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Maize EMBRYO SAC family peptides interact differentially with pollen tubes and fungal cells.

Woriedh M, Merkl R, Dresselhaus T - J. Exp. Bot. (2015)

Bottom Line: Furthermore, peptide fragments were found to bind differently to fungal cells.Mapping of peptide interaction sites identified amino acids differing in pollen tube burst and fungal response reactions.In summary, these findings indicate that residues targeting pollen tube burst in maize are specific to the ES family, while residues targeting fungal growth are conserved within defensins and defensin-like peptides.

View Article: PubMed Central - PubMed

Affiliation: Cell Biology and Plant Biochemistry, Biochemie-Zentrum Regensburg, University of Regensburg, 93053 Regensburg, Germany.

No MeSH data available.


Related in: MedlinePlus

Accumulation of ES-c and ES-d at spores of U. maydis, ROS detection, and spore burst. (A, B) Spores were germinated in PGM for 24h before addition of TAMRA-ES-d at 30 µM. Three hours after application, ES-d appeared in whole spores. (C, D) Three hours after application of 90 µM TAMRA-ES-d, spores showed swelling and enlargement of cells and formation of big vacuoles (arrowheads). (E) ROS were detected by NBT in spores treated for 24h with 60 µM of ES-d (arrowhead). (F, G) Spores were germinated in PGM for 24h before addition of Dabcyl-ES-c at 30 µM. Three hours after application, ES-c appeared in whole spores. (H, I) Three hours after application of 90 µM Dabcyl-ES-c, spores showed swelling, enlargement of cells, and formation of big vacuoles (arrowheads). ES-c accumulated inside vacuoles and fluorescence was not detected at the surface of spores. (J) ROS was detected by NBT in spores treated for 24h with 60 µM ES-c (arrowhead). (K) Six hours after application of 90 µM ES-d there was burst of swelling spores and excretion of vacuoles and cell components (arrowheads). (L) Spores were germinated in PGM for 24h before addition of TAMRA-ES-a at 30 µM. Three hours after application, fluorescence was not detectable and spores showed normal growth behaviour. (M) Traces of ROS were detected in spores treated for 24h with 60 µM ES-a. Micrographs (A, C, F, H, L) show merge of bright field and fluorescence, while micrographs (B, D, G, I) show fluorescence. (K, M) Bright field micrographs. Scales bars are 5 µm in (A-J, L) and 10 µm in (K, M).
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Figure 7: Accumulation of ES-c and ES-d at spores of U. maydis, ROS detection, and spore burst. (A, B) Spores were germinated in PGM for 24h before addition of TAMRA-ES-d at 30 µM. Three hours after application, ES-d appeared in whole spores. (C, D) Three hours after application of 90 µM TAMRA-ES-d, spores showed swelling and enlargement of cells and formation of big vacuoles (arrowheads). (E) ROS were detected by NBT in spores treated for 24h with 60 µM of ES-d (arrowhead). (F, G) Spores were germinated in PGM for 24h before addition of Dabcyl-ES-c at 30 µM. Three hours after application, ES-c appeared in whole spores. (H, I) Three hours after application of 90 µM Dabcyl-ES-c, spores showed swelling, enlargement of cells, and formation of big vacuoles (arrowheads). ES-c accumulated inside vacuoles and fluorescence was not detected at the surface of spores. (J) ROS was detected by NBT in spores treated for 24h with 60 µM ES-c (arrowhead). (K) Six hours after application of 90 µM ES-d there was burst of swelling spores and excretion of vacuoles and cell components (arrowheads). (L) Spores were germinated in PGM for 24h before addition of TAMRA-ES-a at 30 µM. Three hours after application, fluorescence was not detectable and spores showed normal growth behaviour. (M) Traces of ROS were detected in spores treated for 24h with 60 µM ES-a. Micrographs (A, C, F, H, L) show merge of bright field and fluorescence, while micrographs (B, D, G, I) show fluorescence. (K, M) Bright field micrographs. Scales bars are 5 µm in (A-J, L) and 10 µm in (K, M).

Mentions: All of the above described assays for localization and binding were also applied to U. maydis in order to quantify the effect of ES-c and ES-d on Ustilago spores. Similar to Fusarium, binding and activity of Dabcyl-ES-c, TAMRA-ES-d, and Rhodamine-ES4 on germinated Ustilago spores showed a dose- and time-dependent inhibition. Ustilago spores treated with 30 or 60 µM Dabcyl-ES-c and TAMRA-ES-d showed an accumulation of fluorescence. Spores became thicker and bigger than untreated wild-type spores and mycelia germination was inhibited or prevented completely (Fig. 7A, B, F, G). At a concentration of 90 µM, treated spores were malformed within 3h. Spores appeared swollen and enlarged and formed many vacuoles, which later assembled into few large vacuoles. TAMRA-ES-d was detected in whole spores while Dabcyl-ES-c was detected mainly in vacuoles (Fig. 7C, D, H, I). After 6h, swollen and vacuolated cells started to burst, releasing vacuoles and cellular components (Fig. 7K). In contrast, germinated spores, which were treated with the control TAMRA-ES-a, showed normal spore growth behaviour and fluorescence could not be detected (Fig. 7L). Application of Rhodamine-ES4 resulted in the same inhibition behaviour as ES-c and ES-d. Fluorescence was observed in whole spores and in vacuoles. Furthermore, ROS production was also detected by NBT in spores treated with ES-c, ES-d, and Rhodamine-ES-4. Germinated Ustilago spores produced significant ROS amounts after application of 30 and 60 µM of the above-mentioned peptides (Fig. 7E, J). Traces of ROS were detected when ES-a was applied (Fig 7M).


Maize EMBRYO SAC family peptides interact differentially with pollen tubes and fungal cells.

Woriedh M, Merkl R, Dresselhaus T - J. Exp. Bot. (2015)

Accumulation of ES-c and ES-d at spores of U. maydis, ROS detection, and spore burst. (A, B) Spores were germinated in PGM for 24h before addition of TAMRA-ES-d at 30 µM. Three hours after application, ES-d appeared in whole spores. (C, D) Three hours after application of 90 µM TAMRA-ES-d, spores showed swelling and enlargement of cells and formation of big vacuoles (arrowheads). (E) ROS were detected by NBT in spores treated for 24h with 60 µM of ES-d (arrowhead). (F, G) Spores were germinated in PGM for 24h before addition of Dabcyl-ES-c at 30 µM. Three hours after application, ES-c appeared in whole spores. (H, I) Three hours after application of 90 µM Dabcyl-ES-c, spores showed swelling, enlargement of cells, and formation of big vacuoles (arrowheads). ES-c accumulated inside vacuoles and fluorescence was not detected at the surface of spores. (J) ROS was detected by NBT in spores treated for 24h with 60 µM ES-c (arrowhead). (K) Six hours after application of 90 µM ES-d there was burst of swelling spores and excretion of vacuoles and cell components (arrowheads). (L) Spores were germinated in PGM for 24h before addition of TAMRA-ES-a at 30 µM. Three hours after application, fluorescence was not detectable and spores showed normal growth behaviour. (M) Traces of ROS were detected in spores treated for 24h with 60 µM ES-a. Micrographs (A, C, F, H, L) show merge of bright field and fluorescence, while micrographs (B, D, G, I) show fluorescence. (K, M) Bright field micrographs. Scales bars are 5 µm in (A-J, L) and 10 µm in (K, M).
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Figure 7: Accumulation of ES-c and ES-d at spores of U. maydis, ROS detection, and spore burst. (A, B) Spores were germinated in PGM for 24h before addition of TAMRA-ES-d at 30 µM. Three hours after application, ES-d appeared in whole spores. (C, D) Three hours after application of 90 µM TAMRA-ES-d, spores showed swelling and enlargement of cells and formation of big vacuoles (arrowheads). (E) ROS were detected by NBT in spores treated for 24h with 60 µM of ES-d (arrowhead). (F, G) Spores were germinated in PGM for 24h before addition of Dabcyl-ES-c at 30 µM. Three hours after application, ES-c appeared in whole spores. (H, I) Three hours after application of 90 µM Dabcyl-ES-c, spores showed swelling, enlargement of cells, and formation of big vacuoles (arrowheads). ES-c accumulated inside vacuoles and fluorescence was not detected at the surface of spores. (J) ROS was detected by NBT in spores treated for 24h with 60 µM ES-c (arrowhead). (K) Six hours after application of 90 µM ES-d there was burst of swelling spores and excretion of vacuoles and cell components (arrowheads). (L) Spores were germinated in PGM for 24h before addition of TAMRA-ES-a at 30 µM. Three hours after application, fluorescence was not detectable and spores showed normal growth behaviour. (M) Traces of ROS were detected in spores treated for 24h with 60 µM ES-a. Micrographs (A, C, F, H, L) show merge of bright field and fluorescence, while micrographs (B, D, G, I) show fluorescence. (K, M) Bright field micrographs. Scales bars are 5 µm in (A-J, L) and 10 µm in (K, M).
Mentions: All of the above described assays for localization and binding were also applied to U. maydis in order to quantify the effect of ES-c and ES-d on Ustilago spores. Similar to Fusarium, binding and activity of Dabcyl-ES-c, TAMRA-ES-d, and Rhodamine-ES4 on germinated Ustilago spores showed a dose- and time-dependent inhibition. Ustilago spores treated with 30 or 60 µM Dabcyl-ES-c and TAMRA-ES-d showed an accumulation of fluorescence. Spores became thicker and bigger than untreated wild-type spores and mycelia germination was inhibited or prevented completely (Fig. 7A, B, F, G). At a concentration of 90 µM, treated spores were malformed within 3h. Spores appeared swollen and enlarged and formed many vacuoles, which later assembled into few large vacuoles. TAMRA-ES-d was detected in whole spores while Dabcyl-ES-c was detected mainly in vacuoles (Fig. 7C, D, H, I). After 6h, swollen and vacuolated cells started to burst, releasing vacuoles and cellular components (Fig. 7K). In contrast, germinated spores, which were treated with the control TAMRA-ES-a, showed normal spore growth behaviour and fluorescence could not be detected (Fig. 7L). Application of Rhodamine-ES4 resulted in the same inhibition behaviour as ES-c and ES-d. Fluorescence was observed in whole spores and in vacuoles. Furthermore, ROS production was also detected by NBT in spores treated with ES-c, ES-d, and Rhodamine-ES-4. Germinated Ustilago spores produced significant ROS amounts after application of 30 and 60 µM of the above-mentioned peptides (Fig. 7E, J). Traces of ROS were detected when ES-a was applied (Fig 7M).

Bottom Line: Furthermore, peptide fragments were found to bind differently to fungal cells.Mapping of peptide interaction sites identified amino acids differing in pollen tube burst and fungal response reactions.In summary, these findings indicate that residues targeting pollen tube burst in maize are specific to the ES family, while residues targeting fungal growth are conserved within defensins and defensin-like peptides.

View Article: PubMed Central - PubMed

Affiliation: Cell Biology and Plant Biochemistry, Biochemie-Zentrum Regensburg, University of Regensburg, 93053 Regensburg, Germany.

No MeSH data available.


Related in: MedlinePlus