Optimizing Cucumis sativus seedling vigor: the role of pistachio wood vinegar and date palm compost in nutrient mobilization.

BACKGROUND: The goal of this research is to enhance the quality of cucumber seedlings grown in greenhouses by experimenting with various soilless culture mediums (CMs) and the application of pistachio wood vinegar (WV). The experimental setup was designed as a factorial experiment within a randomized complete block design (RCBD), in greenhouse conditions featuring three replications to assess the effects of different culture media (CMs) and concentrations of pistachio wood vinegar (WV) on cucumber seedling growth. Cucumber seeds were planted in three CMs: coco peat-peat moss, coco peat-vermicompost, and date palm compost-vermicompost mixed in a 75:25 volume-to-volume ratio. These were then treated with pistachio WV at concentrations of 0, 0.5, and 1%, applied four times during irrigation following the emergence of the third leaf. RESULTS: The study revealed that treating seedlings with 0.5% WV in the date palm compost-vermicompost CM significantly enhanced various growth parameters. Specifically, it resulted in a 90% increase in shoot fresh mass, a 59% increase in shoot dry mass, an 11% increase in root fresh mass, a 36% increase in root dry mass, a 65% increase in shoot length, a 62% increase in leaf area, a 25% increase in stem diameter, a 41% increase in relative water content (RWC), and a 6% improvement in membrane stability index (MSI), all in comparison to untreated seedlings grown in coco peat-peat moss CM. Furthermore, chlorophyll a, b, total chlorophyll, and carotenoid levels were 2.3, 2.7, 2.6, and 2.7 times higher, respectively, in seedlings treated with 0.5% WV and grown in the date palm compost-vermicompost CM, compared to those treated with the same concentration of WV but grown in coco peat-peat moss CM. Additionally, the Fv/Fm ratio saw a 52% increase. When plant nutrition was enhanced with the date palm compost-vermicompost CM and 1% WV, auxin content rose by 130% compared to seedlings grown in coco peat-peat moss CM and treated with 0.5% WV. CONCLUSIONS: The study demonstrates that using 0.5% WV in conjunction with date palm compost-vermicompost CM significantly betters the quality of cucumber seedlings, outperforming other treatment combinations.

The Mineral Composition of Date Palm Fruits (Phoenix dactylifera L.) under Low to High Salinity Irrigation.

Adaptability to salinity varies between different varieties of date palm trees. This research aims to explore the long-term impact of different salinity irrigation levels on the mineral content of 13 date palm varieties grown in the United Arab Emirates (UAE). Date varieties were grown using three irrigation water salinity levels of 5, 10 and 15 dS m-1. The mineral composition (B, Ca, Cu, Fe, K, Mg, Na, P and Zn) of date palm fruits was determined using Inductively Coupled Plasma-Optical Emission Spectrometry (ICP-OES). High salinity levels showed no effect on the mineral content of Ajwat AlMadinah, Naghal, Barhi, Shagri, Abu Maan, Jabri, Sukkari and Rothan varieties. All date varieties remained good sources of dietary potassium, magnesium, manganese and boron even at high salinity levels. Increased salinity had no effect on the percent Daily Value (%DV) categories of most of the analyzed minerals. While no genotypes showed a general adaptation to different saline environments, Barhi, Ajwat Al Madinah, Khinizi, Maktoumi and Shagri varieties were more stable towards salinity variation. In the UAE, the genotype x saline-environment interaction was found to be high which makes it impossible to attribute the variation in mineral content to a single varietal or salinity effect.

Study of the Expected Impact of Palm Pollen on Human Respiratory Tract Allergy.

<b>Background and Objective:</b> Date-Palm trees (<i>Phoenix dactylifera</i> L.) are the most abundant crop in Saudi Arabia. Date Palm Pollen (DPP) are considered an important allergens. The reasons for the increase in susceptibility to allergy are not clear. This investigation aimed to link the size, shape, moisture contents and biogenic contents of the collected pollens to susceptibility. <b>Materials and Methods:</b> Pollens were collected from three different regions in the Kingdom. Date palm pollen sizes were determined in wet and dry states by Microtrac. A microscope with a camera was used to image the forms of date palm pollens. The biogenic amines were determined after extraction using HPLC. Data were analyzed by one-way analysis of variance (ANOVA), using SPSS version 16.0. <b>Results:</b> There was a variation in the sizes of pollens from different regions ranging between 3.3-704 µm. Most pollen grains are spherical. The concentrations of six biogenic amines were detected in all samples but with different concentrations. They included B-phenyl ethylamine, Putrescine, Cadaverine, Histamine, Tyramine and Spermidine. <b>Conclusion:</b> The shape and size of the pollen grains studied were variable and will not facilitate deep penetration into the respiratory system but their biogenic contents were very high and suggested to cause allergy.

Molecular Characterization of a Date Palm Vascular Highway 1-Interacting Kinase (PdVIK) Under Abiotic Stresses.

The date palm (Khalas) is an extremophile plant that can adapt to various abiotic stresses including drought and salinity. Salinity tolerance is a complex trait controlled by numerous genes. Identification and functional characterization of salt-responsive genes from the date palm is fundamental to understand salinity tolerance at the molecular level in this plant species. In this study, a salt-inducible vascular highway 1-interacting kinase (PdVIK) that is a MAP kinase kinase kinase (MAPKKK) gene from the date palm, was functionally characterized using in vitro and in vivo strategies. PdVIK, one of the 597 kinases encoded by the date palm genome possesses an ankyrin repeat domain and a kinase domain. The recombinant PdVIK protein exhibited phosphotyrosine activity against myelin basic protein (MBP) substrate. Overexpression of PdVIK in yeast significantly improved its tolerance to salinity, LiCl, and oxidative stresses. Transgenic Arabidopsis seedlings overexpressing PdVIK displayed improved tolerance to salinity, osmotic, and oxidative stresses as assessed by root growth assay. The transgenic lines grown in the soil also displayed modulated salt response, compared to wild-type controls as evaluated by the overall plant growth and proline levels. Likewise, the transgenic lines exhibited drought tolerance by maintaining better relative water content (RWC) compared to non-transgenic control plants. Collectively, these results implicate the involvement of PdVIK in modulating the abiotic stress response of the date palm.

Silicon-mediated alleviation of combined salinity and cadmium stress in date palm (Phoenix dactylifera L.) by regulating physio-hormonal alteration.

We investigated the physio-molecular effects of separate and combined cadmium (Cd; 200 µM) and salinity (NaCl; 100 mM) stress on date palm during silicon (Si; 1.0 mM) applications. The results showed that exogenous Si led to significant improvements in plant growth, as well as physiology when compared with non-Si-treated seedling under stressed (Cd/NaCl) conditions. Interestingly, Si application led to lower metal (Cd) uptake and enhanced plant macronutrient uptake under combined stress, in turn, alleviating the combined salinity- and Cd-induced oxidative stress by lowering the lipid peroxidation rate, and peroxidase and catalase activities. Furthermore, ascorbate peroxidase level and the cytosolic Cu/Zn superoxide dismutase expression were significantly enhanced by Si application under combined stress. We further analyzed the effect of Si on modulation of stress-related hormonal crosstalk. Si markedly downregulated endogenous salicylic acid, jasmonic acid, and abscisic acid under NaCl stress and combined NaCl-Cd stress. However, during Cd toxicity alone, Si showed varying accumulation of these phytohormones. The results suggest that hindering the Cd uptake and enhancing silicon accumulation ultimately led to improvement of biomass and efficiency of the antioxidant system for alleviating combined stress. Moreover, higher transcript accumulation of PROLINE TRANSPORTER 2 and GAPDH and downregulation of ABA RECEPTOR by Si treatment under combined stress in date palm seedlings indicate the stress-ameliorative role of Si. The study provides evidence of the positive influence of Si on alleviating the combined toxicity of Cd and NaCl in date palm and can be further extended for field trials in Cd- and salinity-affected areas.

Functional Characterization of Date Palm Aquaporin Gene PdPIP1;2 Confers Drought and Salinity Tolerance to Yeast and Arabidopsis.

Recent studies on salinity tolerance in date palm revealed the discovery of salt-responsive genes including PdPIP1;2, a highly conserved aquaporin gene in plants, which was functionally characterized in this study to investigate its precise role in drought and salinity tolerance. Immunoblot assay showed a high level of PIP1 protein accumulation only in the leaves of date palm plants when grown under drought, an observation which may imply the involvement of PIP1;2 in CO2 uptake. Heterologous overexpression of PdPIP1;2 in yeast (Saccharomyces cerevisiae) improved tolerance to salinity and oxidative stress. While, heterologous overexpression of PdPIP1;2 in Arabidopsis had significantly (p < 0.05) increased biomass, chlorophyll content, and root length under drought and salinity. In addition, a significantly (p < 0.05) higher percentage of transgenic plants could be recovered by rewatering after drought stress, indicating the ability of the transgenic plants to maintain water and viability under drought. Transgenic plants under drought and salinity maintained significantly (p < 0.05) higher K+/Na+ ratios than wild type (WT) plants, an observation which may represent an efficient tolerance mechanism controlled by the transgene. Collectively, this study provided an insight on the mechanism by which PdPIP1;2 conferred tolerance to salt and drought stresses in date palm.

Modelling the potential effects of climate factors on Dubas bug (Ommatissus lybicus) presence/absence and its infestation rate: A case study from Oman.

BACKGROUND: Ommatissus lybicus de Bergevin (Hemiptera: Tropiduchidae) (Dubas Bug, DB) is an insect pest attacking date palms. It occurs in Arab countries including Oman. In this paper, the logistic, ordinary least square, and geographical weighted regressions were applied to model the absence/presence and density of DB against climate factors. A method is proposed for modelling spatially correlated prorations annually over the study period, based on annual and seasonal outbreaks. The historical 2006-2015 climate data were obtained from weather stations located in nine governorates in northern Oman, while dataloggers collected the 2017 microclimate data in eight of these nine governorates. RESULTS: Logistic regression model showed the percentages of correctly predicted values using a cut-off point of 0.5 were 90%, 88% and 84%, indicating good classification accuracy. OLS and GWR models showed an overall trend of strong linear correlation between DB infestation levels and short- and long-term climate factors. The three models suggested that precipitation, elevation, temperature, humidity, wind direction and wind speed are important in influencing the spatial distribution and the presence/absence of dense DB populations. CONCLUSION: The results provide an improved understanding of climate factors that impact DB's spread and is considered useful for managing DB infestations in date palm plantations. © 2019 Society of Chemical Industry.

Impact of Antioxidants on in vitro Rooting and Acclimatization of Two Egyptian Dry Date Palm Cultivars.

BACKGROUND AND OBJECTIVES: Date palm (Phoenix dactylifera L.) is propagated vegetatively via offshoots, which is limited by either the offshoots numbers produced from a superior selected plant, or the occurrence of these offshoots only during the juvenile phase of the date palm life cycle. As a result, the in vitro propagation could be considered an alternative technique for large scale propagation of date palm. Obtaining well-developed root system is considered the most important step in establishment of reliable protocol for successful production of date palm and subsequently successful acclimatization of the in vitro derived plants. The aim of this study is to obtain a well-developed root system through using different antioxidants, with detecting the similarity between the in vitro derived plants and the mother plants using RAPD, long RAPD and ISSR techniques. MATERIALS AND METHODS: Individual plantlets obtained from maturation of somatic embryos in vitro of about 5-7 cm in length with 2-3 leaves were used as plant material. Plantlets were cultured on half strength MS liquid medium supplemented with 0.5 mg L-1 thiamine-HCl+2.0 mg L-1 glycine+0.1 mg L-1 biotin+40 g L-1 sucrose+ 0.1mg L-1 NAA with different concentrations from either AgNO3 or citric or ascorbic acids (0.0, 0.5, 1, 2 mg L-1). Growth development, root number and root length (cm) were evaluated at the end of the second subculture (12 weeks). Data were reported as Mean±Standard deviation (SD). Data were subjected to one way-analysis of variance (p<0.05). Results were processed by Excel (2010). RESULTS: Among the different antioxidants with different concentrations used, generally it was found that 2 mg L-1 of each agent gave the highest values of growth development, roots number and roots length. However, using 2 mg L-1 AgNO3 gave the best results with all parameters. Regarding the response of date palm cultivar, it was remarkable that Bartamoda showed relatively better results than Sakkoty cultivar. According to PCR reactions, the results of RAPD, long RAPD and ISSR profile of tissue culture-derived plantlets grown on a medium supplemented with 2 mg L-1 AgNO3 obviously revealed high similarity to mother plants. CONCLUSION: It could be concluded that using 2 mg L-1 AgNO3 gave the best results for growth development, root numbers and length of the two cultivars but Bartamoda showed relatively better results than Sakkoty cultivar. The tissue culture-derived plantlets on this medium (2 mg L-1 AgNO3) revealed high similarity to mother plant as a result to RAPD, long RAPD and ISSR profiles.

Biological Control Potential of Two Steinernematid Species Against the Date Fruit Stalk Borer (Oryctes elegans Prell, Coleoptera: Scarabaeidae).

The fruit stalk borer (Oryctes elegans) is an important pest of date palm (Phoenix dactylifera) trees in Saudi Arabia. This study was conducted to determine efficacy of using two species of entomopathogenic nematodes, Steinernema kushidai and Steinernema glaseri, against O. elegans under laboratory and field conditions. Under laboratory conditions, both species of nematodes showed a significant effect on the mortality of O. elegans larvae. Significant variations were observed when insects were exposed to nematodes for variable durations under laboratory conditions. They showed no differences in insect larval mortality when tested either in aqueous suspensions or in Galleria-infected cadavers. Insects exposed to nematode aqueous suspension for 4 d and those treated with Galleria-infected cadavers showed the same rates of mortality, which differed when insects were exposed to nematode-infected cadavers under field conditions. Mean percentages of corrected mortality varied between nematode species and number of infected cadavers. S. kushidai caused significantly higher mortality percentages ± SE (72.17 ± 5.57, 95.83 ± 4.17, 94.43 ± 5.57, and 100%) compared with S. glaseri when the fruit stalk borer, O. elegans, was treated for 6 wk with two, four, six, and eight infected cadavers, respectively.

Differential DNA methylation and transcription profiles in date palm roots exposed to salinity.

As a salt-adaptive plant, the date palm (Phoenix dactylifera L.) requires a suitable mechanism to adapt to the stress of saline soils. There is growing evidence that DNA methylation plays an important role in regulating gene expression in response to abiotic stresses, including salinity. Thus, the present study sought to examine the differential methylation status that occurs in the date palm genome when plants are exposed to salinity, and to identify salinity responsive genes that are regulated by DNA methylation. To achieve these, whole-genome bisulfite sequencing (WGBS) was employed and mRNA was sequenced from salinity-treated and untreated roots. The WGBS analysis included 324,987,795 and 317,056,091 total reads of the control and the salinity-treated samples, respectively. The analysis covered about 81% of the total genomic DNA with about 40% of mapping efficiency of the sequenced reads and an average read depth of 17-fold coverage per DNA strand, and with a bisulfite conversion rate of around 99%. The level of methylation within the differentially methylated regions (DMRs) was significantly (p < 0.05, FDR ≤ 0.05) increased in response to salinity specifically at the mCHG and mCHH sequence contexts. Consistently, the mass spectrometry and the enzyme-linked immunosorbent assay (ELISA) showed that there was a significant (p < 0.05) increase in the global DNA methylation in response to salinity. mRNA sequencing revealed the presence of 6,405 differentially regulated genes with a significant value (p < 0.001, FDR ≤ 0.05) in response to salinity. Integration of high-resolution methylome and transcriptome analyses revealed a negative correlation between mCG methylation located within the promoters and the gene expression, while a positive correlation was noticed between mCHG/mCHH methylation rations and gene expression specifically when plants grew under control conditions. Therefore, the methylome and transcriptome relationships vary based on the methylated sequence context, the methylated region within the gene, the protein-coding ability of the gene, and the salinity treatment. These results provide insights into interplay among DNA methylation and gene expression, and highlight the effect of salinity on the nature of this relationship, which may involve other genetic and epigenetic players under salt stress conditions. The results obtained from this project provide the first draft map of the differential methylome and transcriptome of date palm when exposed to an abiotic stress.

Genotype-dependent changes of gene expression during somatic embryogenesis in oil palm hybrids (Elaeis oleifera x E. guineensis).

To understand the molecular processes triggered during the different steps of somatic embryogenesis (SE) in oil palm, the expression of 19 genes associated to SE identified in proteomic and transcriptomic studies was investigated by qRT-PCR. To evaluate the differential expression of these genes, two interspecific hybrid genotypes (Elaeis oleifera x Elaeis guineensis) contrasting for the acquisition of embryogenic competence were used. Aclorophyllated leaves of both hybrids, one responsive (B351733) and the other non-responsive (B352933) to SE were submitted to callus induction and collected at different time points: 0 (before induction), 14, 30, 90 and 150 days of callus induction (doi). The results obtained showed that all evaluated genes were downregulated at 14 doi in the responsive genotype when compared to the non-responsive. It was also possible to observe that most of the genes changed their expression behavior at 30 doi and were upregulated thereafter until 150 doi, with the exception of the pathogenesis-related PRB1-3-like (PRB1-3) gene, which did not show differential expression at 30 doi and was downregulated at 90 and 150 doi when compared to the non-responsive hybrid. These results indicate that 30 doi is a turning point in gene expression, probably associated to embryogenic competence acquisition. We also show that the expression behavior of the responsive genotype is more stable than that of the non-responsive when the different induction time points are compared to 0 doi (before induction). Moreover, the results obtained in this study corroborate our hypothesis that the regulation of genes involved in the control of oxidative stress and energy metabolism are crucial for the acquisition of embryogenic competence in oil palm.

Date Fruit Proteomics During Development and Ripening Stages.

Gel-based comparative proteomics approach is a valuable technique for studying the changes in abundance of proteins in any given system. The combination of this technique with mass spectrometry has provided immense insight into protein dynamics during fruit development and ripening. This chapter describes, informatively, the procedures for carrying out comparative proteomics analysis of date palm (Phoenix dactylifera L.) fruits at different developmental stages using a combination of two-dimensional gel electrophoresis (2-DE) and mass spectrometry. A comparative proteomics approach provides an overview of protein abundances during fruit maturation and insights into proteins that play key roles during fruit maturation. Moreover, 2-DE technique enables the visualization of total protein distribution and abundance in addition to providing a comparative platform following separation of complex proteins based on their molecular weight and isoelectric point. Overall, this chapter describes methodologies for extraction of proteins from a high carbohydrate-containing fruit, protein quality assessment using one-dimensional gel electrophoresis (1-DE), separation using 2-DE, comparative analysis using Delta2D v4.6, processing of spots of interest, and protein identification using mass spectrometry. This protocol is important for studies aiming at comparative proteomics to gain insights into changes of protein abundances in tissues and organs in general and date palm fruits, in particular.

Cultivar-Dependent Direct Organogenesis of Date Palm from Shoot Tip Explants.

A number of public and private laboratories are working on date palm micropropagation to meet the increasing worldwide demand for date palm planting material. A standardized direct organogenesis protocol exists for the production of date palm plantlets to maintain the genetic fidelity of regenerated plants. Organogenesis has the advantage of using low concentrations of plant growth regulators and avoiding the callus phase. In addition, direct regeneration of vegetative buds minimizes the risk of somaclonal variation among plant regenerants. However, in vitro multiplication cycles should be limited in duration by frequent renewal of plant material. This chapter describes a simple and routine organogenesis protocol for date palm multiplication using shoot tip explants.

NAA-Induced Direct Organogenesis from Female Immature Inflorescence Explants of Date Palm.

Micropropagation has great potential for the multiplication of female and male date palms of commercially grown cultivars by using inflorescences. This approach is simple, convenient, and much faster than the conventional method of using shoot-tip explants. We describe here a stepwise micropropagation procedure using inflorescence explants of Iraqi date palm cultivar Maktoom. Cultured explants were derived from 0.5-cm-long spike segments excised from 8 to 10-cm-long spathes. About 70% formed adventitious buds on Murashige and Skoog (MS) medium supplemented with 2 mg/L naphthalene acetic acid (NAA), 4 mg/L benzylaminopurine (BAP), and 40 g/L sucrose and maintained in the dark for 16 weeks before transferring to normal light conditions. The best multiplication rate was achieved with 3 mg/L 2ip and 2 mg/L; for shoot elongation, the best medium is MS containing 0.5 mg/L BAP, 0.5 mg/L 2ip, and 1 mg/L GA3. Well-developed shoots were cultured for rooting in half MS medium amended with 1 mg/L NAA and 45 g/L sucrose. Plantlets with well-developed roots were successfully hardened in the greenhouse. Inflorescence explants proved to be a promising alternative explant source for micropropagation of date palm cultivars.

Direct Organogenesis from Immature Female Inflorescence of Date Palm by Gradual Reduction of 2,4-D Concentration.

Inflorescences represent an alternative explant source for superior date palm trees, especially those that do not produce offshoots. They provide large numbers of explants free of fungal and bacterial contamination for successful tissue culture initiation. Furthermore, they are characterized by the capacity of plant regeneration within a short time as compared to other explant types. This chapter focuses on the procedures employed for plant regeneration by direct organogenesis using immature female inflorescence explants, including initiation of adventitious buds, differentiation, multiplication, shoot elongation, rooting, and acclimatization. Adding 5 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) into the initiation medium and gradually reducing it to 1 and then to 0.5 mg/L in the subsequent 2 subcultures, respectively, are determining factors in direct adventitious bud formation from the inflorescence. Bud differentiation is obtained on MS medium containing 0.25 mg/L kinetin (Kin), 0.25 mg/L benzyladenine (BA), 0.25 mg/L abscisic acid (ABA), 0.1 mg/L naphthaleneacetic acid (NAA), and 0.2 g/L activated charcoal (AC). Regenerated shoots exhibit sufficient root formation on MS medium supplemented with 2 mg/L indole butyric acid (IBA) and 1 mg/L NAA and subsequent survival in the greenhouse.

Optimized Direct Organogenesis from Shoot Tip Explants of Date Palm.

In vitro propagation is an available alternative to produce uniform and good-quality planting material to establish large-scale date palm cultivation in a short time. This study was carried out to achieve organogenesis and multiplication directly from shoot tips without callus formation, thus avoiding any possibility of undesirable genetic variability among the regenerated plants. The shoot tips explants are cultured on Murashige and Skoog (MS) medium supplemented with 1 mg/L naphthaleneacetic acid (NAA), 1 mg/L naphthoxyacetic acid (NOA), 2.5 mg/L benzyladenine (BA), and 2.5 mg/L isopentenyladenine (2iP). Numerous adventitious buds appeared from the shoot tip explants in darkness after six subcultures at 4-week intervals. Vegetative buds pass through three stages: initiation bud formation, vegetative bud differentiation, and shoot bud proliferation. Shoots are transferred onto medium containing low concentrations of growth regulators for shoot multiplication. The organogenesis protocol described herein consists of six steps: initiation of meristematic buds, multiplication, elongation, rooting, pre-acclimatization, and finally plant acclimatization.

Direct Organogenesis and Indirect Somatic Embryogenesis by In Vitro Reversion of Mature Female Floral Buds to a Vegetative State.

This protocol describes in vitro plant regeneration from mature female inflorescence explants of date palm (Phoenix dactylifera L.) by reversion of floral state (reproductive phase) to the vegetative state. The mature female inflorescence (fully developed) is cultured on MS induction medium containing 10 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), 3 mg/L 2-isopentenyladenine (2iP), and 2 mg/L paclobutrazol (PBZ) or 2 mg/L abscisic acid (ABA). The basal part of the petals has meristematic cells, which can be induced to initiate callus or direct shoot formation depending on the plant growth regulator amendments. Callus forms on the induction medium supplemented with PBZ after 12 weeks, whereas it differentiates into somatic embryos on a medium containing 0.1 mg/L naphthaleneacetic acid (NAA). Direct shoots are regenerated on the induction medium amended with ABA after 24 weeks. Procedures for plant regeneration from mature female inflorescence explants are described, and histological changes which occur during the reversion process are presented.

Enhanced Indirect Somatic Embryogenesis of Date Palm Using Low Levels of Seawater.

Date palm tolerates salinity, drought, and high temperatures. Arid and semiarid zones, especially the Middle East region, need a huge number of date palms for cultivation. To meet this demand, tissue culture techniques have great potential for mass production of plantlets, especially using the indirect embryogenesis technique; any improvement of these techniques is a worthy objective. Low levels of salinity can enhance growth and development of tolerant plants. A low level of seawater, a natural source of salinity, reduces the time required for micropropagation processes of date palm cv. Malkaby when added to MS medium. Medium containing seawater at 500 ppm total dissolved solid (TDS) (12.2 mL/L) improves callus proliferation, whereas 1500 ppm (36.59 mL/L) enhances plant regeneration including multiplication of secondary embryos, embryo germination, and rooting.

Enhanced Indirect Somatic Embryogenesis from Shoot-Tip Explants of Date Palm by Gradual Reductions of 2,4-D Concentration.

Shoot-tip explants obtained from offshoots of adult date palms are an excellent source for callus induction and subsequent somatic embryogenesis. In this protocol, the shoot-tip explants are transferred sequentially to a series of media containing gradually reduced concentrations of plant growth hormones: (a) 10 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) and 3 mg/L 2-isopentenyl adenine (2iP), (b) 7 mg/L 2,4-D and 1 mg/L 2iP, (c) 5 mg/L 2,4-D and 1 mg/L 2iP, and (d) 3 mg/L 2,4-D and 1 mg/L 2iP. Embryogenic callus differentiates into somatic embryos upon transfer to MS medium containing 0.5 mg/L abscisic acid (ABA) and 0.1 mg/L naphthalene acetic acid (NAA). Well-matured somatic embryos germinate on a medium containing 0.1 mg/L NAA. Repeated, multiple, and secondary somatic embryos are induced to produce normal well-developed somatic embryos upon transfer to MS medium containing 0.1 mg/L NAA and 0.05 mg/L benzyladenine (BA). This protocol is potentially applicable for commercial micropropagation of date palm.

Indirect Somatic Embryogenesis from Mature Inflorescence Explants of Date Palm.

Due to the limitations associated with shoot tip explants in the micropropagation of date palm, inflorescence explants are an ideal alternative. This chapter focuses on the protocol for the induction of callus from inflorescence tissue, establishment for proliferation of somatic embryos, germination, elongation, rooting, and acclimatization. Female inflorescences, 30-40 cm in length, cv. Shaishee, were used for culture initiation. After disinfection, the outer inflorescence cover (spathe) is cut open, and the spikelet explants, 1 cm long, are cultured on modified Murashige and Skoog (MS) medium containing 100 mg/L 2,4-D, 3 mg/L kinetin, and 3 mg/L 2ip and incubated at 25 ± 2 °C in the dark. Callus obtained after 6-8 months of culturing is transferred to the culture medium to induce somatic embryogenesis and plant regeneration. Well-developed regenerated shoots are cultured on MS medium containing 0.2 mg/L NAA for root induction and plantlets acclimatized in the greenhouse before transfer to the field.