American Journal of Agricultural and Biological Sciences 9 (1): 6-18, 2014 ISSN: 1557-4989 © 2014 A.M.W. AL-Mayahi, This open access article is distributed under a Creative Commons Attribution (CC-BY) 3.0 license doi:10.3844/ajabssp.2014.6.18 Published Online 9 (1) 2014 (http://www.thescipub.com/ajabs.toc) 6 Science Publications AJABS EFFECT OF COPPER SULPHATE AND COBALT CHLORIDE ON GROWTH OF THE IN VITRO CULTURE TISSUES FOR DATE PALM (PHOENIX DACTYLIFERA L.) CV. ASHGAR Ahmed Madi Waheed AL-Mayahi Department of Plant Tissue Culture, Date Palm Research Centre, University of Basrah, Basrah, Iraq Received 2013-09-23; Revised 2013-11-23; Accepted 2013-11-27 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. Keywords: Copper Sulphate, Cobalt Chloride, In Vitro, Ms, N6 Medium, Date Palm 1. INTRODUCTION Date palm (Phoenix dactylifera L.) of the family Arecaceae is a key plantation crop of many countries of arid regions of West Asia and North Africa (Al-Khalifah et al., 2012). The propagation is of both types (sexual and vegetative) Sexual propagation is through seeds and vegetative propagation by offshoot (Al-Khalifah and Shanavaskhan, 2012). The vegetative multiplication of date palm is traditionally achieved by offshoots.This offshoot propagation has limitations such as slow propagation rate, transmission of disease- causing pathogens and insects and production of offshoots in a limited number for a certain period in the life time of a young palm tree (Gueye et al., 2009). Second sources of the propagation are seeds, but it has many limitations like low rate of germination and progeny variations (Chand and Singh, 2004). To overcome the propagation problems and to maintain the germplasm, the in vitro micropropagation (somatic embryogenesis and organogenesis) is the successful technique (Al-Khayri, 2003; Mujib et al., 2004; Bhattacharjee, 2006). Plant tissues and organs are grown in vitro on artificial media, which supply the nutrients necessary for growth. The success of plant tissue culture as a means of plant propagation is greatly influenced by the nature of the culture medium used. For healthy and vigorous growth, intact plants need to take up from the soil. It has become evident that several heavy metals as micro elements play important roles in the regeneration of plant tissue cultures. The metals like cobalt, iron, manganese, copper and zinc are essential for plant life but are required in a very small or trace amounts and become toxic at higher concentrations (Hussein et al., 2010). Copper is a microelement that is essential for normal growth and development of plants. In plant organisms it performs very important physiological and biochemical functions. It takes part in the processes of photosynthesis, respiration, conversion of nitrogen compounds, transport of