Enrichment for microbes living in association with plant tissues

Jiao, Jun-Ying; Wang, H.-X.; Zeng, Yan; Shen, Yuemao

doi:10.1111/j.1365-2672.2006.02830.x

Cited by 58 publications

(46 citation statements)

References 35 publications

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“…Some plants, e.g., Maytenus austroyunnanensis, were aseptically crumbled into small fragments and isolated using the combined enzymatic hydrolysis and differential centrifugation method (44). The homogenate was centrifuged at 200 ϫ g for 20 min at 4°C, and the supernatant was subsequently centrifuged at 3,000 ϫ g for 30 min at 4°C to collect the sediments, which were diluted to 10 Ϫ2 with sterile water and spread-plated onto medium.…”

Section: Methodsmentioning

confidence: 99%

Isolation, Diversity, and Antimicrobial Activity of Rare Actinobacteria from Medicinal Plants of Tropical Rain Forests in Xishuangbanna, China

Qin

Chen

et al. 2009

Appl Environ Microbiol

380

263

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Endophytic actinobacteria are relatively unexplored as potential sources of novel species and novel natural products for medical and commercial exploitation. Xishuangbanna is recognized throughout the world for its diverse flora, especially the rain forest plants, many of which have indigenous pharmaceutical histories. However, little is known about the endophytic actinobacteria of this tropical area. In this work, we studied the diversity of actinobacteria isolated from medicinal plants collected from tropical rain forests in Xishuangbanna. By the use of different selective isolation media and methods, a total of 2,174 actinobacteria were isolated. Forty-six isolates were selected on the basis of their morphologies on different media and were further characterized by 16S rRNA gene sequencing. The results showed an unexpected level of diversity, with 32 different genera. To our knowledge, this is the first report describing the isolation of Saccharopolyspora, Dietzia, Blastococcus, Dactylosporangium, Promicromonospora, Oerskovia, Actinocorallia, and Jiangella species from endophytic environments. At least 19 isolates are considered novel taxa by our current research. In addition, all 46 isolates were tested for antimicrobial activity and were screened for the presence of genes encoding polyketide synthetases and nonribosomal peptide synthetases. The results confirm that the medicinal plants of Xishuangbanna represent an extremely rich reservoir for the isolation of a significant diversity of actinobacteria, including novel species, that are potential sources for the discovery of biologically active compounds.The class Actinobacteria accounts for a high proportion of soil microbial biomass and contains the most economically significant prokaryotes, producing more than half of the bioactive compounds in a literature survey (46), including antibiotics (6), immunosuppressive agents (55), antitumor agents (18), and enzymes (64). Actinobacteria belonging to the genus Streptomyces, in particular, are excellent producers. The emergence of drug resistance in many bacterial pathogens and the current increase in the number of fungal infections has caused a resurgence of interest in finding new reserves of biologically active compounds (63). As the search for novel natural products continues, it becomes apparent that the rate of discovery of new compounds from soil streptomycetes has decreased, whereas the rate of reisolation of known compounds has increased (28). Recently, evidence has accumulated that rare actinomycete species, which are often very difficult to isolate and cultivate, might represent a unique source of novel biologically active compounds (4). On the other hand, new microbial habitats need to be examined in the search for novel bioactive compounds. One biologically important but relatively overlooked niche is the inner tissues of higher plants. Early studies have demonstrated that some actinobacteria can form intimate associations with plants and colonize their inner tissues. Frankia species and Streptomyce...

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Section: Methodsmentioning

confidence: 99%

Isolation, Diversity, and Antimicrobial Activity of Rare Actinobacteria from Medicinal Plants of Tropical Rain Forests in Xishuangbanna, China

Qin

Chen

et al. 2009

Appl Environ Microbiol

380

263

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“…In order to avoid plant host DNA, enrichment of endophytic bacteria prior to DNA amplification should be done (Govindasamy et al, 2014). To exclude interference of plant host DNA, Jiao et al (2006) enriched bacterial endophytes by hydrolysis of the plant cell walls, followed by differential centrifugation. For bacterial DNA ratio enrichment in stems and leaves of soybean and rice, Ikeda et al (2009) used series of differential centrifugation steps followed by density gradient centrifugation.…”

Section: Occurrence and Diversity Of Bacterial Endophytes In Agricultmentioning

confidence: 99%

“…Metagenomic analysis involves direct isolation of bacterial DNA, library construction and functional analysis (Handelsman, 2004;Jiao et al, 2006). To investigate endophytic bacterial diversity, highly specific methods should be used (Sun et al, 2008).…”

Section: Occurrence and Diversity Of Bacterial Endophytes In Agricultmentioning

confidence: 99%

Bacterial endophytes in agricultural crops and their role in stress tolerance: a review

Miliūtė

Buzaitė

Baniulis

et al. 2015

Zemdirbyste-Agriculture

205

112

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Bacterial endophytes are a class of endosymbiotic microorganisms widespread among plants that colonize intercellular and intracellular spaces of all plant compartments and do not cause plant disease or significant morphological changes. Plant and endophytic bacteria association includes vast diversity of bacterial taxa and plant hosts and in this review we present an overview of taxonomic composition of endophytes identified in common agricultural crops. Further, during the last decade, new aspects of the microbial diversity have emerged with application of new metagenomic analysis methods in studies of bacterial endophytes. Endophytic bacteria community structure is influenced by plant genotype, abiotic and biotic factors such as environment conditions, microbe -microbe interactions and plant -microbe interactions. Agricultural practices, such as soil tillage, irrigation, use of pesticides and fertilizers have a major effect on function and structure of soil and endophytic microbial populations. Therefore, the use of agricultural practices that maintain natural diversity of plant endophytic bacteria is becoming an important element of sustainable agriculture that could ensure plant productivity and quality of agricultural production. The diverse endophytic microbial communities play integral and unique role in the functioning of agroecosystems. Endophytic bacteria have been shown to have several beneficial effects on their host plant, including growth promoting activity, modulation of plant metabolism and phytohormone signalling that leads to adaptation to environmental abiotic or biotic stress. Use of endophytic bacteria presents a special interest for development of agricultural applications that ensure improved crop performance under cold, draught or contaminated soil stress conditions or enhanced disease resistance.

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“…The TSA plates were examined for bacterial growth after 5 days of incubation at 30°C, and the grapevine leaves not contaminated were used for further analysis (Ferreira et al, 2008;Sun et al, 2008). Bacterial DNA from grapevine leaf samples was extracted with two strategies: with and without microbe enrichment (Jiao et al, 2006). Bacterial enrichment was done from leaf veins (2 g or 20 g), separated with a sterilized blade, or from whole leaves (40 g).…”

mentioning

confidence: 99%

Endophytic bacterial diversity in grapevine (Vitis vinifera L.) leaves described by 16S rRNA gene sequence analysis and length heterogeneity-PCR

et al. 2009

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Diversity of bacterial endophytes associated with grapevine leaf tissues was analyzed by cultivation and cultivation-independent methods. In order to identify bacterial endophytes directly from metagenome, a protocol for bacteria enrichment and DNA extraction was optimized. Sequence analysis of 16S rRNA gene libraries underscored five diverse Operational Taxonomic Units (OTUs), showing best sequence matches with gamma-Proteobacteria, family Enterobacteriaceae, with a dominance of the genus Pantoea. Bacteria isolation through cultivation revealed the presence of six OTUs, showing best sequence matches with Actinobacteria, genus Curtobacterium, and with Firmicutes genera Bacillus and Enterococcus. Length Heterogeneity-PCR (LH-PCR) electrophoretic peaks from single bacterial clones were used to setup a database representing the bacterial endophytes identified in association with grapevine tissues. Analysis of healthy and phytoplasma-infected grapevine plants showed that LH-PCR could be a useful complementary tool for examining the diversity of bacterial endophytes especially for diversity survey on a large number of samples.

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Enrichment for microbes living in association with plant tissues

Cited by 58 publications

References 35 publications

Isolation, Diversity, and Antimicrobial Activity of Rare Actinobacteria from Medicinal Plants of Tropical Rain Forests in Xishuangbanna, China

Isolation, Diversity, and Antimicrobial Activity of Rare Actinobacteria from Medicinal Plants of Tropical Rain Forests in Xishuangbanna, China

Bacterial endophytes in agricultural crops and their role in stress tolerance: a review

Endophytic bacterial diversity in grapevine (Vitis vinifera L.) leaves described by 16S rRNA gene sequence analysis and length heterogeneity-PCR

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