تأثير استخدام UV وحامض السالسليك في انتاج بعض المركبات الفعالة لنبات إكليل الجبل Rosmarinus officinalis L. خارج الجسم الحي

Rp-Effect of UV ray and Salicylic acid to increase the formation of active compound in Rosmarinus officinalis L. in vitro.pdf

This study was carried out in the Plant Tissue Culture Laboratory at the College of Agriculture – University of Karbala. The HPLC analyses were done in the Alhuqul Albayda Company for Investments, Environmental and Engineering Studies/ Baghdad / Iraq, for the period from 2021 to 2022. The study used plant tissue culture technology to establish vegetative shoots and callus cultures for rosemary plants, and stimulate them to increase the production of volatile oils and phenolics compounds of medical importance. It was carried out in two phases after conducting the sterilization process: the first included the establishment of vegetative shoots and callus cultures by using different combinations of plant growth regulators (Auxins and Cytokinines). ultraviolet light and a concentration of (0-10-20-30) mg L-1 of salicylic acid to stimulate it to increase the production of secondary metabolites in independent factorial experiments using the complete randomized design (CRD). The results can be summarized as follows:

The best treatment for sterilizing explants is sodium hypochlorite (NaOCl) at a concentration of 2% for 15 minutes.
The shoots tip as a explants used in the establishment stage gave the highest response of 90% at a concentration of 1 mg-1 of BA, while the lateral buds gave the highest response of 70% at the same concentration.
The media supplemented with BA and with a concentration of 2 mg l-1 was the best in increasing the number of branches and their length reached 4.74 branch-1 and 4.12 cm, respectively, while the concentration 3 mg l-1 achieved the highest mean of the number of leaves and the fresh and dry weight of the shoot was 15.24 leaf-1, 2986 mg and 1823 mg, respectively.
Concentration 0.2 mg l-1 of NAA achieved the highest rate of number of shoots and leaves and dry weight reached 4.16 branch plant-1, 14.21 leaf plant-1 and 1594 mg respectively, the concentration of 0.4 mg l-1 which achieved the highest average of shoot length reached 3.74 cm, while the concentration of 0.1 mg l-1 achieved the highest average fresh weight was 2606 mg.
The treatment of shoots culture with salicylic acid led to a superior concentration of 10 mg l-1 of it in achieving the highest rate of characters , which included the number and length of vegetative branches, reaching 7.45 plant branches-1 and 3.63 cm, respectively, and the concentration exceeded 20 mg l-1 of it in The average number of leaves, fresh and dry weight of shoots, as well as the average concentration of chlorophyll and carbohydrates were 16.86 leaf-1, 3424 and 2373 mg, 3.57 and 4.28 mg -1 respectively.
The UV irradiation treatment at the 20th minute achieved the highest rate of number and length of shoots, number of leaves, fresh and dry weight of the vegetative cultures, as well as the average weight of chlorophyll and carbohydrates were 7.24 branches -1, 4.04 cm, 16.89 leaves -1, 3374 and 2141 mg, 3.67 and 4.32 mg -1 respectively.
Treatment of shoots culture with different concentrations of salicylic acid resulted in significant differences in the amount of active compounds that were measured, where the concentration of 10 mg l-1 achieved the highest average concentration of the Geranal compound amounted to 30.677 μg gm-1, and the concentration of 20 mg l-1 achieved the highest rate. The concentration of pinene-ά, Camphor, Verbene and Broneal reached 33.28, 30.443, 31.933 and 29.820 μg mg-1, respectively, while the concentration of 30 mg l-1 achieved the highest average concentration of 1-8cinole, Rosmaric acid, Linolool, Cymene and Terpinol were 30.130, 62.82, 34.430, 31.3400, and 33.377 μg g-1, respectively.
The irradiation treatment at the 20th minute gave the highest rate for the compounds a- pinene, 1-8cinole, Camphor, Verbene, Rosmaric acid, Broneal, Geranal, Linolool, Cymene and ά- Terpinol 39.60, 36.495, 33.548, 34.745, 62.75, 28.582 and 29.657, 31.102, 35.0375, and 32.743 μg g-1, respectively.
The shoot tip was significantly superior compared with the leaves in achieving the highest rate of response to callus induction of 100% in all combinations of 2,4-D with BA.
The concentration of 2 mg l-1 of 2,4-D achieved the highest average fresh and dry weight of the callus induced from the growing apex, which reached (156.78 and 15.66) mg, respectively, while the concentration of 0.2 mg l-1 BA achieved the highest average weight for the traits. The same amounted to 150.19 and 12.464 mg respectively.
The treatment of induced callus from the shoot tip with different concentrations of salicylic acid, where the concentration of 20 mg l-1 achieved the highest rate of fresh and dry weight of callus amounted to 308.47 and 27.30 mg respectively. Also, the irradiation treatment at 20 minutes achieved the highest rate for the same characteristics amounted to 291.44 and 25.44 mg respectively.
When treating callus cultures with different concentrations of salicylic acid, the concentration of 10 mg l-1 achieved the highest average concentration of the Geranal compound amounted to 26.8967 µg g-1, and the concentration of 20 mg l-1 achieved the highest average concentration of the Verbene compound amounted to 28.693 µg g-1, in When the concentration of 30 mg l-1 achieved the highest average concentration of ά- pinene, 1-8cinole, Camphor, Rosmaric acid, Broneal, ά-Terpinol, Linolool and Cymene were 30.07, 28.99, 28.86, 49.68, 25.09, 29.753, 26.77, and 28.263 µg g-1respectively.
The irradiation treatment at the 20th minute achieved the highest rate for the compounds ά-pinene, 1-8cinole, Camphor, Verbene, Rosmaric acid, Broneal, Geranal, Linolool, Cymene and ά- Terpinol were 34.80, 34.50, 29.450, 29.112, 45.240, 25.342 and 23.5475, 28.305, 28.26, and 25.878 µg g-1 respectively.