The Date Palm (Phoenix dactylifera L.) leaf proteome: identification of a gender biomarker to screen male parents

Abstract To establish a proteomic reference map of date palm leaves (Deglet Nour cultivar), we separated and identified leaf proteins using two-dimensional polyacrylamide gel electrophoresis and mass spectrometry, respectively. In total, 284 spots were excised from gel and analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). Among them, 158 were successfully identified (i.e, a success rate of 55.6%) conducting to the identification of 126 unique proteins. These proteins were then clustered according to their functional annotations. Identified proteins were involved in metabolism, electron transport, photosynthesis, protein synthesis, cell structure or defence. However, 29.4 % of the identifications gave unknown function. We then compared the proteome map of female and male trees. Only one discriminated spot was found to be specific of the gender. We identified the corresponding protein as an ABC superfamily ATP binding cassette transporter, ABC protein, a protein whose an ortholog in Arabidopsis thaliana was already reported as required for male fertility and pollen formation. The relevance of this protein as gender biomarker was then confirmed in four other cultivars, i.e., Aligue, Khouet Aligue, Kentichi and Kenta. Such biomarker should be helpful in rapidly distinguishing date palm gender of immature trees

Effects of salt on lipid peroxidation and antioxidant enzyme activities of Catharanthus roseus suspension cells

Cells of Catharanthus roseus (L.) G. Don were maintained and sub-cultured in the absence of NaCl (control) or were sub-cultured for 7 days in the presence of 50 mM (“50”) or 100 mM (“100”) NaCl, or were maintained and sub-cultured for 8 months in the continuous presence of 50 mM NaCl (“50T” or salt-adapted cells). Exposure of cells to salt treatment reduced growth in a concentration-dependent manner. Salt-treated cells strongly accumulated Na+ and, to a lesser extent, Cl−. However, no significant change in Cl− accumulation was observed in 50T cells. Salt progressively increased superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities whereas a strong increase in catalase (CAT) activity was only obtained in 50 and 50T cells. SOD activity was lower in 50T than in 50 cells, whereas the opposite was observed for APX activity. After native polyacrylamide gel electrophoresis (PAGE) analysis, five SOD isoenzymes were detected and the increase in SOD activity observed in salt-treated cells seemed to be mainly due to isoenzymes (SOD 1, 2 and 3). Three APX isoenzymes (APX 1, 2 and 3) were detected markedly, especially in salt-adapted cells. Salt strongly induced CAT 2 isoenzyme in 50 cells and CAT 1 isoenzyme in 50T cells. These data suggest that salt treatment provoked an oxidative stress in C. roseus cells, as shown by the increase in lipid peroxidation, in spite of the induction of antioxidant enzymes. This increase in lipid peroxidation was paralleled by a rise in lipoxygenase (LOX) activity. Increases in antioxidant activities could also be a response to the cellular damage provoked by NaCl. Probably, this increase could not stop the deleterious effects of salt, but reduced stress severity thus allowing cell growth to occur.The Sécrétariat d’état à la recherche Scientifique et à la Technologie of Tunisia is gratefully acknowledged for supporting this research.Peer reviewe

Study of the involvement of H+-ATPase and osmotic adjustment in the resistance of Catharanthus roseus suspension cells to salt stress

The salt-induced H+-ATPase activity and osmotic adjustment responses of Catharanthus roseus (L.) G. Don suspension cultures were studied. Cells were treated with 0, 50 or 100mM NaCl for 7days or were maintained for 8 months with 50 mM NaCl (50T cells). Growth, osmotic potential (), ions content, soluble sugars, proline and total amino acids were determined in the sap of control and salt-treated cells. Salinity reduced cell growth and . The higher decrease in the in salt-treated cells was due to higher accumulation of Na+ and Cl–. The levels of organic solutes, such as soluble sugars, free proline and total amino acids, increased with salt treatment. These results suggest that salt-tolerant cells are able to osmotically adjust. Salinity treatments stimulated H+-ATPase activity. Immunodetection of the enzyme showed that the increased activity was due to an increased amount of protein in the plasmalemma. The induction by NaCl, especially at 100 mM NaCl and for 50T cells, could account for the K+ and Cl– uptake but not for higher or lower tolerance.The Secretariat of State for Scientific Research and Technology from Tunisia is gratefully acknowledged for supporting this research.Peer reviewe