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Localization and expression of EDS5H a homologue of the SA transporter EDS5.

Parinthawong N, Cottier S, Buchala A, Nawrath C, Métraux JP - BMC Plant Biol. (2015)

Bottom Line: An important signal transduction pathway in plant defence depends on the accumulation of salicylic acid (SA).Both transporters are located at the envelope of the chloroplast, the compartment of SA biosynthesis.EDS5H most likely transports related substances such as for example phenolic acids, but unlikely SA.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, University of Fribourg, 1700, Fribourg, Switzerland. kpnongla@kmitl.ac.th.

ABSTRACT

Background: An important signal transduction pathway in plant defence depends on the accumulation of salicylic acid (SA). SA is produced in chloroplasts and the multidrug and toxin extrusion transporter ENHANCED DISEASE SUSCEPTIBILITY5 (EDS5; At4g39030) is necessary for the accumulation of SA after pathogen and abiotic stress. EDS5 is localized at the chloroplast and functions in transporting SA from the chloroplast to the cytoplasm. EDS5 has a homologue called EDS5H (EDS5 HOMOLOGUE; At2g21340) but its relationship to EDS5 has not been described and its function is not known.

Results: EDS5H exhibits about 72% similarity and 59% identity to EDS5. In contrast to EDS5 that is induced after pathogen inoculation, EDS5H was constitutively expressed in all green tissues, independently of pathogen infection. Both transporters are located at the envelope of the chloroplast, the compartment of SA biosynthesis. EDS5H is not involved with the accumulation of SA after inoculation with a pathogen or exposure to UV stress. A phylogenetic analysis supports the hypothesis that EDS5H may be an H(+)/organic acid antiporter like EDS5.

Conclusions: The data based on genetic and molecular studies indicate that EDS5H despite its homology to EDS5 does not contribute to pathogen-induced SA accumulation like EDS5. EDS5H most likely transports related substances such as for example phenolic acids, but unlikely SA.

No MeSH data available.


Related in: MedlinePlus

Subcellular localization of EDS5H-3myc in transgenic Arabidopsis. Mesophyll cells of transgenic plants carrying the CaMV35S::EDS5H::3xmyc construct. The two panes show the cMyc-epitope labelled with Alexa Fluor 488 excited at the wavelength of 488 nm and detected using the emission filter 522 DF32 (green) and Red chlorophyll catalase reductase labelled with Alexa Fluor 568 excited at a wavelength of 568 nm and detected using the emission filters 605 DF32 and 585 EFLP (red). Bar = 5 μm. A representative picture out of 11 transformed lines is presented
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Fig4: Subcellular localization of EDS5H-3myc in transgenic Arabidopsis. Mesophyll cells of transgenic plants carrying the CaMV35S::EDS5H::3xmyc construct. The two panes show the cMyc-epitope labelled with Alexa Fluor 488 excited at the wavelength of 488 nm and detected using the emission filter 522 DF32 (green) and Red chlorophyll catalase reductase labelled with Alexa Fluor 568 excited at a wavelength of 568 nm and detected using the emission filters 605 DF32 and 585 EFLP (red). Bar = 5 μm. A representative picture out of 11 transformed lines is presented

Mentions: The ORF of EDS5H was placed under the control of the CaMV 35S promoter upstream of a triple cMyc epitope tag. In tissue sections of transgenic plants carrying the CaMV35S::EDS5H::3xcMyc construct, the cMyc-epitope tag was labelled with a primary anti-myc antibody that was identified by a goat anti-mouse IgG conjugated with Alexa Fluor 488 (green). RCCR (red chlorophyll catalase reductase) was used as an example for a protein targeted to the chloroplast [21]. Tissues were labelled with a primary antibody against RCCR and secondary antibody goat anti-rabbit IgG conjugated with Alexa Fluor 568 (red). Fig. 4 shows an intense and fine green labelling localized around the chloroplast, while the red label was in the middle of the chloroplast. The co-localization of the EDS5H with RCCR shows that EDS5H is located to the chloroplast. cMyc-tag could not be detected in plant cells when the natural EDS5H promoter was used, although the transcription of the EDS5H::cMyc could be determined by RT-PCR (data not shown). This suggests that the EDS5H promoter, which is constitutive, was too weak to drive the whole cassette of the EDS5H::3xcMyc to its target organelle or the amount was below detection limits. Taken together, these studies show that EDS5H is localized at the chloroplast as was shown previously for EDS5 [11].Fig. 4


Localization and expression of EDS5H a homologue of the SA transporter EDS5.

Parinthawong N, Cottier S, Buchala A, Nawrath C, Métraux JP - BMC Plant Biol. (2015)

Subcellular localization of EDS5H-3myc in transgenic Arabidopsis. Mesophyll cells of transgenic plants carrying the CaMV35S::EDS5H::3xmyc construct. The two panes show the cMyc-epitope labelled with Alexa Fluor 488 excited at the wavelength of 488 nm and detected using the emission filter 522 DF32 (green) and Red chlorophyll catalase reductase labelled with Alexa Fluor 568 excited at a wavelength of 568 nm and detected using the emission filters 605 DF32 and 585 EFLP (red). Bar = 5 μm. A representative picture out of 11 transformed lines is presented
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4459457&req=5

Fig4: Subcellular localization of EDS5H-3myc in transgenic Arabidopsis. Mesophyll cells of transgenic plants carrying the CaMV35S::EDS5H::3xmyc construct. The two panes show the cMyc-epitope labelled with Alexa Fluor 488 excited at the wavelength of 488 nm and detected using the emission filter 522 DF32 (green) and Red chlorophyll catalase reductase labelled with Alexa Fluor 568 excited at a wavelength of 568 nm and detected using the emission filters 605 DF32 and 585 EFLP (red). Bar = 5 μm. A representative picture out of 11 transformed lines is presented
Mentions: The ORF of EDS5H was placed under the control of the CaMV 35S promoter upstream of a triple cMyc epitope tag. In tissue sections of transgenic plants carrying the CaMV35S::EDS5H::3xcMyc construct, the cMyc-epitope tag was labelled with a primary anti-myc antibody that was identified by a goat anti-mouse IgG conjugated with Alexa Fluor 488 (green). RCCR (red chlorophyll catalase reductase) was used as an example for a protein targeted to the chloroplast [21]. Tissues were labelled with a primary antibody against RCCR and secondary antibody goat anti-rabbit IgG conjugated with Alexa Fluor 568 (red). Fig. 4 shows an intense and fine green labelling localized around the chloroplast, while the red label was in the middle of the chloroplast. The co-localization of the EDS5H with RCCR shows that EDS5H is located to the chloroplast. cMyc-tag could not be detected in plant cells when the natural EDS5H promoter was used, although the transcription of the EDS5H::cMyc could be determined by RT-PCR (data not shown). This suggests that the EDS5H promoter, which is constitutive, was too weak to drive the whole cassette of the EDS5H::3xcMyc to its target organelle or the amount was below detection limits. Taken together, these studies show that EDS5H is localized at the chloroplast as was shown previously for EDS5 [11].Fig. 4

Bottom Line: An important signal transduction pathway in plant defence depends on the accumulation of salicylic acid (SA).Both transporters are located at the envelope of the chloroplast, the compartment of SA biosynthesis.EDS5H most likely transports related substances such as for example phenolic acids, but unlikely SA.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, University of Fribourg, 1700, Fribourg, Switzerland. kpnongla@kmitl.ac.th.

ABSTRACT

Background: An important signal transduction pathway in plant defence depends on the accumulation of salicylic acid (SA). SA is produced in chloroplasts and the multidrug and toxin extrusion transporter ENHANCED DISEASE SUSCEPTIBILITY5 (EDS5; At4g39030) is necessary for the accumulation of SA after pathogen and abiotic stress. EDS5 is localized at the chloroplast and functions in transporting SA from the chloroplast to the cytoplasm. EDS5 has a homologue called EDS5H (EDS5 HOMOLOGUE; At2g21340) but its relationship to EDS5 has not been described and its function is not known.

Results: EDS5H exhibits about 72% similarity and 59% identity to EDS5. In contrast to EDS5 that is induced after pathogen inoculation, EDS5H was constitutively expressed in all green tissues, independently of pathogen infection. Both transporters are located at the envelope of the chloroplast, the compartment of SA biosynthesis. EDS5H is not involved with the accumulation of SA after inoculation with a pathogen or exposure to UV stress. A phylogenetic analysis supports the hypothesis that EDS5H may be an H(+)/organic acid antiporter like EDS5.

Conclusions: The data based on genetic and molecular studies indicate that EDS5H despite its homology to EDS5 does not contribute to pathogen-induced SA accumulation like EDS5. EDS5H most likely transports related substances such as for example phenolic acids, but unlikely SA.

No MeSH data available.


Related in: MedlinePlus