Construction of EGFP-Labeling System for Visualizing the Infection Process of Xanthomonas axonopodis pv. citri in planta

Liu, Liping; Deng, Zhili; JinWang, Qu; Yan, Jiawen; Catara, Vittoria; Li, Dazhi; Long, Guiyou; Li, Na

doi:10.1007/s00284-012-0155-y

Cited by 14 publications

(13 citation statements)

References 23 publications

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“…To obtain a stable genetic transformation, the target gene was introduced into YL6 by a triparental mating (Liu et al, 2012). The GFP-expressing YL6 strain (YL6-GFP) was similar to the original strain (YL6), both in its growth and ability to dissolve P. The ability of PSB survival in the soil is the key to determining its role in the natural environment and is an important basis on which to evaluate PSB colonization ability in the rhizosphere (Shen et al, 2013; Hanif et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

Isolation and Characterization of a Phosphorus-Solubilizing Bacterium from Rhizosphere Soils and Its Colonization of Chinese Cabbage (Brassica campestris ssp. chinensis)

Wang

et al. 2017

Front. Microbiol.

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Phosphate-solubilizing bacteria (PSB) can promote the dissolution of insoluble phosphorus (P) in soil, enhancing the availability of soluble P. Thus, their application can reduce the consumption of fertilizer and aid in sustainable agricultural development. From the rhizosphere of Chinese cabbage plants grown in Yangling, we isolated a strain of PSB (YL6) with a strong ability to dissolve P and showed that this strain promoted the growth of these plants under field conditions. However, systematic research on the colonization of bacteria in the plant rhizosphere remains deficient. Thus, to further study the effects of PSB on plant growth, in this study, green fluorescent protein (GFP) was used to study the colonization of YL6 on Chinese cabbage roots. GFP expression had little effect on the ability of YL6 to grow and solubilize P. In addition, the GFP-expressing strain stably colonized the Chinese cabbage rhizosphere (the number of colonizing bacteria in the rhizosphere soil was 4.9 lg CFU/g). Using fluorescence microscopy, we observed a high abundance of YL6-GFP bacteria at the Chinese cabbage root cap and meristematic zone, as well as in the root hairs and hypocotyl epidermal cells. High quantities of GFP-expressing bacteria were recovered from Chinese cabbage plants during different planting periods for further observation, indicating that YL6-GFP had the ability to endogenously colonize the plants. This study has laid a solid and significant foundation for further research on how PSB affects the physiological processes in Chinese cabbage to promote plant growth.

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Section: Discussionmentioning

confidence: 99%

Isolation and Characterization of a Phosphorus-Solubilizing Bacterium from Rhizosphere Soils and Its Colonization of Chinese Cabbage (Brassica campestris ssp. chinensis)

Wang

et al. 2017

Front. Microbiol.

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“…Enhanced green fluorescent protein (EGFP) and red fluorescent protein (Red) expressed in prokaryotic and eukaryotic cells are capable of producing strong green and red fluorescence when excited by illumination with ultraviolet, respectively, which makes them excellent reporters for in vivo observation [33,34]. Fusion expression of fluorescent protein for a target protein has been extensively used for direct visualization of a target protein in various cells and organs using confocal microscopy.…”

Section: Efficient Dna Delivery In Cultured Silkworm Cells Using G5-pmentioning

confidence: 99%

Efficient Delivery of dsRNA and DNA in Cultured Silkworm Cells for Gene Function Analysis Using PAMAM Dendrimers System

Lü

Chang

et al. 2019

Insects

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Polyamidoamine (PAMAM) dendrimers are emerging as intriguing nanovectors for nucleic acid delivery because of their unique well-defined architecture and high binding capacity, which have been broadly applied in DNA-and RNA-based therapeutics. The low-cost and high-efficiency of PAMAM dendrimers relative to traditional liposomal transfection reagents also promote their application in gene function analysis. In this study, we first investigated the potential use of a PAMAM system in the silkworm model insect. We determined the binding property of G5-PAMAM using dsRNA and DNA in vitro, and substantially achieved the delivery of dsRNA and DNA from culture medium to both silkworm BmN and BmE cells, thus leading to efficient knockdown and expression of target genes. Under treatments with different concentrations of G5-PAMAM, we evaluated its cellular cytotoxicity on silkworm cells, and the results show that G5-PAMAM had no obvious toxicity to cells. The presence of serum in the culture medium did not affect the delivery performance of DNA and dsRNA by G5-PAMAM, revealing its convenient use for various purposes. In conclusion, our data demonstrate that the PAMAM system provides a promising strategy for delivering dsRNA and DNA in cultured silkworm cells and promote its further application in individuals.

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“…; Liu et al. ). Both metabolically active and starving bacterial cells can be detected by the expression of the GFP marker (Unge and Jansson ; Campos‐Soriano and Segundo ).…”

Section: Introductionmentioning

confidence: 98%

“…citri, Herbcrspirillztm sp. and Klebsiella oxytoca Zakria et al 2007;Liu et al 2012). Both metabolically active and starving bacterial cells can be detected by the expression of the GFP marker (Unge and Jansson 2001;Campos-Soriano and Segundo 2009).…”

Section: Introductionmentioning

confidence: 99%

Infection Process of Burkholderia glumae in Rice Spikelets

Wang

Liu

et al. 2016

Journal of Phytopathology

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Burkholderia glumae, which causes bacterial panicle blight of rice (BPBR), is a well‐known pathogen. The pathogen‐induced symptoms include seedling rot, grain rot and leaf‐sheath browning in rice plants. B. glumae can incubate in rice plants as endophytes before booting stage of rice. In this study, we constructed a gfp‐labelled system of B. glumae LMG 2196 and used SEM to clarify the colonization course of B. glumae at the heading stage. New locations of B. glumae were found. The pathogens initially distributed on the surface of the glumes and colonized in the glume hairs and cells of the edge of sterile lemma, palea and lemma. The base of glume hairs was the initial position for colonization. Bacterial population raised around glume hairs, penetrated into the inner surface of the palea and lemma, and spread on the gynoecium and stamens through contact. The spreading of B. glumae among the panicles mainly occurred through the contact or friction among glumes or leaf sheaths, but the inner spread of the stamens mainly occurred through the connective tissue of anther. We also detected the differences of bacterial content in stamens, gynoecia and glumes. The growing stage of B. glumae in spikelets could be divided into two sections. The biomass of all parts continued to increase to nearly 108 CFU/g at 10 DAI. This caused wilt symptoms and stopped the pollination. This work showed that glume hairs played an important role in the initial colonization of B. glumae, and provides a foundation for further studies of the infection manner of B. glumae and other pathogenic bacteria.

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Construction of EGFP-Labeling System for Visualizing the Infection Process of Xanthomonas axonopodis pv. citri in planta

Cited by 14 publications

References 23 publications

Isolation and Characterization of a Phosphorus-Solubilizing Bacterium from Rhizosphere Soils and Its Colonization of Chinese Cabbage (Brassica campestris ssp. chinensis)

Isolation and Characterization of a Phosphorus-Solubilizing Bacterium from Rhizosphere Soils and Its Colonization of Chinese Cabbage (Brassica campestris ssp. chinensis)

Efficient Delivery of dsRNA and DNA in Cultured Silkworm Cells for Gene Function Analysis Using PAMAM Dendrimers System

Infection Process of Burkholderia glumae in Rice Spikelets

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