Kufa University College of Agriculture Horticulture & Landscape Department

Micropropagation of Carnation Dianthus

caryophyllus L. and Studying biochemical

changes in microcuttings during rooting Stage

A thesis Submitted to the Council of the College of Agriculture at the University of Kufa in Partial Fulfillment of the Requirements for the Degree of Master in Horticulture Science

By Riyadh Abdel Amir Hassan AL-Ibrahemy

Advised by Muslim Abd Ali Abdulhussein

2011

Abstract This study was conducted at Plant Tissue Culture Laboratory / Horticulture Department / Agriculture College / Kufa University during 2009-2010. Several Experiments were conducted to study the the effects of plant growth regulators (cytokinine and auxin) in intiation and shoot multiplication stages, besides the effect of auxins , MS salt

concentrations and sucrose concentrations on rooting parameters, also studying the biochemical changes during in vitro adventitious root formation on microcuttings of 'Chabaud' carnation in vitro . Results are summarized as follows : In initiation stage, shoot tip gave a better response, shoot length and node number per shoot than single node. BA was better than Kin in these characteristics. Explants which cultured on MS medium gave a differences response according to type and concentrations of

cytokinine. Best response for shoot tips and single node explants which gave ( 80-90 % ) was obtained with MS medium supplied with 0.5 mg\1 BA compared with (20-40 %) respectively in free hormone medium. In multiple stage the highest shoot number and shoot length ( (13.50 shoot/explant and 2.83 cm) respectively was achieved when shoot were cultured on MS medium containing both 2 mg/L BA and 0.1 mg/L NAA . Best rooting Percentage, root number and length were 90% , 09.4 root/shoot and 2.27 cm when the medium was supplemented with 1 mg/L IAA compared to control (30%,0.88 and 0.35 respectively). Reducing the strength of MS medium salts to a half elevated rooting percentage, root number and length to 95% , 8.7 and 5.3cm. Also

addition 30 g/L sucrose to half MS salts strength gave 90% rooting percentage with highest root number/ explant and length of root ,while increaced sucrose concentration to 50 g/L reduced rooting percentage , root number/ explant and length of root . Total reducing sugar, endogenous IAA level and peroxidase activity was determined during root induction (0-day to the 15th day at every 3 day interval) from the base of microshoots derived in vitro. The level and activity was at the maximum level during the the first three days and minimized after the sixth day These finding may be useful in monitoring the rooting behaviour in microshoots and peroxidase activity as a marker for root initiation.