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Endopathogenic lifestyle of Pseudomonas savastanoi pv. savastanoi in olive knots.

Rodríguez-Moreno L, Jiménez AJ, Ramos C - Microb Biotechnol (2009)

Bottom Line: The endophytic phase of Pseudomonas savastanoi pv. savastanoi in olive stems and the structural and ultrastructural histogenesis of olive knots have been studied.Hypertrophy of the stem tissue was concomitant with the formation of bacterial aggregates, microcolonies and multilayer biofilms, over the cell surfaces and the interior of plasmolysed cells facing the air-tissue interface of internal opened fissures, and was followed by invasion of the outer layers of the hypertrophied tissue.This is the first real-time monitoring of P. savastanoi disease development and the first illustrated description of the ultrastructure of P. savastanoi-induced knots.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biología Celular, Genética y Fisiología, Universidad de Málaga, Campus de Teatinos s/n, E-29071, Málaga, Spain.

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Schematic drawing of plasmid pLRM1‐GFP. The GFP expression cassette from plasmid pJBA29 (Andersen et al., 1998), shown in black, was cloned into SalI/EcoRI‐digested plasmid pBBR1‐MCS5 (Kovach et al., 1995). pLRM1‐GFP consists of a 2.05 kb fragment carrying a fusion of the IPTG‐inducible promoter PA1/04/03(Lanzer and Bujard, 1988) to the gfpmut3* gene, encoding GFPmut3 (Cormack et al., 1996), a synthetic ribosome binding site (RBSII), translational stop codons in all three reading frames, and two strong transcriptional terminators, T0 (derived from phage λ) and T1 (derived from the rrnB operon of E. coli).
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f1: Schematic drawing of plasmid pLRM1‐GFP. The GFP expression cassette from plasmid pJBA29 (Andersen et al., 1998), shown in black, was cloned into SalI/EcoRI‐digested plasmid pBBR1‐MCS5 (Kovach et al., 1995). pLRM1‐GFP consists of a 2.05 kb fragment carrying a fusion of the IPTG‐inducible promoter PA1/04/03(Lanzer and Bujard, 1988) to the gfpmut3* gene, encoding GFPmut3 (Cormack et al., 1996), a synthetic ribosome binding site (RBSII), translational stop codons in all three reading frames, and two strong transcriptional terminators, T0 (derived from phage λ) and T1 (derived from the rrnB operon of E. coli).

Mentions: The stability of plasmid pLRM1‐GFP (Fig. 1) in P. savastanoi pv. savastanoi NCPPB 3335‐GFP was examined in vitro. As judged by resistance to the appropriate antibiotics, the strain stably maintained the plasmid (between 90% and 100% of the cells) for at least 40 generations (Fig. 2A). Furthermore, 100% of the colonies isolated at the end of the experiment from media not containing antibiotics exhibited uniform green fluorescence. The stability of the plasmid was also assessed in planta by infecting in vitro olive plants with NCPPB 3335‐GFP. During the 28 days of the experiment, estimated colony‐forming units (cfu) counts in media with and without antibiotics were similar (Fig. 2B). Moreover, 100% of the colonies isolated at 28 days post‐inoculation (dpi) were fluorescent green, further confirming the stability of the plasmid in P. savastanoi pv. savastanoi during bacterial growth in olive plants.


Endopathogenic lifestyle of Pseudomonas savastanoi pv. savastanoi in olive knots.

Rodríguez-Moreno L, Jiménez AJ, Ramos C - Microb Biotechnol (2009)

Schematic drawing of plasmid pLRM1‐GFP. The GFP expression cassette from plasmid pJBA29 (Andersen et al., 1998), shown in black, was cloned into SalI/EcoRI‐digested plasmid pBBR1‐MCS5 (Kovach et al., 1995). pLRM1‐GFP consists of a 2.05 kb fragment carrying a fusion of the IPTG‐inducible promoter PA1/04/03(Lanzer and Bujard, 1988) to the gfpmut3* gene, encoding GFPmut3 (Cormack et al., 1996), a synthetic ribosome binding site (RBSII), translational stop codons in all three reading frames, and two strong transcriptional terminators, T0 (derived from phage λ) and T1 (derived from the rrnB operon of E. coli).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815908&req=5

f1: Schematic drawing of plasmid pLRM1‐GFP. The GFP expression cassette from plasmid pJBA29 (Andersen et al., 1998), shown in black, was cloned into SalI/EcoRI‐digested plasmid pBBR1‐MCS5 (Kovach et al., 1995). pLRM1‐GFP consists of a 2.05 kb fragment carrying a fusion of the IPTG‐inducible promoter PA1/04/03(Lanzer and Bujard, 1988) to the gfpmut3* gene, encoding GFPmut3 (Cormack et al., 1996), a synthetic ribosome binding site (RBSII), translational stop codons in all three reading frames, and two strong transcriptional terminators, T0 (derived from phage λ) and T1 (derived from the rrnB operon of E. coli).
Mentions: The stability of plasmid pLRM1‐GFP (Fig. 1) in P. savastanoi pv. savastanoi NCPPB 3335‐GFP was examined in vitro. As judged by resistance to the appropriate antibiotics, the strain stably maintained the plasmid (between 90% and 100% of the cells) for at least 40 generations (Fig. 2A). Furthermore, 100% of the colonies isolated at the end of the experiment from media not containing antibiotics exhibited uniform green fluorescence. The stability of the plasmid was also assessed in planta by infecting in vitro olive plants with NCPPB 3335‐GFP. During the 28 days of the experiment, estimated colony‐forming units (cfu) counts in media with and without antibiotics were similar (Fig. 2B). Moreover, 100% of the colonies isolated at 28 days post‐inoculation (dpi) were fluorescent green, further confirming the stability of the plasmid in P. savastanoi pv. savastanoi during bacterial growth in olive plants.

Bottom Line: The endophytic phase of Pseudomonas savastanoi pv. savastanoi in olive stems and the structural and ultrastructural histogenesis of olive knots have been studied.Hypertrophy of the stem tissue was concomitant with the formation of bacterial aggregates, microcolonies and multilayer biofilms, over the cell surfaces and the interior of plasmolysed cells facing the air-tissue interface of internal opened fissures, and was followed by invasion of the outer layers of the hypertrophied tissue.This is the first real-time monitoring of P. savastanoi disease development and the first illustrated description of the ultrastructure of P. savastanoi-induced knots.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biología Celular, Genética y Fisiología, Universidad de Málaga, Campus de Teatinos s/n, E-29071, Málaga, Spain.

Show MeSH
Related in: MedlinePlus