Variability in inflorescence and offshoot productions of date palm clones derived from tissue culture

Variability in inflorescence and offshoot productions of date palm clones derived from tissue culture

Variability in inflorescence and offshoot productions of date palm clones derived from tissue culture', presented as oral contribution in the Conference 'New Prospects for Science and Education in the MENA region', March, 9-11, 2012, Marrakech, Mrocco. See presentation 'date palm clones' as pdf file)
Date palm (Phoenix dactylifera L.), a long lived dioecious monocotyledon, is an economically crop for many populations in the Maghreb countries. Traditionally, this species is propagated by offshoots produced by trees (average of 30 offshoots by tree during its live).

date palm clones from tissue culture

Considering the negative effect of many abiotic and biotic stresses (such as Bayoud disease) on date palm culture, a need for accelerating date palm propagation is an urgent action. The tissue culture of date palm includes organogenesis and embryogenesis techniques. In Morocco, vitro plants produced through organogenesis and planted in Errachidia region, were checked for their variations on two morphological characters regarding frequencies of inflorescence and offshoots produced by plant clones. Isoenzyme markers including peroxidases, polyphenooxidase, transaminases (GOT), esterases and endopeptidases, were used to characterize morphological traits. Plant clones derived from two selected seedling genotypes (S16 and S35) showed more inflorescence formation than the clones obtained from Jihel (JHL) and Bou-Skri (BSK) date palm cultivars. Using the factorial component analysis (ACP), date palm clones typified respectively by high inflorescences and high offshoots formed two separate groups showing each one some isoenzyme characteristics. The possible occurrence of epigenetic variations in date palm clones is discussed.

Team Publications related to date palm (Phoenix dactylifera L.) propagation through tissue culture

ABOHATEM, M., CHAKRAFI, F., JAITI, F., DIHAZI, A & BAAZIZ, M. 2011. Arbuscular mycorrhizal fungi limit incidence of Fusarium oxysporum f.sp. albedinis on date palm seedlings by increasing nutrient contents, total phenols and peroxidase activities. The Open Horticulture Journal 4, 10-16.
ABOHATEM, M., ZOUINE, J. & EL HADRAMI, I. 2011. Low concentrations of BAP and high rate of subcultures improve the establishment and multiplication of somatic embryos in date palm suspension cultures by limiting oxidative browning associated with high levels of total phenols and peroxidase activities. Scientia Horticulturae 130, 344-348.
BAAZIZ, M. 2011. Reconstitution des palmeraies. Rôle de la recherche scientifique .
إعادة إعمار واحات النخيل بالمغرب. دور البحث العلمي في مواكبة تنمية الواحات
Séminaire de sensibilisation et valorisation des écosystèmes oasiens pour un développement durable, 27-28 Septembre 2011, Foum El Hisn, Tata, Maroc, Morocco.
AZEQOUR, M., AMSSA, M. & BAAZIZ, M.2002. Identification de la variabilité intra-clonale des vitro-plants de palmier dattier issus de culture in vitro par organogenèse. Etude morphologique. Comptes Rendus Biologie 325, 947-956.
AZEQOUR, M., MAJOURHAT, K. & BAAZIZ, M. 2002. Morphological variations and isoenzyme polymorphism of date palm clones from in vitro culture acclimatized and established on soil in south Morocoo. Euphytica 123, 57-66.
EL HADRAMI, I., CHEIKH, R.& BAAZIZ, M. 1995. Somatic embryogenesis and plant regeneration from shoot-tip explants in Phoenix dactylifera L.. Biologia Plantarum, 37, 205-211.
EL HADRAMI, I.& BAAZIZ, M. 1995. Somatic embryogenesis and analysis of peroxidases in Phoenix dactylifera L.. Biologia Plantarum, 37, 197-203.
BAAZIZ, M., AISSAM, F., BRAKEZ, Z., BENDIAB, K., EL HADRAMI, I.& CHEIKH, R. 1994. Electrophoretic patterns of acid soluble proteins and active isoforms of peroxidase and polyphenoloxidase typifying calli and somatic embryos of two reputed date palm cultivars in Morocco. Euphytica, 76, 159-168.


MAJOURHAT, K. 2002. Etudes des aspects quantitatif et qualitatif des peroxydases et des polyphénoloxydases du palmier dattier (Phoenix dactylifera L.). Contribution à l'étude de la diversité génétique de la palmeraie de Marrakech et des nouvelles plantations. thèse de Doctorat en Biologie. Université Cadi Ayyad, Marrakech (Morocco).
EL HADRAMI, I. 1995. L'embryogenèse somatique chez Phoenix dactylifera L.: quelques facteurs limitants et marqueurs biochimiques. thèse de Doctorat d'Etat ès-Sciences. Université Cadi Ayyad, Marakech (Morocco).
AISSAM, F. 1993. embryogenèse somatique et régénération du palmier dattier (Phoenix dactylifera L.). Contribution à l'étude de la régulation physiologique de la nutrition azotée durant la phain vitro. diplôme d'Etudes Supérieures,Université Cadi Ayyad, Marrakech (Maroc).

Other Publications on date palm tissue culture

ABASS M.H. 2013. Microbial contaminants of date palm (Phoenix dactylifera L.) in Iraqi tissue culture laboratories. In Emir. J. Food Agric. 2013. 25 (11): 875-882.
Abstract. The date palm is one of the most important economic species of the palm family, grown mainly for its fruits (dates). Nowadays there is increased demand for date palm fruits around the world. To meet this demand, several propagation methods have been utilized, among them micropropagation which has been used in Iraq and many other countries for large-scale multiplication of date palm. Micropropagation faces several constraints; one is microbial contamination which represents a major challenge to the initiation and maintenance of date palm micropropagation laboratories. In recent years, two major groups of contaminants have been identified and isolated from different date palm tissue culture laboratories in Iraq. The first group is fungi. Several fungal species have been isolated and identified as contaminants; most predominant are: Aspergillus niger, Alternaria alternata and Penicillium spp. The second group is bacteria; predominantly of the genera Bacillus, Staphylococcus and Proteus.
ABAHMANE L. 2013. Recent achievements in date palm (Phoenix dactylifera L.) micropropagation from inflorescence tissues. In: Emir. J. Food Agric. 2013. 25 (11): 863-874.
Abstract. The Moroccan demand for date palm plants over the next decade is estimated to be 2.9 million. To fulfill this huge demand, the use of rapid micropropagation techniques is an objective of great interest. Generally, date palm micropropagation is performed from offshoot tissue. However, in the case of some rare genotypes, this technique is inefficient because only limited numbers of offshoots are available. To overcome this problem, a new technique based on the use of emerged inflorescence tissue was developed. The research activities resulted in a completely successful process starting from plant material excision to plant acclimatization and field planting. The technique was used for the micropropagation of 16 date palm genotypes with good fruit quality. To date, hundreds of well-acclimatized plants belonging to 9 genotypes have been produced. In addition, clusters of buds of 3 selected genotypes and Medjool cv. were produced and transferred to a private laboratory for mass propagation. In vitro plants needed to test for bayoud disease resistance of 6 selected genotypes were produced using this technique. Since 2001, well-acclimatized in vitro plants were gradually transferred to the field to study their behavior. Inflorescence-derived plants have shown normal growth and no abnormalities. Fruit set of planted palms started in 2005. In the present paper, micropropagation with inflorescence tissue as well as the related major achievements are presented.
AL-MAYAHI A. M. W. 2014. Effect of copper sulphate and cobalt chloride on growth of the in vitro culture tissues for date palm (Phoenix dactylifera L.) Cv. Ashgar. In: American Journal of Agricultural and Biological Sciences 9 (1): 6-18, 2014. Link:
Abstract. This study was carried out to investigate the effects of copper sulphate and cobalt chloride on propagation of date palm cv. Ashgar in vitro. The rate of callus proliferation was significantly higher in the medium supplemented with 2 µM copper sulphate and 2 µM cobalt chloride together (p<0.05). Addition of copper sulphate and Cobalt chloride to the medium was most effective for shoots regeneration from callus and enhanced regeneration frequency as well as number of shoots obtained per explant, the best result (7.12 shoot/explant) was obtained by using copper sulphate and Cobalt chloride at 2.0 µM. Also the results of this experiment indicates that the maximum induction of roots can be achieved in the media containing both additives at 0.5 µM. The callus exhibited a decline in carbohydrate contents, proteins, whereas total phenol content increased at high concentration of Cu and Co. The greatest formation of chlorophylls a, b and total chlorophyll was obtained in plantlets grown in the media containing both additives at 0.5 µM. As well as Maximum survivability was noticed for plants cultured in media that containing both copper sulphate and Cobalt chloride at 0.5 µM.
AZAD M. A. K., AREFIN H., HOSSAIN A. 2013. In vitro Morphogenesis of Arabian Date Palm (Phoenix dactylifera L.). In: Plant Tissue Cult. & Biotech. 23(2): 211-219, 2013. Link:
Abstract. After inoculation of young leaves of date palm offshoot required about six months to complete the morphogenesis process. Fourteen weeks were required for embryogenic callus formation under continuous dark condition and nine weeks for shoot initiation (under 16/8 h light/dark). The highest number of explants (80%) produced callus in modified MS containing 5 mg/l 2,4-D + 2 mg/l 2ip. Sixty per cent of explants produced callus in the modified medium containing 5 mg/l 2,4-D + 5 mg/l NAA. While only 50 per cent of the explants formed callus in the same medium when supplemented with only 5 mg/l 2,4-D. The induced calli were transferred to modified MS for shoot proliferation. A combination of two cytokines showed better performance than single ones in shoot induction. The highest percentage (70) of shoot developed in modified MS containing 2 mg/l BAP + 1 mg/l Kn. Forty per cent shoot induction was found in the same medium supplemented with 2 mg/l of BAP. Thirty per cent shoot formed in MS containing 1 mg/l of Kn. The shoots were subcultured at three- four week intervals throughout culture duration.
EL-ASHRY A.A., SHALTOUT A.D., EL-BAHR M.K., EL HAMID A., M.A. MATTER, BEKHEET S.A. 2013. In vitro preservation of embryogenic cultures of two Egyptian dry date palm cultivars at darkness and low temperature conditions. In : Journal of Applied Sciences Research, 9(3): 1985-1992, 2013. Link:
Abstract. This study aimed to investigate the effect of low temperature (5°C) and darkness conditions on preservation of embryogenic cultures of Bartamoda and Sakkoty date palm cultivars. In vitro cultures of those cultivars were obtained by culturing shoot tips excised from offshoots. Embryogenic cultures were proliferated on MS medium supplemented with 10 mg/ l 2,4-D + 3 mg/l 2iP. For low temperature preservation, cultures of the two cultivars were incubated at 5°C for twelve months. Generally, survival percentage was decreased as storage period increased in both two cultivars. After twelve months Sakkoty registered 88. 8 % and Bartamoda registered 87.7 % of survival with considerable browning specially with Bartamoda cultivar. At low temperature, rate of the new proliferated cultures of the two cultivars was decreased as increasing of storage period. Also, the effect of preservation in complete dark at room temperature conditions was investigated. It was found that survival percentage significantly declined after twelve months of preservation. Sakkoty registered high percentage of survival compared with Bartamoda cultivar. It was found that dark condition had negative effects on proliferation of embryogenic cultures in both two cultivars of date palm. Genetic stability was tested using RAPD - PCR analysis. PCR products revealed that the persevered cultures were relatively genetically stable. Bartamoda gave high similarity compared with sakkoty cultivar.

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Other publications on date palm

BALLARDINI M, MERCURI A, LITTARDI C, ABBAS S, COUDERC M, LUDEÑA B, PINTAUD JC 2013. The chloroplast DNA locus psbZ-trnfM as a potential barcode marker in Phoenix L. (Arecaceae). In: Nagy ZT, Backeljau T, De Meyer M, Jordaens K (Eds) DNA barcoding: a practical tool for fundamental and applied biodiversity research. ZooKeys 365: 71–82. Link :
Abstract. The genus Phoenix (Arecaceae) comprises 14 species distributed from Cape Verde Islands to SE Asia. It includes the economically important species Phoenix dactylifera. The paucity of differential morphological and anatomical useful characters, and interspecific hybridization, make identification of Phoenix species difficult. In this context, the development of reliable DNA markers for species and hybrid identification would be of great utility. Previous studies identified a 12 bp polymorphic chloroplast minisatellite in the trnG (GCC)-trnfM (CAU) spacer, and showed its potential for species identification in Phoenix. In this work, in order to develop an efficient DNA barcode marker for Phoenix, a longer cpDNA region (700 bp) comprising the mentioned minisatellite, and located between the psbZ and trnfM (CAU) genes, was sequenced. One hundred and thirty-six individuals, representing all Phoenix species except P. andamanensis, were analysed. The minisatellite showed 2-7 repetitions of the 12 bp motif, with 1-3 out of seven haplotypes per species. Phoenix reclinata and P. canariensis had species-specific haplotypes. Additional polymorphisms were found in the flanking regions of the minisatellite, including substitutions, indels and homopolymers. All this information allowed us to identify unambiguously eight out of the 13 species, and overall 80% of the individuals sampled. Phoenix rupicola and P. theophrastihad the same haplotype, and so had P. atlantica, P. dactylifera, and P. sylvestris (the “date palm complex” sensu Pintaud et al. 2013). For these species, additional molecular markers will be required for their unambiguous identification. The psbZ-trnfM (CAU) region therefore could be considered as a good basis for the establishment of a DNA barcoding system inPhoenix, and is potentially useful for the identification of the female parent in Phoenix hybrids.
GROS BALTHARD M. 2013. Hybridization in the genus Phoenix. A review. In: Emir. J. Food Agric. 2013. 25 (11): 831-842. Link:
Abstract. The genus Phoenix is composed of 14 species naturally distributed in the Old World. This genus comprises the date palm, Phoenix dactylifera L., cultivated for its fruits, the dates, while other species are grown for food, ornament and religious purposes. Phoenix species were, for these reasons, spread out of their natural distribution area. It is therefore common to find species not naturally sympatric, growing together, in cultivation or in the wild. Phoenix species are interfertile and crossing distinct species leads to fertile hybrid offspring (interspecific hybridization). The introduction of a species in the wild generates gene flows leading to the creation of new hybrids and has conservation implications. In cultivation, such crossings may be spontaneous or are the result of artificial pollination, as several reasons impel doing so. Crossing gives rise to beautiful hybrids and is also useful for the conservation of old palm groves threatened by pests. Moreover, artificial pollination of date palms using another Phoenix species can be of interest given the metaxenic pollen effects. In addition, this process may have some potential benefits in date palm improvements, by the creation of hybrid cultivars. Thus, an increasing need of hybrid detection and characterization exists, particularly as morphology alone is not sufficient for this task. Besides new methods such as traditional and geometric morphometrics that may bring new clues, the advent of genetic and molecular markers helps to detect hybrids, especially based on the combination of nuclear and chloroplastic data. The application of methods such as near-infrared reflectance spectroscopy is currently under examination to estimate their potential use for hybrid characterization.
HAIDER M. S., KHAN I. A., NAQVI S. A., JASKANI M.J., KHAN R. W., NAFEES M., PASHA M. I. 2013. Fruit developmental stages effects on attributes in date palm. In: Pak. J. Agri. Sci., Vol. 50(4), 577-583; 2013. Link:
Abstract. Some date palm cultivars grown in Pakistan were biochemically characterized and the effect of fruit maturity on radical scavenging capacity (DPPH), total phenolic contents (TPC), specific activity of antioxidant enzymes, sugars profile and soluble protein contents was assessed. Higher range of differences in composition of studied phytochemicals was recorded among different cultivars. Antiradical efficiency (AE), TPC, antioxidant enzymes and soluble protein contents were recorded higher at khalal stage thereafter, declined at rutab then finally at tamar stage. The amount of glucose (11.32-32.50%) and fructose (10.95-32.41%) started accumulation from khalal stage and were in higher composition at tamar stage due to hydrolysis and inversion of sucrose (10.82-3.1%) contents. The results concluded that variation in biochemical attributes primarily influenced by type of cultivars and different fruit developmental stages. Keywords: Dates, fruit maturity, antioxidants, sugars, total phenolic contents
HAMZA A. M., COLLINS A., ADO S. G., IKUENOBE C. E., ATAGA C. D., ODEWALE J. O. 2014. Proximate Compositions Evaluation and Variability among Cultivars of Date Palm (Phoenix dactylifera L.) in Nigeria. In: International Journal of Plant & Soil Science 3(3). Link:
Abstract. Genetic variability among the fruits of 22 date palm cultivars was studied using 6 nutritional characters to enable us classify the available germplasm into distinct groups on the basis of their genetic diversity using their nutritional characteristics from proximate composition. The main date palm-growing areas of Nigeria were surveyed in 2011 with the objective of characterizing cultivars as to the quality and economic value of their fruits. Descriptive statistics and some multivariate analysis techniques were used to classify the 22 date palm cultivars. The genotypes based on studied traits were grouped into three clusters. Discriminant function analysis was used to confirm the accuracy of grouping that was produced by cluster analysis. Sugar and crude protein content were identified as important traits that could be used to differentiate the genotypes as revealed by both principal component and discriminant analysis. Genetic distance between Daushenga 1 and Saberari 2 exhibited the greatest dissimilarity followed by Daushenga 1 and Hausawa. Hence the use of these parents for hybridization should be given greater emphasis for the production of transgressive segregants with high nutritional potential. The level of variability observed suggested a high diversity among the cultivars. The result of the principal component analysis indicated that the contribution of the first three factors with Eigen value greater than one accounted for 73.03% of the total variation. The moderately high ash content shows that date palm fruit can effectively serve as a source of inorganic minerals and good adsorbent in the removal of metallic ions, odour, colours and other particulate matter from aqueous medium of water and waste water thus making date palm fruit an effective material precursor in water and waste water treatment among other uses. The high soluble carbohydrate content indeed gives an indication that it compares favourably with other fast energy giving food stuffs and thus it can be added to some food content as an additive .This work identified the existence of inherent variability in the local germplasm collections, and the traits that could be used to exploit the observed variability, eliciting important relationships among the traits in the process.
RIAHI K., CHAABANE S., BEN THAYER B. 2013. A kinetic modeling study of phosphate adsorption on to Phoenix dactylifera L. date palm fibers in batch mode. In: Journal of Saudi Chemical Society. Available online 17 December 2013.
Abstract. The removal of phosphates from aqueous solutions by adsorption onto date palm fibers (DPF) has been studied in batch mode. The aim of this study was to understand the mechanisms that govern phosphate sorption and find an appropriate model for the kinetics of removal. In order to investigate the mechanism of sorption and potential rate controlling steps, pseudo first-order, pseudo second-order, intra-particle diffusion and the Elovich equations have been used to test experimental data. Kinetic analysis of the four models has been carried out for initial phosphate concentration in the range of 30–110 mg/L. The rate constants for the four models have been determined and the correlation coefficients have been calculated in order to assess which model provides the best fit predicted data with experimental results. Seven statistical functions were used to estimate the error deviations between experimental and theoretically predicted kinetic adsorption values, including the average relative error deviation (ARED), Marquardt’s percent standard error deviation (MPSED), the hybrid fractional error function (HYBRID), the sum of the squares of the errors (SSE) and three alternative statistical functions, including the Chi-square test, the F-test and Student’s T-test. The results showed that, both Elovich equation and pseudo second-order equation provide the best fit to experimental data for different initial phosphate concentrations.
LIA M., BERNIER U.R., AGRAMONTE N. M., ALQASOUMI S. I., AL-YAHYA M. A., AL-REHAILY A.J., YUSUFOGLU H. S., DEMIRCI F., CAN BASER K. H., KHAN I. A , TABANCA N. 2013. Phoenix dactylifera L. spathe essential oil. Chemical composition and repellent activity against the yellow fever mosquito. In : Acta Tropica 128 (2013) 557–560. Link:
Abstract. Date palm, Phoenix dactylifera L. (Arecaceae), grows commonly in the Arabian Peninsula and is tradition-ally used to treat various diseases. The aim of the present study was to identify chemical composition ofthe essential oil and to investigate the repellent activity. The essential oil of P. dactylifera was obtained byhydrodistillation from the spathe, a specialized leaf structure that surrounds the pollinating organs of thepalm. The oil was subsequently analyzed by GC–FID and GC–MS. The oil showed promising repellent activity against yellow fever mosquito – Aedes aegypti. Sixteen components were characterized, constituting99% of the oil. The main components were 3,4-dimethoxytoluene (73.5%), 2,4-dimethoxytoluene (9.5%),-caryophyllene (5.5%), p-cresyl methyl ether (3.8%), and caryophyllene oxide (2.4%). The minimumeffective dosage (MED) for repellency for the P. dactylifera oil was 0.051 mg/cm 2 , which had moderatelylower potency compared to reference standard N,N-diethyl-3-methylbenzamide, DEET (0.018 mg/cm 2 )in the “cloth patch assay”. The five major compounds were individually assayed for repellency todetermine to what extent each is responsible for repellency from the oil. 3,4-Dimethoxytoluene and2,4-dimethoxytoluene showed the best repellent activity with the same MED value of 0.063 mg/cm 2 ,respectively. The results indicate that these two constituents which comprise a large proportion of theP. dactylifera oil (83%) are likely responsible for the observed repellent activity. In this aspect, the P.dactylifera spathe oil is a sustainable, promising new source of natural repellents.
SHABANI F, KUMAR L 2013. Risk Levels of Invasive Fusarium oxysporum f. sp. in Areas Suitable for Date Palm (Phoenix dactylifera) Cultivation under Various Climate Change Projections. In: PLoS ONE 8(12): e83404. doi:10.1371/journal.pone.0083404. Link:
Abstract. 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.


- Variétés (cultivars) de palmier dattier
- Culture des tissus chez le palmier dattier
- vitro-clones de palmier dattier au Maroc
- Palmier dattier au Maghreb
- Sauver les palmiers au Maroc
- Palmier dattier au Maghreb. principaux stress
- Embryogenèse somatique, mycorhization du palmier dattier
- Date palm (Phoenix dactylifera L. (Ar)
- Marquage en vitro-culture
- QCM-culture in vitro
- Embryogenèse somatique che le palmier dattier


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    Date palm clones-tissue culture