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Article(id=1226236834167042710, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226236828399878330, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20240803, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1733932800000, receivedDateStr=2024-12-12, revisedDate=null, revisedDateStr=null, acceptedDate=1735228800000, acceptedDateStr=2024-12-27, onlineDate=1770287243643, onlineDateStr=2026-02-05, pubDate=1746288000000, pubDateStr=2025-05-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1770287243643, onlineIssueDateStr=2026-02-05, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1770287243643, creator=13701087609, updateTime=1770287243643, updator=13701087609, issue=Issue{id=1226236828399878330, tenantId=1146029695717560320, journalId=1192105938417971205, year='2025', volume='65', issue='5', pageStart='1831', pageEnd='2319', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1770287242269, creator=13701087609, updateTime=1770344517883, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1226477059812274835, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226236828399878330, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1226477059816469140, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226236828399878330, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=2034, endPage=2048, ext={EN=ArticleExt(id=1226236834494198444, articleId=1226236834167042710, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Microbacterium aurantiacum GX14001 promotes the growth of tomato plants by altering the microbial community structure in rhizosphere soil, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

As autotoxic substances secreted by plant roots, phenolic acids such as p-hydroxybenzoic acid (PHBA), are the main factors causing continuous cropping obstacles. [Objective] To study the effects of Microbacterium aurantiacum on the growth of tomato plants and the microbial community structure in rhizosphere soil under PHBA stress. [Methods] We irrigated tomato rhizosphere with the suspension of M. aurantiacum GX14001 and then measured the growth traits of tomato plants and the soil microbial community changes in the rhizosphere soil. [Results] Under the PHBA treatment, GX14001 significantly promoted the growth of tomato plants, increasing the leaf area, stem diameter, and plant height by 244.0%, 156.5%, and 128.0%, respectively. GX14001 increased the richness but did not cause changes in the diversity of bacteria in the rhizosphere soil. Meanwhile, it decreased the richness and diversity of fungi in the rhizosphere soil. At the phylum level, compared with the control group, the GX14001 group showed increased relative abundance of Actinobacteriota, Chloroflexi, and Proteobacteria, with Ascomycetes as the dominant fungal phylum. [Conclusion] M. aurantiacum GX14001 promotes the growth of tomato plants by changing the microbial community structure in the rhizosphere soil. It increases the relative abundance of beneficial microorganisms in the soil to create a favorable environment for tomato growth.

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E-mail:
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植物根系分泌的自毒物质，如对羟基苯甲酸(p-hydroxybenzoic acid, PHBA)等酚酸类物质，是造成植物连作障碍的主要因素。【目的】为探究在PHBA胁迫下，橙色微杆菌(Microbacterium aurantiacum)对番茄生长的促生效果及其对番茄根际微生物结构的影响。【方法】将橙色微杆菌GX14001菌悬液浇灌番茄根际，测定番茄的生长发育状况及其根际土壤微生物群落的变化。【结果】在PHBA处理下，GX14001对番茄生长具有显著的促进的作用，叶片面积、茎粗和株高分别增加了244.0%，156.5%和128.0%。番茄根际土壤的细菌丰富度增加，但多样性无显著变化；真菌的丰富度和多样性显著降低。在门水平上，与对照组相比，细菌菌门中的放线菌门(Actinobacteriota)、绿弯菌门(Chloroflexi)和变形菌门(Proteobacteria)的相对丰度显著升高，子囊菌门(Ascomycota)为绝对优势真菌门。【结论】橙色微杆菌GX14001可能通过改变番茄根际土壤的微生物群落结构，提高土壤中有益微生物的相对丰度，从而为植物提供有利的生长环境，进而促进番茄的生长发育。

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作者贡献声明

吴佩佩：实验分析、数据整理、数据分析、初稿撰写和文稿修改；叶欣乐：实验分析；熊利群：实验分析；丰景：概念构思和文稿修改。

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orderNo=1, keyword=橙色微杆菌), Keyword(id=1226592756949234048, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=CN, orderNo=2, keyword=番茄), Keyword(id=1226592757096034700, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=CN, orderNo=3, keyword=根际土壤), Keyword(id=1226592757213475221, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=CN, orderNo=4, keyword=微生物群落结构)], refs=[Reference(id=1226592762334720644, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, doi=null, pmid=null, pmcid=null, year=2024, volume=51, issue=8, pageStart=2844, pageEnd=2856, url=null, language=null, rfNumber=[1], rfOrder=0, authorNames=白雪飞, 李倩, 丁丽丽, 张岱, 赵益, 杨志辉, 朱杰华, journalName=微生物学通报, refType=null, unstructuredReference=白雪飞, 李倩, 丁丽丽, 张岱, 赵益, 杨志辉, 朱杰华. 贝莱斯芽孢杆菌HN-Q-8对马铃薯根际土壤微生物群落结构及理化性质的影响[J]. 微生物学通报, 2024, 51(8): 2844-2856., 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A: Growth of potted tomatoes under normal soil and GX14001 addition treatment; B: Growth of potted tomato under PHBA and PHBA+GX14001 treatments; C: Growth of potted tomatoes under four treatments. The ruler indicates the plant height., figureFileSmall=xsq3RfBWjd5/0zfwU5ZvQQ==, figureFileBig=POiN0FzPwr7dJ6NyCRLyVQ==, tableContent=null), ArticleFig(id=1226592757611934131, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=CN, label=图1, caption=菌株GX14001缓解PHBA对番茄的毒害。A：正常土壤及添加GX14001处理下盆栽番茄的生长状况；B：PHBA及PHBA+GX14001处理下盆栽番茄的生长状况；C：4种处理下盆栽番茄的生长状况。标尺表示株高。, figureFileSmall=xsq3RfBWjd5/0zfwU5ZvQQ==, figureFileBig=POiN0FzPwr7dJ6NyCRLyVQ==, tableContent=null), ArticleFig(id=1226592757792289218, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=EN, label=Figure 2, caption=Effects of GX14001 and exogenous PHBA treatments on tomato growth characteristics. A: Comparison of tomato leaf size under different treatments; B: Comparison of stem thickness of tomato under different treatments; C: Comparison of plant height of tomato under different treatments. *: P<0.05., figureFileSmall=2aFU82MPQKVCSLb4KXFAhA==, figureFileBig=W9cxer6iJXXxgd5mH58WKg==, tableContent=null), ArticleFig(id=1226592757897146823, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=CN, label=图2, caption=GX14001及外源PHBA处理对番茄生长特征的影响。A：不同处理下番茄叶片大小的比较；B：不同处理下番茄茎粗的比较；C：不同处理下番茄株高的比较。, figureFileSmall=2aFU82MPQKVCSLb4KXFAhA==, figureFileBig=W9cxer6iJXXxgd5mH58WKg==, tableContent=null), ArticleFig(id=1226592758048141780, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=EN, label=Figure 3, caption=Effects of GX14001 and PHBA on tomato leaf activity indexes. A: Comparison of SOD activity in tomato leaves; B: Comparison of CAT activity in tomato leaves; C: Comparison of POD activity in tomato leaves. *: P<0.05; **: P<0.01., figureFileSmall=slutBQrBQZFgoIrpzTcuwA==, figureFileBig=v9YcF8EU4KsxTDMwmChxGw==, tableContent=null), ArticleFig(id=1226592759423873496, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=CN, label=图3, caption=GX14001和PHBA对番茄叶片活性指标的影响。A：番茄叶片SOD活性的比较；B：番茄叶片CAT活性的比较；C：番茄叶片POD活性的比较。, figureFileSmall=slutBQrBQZFgoIrpzTcuwA==, figureFileBig=v9YcF8EU4KsxTDMwmChxGw==, tableContent=null), ArticleFig(id=1226592759587451363, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=EN, label=Figure 4, caption=Venn diagram at the genus level of soil bacteria (A) and fungi (B) in the rhizosphere of tomato., figureFileSmall=mPy4BvFs5/2KHYMcctr27A==, figureFileBig=8DlQM6Jza74jcyQtRSkNQw==, tableContent=null), ArticleFig(id=1226592759704891882, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=CN, label=图4, caption=番茄根际土壤样品细菌(A)和真菌(B)属水平韦恩图, figureFileSmall=mPy4BvFs5/2KHYMcctr27A==, figureFileBig=8DlQM6Jza74jcyQtRSkNQw==, tableContent=null), ArticleFig(id=1226592759818138098, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=EN, label=Figure 5, caption=Analysis of alpha diversity of soil microbial communities in tomato rhizosphere. A: ACE index of bacteria; B: Shannon index of bacteria; C: ACE index of fungi; D: Shannon index of fungi. *: P<0.05; **: P<0.01., figureFileSmall=rN4LwI+OwPVaxn9K+USFTg==, figureFileBig=5S0fg9ypQWxJF/kGpazelQ==, tableContent=null), ArticleFig(id=1226592759897829885, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=CN, label=图5, caption=番茄根际土壤微生物群落的α多样性分析。A：细菌的ACE指数；B：细菌的Shannon指数；C：真菌的ACE指数；D：真菌的Shannon指数。, figureFileSmall=rN4LwI+OwPVaxn9K+USFTg==, figureFileBig=5S0fg9ypQWxJF/kGpazelQ==, tableContent=null), ArticleFig(id=1226592760002687493, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=EN, label=Figure 6, caption=Main coordinates (PCoA) diagram of the soil microbial community in tomato rhizosphere. A: Bacterial PCoA diagram; B: Fungal PCoA plot., figureFileSmall=VQHztIHTpoc4e+aWNKfqbA==, figureFileBig=vYxecNvgr6OuFpMi8RAGPA==, tableContent=null), ArticleFig(id=1226592760103350798, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=CN, label=图6, caption=番茄根际土壤微生物群落的主坐标(PCoA)图。A：细菌PCoA图；B：真菌PCoA图。, figureFileSmall=VQHztIHTpoc4e+aWNKfqbA==, figureFileBig=vYxecNvgr6OuFpMi8RAGPA==, tableContent=null), ArticleFig(id=1226592760224985626, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=EN, label=Figure 7, caption=Linear discriminant analysis (LDA) value distribution of soil microbial communities in tomato rhizosphere. A: Bacterial LDA value distribution map; B: Distribution of LDA values in fungi., figureFileSmall=xEuPC3DJkZj2JrvxcS1K7g==, figureFileBig=7ELo+h3TYXAfq0b/Sd3trA==, tableContent=null), ArticleFig(id=1226592760346620451, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=CN, label=图7, caption=番茄根际土壤微生物群落线性判别分析(LDA)值分布图。A：细菌LDA值分布图；B：真菌LDA值分布图。, figureFileSmall=xEuPC3DJkZj2JrvxcS1K7g==, figureFileBig=7ELo+h3TYXAfq0b/Sd3trA==, tableContent=null), ArticleFig(id=1226592760526975529, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=EN, label=Figure 8, caption=Composition of soil microbial phylum at tomato rhizosphere. A: Composition of soil bacterial communities; B: Composition of soil fungal communities., figureFileSmall=pomctDfbiO63En9+zz7W+w==, figureFileBig=E9yvepKfK9Z7CH21oABgnw==, tableContent=null), ArticleFig(id=1226592760623444526, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=CN, label=图8, caption=番茄根际土壤微生物门水平群落组成。A：土壤细菌群落的构成；B：土壤真菌群落的构成。, figureFileSmall=pomctDfbiO63En9+zz7W+w==, figureFileBig=E9yvepKfK9Z7CH21oABgnw==, tableContent=null), ArticleFig(id=1226592760799605300, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=EN, label=Figure 9, caption=Tomato rhizosphere soil microbial composition is composed of horizontal communities. A: Composition of soil bacterial communities; B: Composition of soil fungal communities., figureFileSmall=eEmQUtT6ib1t+2fcwhDPGQ==, figureFileBig=MxorX0MbPOcl/uwaM/FnFQ==, tableContent=null), ArticleFig(id=1226592760954794557, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=CN, label=图9, caption=番茄根际土壤微生物属水平群落组成。A：土壤细菌群落的构成；B：土壤真菌群落的构成。, figureFileSmall=eEmQUtT6ib1t+2fcwhDPGQ==, figureFileBig=MxorX0MbPOcl/uwaM/FnFQ==, tableContent=null), ArticleFig(id=1226592761101595205, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=EN, label=Table 1, caption=

Statistics of effective sequences

, figureFileSmall=null, figureFileBig=null, tableContent=

样品

Samples

序列数

Number of sequences

碱基

Bases (bp)

平均序列长度

Average sequence length (bp)

细菌

Bacteria

真菌

Fungi

细菌

Bacteria

真菌

Fungi

细菌

Bacteria

真菌

Fungi

CG2-136 56035 00115 202 2498 429 994415.82240.85
CG2-241 49435 46017 131 2318 509 542412.86239.98
CG2-333 67340 68813 895 2039 866 108412.65242.48
OG-138 16633 22815 899 8057 877 402416.60237.07
OG-233 94037 21314 111 0888 813 576415.77236.84
OG-333 87830 06514 110 0787 157 468416.50238.07
TG1-134 82841 01514 345 61810 021 173411.90244.33
TG1-236 08338 50014 911 0229 340 983413.24242.62
TG1-341 64134 37417 252 1238 466 107414.31246.29
), ArticleFig(id=1226592761227424328, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226236834167042710, language=CN, label=表1, caption=

有效序列统计

, figureFileSmall=null, figureFileBig=null, tableContent=

样品

Samples

序列数

Number of sequences

碱基

Bases (bp)

平均序列长度

Average sequence length (bp)

细菌

Bacteria

真菌

Fungi

细菌

Bacteria

真菌

Fungi

细菌

Bacteria

真菌

Fungi

CG2-136 56035 00115 202 2498 429 994415.82240.85
CG2-241 49435 46017 131 2318 509 542412.86239.98
CG2-333 67340 68813 895 2039 866 108412.65242.48
OG-138 16633 22815 899 8057 877 402416.60237.07
OG-233 94037 21314 111 0888 813 576415.77236.84
OG-333 87830 06514 110 0787 157 468416.50238.07
TG1-134 82841 01514 345 61810 021 173411.90244.33
TG1-236 08338 50014 911 0229 340 983413.24242.62
TG1-341 64134 37417 252 1238 466 107414.31246.29
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橙色微杆菌GX14001对番茄的促生及根际土壤微生物群落结构的影响
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吴佩佩 1 , 叶欣乐 2 , 熊利群 2 , 丰景 1
微生物学报 | 研究报告 2025,65(5): 2034-2048
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微生物学报 | 研究报告 2025, 65(5): 2034-2048
橙色微杆菌GX14001对番茄的促生及根际土壤微生物群落结构的影响
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吴佩佩1, 叶欣乐2, 熊利群2, 丰景1
作者信息
  • 1.广西民族大学 海洋与生物技术学院，广西多糖材料与改性重点实验室，广西 南宁
  • 2.广西民族大学 海洋与生物技术学院 广西 南宁
Microbacterium aurantiacum GX14001 promotes the growth of tomato plants by altering the microbial community structure in rhizosphere soil
Peipei WU1, Xinle YE2, Liqun XIONG2, Jing FENG1
Affiliations
  • 1.Guangxi Key Laboratory for Polysaccharide Materials and Modification, School of Marine Sciences and Biotechnology, Guangxi Minzu University, Nanning, Guangxi, China
  • 2.School of Marine Sciences and Biotechnology, Guangxi Minzu University, Nanning, Guangxi, China
出版时间: 2025-05-04 doi: 10.13343/j.cnki.wsxb.20240803
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摘要
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植物根系分泌的自毒物质，如对羟基苯甲酸(p-hydroxybenzoic acid, PHBA)等酚酸类物质，是造成植物连作障碍的主要因素。【目的】为探究在PHBA胁迫下，橙色微杆菌(Microbacterium aurantiacum)对番茄生长的促生效果及其对番茄根际微生物结构的影响。【方法】将橙色微杆菌GX14001菌悬液浇灌番茄根际，测定番茄的生长发育状况及其根际土壤微生物群落的变化。【结果】在PHBA处理下，GX14001对番茄生长具有显著的促进的作用，叶片面积、茎粗和株高分别增加了244.0%，156.5%和128.0%。番茄根际土壤的细菌丰富度增加，但多样性无显著变化；真菌的丰富度和多样性显著降低。在门水平上，与对照组相比，细菌菌门中的放线菌门(Actinobacteriota)、绿弯菌门(Chloroflexi)和变形菌门(Proteobacteria)的相对丰度显著升高，子囊菌门(Ascomycota)为绝对优势真菌门。【结论】橙色微杆菌GX14001可能通过改变番茄根际土壤的微生物群落结构，提高土壤中有益微生物的相对丰度，从而为植物提供有利的生长环境，进而促进番茄的生长发育。

橙色微杆菌  /  番茄  /  根际土壤  /  微生物群落结构

As autotoxic substances secreted by plant roots, phenolic acids such as p-hydroxybenzoic acid (PHBA), are the main factors causing continuous cropping obstacles. [Objective] To study the effects of Microbacterium aurantiacum on the growth of tomato plants and the microbial community structure in rhizosphere soil under PHBA stress. [Methods] We irrigated tomato rhizosphere with the suspension of M. aurantiacum GX14001 and then measured the growth traits of tomato plants and the soil microbial community changes in the rhizosphere soil. [Results] Under the PHBA treatment, GX14001 significantly promoted the growth of tomato plants, increasing the leaf area, stem diameter, and plant height by 244.0%, 156.5%, and 128.0%, respectively. GX14001 increased the richness but did not cause changes in the diversity of bacteria in the rhizosphere soil. Meanwhile, it decreased the richness and diversity of fungi in the rhizosphere soil. At the phylum level, compared with the control group, the GX14001 group showed increased relative abundance of Actinobacteriota, Chloroflexi, and Proteobacteria, with Ascomycetes as the dominant fungal phylum. [Conclusion] M. aurantiacum GX14001 promotes the growth of tomato plants by changing the microbial community structure in the rhizosphere soil. It increases the relative abundance of beneficial microorganisms in the soil to create a favorable environment for tomato growth.

Microbacterium aurantiacum  /  tomato  /  rhizosphere soil  /  microbial community structure
吴佩佩, 叶欣乐, 熊利群, 丰景. 橙色微杆菌GX14001对番茄的促生及根际土壤微生物群落结构的影响. 微生物学报, 2025 , 65 (5) : 2034 -2048 . DOI: 10.13343/j.cnki.wsxb.20240803
Peipei WU, Xinle YE, Liqun XIONG, Jing FENG. Microbacterium aurantiacum GX14001 promotes the growth of tomato plants by altering the microbial community structure in rhizosphere soil[J]. Acta Microbiologica Sinica, 2025 , 65 (5) : 2034 -2048 . DOI: 10.13343/j.cnki.wsxb.20240803
正文
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番茄(Solanum lycopersicum L.)作为茄科的一员，是一种富含多种维生素的草本植物，具有较高的营养价值，在中国农业中占据重要地位。近年来，番茄栽培面积在农业产业结构转型升级的同时大幅提高。然而，为了追求更高的产量，一些农户倾向于大量施用化肥。这种做法虽然能在短期内促进番茄生长，但长期来看会引发一系列环境问题，如土壤酸化、硬化及土壤生物活性降低等。因此，减少对化学肥料的依赖，采用更绿色、科学的方法来提高番茄产量，对农业可持续发展具有重要意义。
在植物的整个生命周期中，根系起着关键作用。作为植物与土壤直接交互的部分，根系不仅负责从土壤中吸收并转运养分[1]，还能通过其分泌物为土壤中的微生物提供必要的营养成分。此外，植物根系还会释放一些低分子量的有机化合物，如酚类和萜类物质，从而改变根际微生物群落结构。对羟基苯甲酸(p-hydroxybenzoic acid, PHBA)是一种常见的酚酸类自毒物质，广泛存在于植物根部分泌物中，其积累会对多种作物产生负面影响，包括降低土壤肥力、抑制作物生长发育，还可能通过改变土壤微生物群落结构进一步加剧连作障碍。自然界中存在多种降解酚酸类物质的菌株，利用微生物降解是一种安全且环保的方法，能够有效降低农作物生长环境中自毒物质含量，促进农业可持续发展[2]。王丹丹等[3]从大豆根际土壤样品中筛选出6株具有PHBA降解及利用能力的根际促生菌，其中假交替单胞菌(Pseudoalteromonas agarivorans) H3和待定布罗达氏阶梯烷菌(Candidatus Scalindua brodae) PA17不仅能降解PHBA，还能促进大豆种子萌发及幼苗生长。张锋等[4]研究发现，不同蔬菜品种(如‘京茄9号’ ‘超人’ ‘京旋1号’)接种解淀粉芽孢杆菌(Bacillus amyloliquefaciens) L-S60、LPL-117、L-H15后，蔬菜幼苗胚轴及根际微生物群落多样性发生了显著变化，表明促生菌能够改变蔬菜穴盘苗的根际微生物群落结构和组成。以上研究表明，具有PHBA降解能力的菌株不仅能促进植物生长，还能影响土壤微生物的多样性和群落结构。然而，目前针对橙色微杆菌(Microbacterium aurantiacum)如何影响番茄连作条件下根际微生物多样性的系统性研究仍显不足。
本研究使用橙色微杆菌GX14001菌剂对番茄进行灌根处理，发现其具有良好的促生效果。影响植物生长的因素很多，其中植物根际土壤微生物是重要因素之一，但针对橙色微杆菌对番茄促生及根际土壤微生物影响的相关报道较为罕见[5]。因此，本研究利用橙色微杆菌GX14001菌剂对番茄进行灌根处理，并采用新一代测序技术对番茄根际土壤细菌和真菌进行测定[5]，对比分析根际土壤微生物群落的变化规律，旨在揭示橙色微杆菌促进番茄生长的具体机制，并为解决因连作而导致的番茄生产问题提供科学指导。
1 材料与方法
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1.1 材料
1.1.1 菌株和盆栽材料
橙色微杆菌由实验室前期从海洋及海岸红树林土壤中筛选分离得到，并命名为GX14001[6]。
供试番茄品种为华煜贵妃樱桃番茄，其育苗过程采用的基质由南宁桂裕鑫农业科技有限公司提供[7]。
1.1.2 主要试剂和仪器
对羟基苯甲酸，上海阿拉丁生化科技股份有限公司。
超净工作台，上海智诚分析仪器制造有限公司；全自动立式高压灭菌锅，致微(厦门)仪器有限公司；生化培养箱，上海博迅医疗生物仪器股份有限公司；摇床，上海旻泉仪器有限公司；酶标仪，伯腾仪器有限公司[8]。
1.2 番茄根际PHBA胁迫盆栽试验
GX14001菌剂为接种于LB肉汤培养基中，并置于30 ℃、180 r/min恒温摇床中培养3 d的菌液。取5 mL菌悬液于8 000 r/min离心2 min后，用5 mL无菌水重悬菌体，稀释至浓度为1×109 CFU/mL的菌剂。配制不同处理的盆栽，包括将PHBA与土壤充分混合，配制含1% PHBA的育苗盆栽和不添加PHBA的正常土壤。每盆种植3株番茄。当番茄生长出2片真叶时，保留每个盆栽中大小及生长状况较为一致的一株番茄。试验包括4个处理，每个处理设3个平行。OG组：正常土壤种植番茄；GX14001组：正常土壤中接种5 mL GX14001菌剂后种植番茄；TG1组：添加1% PHBA后土壤中种植番茄；CG2组：添加1% PHBA后土壤中接种5 mL GX14001菌剂后种植番茄[9]。所有植物在温度控制为25 ℃、光照周期为12 h/12 h (日/夜)的条件下生长[10]。培养25 d后，观察番茄生长情况，测量叶片长度和宽度、株高和茎粗，并测定各组番茄叶片中的超氧化物歧化酶(superoxide dismutase, SOD)、过氧化物酶(peroxidase, POD)和过氧化氢酶(catalase, CAT)等抗氧化酶活性[11]。采集OG组、TG1组和CG2组3个处理下番茄根际土壤样本进行后续处理。
1.3 样品处理
采集3个处理的番茄根际土壤样本，每个样本进行3次重复。使用土壤基因组DNA提取试剂盒，从土壤中提取细菌和真菌的总DNA。具体操作步骤如下：取1 g土壤样品，加入1 mL PBS缓冲液、0.5 mL溶菌酶溶液及20 μL蛋白酶K，在37 ℃水浴保温2 h；添加100 μL含有5 mol/L NaCl和10% CTAB的溶液I，继续在65 ℃下加热10 min；之后通过12000 r/min离心8 min去除沉淀物，并利用酚-氯仿法进行抽提，最终以乙醇沉淀方式获得DNA产物，再用 100 μL无菌水溶解。随后将样品送至上海凌恩生物科技有限公司，通过Illumina PE250测序平台进行高通量测序。PCR扩增体系(20 μL)：5×FastPfu Buffer 4 μL，dNTPs (2.5 mmol/L) 2 μL，上、下游引物(10 µmol/L)各0.8 μL，FastPfu聚合酶(5 U/μL) 0.4 μL，模板DNA 10 ng，ddH2O补足20 μL。PCR反应程序：95 ℃预变性 5 min；95 ℃变性30 s，58 ℃退火30 s，72 ℃延伸2 min，35个循环；72 ℃终延伸10 min。细菌16S rRNA基因所用引物为341F (5′-CCTACGG GNGGCWGCAG-3′)和806R (5′- GGACTACHV GGGTWTCTAAT-3′)，真菌内部转录间隔序列(internal transcribed spacer, ITS) rDNA所用引物为ITS1F (5′-CTTGGTCATTTA GAGGAAGTAA-3′)和ITS2R (5′-GCTGCGTTC TTCATCGATGC-3′)[12]。
1.4 生物信息学统计及数据分析
使用Usearch v10软件对有效序列进行聚类，构建操作分类单元(operational taxonomic unit, OTU)[12]，并通过RDP分类器与UNITE数据库(https://unite.ut.ee)完成物种鉴定。土壤α多样性指数的计算通过Mothur v1.45.3软件实现，而OTU序列的β多样性分析则借助MUSCLE 3.8.31软件完成。此外，通过R语言(v3.31)完成对韦恩图绘制和线性判别分析效应量(linear discriminant analysis effect size, LEfSe)分析。通过SPSS 22.0和GraphPad Prism 10软件进行作图比较[13]。
2 结果与分析
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2.1 GX14001对外源PHBA处理下番茄生长的影响
如图1所示，在盆栽种植25 d时，仅使用GX14001处理的番茄植株生长状态明显优于其他处理组。通过单因素均值比较分析显示，处理组与空白对照组之间的差异达到显著水平，表明GX14001的应用能够促进番茄的成长，具体体现在叶片面积、茎粗和株高方面，与空白对照组相比，GX14001处理组的叶片面积增加了173.0%，茎粗增加了150.0%，株高增加了131.0% (图2)。此外，当同时施加外源GX14001和PHBA时，与仅用PHBA处理的组相比，植物叶面积、茎粗和株高均显著提高，分别增加了244.0%、156.6%和128.0%。
2.2 GX14001对外源PHBA处理下番茄叶片抗氧化酶及相关指标的影响
如图3所示，在番茄处理生长25 d时，GX14001处理组的SOD、CAT和POD活性显著高于正常土壤处理组及GX14001+PHBA处理组。相比之下，PHBA处理组的叶片抗氧化酶活性明显低于对照组，而经过GX14001灌根处理的组中，番茄叶片的抗氧化酶活性有所提升。
2.3 微生物群落有效序列及Venn分析
经过高通量测序和数据优化处理，分别获得了330 263条细菌序列和325 544条真菌序列。其中，细菌序列和真菌序列的平均长度分别为414 bp和240 bp。各组样品测序数据情况见表1。基于97%的序列相似度进行聚类分析后，共鉴定出3 758个细菌OTUs和511个真菌OTUs。每个样本的OTUs覆盖率均超过98%，表明本研究的检测结果涵盖了土壤中绝大多数的细菌和真菌，测序深度较为适宜。
利用韦恩图对土壤中细菌和真菌群落的组成进行了分析。结果显示，在正常土壤(OG)、PHBA处理土壤(TG1)和PHBA+GX14001处理土壤(CG2)中，分别检测到2 257、2 646和2 516个OTUs。三者共有OTUs为1 330个，而各自特有的OTUs分别为647、414和366个，分别占各组样品总OTUs的17.2%、11.0%和9.74%。真菌OTUs的数量分别为429、355和406个，它们之间的共有OTUs为261个。此外，每个样本独有的真菌OTU数量分别为43、13和37个，分别占各自总数的8.41%、2.54%和7.24% (图4)。上述结果表明，不同类型的土壤样本中细菌和真菌群落的组成存在一定差异，尤其是在PHBA处理后的土壤中，这种差异在真菌群落中表现得尤为明显。
2.4 微生物群落的α多样性分析
α多样性分析是评估微生物群落丰富度和多样性特征的重要工具。如图5所示，对于细菌群落，TG1组和CG2组的ACE指数分别是OG组的1.25倍(P<0.01)和1.22倍(P<0.05)，表明这2组实验中的细菌种类相对更加丰富。对于真菌群落，TG1组和CG2组的ACE指数分别是OG组的84.2% (P>0.05)和90.4% (P>0.05)。研究表明，番茄连作土壤中细菌群落的物种丰度有所提升，但真菌群落的物种丰度呈现下降趋势。进一步基于Shannon指数的分析显示，细菌群落方面，各处理之间的差异未达到显著水平；然而，OG组的真菌多样性显著高于TG1组(P>0.05)和CG2组(P>0.05)。番茄连作对细菌多样性无显著影响，但显著降低了真菌群落的多样性。
2.5 微生物群落的β多样性分析
PCoA是一种重要的β多样性分析方法，用于评估样本间群落结构的异同[13]。研究表明，番茄根际土壤中细菌和真菌群落的总变异程度分别为56%和76%。如图6所示，在细菌群落中，TG1组和CG2组在PCoA1轴上与OG组表现出明显的分离趋势；在真菌群落中，OG组与TG1组和CG2组在PC1轴上同样存在显著差异。PCoA图上样本间的距离表示样本物种组成之间的差异大小，距离越远物种组成差异越显著。因此，上述结果表明，连作种植是影响番茄根际土壤微生物群落的重要因素[14]。
2.6 不同组分间土壤微生物群落的LEfSe分析
LEfSe分析是一种结合统计检验与差异显著性检测的方法，采用线性判别分析(linear discriminant analysis, LDA)来衡量不同组分内物种丰度对整体差异的贡献程度。以LDA≥2作为临界值，在番茄连作种植土壤系统中，细菌和真菌群落表现出显著差异。具体而言，细菌群落包括2门8目13科2属，真菌群落则有3纲6目6科，表现出显著差异(P<0.05)，见图7。在属水平上，与另外2种样品相比，TG1组中有41个细菌属显著存在，其中LDA评分最高的3个差异细菌属分别为红杆菌目(Rhodobacterales)、水道球菌属(Amaricoccus)和多囊菌纲(Polyangia)；同时存在显著差异的真菌属有14个，评分前3的分别为散囊菌目(Eurotiomycetes)、曲霉科(Aspergillaceae)未知属和青霉菌属(Penicillium)。这表明在TG1组中，细菌红杆菌目(Rhodobacterales)和真菌散囊菌目(Eurotiomycetes)发挥着重要作用。在OG组中，与其他2种样品相比，有15个细菌属显著存在，其中LDA评分最高的差异细菌属为伯克氏菌科(Burkholderiaceae)未知属；显著存在的真菌有17个，评分最高的是丛赤壳属(Nectriaceae)。在CG2组中，与另外2种样品相比，有8个显著存在的细菌属，其中LDA评分排名前3位的差异细菌属分别为Tahibacter、短杆菌属(Brevibacterium)和假单胞菌属(Pseudomonas)；与其他2种样品相比，显著存在的真菌有6个，LDA评分排名最高的为刺盾炱目(Chaetothyriales)。
2.7 番茄根际土壤微生物群落在门水平的丰度
在门分类水平上，TG1组、CG2组和OG组的根际微生物群落展现出一定程度的相似性，见图8。这些群落中，丰度较高的细菌门包括拟杆菌门(Bacteroidota)、酸杆菌门(Acidobacteriota)、绿弯菌门(Chloroflexi)、糖菌门(Patescibacteria)、放线菌门(Actinobacteriota)以及变形菌门(Proteobacteria)[15]，它们的平均相对丰度依次为4.11%、6.39%、11.34%、5.36%、20.33%和44.46%。子囊菌门(Ascomycota)在真菌群落中占据主导地位，其平均相对丰度为97.06%。TG1、CG2和OG组根际微生物群落组成存在一定差异。在TG1样品中，担子菌门(Basidiomycota)相对丰度较为显著；在OG中，毛霉门(Mucoromycota)丰度最高；在CG2中，担子菌门(Basidiomycota)和毛霉门(Mucoromycota)的相对丰度则明显低于OG和TG1。这表明GX14001菌液在CG2根际土壤中对真菌群落具有一定影响作用。
2.8 番茄根际土壤微生物群落在属水平的丰度
在属水平上，3种土壤样品中微生物群落相对丰度靠前的3个细菌属分别为嗜甲基菌属(Methylophilus)、TM7菌属(TM7a)和甲基菌属(Methylobaciillus)，其平均相对丰度分别为4.20%、3.58%和3.04%；丰度最高的真菌属是镰孢菌属(Fusarium)，平均相对丰度为33.44%，其次是青霉属(Penicillium)平均相对丰度为22.81%，腐质霉属(Humicola)平均相对丰度为12.54%。
利用高通量测序技术，最终共检测到细菌属753个和真菌属235个。在不同处理条件下，有10个细菌属和10个真菌属表现出显著差异(图9)。细菌属中，其相对丰度主要体现在OG与TG1之间的变化，部分变化出现在OG与CG2之间。真菌属的差异物种相对丰度变化也是如此。上述结果表明，番茄连作种植对土壤根际微生物群落的组成有显著影响[16]。
3 讨论与结论
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根际土壤微生物主要分布在植物根系周围，对作物生长及病害防控具有重要作用，并在生态系统中发挥着关键作用[17]。本研究发现，橙色微杆菌不仅能够促进番茄生长，还能有效降解连作土壤中的自毒物质PHBA，并改变根际土壤微生物群落结构。因此，通过分析根际土壤微生物群落的变化，有助于揭示连作对作物生长和品质的影响机制[18]。已有大量研究表明，连作障碍问题与土壤微生物群落多样性的变化息息相关。孙文庆等[18]研究了不同连作年份加工番茄根际微生物组成与多样性变化，发现随着连作时间的增长，根际土壤中细菌的物种丰富度及Shannon-Wiener指数均呈现下降趋势，而真菌的丰度与多样性则显著上升。王丹丹等[19]针对花生长期连作问题进行了研究，筛选出7株高效降解菌株，这些菌株不仅降解酚酸类物质，还能产生吲哚乙酸、固氮分解磷(包括有机磷和无机磷)以及溶解钾。特别是Bacillus sp. B28，该菌株显著减轻了酚酸类物质对花生种子萌发的抑制效应，并极大地促进了幼苗生长[20]。吴凤慧[21]研究了GLY-P2菌株在降低土壤中酚酸含量、分泌ACC脱氨酶等有益物质方面的作用机制，结果表明GLY-P2不仅能够降低FA和PHBA的浓度，还可以激活植物的抗氧化酶系统，增强土壤酶的活性，改善土壤的理化性质及微生物群落结构，从而有效缓解FA和PHBA对黄瓜生长的不利影响。本研究则聚焦于GX14001菌剂对番茄生长及连作根际土壤微生物群落的影响，探讨其对番茄生长发育的作用机制。
本研究选取了施用橙色微杆菌GX14001菌剂后的番茄植株及其根际土壤作为研究对象，比较了不同处理下番茄的生长状况，并结合高通量测序分析了正常土壤、PHBA处理和PHBA+GX14001处理组的根际土壤微生物群落变化规律。结果表明，GX14001菌剂显著促进了番茄生长，具体体现在叶片面积、茎粗、株高及叶片活性等方面。植物叶片的生理指标对其生理状态具有重要影响，进而影响植物的品质。研究表明，植物体内的渗透调节物质、膜脂过氧化产物和抗氧化酶活性与植物的抗逆性和品质密切相关。超氧化物歧化酶、过氧化氢酶等抗氧化酶在植物应对逆境胁迫和适应环境变化中起着关键作用，能够有效平衡体内的活性氧含量，维持植物的活性氧稳态。施用微生物菌肥能够调节植物的生理指标，从而保持植物内部环境的相对稳定，促进次生代谢成分的合成，进而提升植物的生长和品质[22]。本研究中施用GX14001菌液后，番茄叶片中SOD、CAT和POD的活性显著增加，这不仅增强了番茄的抗逆性，还促进了其生长发育，提高了番茄的品质。
添加微生物菌液可在不同程度上改变土壤微生物群落结构，其改变程度受土壤理化性质和微生态的影响[22]。在本研究中，放线菌门、绿弯菌门和变形菌门是3种处理土壤样品中的主要细菌群落，并且与土壤pH呈正相关。绿弯菌门是土壤中的优势菌门，能够降解有机物，参与碳循环，从而促进土壤健康。研究发现，经过PHBA处理的土壤中绿弯菌门和酸杆菌门相对丰度降低，与土壤EC、AN、AK呈正相关关系，这表明植物连作导致的化感酚酸类自毒物质在一定程度上降低了土壤质量。放线菌门在土壤中起到多种作用，多与病原体拮抗相关，对维护土壤生态平衡具有一定作用，存在于所有样品中，且相对丰度无明显差异。变形菌门平均相对丰度达到44.46%，是最具优势的细菌门，在土壤细菌群落中占据主导地位，这可能与该门内其他细菌共同参与植物的生长和代谢过程，从而促进植物生长有关[23]。子囊菌门是植物根际土壤中的优势真菌门，平均相对丰度达到97.06%。该菌门在土壤有机酸降解及动植物残体分解中具有重要作用。与TG1相比，CG2组土壤中子囊菌门相对丰度显著上升，说明GX14001菌液对连作土壤中PHBA具有一定的降解作用，从而缓解了根际土壤微生物真菌群落的变化，促进了植物生长。
本研究对3种不同处理的土壤(TG1、CG2及OG组)进行了微生物群落组成的比较与分析。结果显示，CG2与OG在细菌群落的组成及相对丰度上更为相似。尽管TG1和CG2的根际土壤中细菌种类数量有所增加，但其多样性水平并未出现显著差异。与此同时，真菌群落无论是在丰富度还是多样性上都出现了明显的下降趋势。特别是经过PHBA处理的2组土壤样本，观察到真菌群落丰富度和多样性的显著降低，这与卢雨欣等[23]对连作芹菜根际土壤微生物多样性的研究结果一致。由此可以推断，连续种植同一作物可能引起真菌群落数量减少及组成变化，这或许构成了连作障碍的一个关键因素。
PHBA是连作障碍土壤中积累的主要自毒化感物质之一，严重影响了作物的正常发育，成为农业可持续发展面临的一大挑战[24]。橙色微杆菌GX14001能够促进番茄生长，分解连作土壤中的PHBA，降低土壤中真菌的丰富度，并改变番茄根际土壤的微生物群落结构。已有研究表明，利用功能性的微生物制剂和生物肥料可以调节植物与土壤微生物间的相互作用，从而有助于缓解土壤连作带来的障碍。因此，持续寻找既能促进植物生长又能降解酚酸类化合物的有效菌株，并深入探讨连作障碍背后的作用机理仍具有重要意义。
基金
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  • 国家自然科学基金(32360024)
  • 广西壮族自治区自然科学基金(2023GXNSFBA026041)
  • 广西民族大学相思湖青年学者创新团队(2023GXUNXSHQN03)
文献
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2025年第65卷第5期
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doi: 10.13343/j.cnki.wsxb.20240803
  • 接收时间：2024-12-12
  • 首发时间：2026-02-05
  • 出版时间：2025-05-04
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  • 收稿日期：2024-12-12
  • 录用日期：2024-12-27
基金
National Natural Science Foundation of China(32360024)
国家自然科学基金(32360024)
Natural Science Foundation of Guangxi Zhuang Autonomous Region(2023GXNSFBA026041)
广西壮族自治区自然科学基金(2023GXNSFBA026041)
Xiangsi Lake Youth Scholar Innovation Team of Guangxi Minzu University(2023GXUNXSHQN03)
广西民族大学相思湖青年学者创新团队(2023GXUNXSHQN03)
作者信息
    1.广西民族大学 海洋与生物技术学院，广西多糖材料与改性重点实验室，广西 南宁
    2.广西民族大学 海洋与生物技术学院 广西 南宁
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https://castjournals.cast.org.cn/joweb/wswxb/CN/10.13343/j.cnki.wsxb.20240803
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
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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