Diversity Of Endophytic Bacterial Communities Research Articles

Diversity of endophytic bacterial community in rice roots and their roles in phosphate solubilization and plant growth

Root endophytic bacteria have significant beneficial effects on plant growth and health, while specific coexisting pathogenic bacteria can cause various diseases. In this study, we assessed the diversity and cultivability of the endophytic bacterial community in rice roots with a focus on their capacity for phosphate solubilization and impact on plant growth. Employing PacBio Single Molecule Real-Time (SMRT) and Sanger sequencing, we generated 16S rRNA gene libraries using metagenomic DNA extracted from the roots of three field-grown rice plants, along with genomic DNA from 156 associated bacterial isolates. QIIME 2-based microbiome analysis revealed dominant classified species at diverse taxonomic levels, including Arcobacter marinus, Paraburkholderia kururiensis, Lactobacillus plantarum, and Lactobacillus brevis. Furthermore, we effectively isolated 25.97% of rice root endophytic bacteria, with a notable 33.33% of pure culture endophytic bacteria exhibiting phosphate solubilization abilities. Finally, we revealed the inhibitory role of Burkholderia vietnamiensis and the growth promoting role of Paraburkholderia kururiensis subsp. Kururiensis in both the monocot model Brachypodium distachyon and the dicot Arabidopsis thaliana. Our study provided novel insight into the microbiome structure in rice roots and its potential to benefit rice cultivation and enhance overall crop health.

Endophytic bacterial community diversity in genetically related hybrid rice seeds.

The Food and Agriculture Organization of the United Nations (FAO) has identified hybrid rice as ideal for addressing food scarcity in poor nations. A comprehensive investigation of the endophytic bacteria in hybrid rice seeds is essential from a microecological perspective to illuminate the mechanisms underlying its high yield, high quality, and multi-resistance. The endophytic bacterial diversity and community structures of 11 genetically correlated hybrid rice seeds with different rice blast resistance levels were studied using high-throughput sequencing (HTS) on the Illumina MiSeq platform to reveal their "core microbiota" and explore the effect of genotypes, genetic relationships, and resistance. Proteobacteria (78.15-99.15%) represented the most abundant group in the 11 hybrid rice cultivars, while Pantoea, Pseudomonas, and Microbacterium comprised the "core microbiota." Hybrid rice seeds with different genotypes, genetic correlations, and rice blast resistance displayed endophytic bacterial community structure and diversity variation. In addition, the network relationships between the rice seed endophytic bacteria of "the same female parent but different male parents" were more complex than those from "the same male parent but different female parents." Matrilineal inheritance may be the primary method of passing on endophytic bacteria in rice from generation to generation. The endophytic bacterial interaction network in rice blast-resistant hybrid rice seed varieties was more complicated than in susceptible varieties. In summary, this study demonstrated that the genotype, genetic relationship, and rice blast resistance were important factors affecting the community structures and diversity of endophytic bacteria in hybrid rice seeds, which was vital for revealing the interaction between endophytic bacteria and the host. KEY POINTS: • Pantoea, Pseudomonas, and Microbacterium represent the main endophytic bacteria in hybrid rice seeds. • Genotype is the primary factor affecting endophytic bacterial diversity in hybrid rice seeds. • The diversity of the endophytic bacterial community in hybrid rice seeds is related to their genotypes, genetic relationships, and rice blast resistance.

Physiological response, microbial diversity characterization, and endophytic bacteria isolation of duckweed under cadmium stress

Duckweed is a cadmium (Cd) hyperaccumulator. However, its enrichment characteristics and physiological responses to Cd have not been systematically studied. The physiological responses, enrichment characteristics, diversity of endophytic bacterial communities, and isolation of Cd-resistant endophytes in duckweed (Lemna minor 0014) were studied for different durations and Cd concentrations. The results indicated that peroxidase (POD) and catalase (CAT) activities decreased while superoxide dismutase activity first increased and then decreased with increasing Cd stress duration. POD activities, CAT activities, and O2− increased as Cd concentrations increased. Malondialdehyde content and Cd accumulation in duckweed increased with increasing concentrations and time. This endophytic diversity study identified 488 operational taxonomic units, with the dominant groups being Proteobacteria, Firmicutes, and Actinobacteria. Paenibacillus sp. Y11, a strain tolerant to high concentrations of Cd and capable of significantly promoting duckweed growth, was isolated from the plant. Our study revealed the effects of heavy metals on aquatic plants, providing a theoretical basis for the application of duckweed in water pollution.

Spread and driving factors of antibiotic resistance genes in soil-plant system in long-term manured greenhouse under lead (Pb) stress

Livestock manure is often used as fertilizer in greenhouses, resulting in simultaneous enrichment of heavy metals and antibiotic resistance genes (ARGs) in soils. The soil-plant system is a non-negligible way to spread ARGs; however, the effects of lead (Pb) on the spread of ARGs and their driving factors in the greenhouse soil-plant system remain unclear. In this present study, the occurrence of ARGs in greenhouse soils and their spread into plants under Pb stress were studied. Overall, Pb promoted the accumulation of ARGs in root endophytes at 10, 50, and 100 mg/kg as well as in soils at 10 and 200 mg/kg, but reduced the total relative abundance of ARGs in leaf endophytes. Particularly, Pb increased the mobile genetic elements (MGEs) relative abundance and endophytic bacterial community diversity in roots, consistent with the change in the total relative abundance of ARGs. Network analysis revealed that bacterial community and MGEs may jointly affect the migration of ARGs in the soil-plant system of greenhouses. Overall, this study extended our knowledge of how Pb can promote the transmission of ARGs to plant roots from greenhouse soils receiving long-term manure applications, which must be considered when assessing the risk of ARGs to public health.

Composition and Diversity of Endophytic Bacterial Communities in the Seeds of Upland Rice Resources from Different Origin Habitats in China

Composition and Diversity of Endophytic Bacterial Communities in the Seeds of Upland Rice Resources from Different Origin Habitats in China

Metagenomic study of the communities of bacterial endophytes in the desert plant Senna Italica and their role in abiotic stress resistance in the plant.

Plant leaves and roots are home to diverse communities of bacteria, which play a significant role in plant health and growth. Although one of the most unfriendly environments for plant growth is deserts, desert plants can influence their surrounding microbial population and choose favorable bacteria that encourage their growth under these severe circumstances. Senna italica is known for its excellent medicinal values as a traditional medical plant, but little is known about its associated endophytic bacterial community under extreme conditions. In the present study, metagenomic sequencing of 16S rRNA was used to report the diversity of endophytic bacterial communities associated with the leaves and roots of the desert medicinal plant Senna italica that was collected from the Asfan region in northeast Jeddah, Saudi Arabia. Analyses of the 16S rRNA sequences at the taxonomic phylum level revealed that bacterial communities in the roots and leaves samples belonged to five phyla, including Cyanobacteria, Proteobacteria, Actinobacteria, Firmicutes, and unclassified phyla. Results indicated that the most common phyla were Cyanobacteria/Chloroplast and Actinobacteria. Analysis of the 16S rRNA sequences at the taxonomic phylum level revealed that bacterial communities in the roots and leaves samples belonged to twelve genera at the taxonomic genus level. The most abundant ones were highlighted for further analysis, including Okibacterium and Streptomyces found in Actinobacteria, which were the dominant genus in roots samples. However, Streptophyta found in Cyanobacteria/Chloroplast was the dominant genus in leaf samples. Metagenomic analysis of medicinal plants leads to identifying novel organisms or genes that may have a role in abiotic stress resistance in the plant. The study of endophytic microbiome taxonomic, phylogenetic, and functional diversity will better know innovative candidates that may be selected as biological agents to enhance agricultural and industrial processes, especially for crop desert agricultural improvement.

Plant growth-promoting abilities and community structure of culturable endophytic bacteria from the fruit of an invasive plant Xanthium italicum.

Diversity of endophytic bacterial communities of capsular fruit, upper and lower seeds of an invasive plant Xanthium italicum growing in Xinjiang, China, was investigated. All isolates from the seed capsules, the upper seeds, and the lower seeds were identified by 16S rRNA gene sequencing, and sequences were compared to bacterial databases to define operational taxonomic units (OTUs). Finally, we obtained 316 endophytic isolates corresponding to 58 OTUs based on 16S rRNA gene sequences. The most common OTU corresponded most closely to Bacillus zhangzhouensis and comprised 9.49% of all bacterial isolates. The richness and diversity of endophytes in lower seeds were higher than that of the upper seeds; moreover, the Chao estimator and Shannon index of endophytes in the lower seeds were approximate to that in the seed capsules. Bacillus and Staphylococcus were found as the common taxa in three different tissues that were investigated (OTUs belong to these genera constituted > 70% of the total community). The bacterial endophytic communities differed significantly among these three fruit tissues, especially Bacillus strains, which have been reported to contribute to plant growth promotion and stress resilience to their hosts in harsh environment; abundance of Bacillus species was in the following order: capsules (78 OTUs) > lower seeds (55 OTUs) > upper seeds (40 OTUs). The lower seeds harboring more Bacillus species might be responsible for their earlier seed germination compared with the upper seeds.

Endophytic bacterial community diversity in two citrus cultivars with different citrus canker disease resistance.

The selective infection of Xanthomonas citri pv. citri (Xcc) to citrus cultivars is universally known, but the relationship between endophytic bacteria and the resistance of host variety to canker disease remains unclear. In this study, endophytic bacterial populations of two citrus cultivars-the resistant satsuma mandarin and the susceptible Newhall navel orange-were analyzed through high-throughput sequencing. The results showed that endophytic bacterial community of satsuma mandarin was more abundant than that of Newhall navel orange. In addition, bacterial abundance was the highest in the spring samples, followed by that in summer and winter samples, in both the varieties. In all samples, the predominant phyla were Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes; the major genera were Bacillus and Stenotrophomonas, and the main species was Bacillus subtilis. According to the analysis of the predominant bacteria in the two citrus cultivars, B. subtilis with potential antagonistic characteristics against Xcc existed universally in all samples. However, the susceptible Newhall navel oranges were abundant in Bacillus subtilis and had a relatively large number of canker-causing cooperative bacteria such as Stenotrophomonas. The results suggested that endophytic bacterial community of the two citrus cultivars had some differences based on the season or plant tissue, and these differences were mainly in the quantity of bacteria, affecting citrus canker disease occurrence. In conclusion, the differences in endophytic bacteria on citrus cultivars might be related to host resistance or susceptibility to citrus canker disease.

High-throughput sequencing-based analysis of the composition and diversity of endophytic bacterial community in seeds of saline-alkali tolerant rice

The study of endophytic bacteria in saline-alkali tolerant rice seeds is of great help to the follow-up study of saline-alkali tolerant mechanism and the exploitation of saline-alkali tolerant microbial resources. In this study, high-throughput sequencing technology based on the Illumina Miseq platform was used to reveal the “core microbiota” by examining the diversity and community structures of seed endophytic bacteria in saline-alkali tolerant rice grown under different salt concentrations and explore the effect of salt concentration on its endophytic bacteria. Here, 49 endophytic OTUs were found to coexist in all samples. At the phylum level, the dominant phyla were Proteobacteria (83.90 %–99.87 %). At the genus level, Pantoea (44.65–94.76 %) which represents the core microbiota in saline-alkali tolerant rice seeds, served as the dominant genus that coexisted in all samples tested. Through further analysis, we found that the abundance of Pantoea in saline-alkali tolerant rice seeds was positively proportional to the level of salt concentration. Overall, this study showed that the core microbiota of saline-alkali tolerant rice seeds is Pantoea, and the change of salt concentration is a key factor in the formation of endophytic bacteria in saline-alkali tolerant rice seeds.

Endogenous bacterial community structure of Pinus massoniana with differing resistance to pine wilt disease

Summary Pine wilt disease (PWD) is a devastating pine disease caused by Bursaphelenchus xylophilus and its main host in China is Pinus massoniana. The relationship between endophytic bacteria and disease resistance in P. massoniana remains unclear. In this paper, the leaves, roots, stems and treetops of different disease-resistant P. massoniana were studied as the research objective and Illumina MiSeq sequencing was used to analyse whether there were significant differences in the composition and diversity of endophytic bacterial communities between different disease-resistant P. massoniana. The results showed that at the genus level there were no obvious differences in the composition of the endophytic bacterial community of different disease-resistant P. massoniana in the leaves, but there were obvious differences in the roots, stems and treetops. The richness and diversity of endophytic bacteria in P. massoniana had no significant impact on its disease resistance, whilst the structure of endophytic bacterial community in stems and treetops may be related to its disease resistance.

Peptide Nucleic Acid (PNA) Clamps to Reduce Co-amplification of Plant DNA During PCR Amplification of 16S rRNA Genes from Endophytic Bacteria.

High-throughput sequencing of universal bacterial 16S rRNA gene (16S rDNA) amplicons is a routine method for characterizing bacterial diversity in a range of environments. For eukaryotic host-associated communities, however, plastid and mitochondrial genes are often co-amplified with, and greatly outnumber, bacterial 16S rDNA. This makes it difficult to obtain sufficient numbers of target 16S rDNA sequences to characterize the diversity of endophytic bacterial communities. This chapter describes a method that improves the amplification of bacterial 16S rDNA from plant tissues by using a peptide nucleic acid (PNA) PCR clamp. The PNA clamp selectively binds to a targeted region of the plant genome and inhibits its amplification during PCR. PNA clamps are especially useful for characterizing bacterial communities on plant tissues with lower levels of microbial colonization such as the root tips and leaves.

High-throughput sequencing-based analysis of the composition and diversity of endophytic bacterial community in seeds of upland rice

Upland rice is an ecotype crop resulting from the long-term domestication and evolution of rice in dry land without a water layer. Generally, the stems and leaves are thick and luxuriant, while the leaves also typically broad and light. The root system is developed with abundant root hair, and the osmotic pressure of the root and cell juice concentration in the leaves is high, while this plant is drought-resistant, heat-resistant, and water absorbent. This study aims to reveal the "core flora" of the endophytes in upland rice seeds by examining their diversity and community structures. It further intends to reveal the impact of the soil environment on the formation of endophyte community structures in upland rice seeds by comparing the environmental soil microorganisms in upland rice habitats. In this study, high-throughput sequencing technology based on the Illumina Hiseq 2500 platform was used to investigate the structure and diversity of endophytic bacterial communities using upland rice varieties collected from different locations and soil samples from unified planting sites as materials. Here, 42 endophytic OTUs were found to coexist in the 14 samples. At the phylum level, the first dominant phyla in all the samples were Proteobacteria (93.81-99.99%). At the genus level, Pantoea (8.77-87.77%), Pseudomonas (1.15-61.58%), Methylobacterium (0.40-4.64%), Sphingomonas (0.26-3.85%), Microbacterium (0.01-4.67%) and Aurantimonas (0.04-4.34%), which represent the core microflora in upland rice seeds, served as the dominant genera that coexisted in all the upland rice seeds tested. This study significant for the isolation, screening, functional evaluation, and re-action of various functional microorganisms in upland rice to improve its agronomic traits. It also provides a specific reference for the interaction between microorganisms and plants.

Effects of calcium application on the structural diversity of endophytic bacterial community in peanut roots under acidic red soil cultivation

Effects of calcium application on the structural diversity of endophytic bacterial community in peanut roots under acidic red soil cultivation

Graphene oxide enters the rice roots and disturbs the endophytic bacterial communities

The environmental release of graphene oxide (GO) will certainly induce the GO exposure to plants. To date, the influence of GO on the intracellular structures and the endophytic bacterial ecology of plants have been rarely reported. In the present study, the rice seedlings were exposed to GO (5mg/L) under hydroponic condition for fifteen days with periodic stir. The cellular structures damage, GO deposition and oxidative stress were found in rice root after GO exposure. A Illumina analysis based on the bacterial 16S rRNA gene showed that the richness, evenness and diversity of endophytic bacterial communities of rice root decreased due to GO exposure. The relative abundance of beneficial endophytic bacterial populations decreased after GO exposure. Out of potential phenotypes predicted by BugBase, the relative abundance of Gram negative, stress-tolerant and biofilm-forming phenotypes, presented an increase trend after GO exposure.

Investigating the endophytic bacterial diversity and community structures in seeds of genetically related maize (Zea mays L.) genotypes.

This research aimed to investigate the composition and diversity of endophytic bacterial community in seeds of four hybrid maize and their parental lines, which was used to reveal the potential relationship and association of endophytic bacteria between maize genotypes and their genetic relevance. High-throughput sequencing (HTS) technology showed that a total of 1419 OTUs (46.6%) were parental lines unique and 1052 OTUs (34.5%) were hybrid varieties unique, with only 575 core OTUs revealed in all the samples. Most OTUs belonged to Proteobacteria. Enterobacter (23.2%), Shigella (21.2%), Pseudomonas (15.8%) and Achromobacter (10.1%) were the major genera; the bacterial community composition and diversity of endophytic bacteria were inconsistent among different seed genotypes. Based on principal component analysis (PCA), the results referred that the endophytic composition of hybrid sample showed obvious correlation with their female parental lines, and in 'Jingke968' and 'MC738' with the same female line the endophytic community was more similar than other hybrid samples. This was the first ever use of HTS technology for investigating the endophytic bacterial diversity and community structures in seeds of genetically related maize genotypes, it was shown that, there were core microbes shared among all genotypes of seed samples, and the female parental line was more significant to impact on the composition of their hybrid seeds than male parental line. This study would provide scientific clues for the future research on the vertical transmission of endophytes among maize generations.

Effects of endophytes inoculation on rhizosphere and endosphere microecology of Indian mustard (Brassica juncea) grown in vanadium-contaminated soil and its enhancement on phytoremediation

We investigated the effects of endophytes inoculation on ecological factors such as root morphology, rhizosphere soil properties, heavy metal speciation, and rhizosphere and endophytic bacterial communities and their role on phytoremediation. Indian mustards were grown for two months in V-contaminated soil with three treatments (control, inoculation with Serratia PRE01 or Arthrobacter PRE05). Inoculation with PRE01 and PRE05 increased organic matter content by 6.94% and 4.6% respectively and significantly increased bioavailability of heavy metals in rhizosphere soils. Despite the endophyte inocula failed to flourish as stable endophytes, they significantly affected the specific composition and diversity of endophytic bacterial communities in roots, with no significant effect on rhizosphere bacterial communities. The test strains could greatly increase plant growth promotion-related biomarkers in the endosphere, especially those associated with Pseudomonas and Microbacterium genera. PICRUSt analysis predicted high relative abundances of functional genes related to environmental information processing especially in the endophytic microbiota. More biomass production (12.0%–17.4%) and total metals uptake (24.2%–32.0%) were acquired in inoculated treatments. We conclude that endophyte PRE01 or PRE05 inoculation could effectively enhance phytoremediation of V-contaminated soil by improving the rhizosphere and endosphere microecology without causing any ecological damage.

Diversity of endophytic bacterial community inhabiting in tropical aerobic rice under aerobic and flooded condition.

The role and activity of bacterial endophytes remains largely unexplored and detail insight into Indian rice agro ecosystem is still little explored. In this study, we examined the diversity of endophytic bacteria in aerobic rice (variety ARB6) under aerobic and flooded field conditions. Based on 16S rRNA gene RFLP cloning sequencing, 900 clones with 144 representatives (72 aerobic and 72 flooded) revealed majority of clones affiliated toGammaproteobacteria (64.58%),Betaproteobacteria (9.72%),Alphaproteobacteria (17.36), Firmicutes (6.26%) and Bacteroidetes (2.08). The study suggests that the aerobic rice variety harbours plant growth promoting (PGP) genera (viz. Pantoea, Enterobacter, Paenibacillus, etc). Investigations on aerobic rice under aerobic and flooded conditions revealed high richness and diversity of endophytic bacteria under aerobic condition inferring that the endophytic bacteria are beneficial for rice growth and productivity, and hence, would be helpful in designing better strategies for rice cultivation under drought or water scarce conditions.

Composition and diversity of endophytic bacterial community in seeds of super hybrid rice ‘Shenliangyou 5814’ (Oryza sativa L.) and its parental lines

Super hybrid rice is one of the important research achievements in the field of rice super-high yield breeding in China. Compared with general high-yield rice, the yield potential of super hybrid rice is relatively high, and the average yields per hectare can exceed 10.5 t. This study aimed to study the endophytic bacterial community among a super hybrid rice combination and its parental lines and to reveal the potential relationship and association of endophytic bacteria between rice genotypes and their genetic relevance. In this research, through high-throughput sequencing based on Illumina Hiseq 2500 platform, the seeds of super hybrid rice variety ‘Shenliangyou 5814’ (sample C) and its parental lines ‘Y58S’ (sample M) and ‘C4114’ (sample F) independently cultivated by China were used as plant materials to study their endophytic bacterial structure and diversity. The number of OTUs for sample M, F and C was 327, 288 and 283 respectively, and among them 218 endophytic OTUs coexisted in the 3 samples. Pantoea (18.6–31.1%) was the first dominant groups shared in all three samples, and other dominant shared groups belonged to Methylobacterium (4.48%~17.65%), Sphingomonas (4.0%~11.4%), Rhizobium (5.69–8.78%), Microbacterium (3.85%~9.41%) and Pseudomonas (4.13%~5.68%). Although Principal Coordinates Analysis (PCoA) and Non-metric Multidimensional Scaling (NMDS) analysis showed that there were obvious differences in endophytic bacterial community composition and structure among the 3 samples, the dominant endophytic bacterial genus sample C and its parental lines remained consistency. This study would provide scientific clues for the future research on the vertical transmission of endophytes between rice generations.

The Endophytic Bacterial Microbiota Associated with Sweet Sorghum (Sorghum bicolor) Is Modulated by the Application of Chemical N Fertilizer to the Field.

Sweet sorghum (Sorghum bicolor) is a multipurpose crop used as a feedstock to produce bioethanol, sugar, energy, and animal feed. However, it requires high levels of N fertilizer application to achieve the optimal growth, which causes environmental degradation. Bacterial endophytes, which live inside plant tissues, play a key role in the health and productivity of their host. This particular community may be influenced by different agronomical practices. The aim of the work was to evaluate the effects of N fertilization on the structure, diversity, abundance, and composition of endophytic and diazotrophic bacterial community associated with field-grown sweet sorghum. PCR-DGGE, quantitative PCR, and high-throughput sequencing were performed based on the amplification of rrs and nifH genes. The level of N fertilization affected the structure and abundance but not the diversity of the endophytic bacterial communities associated with sweet sorghum plants. This effect was pronounced in the roots of both bacterial communities analyzed and may depend on the physiological state of the plants. Specific bacterial classes and genera increased or decreased when the fertilizer was applied. The data obtained here contribute to a better understanding on the effects of agronomical practices on the microbiota associated with this important crop, with the aim to improve its sustainability.

Hydrocarbon degradation potential and plant growth-promoting activity of culturable endophytic bacteria of Lotus corniculatus and Oenothera biennis from a long-term polluted site

Many endophytic bacteria exert beneficial effects on their host, but still little is known about the bacteria associated with plants growing in areas heavily polluted by hydrocarbons. The aim of the study was characterization of culturable hydrocarbon-degrading endophytic bacteria associated with Lotus corniculatus L. and Oenothera biennis L. collected in long-term petroleum hydrocarbon-polluted site using culture-dependent and molecular approaches. A total of 26 hydrocarbon-degrading endophytes from these plants were isolated. Phylogenetic analyses classified the isolates into the phyla Proteobacteria and Actinobacteria. The majority of strains belonged to the genera Rhizobium, Pseudomonas, Stenotrophomonas, and Rhodococcus. More than 90% of the isolates could grow on medium with diesel oil, approximately 20% could use n-hexadecane as a sole carbon and energy source. PCR analysis revealed that 40% of the isolates possessed the P450 gene encoding for cytochrome P450-type alkane hydroxylase (CYP153). In in vitro tests, all endophytic strains demonstrated a wide range of plant growth-promoting traits such as production of indole-3-acetic acid, hydrogen cyanide, siderophores, and phosphate solubilization. More than 40% of the bacteria carried the gene encoding for the 1-aminocyclopropane-1-carboxylic acid deaminase (acdS). Our study shows that the diversity of endophytic bacterial communities in tested plants was different. The results revealed also that the investigated plants were colonized by endophytic bacteria possessing plant growth-promoting features and a clear potential to degrade hydrocarbons. The properties of isolated endophytes indicate that they have the high potential to improve phytoremediation of petroleum hydrocarbon-polluted soils.