Identification and expression analysis of the NAC transcription factor family in durum wheat (Triticum turgidum L. ssp. durum).

The NAC (NAM, ATAF and CUC) proteins belong to one of the largest plant-specific transcription factor (TF) families and play important roles in plant development processes, response to biotic and abiotic cues and hormone signaling. Our analysis led to the identification of 168 NAC genes in durum wheat, including nine putative membrane-bound TFs and 48 homeologous genes pairs. Phylogenetic analyses of TtNACs along with their Arabidopsis, grape, barley and rice counterparts divided these proteins into 8 phylogenetic groups and allowed the identification of TtNAC-A7, TtNAC-B35, TtNAC-A68, TtNAC-B69 and TtNAC-A43 as homologs of OsNAC1, OsNAC8, OsNTL2, OsNTL5 and ANAC025/NTL14, respectively. In silico expression analysis, using RNA-seq data, revealed tissue-specific and stress responsive TtNAC genes. The expression of ten selected genes was analyzed under salt and drought stresses in two contrasting tolerance cultivars. This analysis is the first report of NAC gene family in durum wheat and will be useful for the identification and selection of candidate genes associated with stress tolerance.

Genome-wide identification and expression profiling of the late embryogenesis abundant genes in potato with emphasis on dehydrins.

Late embryogenesis abundant (LEA) proteins were first described as accumulating late in plant seed development. They were also shown to be involved in plant responses to environmental stress and as well as in bacteria, yeast and invertebrates. They are known to play crucial roles in dehydration tolerance. This study describes a genome-wide analysis of LEA proteins and the corresponding genes in Solanum tuberosum. Twenty-nine LEA family members encoding genes in the Solanum genome were identified. Phylogenetic analyses allowed the classification of the potato LEA proteins into nine distinct groups. Some of them were identified as putative orthologs of Arabidopsis and rice LEA genes. In silico analyses confirmed the hydrophilicity of most of the StLEA proteins, whereas some of them can be folded. The in silico expression analyses showed that the identified genes displayed tissue-specific, stress and hormone-responsive expression profiles. Five StLEA classified as dehydrins were selected for expression analyses under salt and drought stresses. The data revealed that they were induced by both stresses. The analyses indicate that several factors such us developmental stages, hormones, and dehydration, can regulate the expression and activities of LEA protein. This report can be helpful for the further functional diversity studies and analyses of LEA proteins in potato. These genes can be overexpressed to improve potato abiotic stress response.

Alterations in lignin content and phenylpropanoids pathway in date palm (Phoenix dactylifera L.) tissues affected by brittle leaf disease.

Brittle leaf disease or Maladie de la Feuille Cassante (MFC) is a lethal disorder of date palm that has assumed epidemic proportions in the oases of Tunisia and Algeria. No pathogen could ever be associated with the disease, while leaflets of affected palms have been previously shown to be deficient in manganese. The work reported here aims to understand the biochemical basis of the date palm response to this disorder. Since the typical disease symptom is the leaf fragility, we have investigated lignin content in leaves and roots. Strong decrease in total lignin content was observed in affected leaves, while lignin content increased in affected roots. Histochemical analyses showed hyperlignification thicker suberin layer in roots cortical cells. The phenylpropanoids pathway was also disrupted in leaves and roots, cinnamoyl-CoA reductase and cinnamyl-alcohol dehydrogenase gene expression was affected by the disease which severely affects the cell wall integrity.

Proteomics analysis of date palm leaves affected at three characteristic stages of brittle leaf disease.

Proteomics analysis has been performed in leaf tissue from field date palm trees showing the brittle leaf disease (BLD) or maladie des feuilles cassantes, the main causal agent of the date palm decline in south Tunisia. To study the evolution of the disease, proteins from healthy and affected leaves taken at three disease stages (S1, S2 and S3) were trichloroacetic acid acetone extracted and subjected to two-dimensional gel electrophoresis (5-8 pH range). Statistical analysis showed that the protein abundance profile is different enough to differentiate the affected leaves from the healthy ones. Fifty-eight variable spots were successfully identified by matrix-assisted laser desorption ionization time of flight, 60 % of which corresponded to chloroplastic ones being involved in the photosynthesis electronic chain and ATP synthesis, metabolic pathways implicated in the balance of the energy, and proteases. Changes in the proteome start at early disease stage (S1), and are greatest at S2. In addition to the degradation of the ribulose-1.5-bisphosphate carboxylase oxygenase in affected leaflets, proteins belonging to the photosynthesis electronic chain and ATP synthesis decreased following the disease, reinforcing the relationship between BLD and manganese deficiency. The manganese-stabilizing proteins 33 kDa, identified in the present work, can be considered as protein biomarkers of the disease, especially at early disease step.

Brittle leaf disease induces an oxidative stress and decreases the expression of manganese-related genes in date palm (Phoenix dactylifera L.).

In Tunisia, date orchards are being decimated by a disease called brittle leaf disease of unknown origin. Previous studies reported that affected soils, roots and leaves were manganese deficient. In this study, we investigated the biochemical and molecular response of MFC-affected date palms to the oxidative stress generated by manganese deficiency. Both the malondialdehyde (MDA) content which is indicative of lipid peroxidation and the activities of antioxidant enzyme were measured in affected leaves and roots. The expression profiles of oxidative stress-related genes encoding superoxide dismutases and peroxidases were also investigated. The data show that the MDA concentration increased but not significantly in affected leaves. However, such MDA increase was significant in roots of MFC-affected plants. The total superoxide dismutase (SOD) activity increased in affected leaves and roots, while RT-PCR experiments showed that MnSOD RNA decreased in affected leaves and roots unlike FeSOD and Cu/Zn-SOD RNA expression increased in these organs. In addition ascorbate peroxidase (APx) and glutathione peroxidase (GPx) RNA expression increased in diseased leaves and roots.

Optimization of RNA isolation from Brittle Leaf Disease affected date palm leaves and construction of a subtractive cDNA library.

A simple and efficient method was described here for the isolation of high-quality RNA from date palm leaves affected with Brittle Leaf Disease (BLD) and containing high amount of phenolic compounds. The procedure was based on the use of a non-ionic detergent Nonidet-P40 (NP-40), Polyvinylpyrrolidone (PVP), and beta-mercaptoethanol in the extraction buffer in order to isolate cytoplasmic RNA and to prevent the oxidation of phenolic compounds. This method allowed the isolation of intact RNA, suitable for cDNA synthesis and library construction. Differential screening of the subtractive cDNA library from affected leaf RNA led to the identification of some BLD-induced genes.