IN VITRO STUDIES ON PROPAGATION OF DIFFERENT VARIETIES OF Rosmarinus officinalis (L.), PLANT AND EFFECT OF SOME TREATMENTS ON GROWTH AND ACTIVE INGREDIENTS

Abstract

This study aimed to: studying and recording the morphological and chemical differences between all rosemary cultivars under study Rosemarinus officinalis L. C1, R. officinalis L. Pyramidalis" C2 and R. officinalis L. Angustifolius C3 by measuring some variability's of all cultivars, determine successful cutting percentage, evaluation essential oil percentage, determine volatile oil composition. it was found that C2 had the highest leave length, leave width and the highest number of axillary shoots also it had the highest percentage of succeed cuttings with 100%. The highest volatile oil was 0.95% for C3 and the main components was verbenon. Study the most suitable protocol of micropropagation for each cultivar by examine the best sterilization method then evaluate the best type of PGR on callus and shoot formation when using alone or in combination then evaluate the best medium type and leave position for production then evaluate using mTR on elongation and investigate the best auxins with medium concentration for rooting then evaluate the proper medium for plant adaptation. The most efficient sterilization method was by using Lemon juice and H2O2 which resulted in 100% sterilized explants with C2. It was found that, the highest callus weight was obtained from C1 by 7.22 g resulted from 2.0 mg/l CPPU while the highest No. of shoots was obtained from C3 by 12shoot/explants resulted from 2.0mg/l CPPU. Concerning combined PGRs it was found that the highest callus weight was obtained from C3 by 7.91 g at 2.0 K+0.1 2,4-D mg/l while the highest No. of shoots was produced from C3 at 2.0 CPPU + 0. 1 2,4-D mg/l by 12 shoot/explants. For medium type, it was found that the highest callus weight was obtained from C2 by 9.02g from MS medium+1 mg/l CPPU while the highest No. of shoots was obtained from C3 by 18.3shoot/explant from WPM+1 mg/l CPPU. For leave position the highest callus weight for C1 was 6.29 g obtained from terminal leaves explants cultured on MS media supplemented with 2mg/l CPPU while the highest number of shoots produced from C3 by 7.0 shoot/explants from basal leaves cultured on MS medium supplemented with 3.0 mg/l CPPU. For shoot elongation experiment it was found that 0.5 mg\l mTR showed highest rate of growth for C3 by 2.1cm with healthy green shootlets. For rooting experiment, it was found that both C1 and C2 can produce roots without need to auxines while half MS supplemented with 1.0 mg/l NAA was suitable to enhance root formation for C3. The best acclimatization medium was sand medium. Determine the salt stress effect on callus formation, shoot formation and volatile oil composition all cultivars, the highest callus weight was 8.55g for C2 at 1000 µM/l, the highest No. of shoot 21.76 for C3 the highest volatiles component was Bornyl acetate for all. Investigate the ability producing polyploidy plant by using Colchicine to improve C3 it was found 0.3 mg/l colchince for 14 days can produce polyploidy plant