Current research work in the field of palm tissue culture is devoted to His achievements in the development of the date sector are a credit and honor to the United Arab Emirates and to all developing countries. Date cultivation (Phoenix dactylifera L.) in the United Arab Emirates (UAE) is considered one of the most important economic supports for the agricultural sector.
Propagation of date palm by scions cannot satisfy the increasing demand for date palm trees and high propagation can only be achieved using tissue culture techniques. The objective of this study was to develop a reliable disinfection technique of date palm explants at the initiation stage that provides a contamination rate of less than 5%, to identify tissue culture contaminants and suggest respective control agents.
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INTRODUCTION
- Overview
- Date palm importance
- Date nutritional value
- Geographical distribution
- World production and trade
- Date palm importance in UAE
- Date palm propagation
- Seed propagation
- Offshoot propagation
- Tissue culture techniques
- Date palm tissue culture techniques
- Problems in date palm tissue culture
The exact origin or gene center of the date palm has been lost in ancient times. Much later, the date palm became associated with three of the world's major religions: Islam, Judaism and Christianity. With its simple structure, it is difficult to imagine all the purposes that a date palm can serve.
The remaining GCC countries (Bahrain Qatar and Kuwait) have fewer date palms and lower production. The remaining GCC countries (Bahrain, Qatar and Kuwait) have fewer date palms and lower production. Cultivation of date palm embryo - cotyledon sheath tissues in media containing naphthaleneacetic acid (NAA) produced callus and roots (19).
Injury from cutting the date palm tissue is accompanied by secretion of discolored substance(s) into the environment.
OBJECTIVES
The expected results of this study will influence the success of tissue culture of date palm varieties as well as improve the reproducibility of in vitro date palm introduction protocols.
MATERIALS & METHODS
The base of shoots was cleaned with running water and the outer large leaves and fibers were carefully and gradually removed with a sharp knife until the soft white meristematic tissues emerged (Figure 3). After soaking the explant in an antioxidant solution and before starting the disinfection experiment, a sample (1 g) was taken from the surface of the shoot tip using a cleaned scalpel and forceps. The explant was then surface sterilized by immersion in 30% NaOCI and 1 µl Al iette solution and shaken well for 20 min.
One set of leaves was removed after rinsing the explant in two consecutive changes of sterile water, and then another sample was taken. The explant was subjected to frequent changes in pressure using a vacuum pump (high-performance vacuum pump; . model SPX Robinair Corp.) to ensure that the disinfection. The emulsifier (Tween 20) was used at a rate of 10 drops/liter of disinfecting solution to increase the surface contact between the explant and the disinfecting solution.
One of these pieces was used as a second sample and the remaining was cultured on a 20 ml initiation 24 x 200 mm test tubes. The infected explants were then soaked in a pre-sterilized container with an oxidant solution to minimize the production of phenols that cause browning, and to protect the explant from desiccation. The disinfected explants were then soaked in a pre-sterilized container containing an antioxidant solution to minimize the production of browning-causing phenols and to protect the explant from desiccation.
T he primary ylem and base of leaves of the explant were cut off and the rest of the explant was cut into half rectangles around the apical dome. Experiment m: Effect of time on the efficiency of the best combinations obtained from experiment I and I I. The results were analyzed by the API computer identification software and further analysis was performed by matching the 1 6S ribosomal DNA sequence of the unknown isolates with the bacterial database at the Advance Biotechnology Lab in Dubai UAE.
RESULTS & DISCUSSION
- Identification of bacteria
- Veil bacteria occu r usual ly at the surrounding area of the explants base. It appears mostly at the multipl ication, e longation and rooting stages
To the best of our knowledge, most of the date palm tissue culture laboratories benefit from the fact that they guarantee the external sterilization of the explant. Table (4) illustrates the effect of combination of different concentrations of NaOCl with different amounts of Aliette on total bacterial count (lo�) in date palm shoot tip culture on Khenezi cv. The highest reduction rate of bacterial number, represented by 100 %, was achieved in four different concentrations of the disinfection solution.
The total number of bacteria on the surface of the Khissab explant before the disinfection test ranged from 3.76 to 4.7 1 10 glO cfu. While most yeasts and fungal microorganisms are eliminated during surface sterilization (132), endophytic bacteria survive by successive reproduction, as. Others are non-latent endophytic bacteria that are initially detected from the halo or c volume around the base of the explant in the surface-treated medium.
Before starting experiment II, the total number of encophytic bacteria was determined in surface-sterilized shoot tips of both date palm cultivars. The results of the effect of the I S[ disinfection solution (NaO I + Al iettes) on the surface bacteria count during different time periods are shown in Figures 6, 7 and 8. The effect of time on the endophytic contaminations while applying the best combination of NaOCI and KMn04 solution in both date palm cultivars is illustrated in Figure 9, 1 0 and 1 1.
In conclusion, for both date palm explants, the highest level of reduction was achieved at 15 min exposure to disinfection. Creamy bacteria, usually found on the surface of the culture medium, but rapidly penetrate cultured explants. Due to the above physiological abnormalities, plants will not survive in vitro when directly transferred from the tissue culture laboratory to the field.
B: Each stage is having a special irrigation and nutritional req u i rement
- REFERENCES
- Erskine W, Ahmed TM, Ahmed EO, Zaki L, Arash N, Tamer B and Subhy MR
- F AO Statistics, Global annual date production, 2004
Vitro installation 1 (VP I) at the first acclimatization outside the test tubes (6 months). The objectives of this study were to develop a reliable disinfection technique of date palm explants in the initiation stage that yields a contamination rate of less than 5%, and to identify any bacterial contaminants in order to propose a better control mean. The best two combinations from experiments I and II that produced the least contamination were retested for 1.5, 20 and 25 minutes.
In this study, two date cultivars (Khissab and Khenezi) were used in a total of 1 26 branches. The best disinfectant concentration that can eliminate surface contamination is 40% NaOCI and 3 gil Al iette for 25 min. The best disinfectant concentration that can reduce endophytic contamination is 30% NaOCI and 400 mgll KMn04- along 15 mm.
The future goals of this study are to apply the current selected disinfection technique to all date palm cultivars propagated in the UAE University Date Research and Development Unit and to strengthen the achievement of less than 5%. Date production and conservation with special emphasis on North Africa and the Middle East FAO Technical Bulletin no. Multiplication vegetative du palmier dattier (Phoenix dactylifera L.) par la culture de tissus de jeunes plantes issues de semis. Serie D, dactylifera L.) par la culture de tissus de jeunes plantes issues de semis.
A histological study of adventitious embryo development in organ cultures of Phoenix dactyli/era L.
- APPENDICES
Une bibliographie mise à jour sur la culture tissulaire et le flétrissement vasculaire du palmier dattier et de certaines autres espèces de palmiers. Dri ra N. Multiplication végétative du palmier dattier [Phoenix dactylifera L) par néoformations induites en culture in vitro sur des organes végétatifs et floraux prélevés à la phase mature. Actes du Deuxième Séminaire maghrébin sur la propagation du palmier par les techniques in vitro.
Vitrification: morphological and physiological disorders of in vitro plants. In Debergh PC and Zimmerman RH, editors, Micropropagation Technology and Application. The application of in vitro technology to Australian plants (with particular emphasis on nutrient requirements). Aeration: a simple method to control vitrification and improve in vitro culture of rare Australian plants.
Ecology of Microbial Saprophytes and Pathogens in Tissue Culture and Field-Grown Plants: Causes of Contamination Pathogens in Tissue Culture and Field-Grown Plants: Causes of Contamination Problems in Vitro. In vitro decline in plant cultures: Detection of a legion of hidden bacteria as a cause of degeneration of long-term micropropagated triploid waternelon cultures. Xanthomonas campestris on plant tissue cultures of Aster, Cherianthus, Delphinium, Iris and Rosa, disease development due to latent infection in vitro.
2 Cap the tubes and place them in the ThermaJ cycler and run the PCR reaction as. 5 Mix 5/1 1 of the PCR products with 3 µl of the gel loading dye and load the samples into appropriate wells. 6 Load the DNA ladder into one of the wells to localize the product size (mix 5j.1 of the ladder DNA and 2/11 of the gel loading dye).
Close the tubes and place them in the thermal cycler, perform the thermal cycle with the following program. After the thermal cycle is complete, briefly swirl the tubes to collect the liquid at the bottom of the tube. Note: Make sure the EDTA aqd Na-acetate reaches the bottom of the tube.
5 Discard the supernatant by gently inverting the tube and allow the last drop of liquid to soak onto blotting paper. Note: Once the run is complete, the data is automatically saved to the data folder, i.e. Select File>New Project to open the project settings selection dialog in the New Project Wizard.
To evaluate results in the Analysis QC report �select: Analysis>Report Manager, then select QC Report;. To evaluate results in the Library Search Report Select: Aoalize>Report Manager and then select Library Search Report;. Concise Alignment: For each sample with a % match < 1 00, display the position at which each mismatch occurred, the base called into the consensus sequence for that position, and the base present for that position in the library.
