Limits...
A salt-regulated peptide derived from the CAP superfamily protein negatively regulates salt-stress tolerance in Arabidopsis.

Chien PS, Nam HG, Chen YR - J. Exp. Bot. (2015)

Bottom Line: This peptide was found by searching homologues in Arabidopsis using the precursor of a tomato CAP-derived peptide (CAPE) that was initially identified as an immune signal.In searching for a CAPE involved in salt responses, we screened CAPE precursor genes that showed salt-responsive expression and found that the PROAtCAPE1 (AT4G33730) gene was regulated by salinity.We confirmed the endogenous Arabidopsis CAP-derived peptide 1 (AtCAPE1) by mass spectrometry and found that a key amino acid residue in PROAtCAPE1 is critical for AtCAPE1 production.

View Article: PubMed Central - PubMed

Affiliation: Molecular and Biological Agricultural Sciences Program, Taiwan International Graduate Program, Academia Sinica, Taipei 115, Taiwan Agricultural Biotechnology Research Center, Academia Sinica, Taipei 115, Taiwan Graduate Institute of Biotechnology, National Chung-Hsing University, Taichung 402, Taiwan.

No MeSH data available.


Related in: MedlinePlus

Production of AtCAPE1 is mainly derived from root tissues and is regulated by salt. (A) Transcriptional levels of PROAtCAPE1 in different tissues were determined by RT-PCR. Total RNA was extracted from root (R), cotyledon (Co), rosette leaf (RL), cauline leaf (CL), stem (St), flower (F) and silique (Si). 18S rRNA transcripts were used as internal control. (B) Salt response of the transcriptional levels of PROAtCAPE1 in shoots and roots. Ten-d-old seedlings of wild type (Ler) were treated with 125mM NaCl for 0, 3, and 24h. The transcripts of PROAtCAPE1 from the harvested roots and shoots were determined by RT-qPCR. Shown are the average values of (2-∆Ctx1000) from four biological repeats. Error bars, means±SE (PROAtCAPE1 transcripts in shoots versus PROAtCAPE1 transcripts in roots at different time points; Student’s t-test, **P≤0.01). (C) Relative level of endogenous AtCAPE1 in shoots and roots. Seedlings grown for 24h without (1/2 MS) and with 125mM NaCl were subjected to quantitative LC-MS/MS analysis. IS, internal standard. The average values from two biological repeats are shown. Error bars, means±SE. Asterisks indicate statistically significant differences between salt-treated and untreated samples (Student’s t-test; **P≤0.01). (D) Post-translational regulation of AtCAPE1 production. Protein extracts from the transgenic lines (CAPE1oxCNYD) harbouring the AtCAPE1-eYFP fusion grown with (+) and without (-) 125mM NaCl for the indicated times were subjected to western blot analysis. The upper and lower bands with approximate size of 45.7 KDa and 26.3 KDa represent the expected size of the PROAtCAPE1-eYFP fusion protein and the AtCAPE1-eYFP fusion protein, respectively. The fusion proteins were detected by anti-GFP antibody. α-tubulin, loading control.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4526916&req=5

Figure 6: Production of AtCAPE1 is mainly derived from root tissues and is regulated by salt. (A) Transcriptional levels of PROAtCAPE1 in different tissues were determined by RT-PCR. Total RNA was extracted from root (R), cotyledon (Co), rosette leaf (RL), cauline leaf (CL), stem (St), flower (F) and silique (Si). 18S rRNA transcripts were used as internal control. (B) Salt response of the transcriptional levels of PROAtCAPE1 in shoots and roots. Ten-d-old seedlings of wild type (Ler) were treated with 125mM NaCl for 0, 3, and 24h. The transcripts of PROAtCAPE1 from the harvested roots and shoots were determined by RT-qPCR. Shown are the average values of (2-∆Ctx1000) from four biological repeats. Error bars, means±SE (PROAtCAPE1 transcripts in shoots versus PROAtCAPE1 transcripts in roots at different time points; Student’s t-test, **P≤0.01). (C) Relative level of endogenous AtCAPE1 in shoots and roots. Seedlings grown for 24h without (1/2 MS) and with 125mM NaCl were subjected to quantitative LC-MS/MS analysis. IS, internal standard. The average values from two biological repeats are shown. Error bars, means±SE. Asterisks indicate statistically significant differences between salt-treated and untreated samples (Student’s t-test; **P≤0.01). (D) Post-translational regulation of AtCAPE1 production. Protein extracts from the transgenic lines (CAPE1oxCNYD) harbouring the AtCAPE1-eYFP fusion grown with (+) and without (-) 125mM NaCl for the indicated times were subjected to western blot analysis. The upper and lower bands with approximate size of 45.7 KDa and 26.3 KDa represent the expected size of the PROAtCAPE1-eYFP fusion protein and the AtCAPE1-eYFP fusion protein, respectively. The fusion proteins were detected by anti-GFP antibody. α-tubulin, loading control.

Mentions: A series of molecular mechanisms converting the signal into phenotypic resistance to salinity in plants is initiated in the root and systemically transmitted to the shoot (Munns and Tester, 2008; Roy et al., 2014). Thus, the expression of PROAtCAPE1 in different tissues was examined to reveal how PROAtCAPE1 functions in response to salinity. Based on RT-PCR analysis, the PROAtCAPE1 gene was found to be expressed most highly in the roots and was barely detected in above-ground tissues except for the silique (Fig. 6A). In comparison with the wild type under normal conditions, slightly shorter primary roots were observed in proatcape1 seedlings (Supplementary Fig. S5E, available at JXB online). This phenotype suggested the possibility of a functional trade-off of AtCAPE1 between root growth and salt tolerance. When grown to the stage of inflorescence, no growth retardation and no abnormal morphology were observed in siliques in a comparison between the wild type and mutants (Supplementary Fig. S5, available at JXB online). A further investigation on roots by β-glucuronidase (GUS) staining showed that the promoter activity was observed in stelar cells from the elongation zone to the maturation zone and the trichoblast at the maturation zone (Supplementary Fig. S6, available at JXB online).


A salt-regulated peptide derived from the CAP superfamily protein negatively regulates salt-stress tolerance in Arabidopsis.

Chien PS, Nam HG, Chen YR - J. Exp. Bot. (2015)

Production of AtCAPE1 is mainly derived from root tissues and is regulated by salt. (A) Transcriptional levels of PROAtCAPE1 in different tissues were determined by RT-PCR. Total RNA was extracted from root (R), cotyledon (Co), rosette leaf (RL), cauline leaf (CL), stem (St), flower (F) and silique (Si). 18S rRNA transcripts were used as internal control. (B) Salt response of the transcriptional levels of PROAtCAPE1 in shoots and roots. Ten-d-old seedlings of wild type (Ler) were treated with 125mM NaCl for 0, 3, and 24h. The transcripts of PROAtCAPE1 from the harvested roots and shoots were determined by RT-qPCR. Shown are the average values of (2-∆Ctx1000) from four biological repeats. Error bars, means±SE (PROAtCAPE1 transcripts in shoots versus PROAtCAPE1 transcripts in roots at different time points; Student’s t-test, **P≤0.01). (C) Relative level of endogenous AtCAPE1 in shoots and roots. Seedlings grown for 24h without (1/2 MS) and with 125mM NaCl were subjected to quantitative LC-MS/MS analysis. IS, internal standard. The average values from two biological repeats are shown. Error bars, means±SE. Asterisks indicate statistically significant differences between salt-treated and untreated samples (Student’s t-test; **P≤0.01). (D) Post-translational regulation of AtCAPE1 production. Protein extracts from the transgenic lines (CAPE1oxCNYD) harbouring the AtCAPE1-eYFP fusion grown with (+) and without (-) 125mM NaCl for the indicated times were subjected to western blot analysis. The upper and lower bands with approximate size of 45.7 KDa and 26.3 KDa represent the expected size of the PROAtCAPE1-eYFP fusion protein and the AtCAPE1-eYFP fusion protein, respectively. The fusion proteins were detected by anti-GFP antibody. α-tubulin, loading control.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4526916&req=5

Figure 6: Production of AtCAPE1 is mainly derived from root tissues and is regulated by salt. (A) Transcriptional levels of PROAtCAPE1 in different tissues were determined by RT-PCR. Total RNA was extracted from root (R), cotyledon (Co), rosette leaf (RL), cauline leaf (CL), stem (St), flower (F) and silique (Si). 18S rRNA transcripts were used as internal control. (B) Salt response of the transcriptional levels of PROAtCAPE1 in shoots and roots. Ten-d-old seedlings of wild type (Ler) were treated with 125mM NaCl for 0, 3, and 24h. The transcripts of PROAtCAPE1 from the harvested roots and shoots were determined by RT-qPCR. Shown are the average values of (2-∆Ctx1000) from four biological repeats. Error bars, means±SE (PROAtCAPE1 transcripts in shoots versus PROAtCAPE1 transcripts in roots at different time points; Student’s t-test, **P≤0.01). (C) Relative level of endogenous AtCAPE1 in shoots and roots. Seedlings grown for 24h without (1/2 MS) and with 125mM NaCl were subjected to quantitative LC-MS/MS analysis. IS, internal standard. The average values from two biological repeats are shown. Error bars, means±SE. Asterisks indicate statistically significant differences between salt-treated and untreated samples (Student’s t-test; **P≤0.01). (D) Post-translational regulation of AtCAPE1 production. Protein extracts from the transgenic lines (CAPE1oxCNYD) harbouring the AtCAPE1-eYFP fusion grown with (+) and without (-) 125mM NaCl for the indicated times were subjected to western blot analysis. The upper and lower bands with approximate size of 45.7 KDa and 26.3 KDa represent the expected size of the PROAtCAPE1-eYFP fusion protein and the AtCAPE1-eYFP fusion protein, respectively. The fusion proteins were detected by anti-GFP antibody. α-tubulin, loading control.
Mentions: A series of molecular mechanisms converting the signal into phenotypic resistance to salinity in plants is initiated in the root and systemically transmitted to the shoot (Munns and Tester, 2008; Roy et al., 2014). Thus, the expression of PROAtCAPE1 in different tissues was examined to reveal how PROAtCAPE1 functions in response to salinity. Based on RT-PCR analysis, the PROAtCAPE1 gene was found to be expressed most highly in the roots and was barely detected in above-ground tissues except for the silique (Fig. 6A). In comparison with the wild type under normal conditions, slightly shorter primary roots were observed in proatcape1 seedlings (Supplementary Fig. S5E, available at JXB online). This phenotype suggested the possibility of a functional trade-off of AtCAPE1 between root growth and salt tolerance. When grown to the stage of inflorescence, no growth retardation and no abnormal morphology were observed in siliques in a comparison between the wild type and mutants (Supplementary Fig. S5, available at JXB online). A further investigation on roots by β-glucuronidase (GUS) staining showed that the promoter activity was observed in stelar cells from the elongation zone to the maturation zone and the trichoblast at the maturation zone (Supplementary Fig. S6, available at JXB online).

Bottom Line: This peptide was found by searching homologues in Arabidopsis using the precursor of a tomato CAP-derived peptide (CAPE) that was initially identified as an immune signal.In searching for a CAPE involved in salt responses, we screened CAPE precursor genes that showed salt-responsive expression and found that the PROAtCAPE1 (AT4G33730) gene was regulated by salinity.We confirmed the endogenous Arabidopsis CAP-derived peptide 1 (AtCAPE1) by mass spectrometry and found that a key amino acid residue in PROAtCAPE1 is critical for AtCAPE1 production.

View Article: PubMed Central - PubMed

Affiliation: Molecular and Biological Agricultural Sciences Program, Taiwan International Graduate Program, Academia Sinica, Taipei 115, Taiwan Agricultural Biotechnology Research Center, Academia Sinica, Taipei 115, Taiwan Graduate Institute of Biotechnology, National Chung-Hsing University, Taichung 402, Taiwan.

No MeSH data available.


Related in: MedlinePlus