Genetic Diversity and Growth-Promoting Functions of Endophytic Nitrogen-Fixing Bacteria in Apple

Understanding the dominant populations and biological functions of endophytic nitrogen-fixing bacteria in apple plants is of great significance for the healthy growth management and sustainable development of apple cultivation. In this study, we investigated the community diversity and potential pla...

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Main Authors: Hongshan Liu, Huan Cheng, Suwen Xu, Donghua Zhang, Jianrong Wu, Zongyan Li, Benzhong Fu, Li Liu
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Plants
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Online Access:https://www.mdpi.com/2223-7747/14/8/1235
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author Hongshan Liu
Huan Cheng
Suwen Xu
Donghua Zhang
Jianrong Wu
Zongyan Li
Benzhong Fu
Li Liu
author_facet Hongshan Liu
Huan Cheng
Suwen Xu
Donghua Zhang
Jianrong Wu
Zongyan Li
Benzhong Fu
Li Liu
author_sort Hongshan Liu
collection DOAJ
description Understanding the dominant populations and biological functions of endophytic nitrogen-fixing bacteria in apple plants is of great significance for the healthy growth management and sustainable development of apple cultivation. In this study, we investigated the community diversity and potential plant growth-promoting abilities of endophytic nitrogen-fixing bacteria in different tissues of apple trees by combining high-throughput sequencing of the <i>nifH</i> gene with traditional isolation and cultivation techniques. Sequencing results revealed that the endophytic bacteria were affiliated with 10 phyla, 14 classes, 30 orders, 42 families, and 72 genera. <i>Rhizobium</i> was the dominant genus in the roots and twigs, while <i>Desulfovibrio</i> dominated the leaf tissues. The diversity and richness of endophytic bacteria in the roots were significantly higher than those in the leaves. Using four types of nitrogen-free media, a total of 138 presumptive endophytic nitrogen-fixing bacterial strains were isolated from roots, leaves, and twigs. These isolates belonged to 32 taxonomic groups spanning 5 phyla, 8 classes, 11 orders, 13 families, and 18 genera. The <i>nifH</i> gene was successfully amplified from the representative strains of all 32 groups using specific primers. Nitrogenase activity among the isolates ranged from 26.86 to 982.28 nmol/(h·mL). Some strains also exhibited the ability to secrete indole-3-acetic acid (IAA), solubilize phosphate and potassium, and produce siderophores. Six individual strains and three microbial consortia were tested for their plant growth-promoting effects on apple tissue culture seedlings. All treatments showed growth-promoting effects to varying degrees, with the RD01+RC16 consortium showing the most significant results: plant height, number of leaves, and chlorophyll content were 2.4, 3.3, and 4.2 times higher than those of the control, respectively. These findings demonstrate the rich diversity of endophytic nitrogen-fixing bacteria in apple plants and their promising potential for application in promoting host plant growth.
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spelling doaj-art-bb3b4f924d2d4098ab71983e288f3e6e2025-08-20T02:18:15ZengMDPI AGPlants2223-77472025-04-01148123510.3390/plants14081235Genetic Diversity and Growth-Promoting Functions of Endophytic Nitrogen-Fixing Bacteria in AppleHongshan Liu0Huan Cheng1Suwen Xu2Donghua Zhang3Jianrong Wu4Zongyan Li5Benzhong Fu6Li Liu7Key Laboratory of Forest Disaster Early Warning and Control of Yunnan Provincial, Forestry College, Southwest Forestry University, Kunming 650224, ChinaKey Laboratory of Forest Disaster Early Warning and Control of Yunnan Provincial, Forestry College, Southwest Forestry University, Kunming 650224, ChinaKey Laboratory of Forest Disaster Early Warning and Control of Yunnan Provincial, Forestry College, Southwest Forestry University, Kunming 650224, ChinaKey Laboratory of Forest Disaster Early Warning and Control of Yunnan Provincial, Forestry College, Southwest Forestry University, Kunming 650224, ChinaKey Laboratory of Forest Disaster Early Warning and Control of Yunnan Provincial, Forestry College, Southwest Forestry University, Kunming 650224, ChinaLandscape Architecture and Horticulture Faculty, Southwest Forestry University, Kunming 650224, ChinaDepartment of Plant Pathology, College of Agronomy, Xinjiang Agricultural University, 311 East Nongda Road, Urumqi 830052, ChinaKey Laboratory of Forest Disaster Early Warning and Control of Yunnan Provincial, Forestry College, Southwest Forestry University, Kunming 650224, ChinaUnderstanding the dominant populations and biological functions of endophytic nitrogen-fixing bacteria in apple plants is of great significance for the healthy growth management and sustainable development of apple cultivation. In this study, we investigated the community diversity and potential plant growth-promoting abilities of endophytic nitrogen-fixing bacteria in different tissues of apple trees by combining high-throughput sequencing of the <i>nifH</i> gene with traditional isolation and cultivation techniques. Sequencing results revealed that the endophytic bacteria were affiliated with 10 phyla, 14 classes, 30 orders, 42 families, and 72 genera. <i>Rhizobium</i> was the dominant genus in the roots and twigs, while <i>Desulfovibrio</i> dominated the leaf tissues. The diversity and richness of endophytic bacteria in the roots were significantly higher than those in the leaves. Using four types of nitrogen-free media, a total of 138 presumptive endophytic nitrogen-fixing bacterial strains were isolated from roots, leaves, and twigs. These isolates belonged to 32 taxonomic groups spanning 5 phyla, 8 classes, 11 orders, 13 families, and 18 genera. The <i>nifH</i> gene was successfully amplified from the representative strains of all 32 groups using specific primers. Nitrogenase activity among the isolates ranged from 26.86 to 982.28 nmol/(h·mL). Some strains also exhibited the ability to secrete indole-3-acetic acid (IAA), solubilize phosphate and potassium, and produce siderophores. Six individual strains and three microbial consortia were tested for their plant growth-promoting effects on apple tissue culture seedlings. All treatments showed growth-promoting effects to varying degrees, with the RD01+RC16 consortium showing the most significant results: plant height, number of leaves, and chlorophyll content were 2.4, 3.3, and 4.2 times higher than those of the control, respectively. These findings demonstrate the rich diversity of endophytic nitrogen-fixing bacteria in apple plants and their promising potential for application in promoting host plant growth.https://www.mdpi.com/2223-7747/14/8/1235apple treeendophytic diazotrophsgenetic diversitynitrogenase activitygrowth-promoting
spellingShingle Hongshan Liu
Huan Cheng
Suwen Xu
Donghua Zhang
Jianrong Wu
Zongyan Li
Benzhong Fu
Li Liu
Genetic Diversity and Growth-Promoting Functions of Endophytic Nitrogen-Fixing Bacteria in Apple
Plants
apple tree
endophytic diazotrophs
genetic diversity
nitrogenase activity
growth-promoting
title Genetic Diversity and Growth-Promoting Functions of Endophytic Nitrogen-Fixing Bacteria in Apple
title_full Genetic Diversity and Growth-Promoting Functions of Endophytic Nitrogen-Fixing Bacteria in Apple
title_fullStr Genetic Diversity and Growth-Promoting Functions of Endophytic Nitrogen-Fixing Bacteria in Apple
title_full_unstemmed Genetic Diversity and Growth-Promoting Functions of Endophytic Nitrogen-Fixing Bacteria in Apple
title_short Genetic Diversity and Growth-Promoting Functions of Endophytic Nitrogen-Fixing Bacteria in Apple
title_sort genetic diversity and growth promoting functions of endophytic nitrogen fixing bacteria in apple
topic apple tree
endophytic diazotrophs
genetic diversity
nitrogenase activity
growth-promoting
url https://www.mdpi.com/2223-7747/14/8/1235
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