玉米与大豆生长过程中的功能,为农业可持续发展提供微生物层面的理论依据与实践支撑。 方法 采用稀释涂布法,以高氏一号培养基为分离介质,分离玉米根际土壤中的放线菌,并筛选出具备蛋白酶分泌、产铁载体以及固氮能力的促生菌株。运用形态学观察结合16S rRNA基因序列分析技术,对筛选所得菌株进行鉴定。经发酵条件优化和耐胁迫能力测试后,获得复合菌株发酵液合成菌群(synthetic microbial consortium, SMC)。通过促生试验,验证其对玉米和大豆生长的促生作用。 结果 从玉米根际土壤中分离获得105株放线菌,其中5株菌株同时具备产蛋白酶、产铁载体和固氮能力。经鉴定,确定JM-18、JM-21为波卡利水稻节杆菌(Arthrobacter pokkalii),JM-24为水稻节杆菌(Arthrobacter oryzae),JM-47为Arthrobacter ginsengisoli,JM-48为滋养节杆菌(Arthrobacter pascens)。同时发现,这5株菌均具备一定的解无机磷和产NH3能力。之后,按1:1的比例将其复配,构成合成菌群SMC。促生试验结果显示,SMC处理对玉米种子发芽和盆栽玉米的生长发育均具有显著促生作用,玉米种子根长和芽长分别增加120.22%、20.94%,盆栽玉米的株高、根长、鲜重、干重均显著提升。此外,SMC对大豆的生长发育同样表现出显著促进效果,可使大豆种子芽长增长42.08%,盆栽大豆的株高、根长、鲜重、干重分别增长39.40%、93.31%、161.14%、163.57%。 结论 本研究筛选出5株兼具产蛋白酶、产铁载体和固氮能力的放线菌,其构建的合成菌群SMC能够显著促进玉米和大豆的生长,为开发高效、环保的微生物肥料提供了极具潜力的优质菌株资源。

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作者贡献声明

乔梦可:完成本研究的主体试验操作、论文撰写工作;王黎明:数据整理及处理;李庭锋:试验样品的采集任务;张康:指导实验安排并提供相关资源支持;曹宏哲:指导完成图片制作与数据处理;邢继红:进行实验指导,并参与文章审阅与修改;董金皋:指导实验设计,并获取资金支持。

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Yangling: Northwest A&F University, 2025 (in Chinese)., articleTitle=null, refAbstract=null), Reference(id=1259928545590109037, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, doi=null, pmid=null, pmcid=null, year=2024, volume=55, issue=5, pageStart=46, pageEnd=55, url=null, language=null, rfNumber=[28], rfOrder=43, authorNames=侯莹莹, 胡小梅, journalName=东北农业大学学报, refType=null, unstructuredReference=侯莹莹, 胡小梅. 3株玉米根际解磷菌筛选鉴定及促生作用研究[J]. 东北农业大学学报, 2024, 55(5): 46-55., articleTitle=3株玉米根际解磷菌筛选鉴定及促生作用研究, refAbstract=null), Reference(id=1259928545875321710, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, doi=null, pmid=null, pmcid=null, year=2024, volume=55, issue=5, pageStart=46, pageEnd=55, url=null, language=null, rfNumber=[28], rfOrder=44, authorNames=Hou YY, Hu XM, journalName=Journal of Northeast Agricultural University, refType=null, unstructuredReference=Hou YY, Hu XM. Screening and identification of three strains of phosphate-solubilizing bacteria from maize rhizosphere and their roles in plant growth promoting effect[J]. Journal of Northeast Agricultural University, 2024, 55(5): 46-55 (in Chinese)., articleTitle=null, refAbstract=null), Reference(id=1259928546047288176, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, doi=null, pmid=null, pmcid=null, year=2022, volume=49, issue=7, pageStart=2625, pageEnd=2637, url=null, language=null, rfNumber=[29], rfOrder=45, authorNames=白建飞, 韩升才, 高聚林, 于晓芳, 青格尔, 胡树平, 张赛楠, 郭江岸, journalName=微生物学通报, refType=null, unstructuredReference=白建飞, 韩升才, 高聚林, 于晓芳, 青格尔, 胡树平, 张赛楠, 郭江岸. 内生菌莫拉维假单胞菌GF-55促进玉米生长和提高抗倒伏功能分析[J]. 微生物学通报, 2022, 49(7): 2625-2637., articleTitle=内生菌莫拉维假单胞菌GF-55促进玉米生长和提高抗倒伏功能分析, refAbstract=null), Reference(id=1259928546324112243, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, doi=null, pmid=null, pmcid=null, year=2022, volume=49, issue=7, pageStart=2625, pageEnd=2637, url=null, language=null, rfNumber=[29], rfOrder=46, authorNames=Bai JF, Han SC, Gao JL, Yu XF, Qing GE, Hu SP, Zhang SN, Guo JA, journalName=Microbiology China, refType=null, unstructuredReference=Bai JF, Han SC, Gao JL, Yu XF, Qing GE, Hu SP, Zhang SN, Guo JA. Endophytic bacterial strain GF-55 improves the growth and lodging resistance of maize[J]. 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A: Screening of plant growth-promoting strains; B: Phylogenetic tree of five strains constructed based on 16S rRNA gene sequence. The numbers in parentheses are GenBank accession numbers, the scale bar indicates a 1% sequence (or nucleotide) difference., figureFileSmall=IWYzX1U4CU0+RI/wF/afKA==, figureFileBig=JuM43S3Wb6a/kYZY/qonsw==, tableContent=null), ArticleFig(id=1259928491936571782, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=CN, label=图1, caption=促生菌株的促生特性与鉴定, figureFileSmall=IWYzX1U4CU0+RI/wF/afKA==, figureFileBig=JuM43S3Wb6a/kYZY/qonsw==, tableContent=null), ArticleFig(id=1259928493735928219, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=EN, label=Figure 2, caption=Studies on other functions of five strains., figureFileSmall=FMDFLBtUCW5e7mBCzBJEYg==, figureFileBig=36wm+axx9kc2vve1wdnTCg==, tableContent=null), ArticleFig(id=1259928494885167524, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=CN, label=图2, caption=五株菌的其他促生功能研究, figureFileSmall=FMDFLBtUCW5e7mBCzBJEYg==, figureFileBig=36wm+axx9kc2vve1wdnTCg==, tableContent=null), ArticleFig(id=1259928496550306220, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=EN, label=Figure 3, caption=Screening of the optimal medium composition for the five strains. A: Optimal carbon source; B: Optimal nitrogen source; C: Optimal inorganic salts. Different lowercase letters indicate significant differences between groups (P<0.05)., figureFileSmall=PPd1u0/16bV42L42Tm0XpQ==, figureFileBig=i7PLvAzWgOVSLb7OW5afzQ==, tableContent=null), ArticleFig(id=1259928497032651184, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=CN, label=图3, caption=五株菌的最适培养基成分筛选, figureFileSmall=PPd1u0/16bV42L42Tm0XpQ==, figureFileBig=i7PLvAzWgOVSLb7OW5afzQ==, tableContent=null), ArticleFig(id=1259928497905066427, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=EN, label=Figure 4, caption=Fermentation condition optimization of the five strains. A: pH; B: Temperature; C: Fermentation speed; D: Medium volume. Different lowercase letters indicate significant differences between groups (P<0.05)., figureFileSmall=IsdoxCCrYNkO5v/2qCqiFQ==, figureFileBig=e35imBo3joE+CwwWXaA0Ug==, tableContent=null), ArticleFig(id=1259928498701984195, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=CN, label=图4, caption=五株菌的发酵条件优化, figureFileSmall=IsdoxCCrYNkO5v/2qCqiFQ==, figureFileBig=e35imBo3joE+CwwWXaA0Ug==, tableContent=null), ArticleFig(id=1259928500849467854, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=EN, label=Figure 5, caption=Stress tolerance of the five growth-promoting strains. A: Effect of high and low temperature stress on the growth of strains; B: Effect of salt stress on the growth of strains; C: Effect of alkali stress on the growth of strains. Different lowercase letters indicate significant differences between groups (P<0.05)., figureFileSmall=RjVF/99epaXt7gfrPcajfw==, figureFileBig=s6vvoRkQfteU0EcFEYQ5Tw==, tableContent=null), ArticleFig(id=1259928504989245915, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=CN, label=图5, caption=五株促生菌的抗逆性, figureFileSmall=RjVF/99epaXt7gfrPcajfw==, figureFileBig=s6vvoRkQfteU0EcFEYQ5Tw==, tableContent=null), ArticleFig(id=1259928505861661158, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=EN, label=Figure 6, caption=Effect of compound bacteria SMC on the growth of maize. A: Effect of SMC on maize seeds (bar=2 cm); B: Effects of SMC on root length and shoot length of maize seeds; C: Effect of SMC on the growth of potted maize (bar=20 cm); D: Effects of SMC on the plant height, root length, fresh weight and dry weight of potted maize. *P<0.05, the difference between groups was significant; **P<0.01, the difference between groups was extremely significant. The same below., figureFileSmall=oVmE9t+chdLnmPRQ2uYV/g==, figureFileBig=zoejlKHWlQiT+crgsVlyKg==, tableContent=null), ArticleFig(id=1259928506193011175, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=CN, label=图6, caption=合成菌群SMC对玉米生长的影响, figureFileSmall=oVmE9t+chdLnmPRQ2uYV/g==, figureFileBig=zoejlKHWlQiT+crgsVlyKg==, tableContent=null), ArticleFig(id=1259928507023483377, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=EN, label=Figure 7, caption=Effect of compound bacteria SMC on the growth of soybeans. A: Effect of SMC on soybean seeds (bar=2 cm); B: Effects of SMC on root length and plant length of soybean seeds; C: Effect of SMC on the growth of potted soybeans (bar=10 cm); D: Effects of SMC on the plant height, root length, fresh weight, and dry weight of potted soybeans., figureFileSmall=L3A2qNlNA8ZvTMgc5RJcEw==, figureFileBig=1vukmKpVc+2W1eKfRLo9lg==, tableContent=null), ArticleFig(id=1259928507950424573, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=CN, label=图7, caption=合成菌群SMC对大豆生长的影响, figureFileSmall=L3A2qNlNA8ZvTMgc5RJcEw==, figureFileBig=1vukmKpVc+2W1eKfRLo9lg==, tableContent=null), ArticleFig(id=1259928510404092424, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=EN, label=Table 1, caption=

Physiological and biochemical characteristics of five growth-promoting strains

, figureFileSmall=null, figureFileBig=null, tableContent=
Test itemJM-18JM-21JM-24JM-47JM-48
Gram staining+++++
Voges-Proskauer test+++++
Citrate utilization+++++
Propionate utilizationWWWWW
D-xylose+++++
L-arabinose---++
D-mannitol+++++
Gelatin liquefaction+++++
Growth at pH 5.7+++++
Nitrate reduction+++++
Starch hydrolysis--+++
), ArticleFig(id=1259928510857077262, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=CN, label=表1, caption=

五株促生菌的生理生化特征

, figureFileSmall=null, figureFileBig=null, tableContent=
Test itemJM-18JM-21JM-24JM-47JM-48
Gram staining+++++
Voges-Proskauer test+++++
Citrate utilization+++++
Propionate utilizationWWWWW
D-xylose+++++
L-arabinose---++
D-mannitol+++++
Gelatin liquefaction+++++
Growth at pH 5.7+++++
Nitrate reduction+++++
Starch hydrolysis--+++
), ArticleFig(id=1259928511247147541, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=EN, label=Table 2, caption=

Strain compatibility test

, figureFileSmall=null, figureFileBig=null, tableContent=
StrainJM-18JM-21JM-24JM-47JM-48
JM-48++++
JM-47+++
JM-24++
JM-21+
JM-18
), ArticleFig(id=1259928512127951393, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888473876644240, language=CN, label=表2, caption=

菌株相容性试验

, figureFileSmall=null, figureFileBig=null, tableContent=
StrainJM-18JM-21JM-24JM-47JM-48
JM-48++++
JM-47+++
JM-24++
JM-21+
JM-18
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玉米-大豆轮作模式下根际促生放线菌的筛选及合成菌群的促生效果
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乔梦可 1, 2 , 王黎明 1, 2 , 李庭锋 3 , 张康 1, 2 , 曹宏哲 1, 2 , 邢继红 1, 2 , 董金皋 1, 2
微生物学报 | 研究报告 2026,66(5): 2208-2225
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微生物学报 | 研究报告 2026, 66(5): 2208-2225
玉米-大豆轮作模式下根际促生放线菌的筛选及合成菌群的促生效果
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乔梦可1, 2, 王黎明1, 2, 李庭锋3, 张康1, 2, 曹宏哲1, 2, 邢继红1, 2 , 董金皋1, 2
作者信息
  • 1.河北农业大学,华北作物改良与调控国家重点实验室,河北 保定
  • 2.河北省植物生理与分子病理学重点实验室,河北 保定
  • 3.黑龙江省农垦科学院农作物开发研究所,黑龙江 佳木斯
Screening of plant growth-promoting Actinomycetes strains and evaluation of plant growth-promoting effect of the synthetic microbial consortium in a maize-soybean rotation system
Mengke QIAO1, 2, Liming WANG1, 2, Tingfeng LI3, Kang ZHANG1, 2, Hongzhe CAO1, 2, Jihong XING1, 2 , Jingao DONG1, 2
Affiliations
  • 1.State Key Laboratory of North China Crop Improvement and Regulation, Hebei Agricultural University, Baoding, Hebei, China
  • 2.Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology, Baoding, Hebei, China
  • 3.Institute of Crop Development, Heilongjiang Academy of Land Reclamation Sciences, Jiamusi, Heilongjiang, China
出版时间: 2026-05-04 doi: 10.13343/j.cnki.wsxb.20250921
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目的 挖掘玉米-大豆轮作模式下玉米根际土壤中的植物根际促生菌(plant growth promoting rhizobacteria, PGPR)资源,明确其在玉米与大豆生长过程中的功能,为农业可持续发展提供微生物层面的理论依据与实践支撑。 方法 采用稀释涂布法,以高氏一号培养基为分离介质,分离玉米根际土壤中的放线菌,并筛选出具备蛋白酶分泌、产铁载体以及固氮能力的促生菌株。运用形态学观察结合16S rRNA基因序列分析技术,对筛选所得菌株进行鉴定。经发酵条件优化和耐胁迫能力测试后,获得复合菌株发酵液合成菌群(synthetic microbial consortium, SMC)。通过促生试验,验证其对玉米和大豆生长的促生作用。 结果 从玉米根际土壤中分离获得105株放线菌,其中5株菌株同时具备产蛋白酶、产铁载体和固氮能力。经鉴定,确定JM-18、JM-21为波卡利水稻节杆菌(Arthrobacter pokkalii),JM-24为水稻节杆菌(Arthrobacter oryzae),JM-47为Arthrobacter ginsengisoli,JM-48为滋养节杆菌(Arthrobacter pascens)。同时发现,这5株菌均具备一定的解无机磷和产NH3能力。之后,按1:1的比例将其复配,构成合成菌群SMC。促生试验结果显示,SMC处理对玉米种子发芽和盆栽玉米的生长发育均具有显著促生作用,玉米种子根长和芽长分别增加120.22%、20.94%,盆栽玉米的株高、根长、鲜重、干重均显著提升。此外,SMC对大豆的生长发育同样表现出显著促进效果,可使大豆种子芽长增长42.08%,盆栽大豆的株高、根长、鲜重、干重分别增长39.40%、93.31%、161.14%、163.57%。 结论 本研究筛选出5株兼具产蛋白酶、产铁载体和固氮能力的放线菌,其构建的合成菌群SMC能够显著促进玉米和大豆的生长,为开发高效、环保的微生物肥料提供了极具潜力的优质菌株资源。

玉米-大豆轮作  /  植物根际促生菌  /  节杆菌属  /  合成菌群

Objective To explore plant growth promoting rhizobacteria (PGPR) resources from the rhizosphere soil of maize in a maize-soybean rotation system and elucidate their roles in promoting the growth of maize and soybean, thus providing a theoretical basis and practical support at the microbial level for the sustainable development of agriculture. Methods Actinomycetes strains were isolated from the rhizosphere soil of maize via the dilution plating method with Gauze’s Synthetic Medium No. 1. The strains capable of secreting protease, producing siderophores, and fixing nitrogen were selected out. The isolated strains were identified by means of morphological observation and 16S rRNA gene sequence analysis. After optimization of the fermentation conditions and tests of stress tolerance, a synthetic microbial consortium (SMC) was prepared. Its growth-promoting effects on maize and soybean were evaluated through seed germination tests and pot experiments. Results A total of 105 Actinomycetes strains were isolated, five of which simultaneously exhibited the abilities of siderophore production, protease secretion, and nitrogen fixation. These strains were identified as Arthrobacter pokkalii (JM-18, JM-21), A. oryzae (JM-24), A. ginsengisoli (JM-47), and A. pascens (JM-48). They were mixed in equal proportions to form a SMC. Growth promotion assays showed that the SMC significantly improved maize seed germination and maize plant growth in pots. Specifically, the SMC increased the root length and shoot length in the seed germination assay by 120.22% and 20.94%, respectively, and it also significantly increased the plant height, root length, fresh weight, and dry weight of maize plants in pots. Moreover, the SMC markedly promoted soybean development, increasing soybean shoot length by 42.08% during seed germination. For potted soybean plants, the SMC increased the plant height, root length, fresh weight, and dry weight by 39.40%, 93.31%, 161.14%, and 163.57%, respectively. Conclusion We successfully identified five Actinomycetes strains capable of secreting protease, producing siderophores, and fixing nitrogen. The SMC constructed from these strains significantly enhances the growth of both maize and soybean. This study offers promising microbial resources for the development of efficient and environmentally friendly biofertilizers.

maize-soybean rotation  /  plant growth-promoting rhizobacteria  /  Arthrobacter  /  synthetic microbial consortium
乔梦可, 王黎明, 李庭锋, 张康, 曹宏哲, 邢继红, 董金皋. 玉米-大豆轮作模式下根际促生放线菌的筛选及合成菌群的促生效果. 微生物学报, 2026 , 66 (5) : 2208 -2225 . DOI: 10.13343/j.cnki.wsxb.20250921
Mengke QIAO, Liming WANG, Tingfeng LI, Kang ZHANG, Hongzhe CAO, Jihong XING, Jingao DONG. Screening of plant growth-promoting Actinomycetes strains and evaluation of plant growth-promoting effect of the synthetic microbial consortium in a maize-soybean rotation system[J]. Acta Microbiologica Sinica, 2026 , 66 (5) : 2208 -2225 . DOI: 10.13343/j.cnki.wsxb.20250921
推动绿色农业可持续发展的关键在于挖掘能够替代或减量施用化肥的生态友好型技术。轮作作为一种契合可持续发展理念的耕作模式,在农业生产中展现出诸多显著优势。它能够有效改善因土壤连作引发的障碍问题,提升耕地质量,进而提高作物产量[1]。相较于玉米单作模式,轮作模式通过改变根际微生物的群落组成,招募假单胞菌、链霉菌等有益菌群,优化土壤营养成分,为作物生长创造更为有利的条件[2]。基于此,从轮作土壤中挖掘具有潜力的微生物肥料,尤其是植物根际促生菌(plant growth promoting rhizobacteria, PGPR),因其能够促进植物生长、增强抗逆性,对于减少对化学肥料的依赖以及推进农业可持续发展具有重要意义[3]
PGPR能够通过多种途径直接或间接促进植物生长。在直接促进方面,它们可通过营养活化与供给机制发挥作用,例如产生蛋白酶分解土壤中的有机质(如有机氮、含磷有机物),将其转化为植物可直接吸收利用的无机养分(如铵、磷酸盐);具备固氮能力,为植物和土壤提供充足的氮源;还能产生铁载体,以高效螯合铁的方式,释放磷或其他元素,满足植物生长对各种元素的需求[4]。在间接调节方面,PGPR可通过产生抗生素、溶菌酶以及诱导系统抗性等方式,增强植物的抗逆性,助力化肥减施增效,推动农业绿色可持续发展[5]。PGPR种类繁多、分布广泛,涵盖了细菌、真菌、原生动物及藻类等多种微生物类群。其中,假单胞菌属(Pseudomonas)、芽孢杆菌属(Bacillus)、链霉菌属(Streptomyces)、木霉菌属(Trichoderma)等为优势菌群[6]。大量研究已证实不同PGPR对植物生长具有显著促进作用。例如,贾艺伟[7]从平邑甜茶根际土中分离获得一株枯草芽孢杆菌(B. subtilis) T3-5,该菌株对平邑甜茶幼苗的生物量积累具有显著促进作用。杨晓帆等[8]筛选到一株荧光假单胞菌(P. fluorescens),该菌株通过增强桃树的光合作用,并协同提升其根系与叶片中的氮、磷、钾及铁等关键营养元素的含量,实现了对桃树植株生长的全面促进。Contreras-Cornejo等[9]研究发现绿色木霉(T. viride)可以产生吲哚-3-乙酸(indole-3-acetic acid, IAA),促进拟南芥侧根的发育,使其鲜重增加62%。
当前,PGPR的应用研究主要集中在细菌与真菌领域,关于放线菌的研究应用相对较少。鉴于放线菌在促进植物生长方面可能具有独特作用[10],Liu等[11]从森林根际土壤中分离出一株链霉菌YNK-FS0020 (S. olivoreticuli,现分类修订为S. abikoensis),研究发现其能抑制多种植物病原菌,具备溶解无机和有机磷及产铁载体的能力,能够显著促进番茄生长,同时代谢组学检测揭示了该菌株能够调控激素以及分泌抗菌物质等机制,这表明开发利用PGPR放线菌以促进粮油作物的生长发育具有广阔的发展潜力。
本研究立足于玉米-大豆轮作模式,旨在从玉米根际土壤中分离筛选获得具有产蛋白酶、产铁载体、固氮等促生作用的放线菌,并对其发酵条件进行优化。进一步将筛选得到的优良放线菌按照等体积比进行复配,通过种子促生试验和盆栽试验验证其对玉米和大豆生长发育的促进作用。本研究旨在深入探究PGPR放线菌在促进粮油作物生长方面的开发与应用,为微生物菌剂的开发提供菌种资源及技术支撑,进而推动农业绿色可持续发展。
土壤采自黑龙江省佳木斯市玉米-大豆轮作种植区(46°46′48″N,130°25′12″E),于轮作第5年玉米成熟时随机选取长势良好的健康玉米植株5株,采用“抖根法”用无菌袋收集根际土壤,置于4 ℃冰箱中保存。本研究所用玉米品种为‘先玉335’、大豆品种为‘W82’。
采用梯度稀释法,结合高氏一号培养基[12],对土壤样品中的放线菌进行分离培养。挑取形态各异的菌株,经多次划线纯化操作后,将菌株接种于50 mL液体高氏一号培养基中,28 ℃、220 r/min培养3-5 d,至OD600=1.0时备用。取3 μL上述菌液,分别滴加到铬天青S (chrome azurol S, CAS)培养基[13]、阿须贝无氮培养基[14]和蛋白酶检测培养基[15]上,培养5-7 d,检测菌株是否具备产铁载体、固氮和产蛋白酶的能力。
将筛选出的促生菌进行菌落形态特征观察,包括菌落的形状、大小、颜色、表面透明度以及边缘情况等。参照《常见细菌系统鉴定手册》[16]的方法进行革兰氏染色、Voges-Proskauer (V-P)、柠檬酸盐、丙酸盐、D-木糖、L-阿拉伯糖、D-甘露醇、明胶液化、pH 5.7生长、硝酸盐还原、淀粉水解试验,依据《伯杰细菌鉴定手册》[17]对菌株进行初步鉴定。使用细菌基因组DNA快速提取试剂盒(北京博迈德生物技术有限公司)提取5株促生菌株的DNA,利用通用引物27F (5′-AGAGTTTGATCCTGGC TCAG-3′)和1492R (5′-TACGGCTACCTTGTTA CGAGTT-3′)进行16S rRNA基因PCR扩增,反应体系和条件参考李志普[18]的方法。PCR产物经电泳检测后进行测序鉴定,将测得的16S rRNA基因序列结果在EzBioCloud数据库(https://www.ezbiocloud.net)上进行比对。最后使用MEGA 7.0软件,结合邻接(neighbor-joining, NJ)法构建菌株的系统发育树,设置bootstrap值为1 000。
将5株菌分别接种于蒙金娜无机磷培养基和硅酸盐固体培养基中,28 ℃恒温培养箱中倒置培养5-7 d,观察菌落周围是否有透明圈产生[18]
采用Salkowski比色法,将各菌株接种至含有L-色氨酸的LB液体培养基中28 ℃培养24 h,每个菌株取50 μL菌悬液于96孔板中,加入等量Salkowski显色液进行显色反应,以IAA标准液作为阳性对照,于室温、避光条件下放置30 min后观察颜色是否变红[19]
将菌株分别接种至装有10 mL蛋白胨水(10 g/L)的试管中,28 ℃培养2-3 d,各试管加入0.5 mL Nessler’s试剂,观察颜色是否变为褐色或黄色[20]
对促生菌的最适碳源、氮源和无机盐进行筛选,以蔗糖、葡萄糖、可溶性淀粉、甘露糖、半乳糖或木糖作为碳源,以牛肉膏、蛋白胨、胰蛋白胨、大豆蛋白胨或酵母浸粉作为氮源,以氯化钠、氯化镁、磷酸钙、氯化钾、硫酸锌或磷酸氢二钾+磷酸二氢钾(1:1)作为无机盐。在28 ℃、220 r/min培养48 h后,测定OD600,以评估不同营养条件下菌株的生长情况,进而确定各菌株的最适培养基。
在此基础上,进一步开展促生菌的发酵条件优化,包括最适pH (6.0、7.0、8.0、9.0、10.0)、发酵温度(24、28、34、37、42 ℃)、发酵转速(120、160、200、240、280 r/min)以及装液量(30%、40%、50%、60%、70%)等参数。
将各菌株接种至最适培养基中发酵后,分别置于-20、0、20、40、60 ℃的环境中培养8 h,待菌液温度回到室温后再次测定菌液的OD600值,以评估菌株对低温和高温环境的耐受能力。之后将各菌株分别接种至NaCl含量为0.0、0.5%、1.5%、2.5%、3.5%、4.5%、5.5%的培养基,以及pH为6.0、7.0、8.0、9.0、10.0、11.0的培养基中,8 h后测定OD600值,以评估菌株的耐盐碱能力。
将各菌株接种至最适培养基中,获得菌株发酵液后开展菌株相容试验。将各菌株菌液的OD600值调至1.0左右,取100 µL菌液均匀涂布于NA培养基表面,并在平板上放置直径为6 mm的无菌滤纸片。于每张滤纸片上滴加2 µL其他待测菌株发酵液,待晾干后,倒置培养3-5 d,观察是否出现抑菌圈,以评估不同菌株间的相容性,每个处理设置3个重复。若各菌株间均能相容,则将各菌株单独发酵后,按照等体积比复合,构成合成菌群(synthetic microbial consortium, SMC)。
使用1% NaOCl对玉米和大豆种子进行表面消毒处理,然后用200 mL SMC发酵液(OD600约为0.6)对种子进行浸种处理8 h,以灭菌水处理作为对照。将种子均匀摆放在铺有浸湿滤纸片的玻璃皿中,在黑暗条件下进行恒温催芽,每个处理设置5个平行,每个平行3个重复。7 d后,观察并记录种子萌发情况。之后,从各处理中取长势一致的发芽种子,移栽至无菌营养土中进行盆栽试验,每个处理9株幼苗,移栽后每隔7 d进行灌菌处理,每次每株幼苗浇灌菌悬液50 mL (OD600约为0.6),对照组浇灌同等体积无菌水。45 d后,观察并测量盆栽植株的生长情况。
使用Excel 2019软件对试验数据进行整理,运用GraphPad Prism 9.5软件通过单因素方差分析和t检验进行显著性差异分析,其中*P<0.05表明组间差异显著;**P<0.01表明组间差异极显著;ns表示组间差异不显著,不同字母表示组间差异显著(P<0.05)。使用MEGA 11软件构建系统发育树。
从玉米根际土壤中成功筛选出105株放线菌。其中,JM-18、JM-21、JM-24、JM-47和JM-48这5株菌同时具备产蛋白酶、产铁载体和固氮能力(图1A)。对这5株菌测定了11个生理生化指标(表1),同时观察其菌落形态,发现均呈黄色或白色,形状为圆形,表面凸起、圆润且边缘规则(图1B)。16S rRNA基因鉴定结果显示,JM-18和JM-21与波卡利水稻节杆菌(Arthrobacter pokkalii)的相似度最高,分别为99.32%和99.18%;JM-24与水稻节杆菌(A. oryzae)的相似度最高,达到100%;JM-47与A. ginsengisoli的相似度最高,达99.60% (图1B);JM-48与滋养节杆菌(A. pascens)的相似度最高,达到100%。结合形态学、生理生化特征与分子鉴定结果,确定JM-18和JM-21为波卡利水稻节杆菌(A. pokkalii),JM-24为水稻节杆菌(A. oryzae),JM-47为A. ginsengisoli,JM-48为滋养节杆菌(A. pascens)。
对5株菌的其他促生特性进行检测,发现5株菌在蒙金娜无机磷培养基上能够产生透明圈,在产NH3试验中菌液均变为黄色,表明5株菌均具备一定的解无机磷和产NH3能力;然而,5株菌在硅酸盐固体培养基上未能产生透明圈,且在Salkowski显色反应中菌液均未变红,表明其均不具备解钾和产IAA能力(图2)。
对5株菌株的发酵培养基进行优化,发现JM-18在营养条件为蔗糖、牛肉膏、K2HPO4+KH2PO4 (1:1)时OD600值达到最高;JM-21在营养条件为蔗糖、大豆蛋白胨、K2HPO4+KH2PO4 (1:1)时OD600值最高;JM-24在营养条件为蔗糖、酵母浸粉、KCl时OD600值最高;JM-47和JM-48在营养条件为蔗糖、酵母浸粉、K2HPO4+KH2PO4 (1:1)时OD600值最高(图3)。
对5株菌株的发酵条件进行优化,结果显示,JM-18在发酵条件为pH 9.0、28 ℃、280 r/min、装液量40%时OD600值最高;JM-21在pH 7.0、28 ℃、280 r/min、装液量40%时OD600值最高;JM-24在pH 7.0、24 ℃、160 r/min、装液量40%时OD600值最高;JM-47在pH 9.0、24 ℃、280 r/min、装液量50%时OD600值最高;JM-48在pH 7.0、28 ℃、160 r/min、装液量40%时OD600值最高(图4)。
对5株菌株的抗逆性进行检测,结果发现:随着温度升高,各菌株的浓度逐渐下降,但在60 ℃条件下,OD600值仍能分别达到0.327、0.376、0.316、0.363、0.324;随着温度降低,各菌株的OD600值缓慢降低,在-20 ℃条件下仍能分别达到0.466、0.471、0.548、0.418、0.449 (图5A),表明这5株菌在高温60 ℃和低温-20 ℃时仍能生长。同样地,随着盐浓度上升,各菌株的浓度逐渐下降,在盐浓度为4.5%时各菌株仍能够正常生长;盐浓度为5.5%时OD600值出现明显下降(图5B)。此外,各菌株在pH 9.0时仍能保持生长,OD600值能达到0.243-0.315;在pH 10.0时菌株生长明显受到抑制,基本停止生长(图5C)。因此,5株促生菌在pH为9.0、盐浓度为4.5%的盐碱胁迫下仍能维持生长。
菌株相容性试验结果表明,5株促生菌之间均具有良好的相容性,能够进行复配(表2)。将5株菌复合构成合成菌群SMC后,验证其对玉米的促生效果。结果显示,相较于对照组,SMC处理对玉米种子的促生效果显著(图6A)。经SMC发酵液处理后,种子的根长增长120.22% (P<0.01),芽长增长20.94% (P<0.05) (图6B)。对盆栽玉米的促生效果同样表明,相较于对照组,合成菌群SMC处理后的盆栽玉米生长势更佳(图6C),玉米苗的株高、根长、鲜重和干重均高于对照组,分别增长了24.74%、48.21% (P<0.05)、110.08%和64.05% (P<0.01) (图6D)。
为明确SMC对大豆是否同样具有促生作用,试验检测了SMC对大豆生长发育的影响。结果发现,经SMC处理后大豆种子的株高达14.28 cm,与对照组差异极显著,两处理间种子的根长差异不显著(图7A7B)。对盆栽大豆的促生结果显示,相较于对照组,SMC处理后盆栽大豆的生长势更好(图7C),经SMC处理后,苗期大豆的株高、根长、鲜重和干重均显著高于对照组,分别增长了39.40%、93.31%、161.14%和163.57% (图7D)。
与单作相比,轮作模式不仅能增加作物产量,还能通过改变土壤微生物群落的丰度来改善土壤环境[21]。闫非凡[22]对玉米-大豆轮作后大豆产量及土壤微生物变化进行研究,发现轮作不仅使大豆产量明显上升,还使假单胞菌门(Pseudomonadota)、放线菌门(Actinobacteria)、酸杆菌门(Acidobacteria)等土壤优势菌株的丰度显著上升。由此可见,从轮作土壤中挖掘潜力促生菌具有重要意义。本研究从轮作玉米根际土中成功筛选出5株同时具有固氮、产蛋白酶和产铁载体能力的放线菌,并进一步验证了构建合成菌群SMC的应用潜力,为开发针对轮作体系的微生物肥料奠定了菌种资源与理论基础。
节杆菌(Arthrobacter)作为PGPR家族中的重要成员,具有强大的环境适应性和代谢多样性。众多研究对其促生特性进行了深入探索。Jiang等[23]从人参根际土中分离获得菌株JI39 (A. nicotinovorans,现分类修订为Paenarthrobacter nicotinovorans),该菌株能产生高浓度吲哚-3-乙酸(13.1 μg/mL),同时具备解磷(164.2 μg/mL)、解钾(16.1 μg/mL)以及固氮的能力。Chhetri等[24]从水稻根部分离获得一株节杆菌GN70,该菌株不仅高产IAA、铁载体和生物膜,还具有促进作物生长和抗病的双重潜力。李引等[25]从花生根际土中分离获得菌株L4 (A. chlorophenolicus,现分类修订为Pseudarthrobacter chlorophenolicus),在培养24 h时其IAA产生量达135.67 μg/mL,同时具有解磷能力,可显著促进花生的生长发育,增加土壤IAA和有效磷含量。本研究从玉米根际土壤样品中筛选到的5株节杆菌,分别为波卡利水稻节杆菌(A. pokkalii) JM-18和JM-21、水稻节杆菌(A. oryzae) JM-24、A. ginsengisoli JM-47和滋养节杆菌(A. pascens) JM-48,它们均具有一定的产蛋白酶、产铁载体和固氮能力,相较于单一菌株,合成菌群在竞争激烈的根际环境中更能够发挥优势作用,本研究证实了节杆菌作为PGPR的应用潜力,为微生物肥料的开发提供了候选菌株。
不同研究报道中PGPR菌株的促生能力存在一定差异。郭英等[26]筛选获得12株促生菌,对大豆株高、根长、地上部鲜重和干重的促生效果最高可分别达到48.26%、26.33%、66.86%和66.67%。王喆琳[27]将根瘤菌Q8与假单胞菌1-4联合接种,协同增强了大豆的光合能力,与对照组相比,大豆株高、地上部干重、净光合速率及水分利用效率分别提高了23.6%、24.7%、35%和30%。在玉米促生菌的研究方面,侯莹莹等[28]分离得到3株促生菌,接种后使玉米株高、茎粗、干重和鲜重分别增长36.99%、21.32%、170.14%和102.11%。白建飞等[29]筛选的摩拉维亚假单胞菌(P. moraviensis) GF-55能够使玉米地上部株高、苗干重和苗鲜重分别增加43.47%、26.67%和82.44%。本研究中,经合成菌群SMC处理后,玉米的株高、根长、鲜重和干重分别增长了24.74%、48.21%、110.08%和64.05%;大豆的株高、根长、鲜重和干重分别增长了39.40%、93.31%、161.14%和163.57%,促生效果可达到中高等水平,这种显著的促生效果可能源于SMC内菌株间的正向互作,形成了协同增效的微生态循环。同时,相较于单一菌株,本研究开发的合成菌群理论上能够提供更稳定、更全面的促生服务,具有一定的开发应用前景。
尽管本研究验证了合成菌群SMC在种子和盆栽试验中的显著促进作用,但仍存在一定的局限性,其在大田复杂环境下的实际促生效果及稳定性仍有待验证。其次,本研究目前仅停留在表观促生效果层面,对于SMC中各菌株之间的互作机制及其促生机制仍需进一步深入研究。最后,高效菌剂是应用的基础,本研究仍需进一步开展菌剂的规模化发酵生产、剂型加工与施用技术研究,最终形成一套可操作、可推广的田间应用技术方案。
本研究从玉米-大豆轮作模式下玉米根际土壤样品中成功分离获得5株节杆菌,即波卡利水稻节杆菌(A. pokkalii) JM-18和JM-21、水稻节杆菌(A. oryzae) JM-24、A. ginsengisoli JM-47和滋养节杆菌(A. pascens) JM-48,这些菌株同时具备产蛋白酶、产铁载体、固氮、解无机磷和产NH3能力。5株节杆菌在温度为60 ℃与-20 ℃、氯化钠浓度为5.5%、pH 9.0的条件下,仍能保持较高活性。将这5株菌株复合施用后发现,其能够显著促进玉米、大豆种子的萌发以及植株的生长发育。本研究为进一步开发适用于玉米、大豆的微生物肥料,推动农业绿色可持续发展提供了新的研究方向。
  • 国家重点研发计划(2023YFD1400105)
  • 河北省高等学校科学研究项目(CXZX2026057)
  • 保定市科技计划(2494N032)
  • 国家现代农业产业技术体系(CARS-02)
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2026年第66卷第5期
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doi: 10.13343/j.cnki.wsxb.20250921
  • 接收时间:2025-12-11
  • 首发时间:2026-05-09
  • 出版时间:2026-05-04
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  • 收稿日期:2025-12-11
  • 录用日期:2026-02-03
基金
The National Key Research and Development Program of China(2023YFD1400105)
国家重点研发计划(2023YFD1400105)
The Research Project of Science and Technology in Universities of Hebei Province(CXZX2026057)
河北省高等学校科学研究项目(CXZX2026057)
The Baoding Science and Technology Planning(2494N032)
保定市科技计划(2494N032)
The National Modern Agricultural Industry Technology System(CARS-02)
国家现代农业产业技术体系(CARS-02)
作者信息
    1.河北农业大学,华北作物改良与调控国家重点实验室,河北 保定
    2.河北省植物生理与分子病理学重点实验室,河北 保定
    3.黑龙江省农垦科学院农作物开发研究所,黑龙江 佳木斯
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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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