Author name:
عبد الامير رحيم عبيد
Supervisor name:
حسيي خلف زاير | لوى حسين عبد القادر
Abstract:
This study was conducted at the tissue culture laboratory at the Dept. of biology, College of Education pure Sciences from 1/3/2012 - 1/12/2014.With the objective of studing the tolerance of the salt stress of two rice (Oryza sativa L.)cultivars namely Anber and Furat using the technology of tissue culture. . Mature seeds (mature embryo and endosperm) were used as bing parts of explants which were cultivated in a semi - sold( Murashige and Skoog 1962) nutrient medium and some additives such as vitamins and amino acids .The growth regulator Dichlorophenoxy acetic acid (2.4 - D)was used, about 8mg/l , sucrose at 30g.l - 1 . . The study ,also ,tested the effect maltose at 30g.l - 1, and the effect of 0.5g/l of casein hydrolest were tested.the study also included the different of sea water concentrations (0,20,40,60)%on the induction of growth and development of callus ,the effect of green algae extract at Cladophora crispate by adding 1g fresh weight to the nutrient medium with all sea water concentrations.All the features of the callus were tested as well as the effect of the interaction between sea water concentrations and algae extract , as well as interactions among rice cultivar x sea water concentration x algal extract.The study depended on some growth indicators to determine the degree of callus to tolerance towards salt stress .the resistance of callus to salt depended on regeneration through the required of time for the formation of embryos ,number of germinated embryos ,and number of plantlets .The concentration of the major metal ion : sodium, potassium, phosphorus, calcium ,magnesium, potassium/sodium percentage , and the minor metal ions : iron ,copper, manganese were also some chemical components such as protines and amino acid proline measured, were also measured .The randomized block design (C.R.D)and factorial experiments were used depending ten replicates per treatment .The results can be summarized as follows : 1 - The results showed that the addition of sugar di maltose as an energy source and Casein hydrolyses to the nutrient medium had appositive effect on significant decrease in the required time to mitiate the callus ,in the increase of callus induction percentage and in the increase of freshweight of initial callus. .2 - The effect of sea water and algae extract : . A - The current study shaued that the addidtion of 20 and 40 % sea water to the nutrient medium resulted in a significant increase in the required time to induce the callus ,and a significant decrease in the percentage of callus induction and weight of the mitial callus as compared with the control treatment .The concentration 60%0f sea water caused the mature seeds to lose their ability to germinate and induce the callus if no induction occurs at all . The effect of algae extract resulted in a significant decrease in the required time for callus induction ,and a significant increase in the percentage of callus induction ,the weight of the mitial callus and 0,20,40% of sea water as compared with free algae extract medium.B - The results showed that the addition of 20% of sea water to the nutrient medium that is prepared for the growth of the embryo callus led to a significant increase in the fresh weight as compared with the control treatment , whereas the two concentrations 40 and 60%sea water led to a significant decrease in the embryo callus weight .The effect of the algae extracted to a significant increase in the fresh weight of the embryo callus with sea water concentrations as compared with the free algae extract medium.C - The addition of 20,40 and 60%sea water cause a significant decrease in the number of the somatic embryos ,thir germination auerage ,number and germination of the formed plantlets . This addition also caused a significant increase in the required time to stare the germination of the somatic embryos as compared with the control treatment .The addition of the algae extract led to increase the number of formed embryos ,germination percentage ,and number and percentage of plantlets ,wher was the addition led to lower the required time to germination the embryos compared with the free algae extract medium.D - The addition of 20 ,40 and 60% sea water to the nutrient medium resulted in a significant increase in the concentration of sodium ,calcium ,and magnesium ion in the embryo callus ,while a significant decrease occurred in the concentration of potassium and phosphorus ion and thedpotassium/sodium percentage as compared with the control treatment .The abone mentioned concentrations 0f sea water led to significant increase in concentration of ion ,copper, and zinc ions ,and significant decrease in manganese ion ,The addition of the algae extract caused in lowering the concentration of sodium ion .This decrease was significant in the 60 % sea water medium. The ingrease of potassium ,calcium ,phosphorus, and magnesium ion in embryo callus tissue when compared with free algae extract medium and was significant with some sea water concentration.E - The results ,also ,showed that the addition on 20, 40 and 60% concentrations0f sea water resoled in a significant decrease in concentration of some chemical ingredients such as protein and amino acid proline of the embryo callus in comparison with the control treatment . as far as the effect of the algae extract is concerned ,the addition of this extract to the nutrient medium led to a significant increase in the concentration of the proteins and amino acid prolne as well as with all sea water concentrations.F - The results of study showed that the addition of 20,40,60% sea water led to a significant increase in the rate 0f effectiveness 0f the enzyme Super oxide dismutase (S.O.D) and enzyme Ascorbate peroxides (APX),reaching the highest rate of effectiveness of the enzyme at the concentration 0f 60% compared with the comparative treatment , whil the highest average of the effectiveness of enzyme Catalase (CAT)and glutathione piroxidase (GP)and polyphenoloxidase (ppo)was at the concentration 20%sea water then it started to decline when increasing the concentration of sea water from 40 to 60% As for the algae extract it has been shown from the results of the current study that the addition of algae extract to nutrient medium to a high effectiveness of enzyme SOD,CAT ,APX and GP and more significant with all concentration of sea water The impact of algae extract on the effectiveness of the enzyme PPO has led to increasing the effectiveness of this enzyme but did not amount to a significant level except with sea water nutrient medium .G - The results of the current study showed a significant increase of the Amber cutivar in the percentage of the callus induction and the required time of this induction as well as the Wight of the initial and embryo callus ,number and germination percentage of thesomatic embryo , number of plantlets ,as wall as the percentage of the accumulation of potassium ,phosphorus ,calcium, and manganese ions ,as will as in the potassium/sodim ratio focus histidine proline and the effectiveness of the enzyme SOD and CAT enzyme and enzyme GP. While the product was higher than in the accumulation of Firat magnesium ion ,zinc copper and in the effectiveness of the enzyme in APX & PPO . .
