Propagation of Date Palm (Phoenix dactylifera L.) in vitro

Adventitious plantlets were obtained from lateral buds, shoottips, embryos, and pieces of stem and rachilla tissue of Phoenixdactylifera L. cultured on a modified Murashige and Skoog mediumcontaining 3 mg l^–1 N-(²-isopenty)adenine 0?1–100mg l^–1 -naphthaleneacetic acid or 2,4-dichlorophenoxyaceticacid, and 3 g l^–1 activated charcoal. Additions of auxinswere necessary to induce explants to produce callus, adventitiousplantlets, and roots. Plantlets were obtained from explantscultured 3–4 months in vitro. No difference in growthresponses between male and female explants was observed duringculture. Complex addenda of activated charcoal and polyvinylpyrrolidonewere tested in the nutrient media at various concentrationsto prevent explant browning. Activated charcoal fostered satisfactorygrowth by reducing the browning and inhibition of growth ofexplants.     

Review of the vertebrate-mediated dispersal of the Date Palm,Phoenix dactylifera

As a major agricultural crop in the semi-arid and arid zone of North Africa and the Middle East, drupes (“fruits”) of the Date Palm Phoenix dactylifera form with their high carbohydrate content of the flesh and oils in the seed a major part of the diet of resident and migratory bird species and fruit bats. This paper reviews the range of known instances of drupe predation by volant and non-volant vertebrate vectors. While documented dispersal distances range from tens of metres to about 50 km, the efficacy of that dispersal has not been assessed in any of the papers under review. While volant animals, primarily birds, make up the greatest number of dispersal vectors and also account for the greatest number of seeds dispersed, long distance dispersal of a larger quantity of seeds per dispersal event seems to rely on terrestrial animals, primarily canids, but also bears and flightless birds, such as the Emu (Dromaius novaehollandiae).     

In Vitro Flowering Induction in Date Palm (Phoenix dactylifera L.)

This article reports on a histological and morphological study on the induction of in vitro flowering in vegetatively propagated plantlets from different date palm cultivars. The study aimed to further explore the control of in vitro flower induction in relation to the photoperiodic requirements in date palm and to come up with a novel system that may allow for early sex determination through plant cycle reduction. In fact, the in vitro reversion of a shoot meristem from a vegetative to a reproductive state was achieved within 1–5 months depending on the variety considered. This reversion was accompanied by several morphological transformations that affected the apical part of the leafy bud corresponding mainly to a size increase of the prefloral meristem zone followed by the appearance of an inflorescence. The flowers that were produced in vitro were histologically and morphologically similar to those formed in vivo. The histological examination of the in vitro flowering induction process showed that the conversion into inflorescences involved the entire apical vegetative meristem of the plantlet used as a starting material and brought about a change in its anatomical structure without affecting its phyllotaxis and the leaf shape. Through alternating between hormone-free and hormone-containing media under different light/dark conditions, the highest flower induction rates were obtained with a basal Murashige and Skoog medium. A change in the architectural model of date palm was induced because unlike the natural lateral flowering, in vitro flowering was terminal. Such in vitro flower induction allowed a significant reduction in plant cycle and can, therefore, be considered a promising candidate to save time for future improvement and selection programs in date palm.     

Physicochemical and Functional Properties of Typical Tunisian Drink: Date Palm Sap (Phoenix dactylifera L.)

The present work aimed to study the physicochemical characteristics and the functional properties of the male date palm sap (Phoenix dactylifera L.). The surface properties at the air–water interface were studied on the basis of the drop volume method. Foaming properties (foam capacity and stability) were evaluated using bubbling method by optical and conductimetric measurements (foamscan). Composition analysis revealed a high sugar content (92.29% w/w dry matter basis) with dominance of sucrose. Sap contains also 5.14% w/w of proteins and 2.57% w/w of ash. Proteins are probably the source of the surface activity and the observed foam power. At higher contents of dry matter, lyophilized sap solutions showed Newtonian comportment and improved surface activity and foam properties. Results present some interesting functional properties that suggest to deepen research particularly in sap proteins.     

Identification of Reference Genes for Quantitative Real-Time PCR in Date Palm (Phoenix dactylifera L.) Subjected to Drought and Salinity

Date palm is an important crop plant in the arid and semi-arid regions supporting human population in the Middle East and North Africa. These areas have been largely affected by drought and salinity due to insufficient rainfall and improper irrigation practices. Date palm is a relatively salt- and drought-tolerant plant and more recently efforts have been directed to identifying genes and pathways that confer stress tolerance in this species. Quantitative real-time PCR (qPCR) is a promising technique for the analysis of stress-induced differential gene expression, which involves the use of stable reference genes for normalizing gene expression. In an attempt to find the best reference genes for date palm’s drought and salinity research, we evaluated the stability of 12 most commonly used reference genes using the geNorm, NormFinder, BestKeeper statistical algorithms and the comparative ΔC_T method. The comprehensive results revealed that HEAT SHOCK PROTEIN (HSP), UBIQUITIN (UBQ) and YTH domain-containing family protein (YT521) were stable in drought-stressed leaves whereas GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE (GAPDH), ACTIN and TUBULIN were stable in drought-stressed roots. On the other hand, SMALL SUBUNIT RIBOSOMAL RNA (25S), YT521 and 18S ribosomal RNA (18S); and UBQ, ACTIN and ELONGATION FACTOR 1-ALPHA (eEF1a) were stable in leaves and roots, respectively, under salt stress. The stability of these reference genes was verified by using the abiotic stress-responsive CYTOSOLIC Cu/Zn SUPEROXIDE DISMUTASE (Cyt-Cu/Zn SOD), an ABA RECEPTOR, and a PROLINE TRANSPORTER 2 (PRO) genes. A combination of top 2 or 3 stable reference genes were found to be suitable for normalization of the target gene expression and will facilitate gene expression analysis studies aimed at identifying functional genes associated with drought and salinity tolerance in date palm.     

Date Seeds (Phoenix dactylifera): Antioxidant Potential and Profile of Free and Bound Polyphenols from Different Cultivars

In this study, the antioxidant activities and detailed phenolic profiling of extracts from seven cultivars of date seeds were investigated. Significant differences were detected among cultivars. Total phenolic content (TP) ranged between 135.9±12.1 and 284.86±21.9 mg GAE/g DM. The total flavonoid value varied between 34.20±0.34 and 94.46±1.04 mg RE/g DM. The condensed tannin ranged from 24.17±1.13 to 201.60±9.95 mg CTE/g DM. Phloroglucinolysis was used to depolymerize the bound polyphenols. Results show the presence of phenolic acids: Hydroxybenzoic acid, hydroxycinnamic acid, and a high amount of flavan-3ols (monomers, dimers, and trimers). Before depolymerization, the highest amount of total polyphenols was identified in Kenta (8.48 g/kg) and the lowest was detected in Hessa (4.74 g/kg). After depolymerization, the flavan-3-ols increased significantly, ranging between 46.91g/kg in Hessa and 72.38 g/kg in Deglet Nour, with a high degree of polymerization (DP) in all cultivars. It can be concluded that date seeds represent a good source of bioactive compounds.     

Phaeoacremonium spp. and Botryosphaeriaceae spp. Associated with Date Palm (Phoenix dactylifera L.) Decline in Iran

During spring and summer of 2011, a survey was undertaken on some palm groves in the Kerman province (south‐eastern Iran) to determine the fungal pathogens associated with date palm (Phoenix dactylifera L.) decline diseases. Samples were taken from date palm trees showing yellowing, wilting and dieback symptoms. Isolations were made from symptomatic tissues on malt extract agar (MEA) supplemented with 100 mg/l streptomycin sulphate (MEAS). Two species of Phaeoacremonium, Phaeoacremonium aleophilum and Pm. parasiticum, and two species of Botryosphaeriaceae, Botryosphaeria dothidea andDiplodia mutila, were isolated from affected trees and identified on the basis of morphological, cultural and molecular characteristics. Pathogenicity tests were performed on date palm (4‐year‐old potted plants) under greenhouse conditions. Based on the pathogenicity tests, Pm. aleophilum was the most virulent and caused the longest lesions. This is the first report of Pm. aleophilum and B. dothidea and their pathogenicity on date palm tree.     

Isolation of Pathogenic Bacillus circulans from Callus Cultures and Healthy Offshoots of Date Palm (Phoenix dactylifera L.)

Pure cultures of a gram-negative, rod-shaped bacterium, which produced endospores after 3 to 5 days on solid medium, were isolated exclusively from tissue cultures of the date palm Phoenix dactylifera L. Electron microscopic examination of thin sections of the bacteria revealed the bilayer membrane typical of gramnegative bacteria and confirmed the nature of the spores as true endospores. Biochemical and physiological tests indicated that the bacteria were Bacillus circulans. B. circulans was consistently isolated from the internal tissues, including the meristem, of apparently healthy offshoots of date palm. When meristem and embryo callus tissue culture samples were injected with B. circulans isolated from similar tissue culture samples and from offshoots, the majority of the isolates produced a rapid, destructive soft rot of the tissues.     

Photooxidative Stress Modulation of Endogenous Phytohormone and Antioxidant Accumulations and Fruit Maturity in Date Palm (Phoenix dactylifera L.)

A common modern cultivation practice is bagging the fruit bunch if date palm (Phoenix dactylifera L.) which may influence fruit maturity and nutraceutical quality. Exposure of fruits to photooxidative stress induces changes in the endogenous concentrations of plant hormones and other metabolites, which may cause accelerated fruit maturity. This study was conducted to examine the effect of exposure to direct and indirect sunlight on date palm fruit development. The indirect sunlight treatment was simulated by fruit bunch bagging, a common practice in modern date production. The exposure of date palm fruits to direct sunlight-induced photooxidative stress causing an increased concentration of ascorbic acid and decreased content of chlorophyll, anthocyanins, carotenoids, and phenols compared to the fruit bagging treatment. Direct sunlight also reduced the concentration of phytohormones, including indoleacetic acid, gibberellin, and zeatin, but increased abscisic acid accumulation. The directly-exposed fruits reached a partially-mature stage (Rutab) in August, whereas the bagged fruits remained at the immature stage (Khalal). This study is the first to describe the biochemical basis of the observed improvement of date palm fruit development in response to reduced light intensity. Besides, it provides insights into controlling date palm fruit maturity and subsequently prolonging the shelf life dates on the tree; thus, extending the marketing period for the benefit of the farmers.

     

Genetic Diversity and Population Structure of Native and Introduced Date Palm (Phoenix dactylifera) Germplasm in the United Arab Emirates

Date palm (Phoenix dactylifera) has been cultivated since ancient times, but little is known about its genetic diversity and population structure. Examination of 80 date palm accessions grown in the United Arab Emirates, including a collection of varieties from around the world, using 21 microsatellite markers, indicated extensive genetic diversity, with many accessions heterozygous for most markers. The average number of alleles per locus (19), expected heterozygosity (0.7), observed heterozygosity (0.25) and fixation indices (Fst?=?0.6, Rst?=?0.72) demonstrated significant population structure. Analysis with a model-based Baysian method, STRUCTURE 2.4.1, indicated that the 80 accessions could be broadly divided into nine groups. Independent samples of genotypes with the same name, collected from different experimental stations, usually clustered together. The study was enriched for germplasm from the United Arab Emirates (UAE), and one STRUCTURE-derived grouping consisted mainly of UAE accessions. In a few other clusters, several genotypes from the UAE, Iraq and Oman grouped together. Two clusters included accessions from both North Africa and the Middle East. Many accessions in the STRUCTURE-derived populations appeared to be genetic admixtures. The results indicated a broad dissemination of related germplasms across date-palm growing regions of the world, with very few alleles that still correlate with particular regional germplasms.     

Risk Levels of Invasive Fusarium oxysporum f. sp. in Areas Suitable for Date Palm (Phoenix dactylifera) Cultivation under Various Climate Change Projections

Global climate model outputs involve uncertainties in prediction, which could be reduced by identifying agreements between the output results of different models, covering all assumptions included in each. Fusarium oxysporum f.sp. is an invasive pathogen that poses risk to date palm cultivation, among other crops. Therefore, in this study, the future distribution of invasive Fusarium oxysporum f.sp., confirmed by CSIRO-Mk3.0 (CS) and MIROC-H (MR) GCMs, was modeled and combined with the future distribution of date palm predicted by the same GCMs, to identify areas suitable for date palm cultivation with different risk levels of invasive Fusarium oxysporum f.sp., for 2030, 2050, 2070 and 2100. Results showed that 40%, 37%, 33% and 28% areas projected to become highly conducive to date palm are under high risk of its lethal fungus, compared with 37%, 39%, 43% and 42% under low risk, for the chosen years respectively. Our study also indicates that areas with marginal risk will be limited to 231, 212, 186 and 172 million hectares by 2030, 2050, 2070 and 2100. The study further demonstrates that CLIMEX outputs refined by a combination of different GCMs results of different species that have symbiosis or parasite relationship, ensure that the predictions become robust, rather than producing hypothetical findings, limited purely to publication.     

Identification of date (Phoenix dactylifera) cultivars by protein patterns

The water-soluble proteins extracted from the soft tissue of ripe dates (cultivars Rcziz, Marzban, Mowahed, Kholass, Hatmy, Shishy, Shahl and Gir) collected in Saudi Arabia and one cultivar (Muskade) from Iraq were freed from some of their acidic polysaccharides by treatment with 33 % propan-2-ol. The best protein separations and the most characteristic patterns with which to identify the cultivars were obtained after PoroPAGE or PAGIF, with both tube and thin-layer techniques. The native proteins showed characteristic M,s in the range of 20,22,24 and 27 k and difierentiation by isoelectric points either in the range pH 4–5 or pH 8–9. The main subunits (M, 25 and 85 k) were used for final discrimination in SDS-PoroPAGE or in SDS-PAGE.     

Molecular characterization and in silico analysis of RNA polymerase alpha subunit gene (rpoA) in Date Palm (Phoenix dactylifera L.) cv. Deglet Nour

The rpoA gene coding for the ??-subunit of DNA-dependent RNA polymerase located in the chloroplastic genome of date palm has been characterized from the elite cultivar Deglet Nour by cloning and sequencing of the PCR amplification product. The full length of rpoA-Pd (Phoenix dactylifera) gene was 1014 bp. The comparison in Genbank showed that the rpoA gene has a 100% homology with the Khalas cultivar of date palm and a strong homology with Oil Palm (99%). The deduced protein full length is 337 amino acid corresponding to 38,692 Da polypeptide. It contained an Alpha N-terminal domain (alpha-NTD) between 1 to 233 (aa) and Alpha C-terminal domain (alpha-CTD) between 266 to 337 (aa). Initially, we have compared the sequences of the full-length DNA rpoA gene from Date Palm and Oil palm, 15 mutations have been detected, 4 do not affect amino acid sequences. A multiple alignment of protein sequences of Date Palm and other plants shows 6 mutations specific for palms family and one is specific to monocots species. A multiple alignment of 35 nucleotide sequences from different plant species shows 3 SNPs specific to Date Palm, 6 SNPs specific to Palms family and 6 other to monocot species. The phylogenetic analysis performed in this work shows a strong similarity between Pd-rpoA and rpoA genes from other plant species, but it shows a great divergence with the rpoA of E. coli. To explain whether the separation of the two clades was due to selection pressure. We calculate the ratio Ka/Ks for different species. A synteny analyses of rpoA genes was effected, a high genomic synteny is observed for the ropA in all the species included in this study.     

Molecular Clocks and Archeogenomics of a Late Period Egyptian Date Palm Leaf Reveal Introgression from Wild Relatives and Add Timestamps on the Domestication

The date palm, Phoenix dactylifera, has been a cornerstone of Middle Eastern and North African agriculture for millennia. It was first domesticated in the Persian Gulf, and its evolution appears to have been influenced by gene flow from two wild relatives, P. theophrasti, currently restricted to Crete and Turkey, and P. sylvestris, widespread from Bangladesh to the West Himalayas. Genomes of ancient date palm seeds show that gene flow from P. theophrasti to P. dactylifera may have occurred by ∼2,200 years ago, but traces of P. sylvestris could not be detected. We here integrate archeogenomics of a ∼2,100-year-old P. dactylifera leaf from Saqqara (Egypt), molecular-clock dating, and coalescence approaches with population genomic tests, to probe the hybridization between the date palm and its two closest relatives and provide minimum and maximum timestamps for its reticulated evolution. The Saqqara date palm shares a close genetic affinity with North African date palm populations, and we find clear genomic admixture from both P. theophrasti, and P. sylvestris, indicating that both had contributed to the date palm genome by 2,100 years ago. Molecular-clocks placed the divergence of P. theophrasti from P. dactylifera/P. sylvestris and that of P. dactylifera from P. sylvestris in the Upper Miocene, but strongly supported, conflicting topologies point to older gene flow between P. theophrasti and P. dactylifera, and P. sylvestris and P. dactylifera. Our work highlights the ancient hybrid origin of the date palms, and prompts the investigation of the functional significance of genetic material introgressed from both close relatives, which in turn could prove useful for modern date palm breeding.     

Date stones of Phoenix dactylifera and jujube shells of Ziziphus lotus as potential biosorbents for anionic dye removal

This study investigated the biosorption of Congo Red (CR) from aqueous solution by Date stones (DS) of Phoenix dactylifera and jujube shells (JS) of Ziziphus lotus. Batch operations were carried out in the liquid phase to observe the effect of various experimental parameters such as contact time, pH, temperature, and initial dye concentration on removal of CR. The characteristics of the DS and JS were also examined by Fourier transform infrared analysis. The biosorption data of CR on DS and JS samples were well fitted with the Langmuir isotherm and pseudo-second kinetics model with a maximum biosorption amount of 45.08 mg g^?1 for DS and 59.55 mg g^?1 for JS at pH = 4, temperature of 50°C, and an initial concentration of 100–800 mg L^?1 after 90 minutes of contact time. The outcomes indicated that DS and JS can be used as a good low-cost alternative for the treatment of effluents containing CR in water.     

Identification and characterization of gene-based SSR markers in date palm (Phoenix dactylifera L.)

Background

Clp/Hsp100 chaperones are involved in protein quality control. They act as independent units or in conjunction with a proteolytic core to degrade irreversibly damaged proteins. Clp chaperones from plant chloroplasts have been also implicated in the process of precursor import, along with Hsp70 chaperones. They are thought to pull the precursors in as the transit peptides enter the organelle. How Clp chaperones identify their substrates and engage in their processing is not known. This information may lie in the position, sequence or structure of the Clp recognition motifs.

Results

We tested the influence of the position of the transit peptide on the interaction with two chloroplastic Clp chaperones, ClpC2 and ClpD from Arabidopsis thaliana (AtClpC2 and AtClpD). The transit peptide of ferredoxin-NADP⁺ reductase was fused to either the N- or C-terminal end of glutathione S-transferase. Another fusion with the transit peptide interleaved between two folded proteins was used to probe if AtClpC2 and AtClpD could recognize tags located in the interior of a polypeptide. We also used a mutated transit peptide that is not targeted by Hsp70 chaperones (TP1234), yet it is imported at a normal rate. The fusions were immobilized on resins and the purified recombinant chaperones were added. After a washing protocol, the amount of bound chaperone was assessed. Both AtClpC2 and AtClpD interacted with the transit peptides when they were located at the N-terminal position of a protein, but not when they were allocated to the C-terminal end or at the interior of a polypeptide.

Conclusions

AtClpC2 and AtClpD have a positional preference for interacting with a transit peptide. In particular, the localization of the signal sequence at the N-terminal end of a protein seems mandatory for interaction to take place. Our results have implications for the understanding of protein quality control and precursor import in chloroplasts.     

A water-soluble galactomannan from the seeds of Phoenix dactylifera L

A water-soluble polysaccharide, isolated from the seeds of dates, has been investigated using methylation, periodate and CrO(3) oxidation, NMR spectroscopy, and reaction with Bandeiraea simplicifolia lectin and alpha-D-galactosidase. The polysaccharide consists of a backbone composed of (1-->4)-beta-D-mannopyranosyl residues and carries a single (1-->6)-alpha-linked D-galactopyranosyl residue.     

Detection and Molecular Identification of Aster Yellows Phytoplasma in Date Palm in Egypt

Phytoplasma‐like symptoms were detected in date palm trees (Phoenix dactylifera L.) in Al‐Giza Governorate in Egypt. Symptoms varied from leaf chlorotic streaks, stunting and marked reduction in fruit and stalk sizes. Direct and nested PCR of symptomatic samples using P1/P7 and R16F2n/R16R2n primers, respectively, of the 16S rRNA gene, resulted in a DNA amplification product of c. 1.3 kbp. Symptomless samples collected from the same location and the healthy control produced no product upon amplification. Products were cloned into TOPO TA vector for sequencing. Data generated were deposited in the GenBank (Accession KF826615 ). A BLAST search showed that the sequence of the 16SrRNA gene shared ‘Candidatus Phytoplasma asteris’ (16SrI group) with other isolates. Phylogenetic analysis revealed that the isolate clustered with the date palm phytoplasma causing Al‐Wijam disease in Saudi Arabia.