تأثير بعض المحفزات الفيزيائية و الكيميائية في بعض المؤشرات الفسيولوجية وانتاج المركبات الايضية الثانوية في نبات الحلبة Trigonella foenum – graecum L. خارج الجسم الحي

Effect of some physical and chemical stimuli in some physiological indicators and the production of secondary metabolic compounds of the Fenugreek

Abstract
The study was carried out in the plant tissue culture laboratories of the Department of Horticulture and Landscape Engineering / College of Agriculture – University of Karbala for the period from May 2021 until June 2022, with the aim of studying the effect of physical and chemical stimuli on the production of Trigonella and Diosgenine from Trigonella foenum-graecum L. In vitro. The results of the study can be summarized as follows:-
The best treatment for sterilization of fenugreek seeds was using 2% sodium hypochlorite NaOCl for 15 minutes.
The nodes as a explants used in the stimulation stage gave the highest response of 80% at the concentration of 2 mg.L-1 BA, while the shoot tips did not give any significant response.
– The medium prepared with BA at a concentration of 2 mg.l-1 was the best in increasing the number of branches, their length, the number of leaves, and the fresh and dry weight of the vegetative group, reaching (8.65 branches. Plant part-1, 4.00 cm, 26.33 leaves. Plant-1, 4.03 mg, 1.22 mg) respectively.
The concentration of 0.2 mg.l-1 NAA achieved the highest average of the number of branches and leaves and the fresh and dry weight of the vegetative total amounted to (7.49 branches.plant-1, 21.50 leaves.plant-1, 3.64 and 0.95 mg) respectively, while the concentration achieved 0.4 mg.l -1 The highest average length of branches was 3.92 cm.
The treatment of vegetative growths with the amino acid tryptophan led to a higher concentration of 20 mg.l-1 of it in achieving the highest rate of trampled characteristics, which included the number and length of vegetative branches, the number of leaves, the fresh and dry weight of the vegetative group, as well as the average concentration of chlorophyll and carbohydrates, which amounted to (34.05 branches. , 5.05 cm, 63.33 mg. leaf-1, 3.21 and 1.54 mg, 3.72 and 4.25 mg. g-1) respectively. While the UV irradiation treatment at minute 20 achieved the highest rate for the same traits, which amounted to (32.64 branches. Plant-1, 5.01 cm, 50.26 leaves.plant-1, 2.64 and 1.16 mg, 3.16 and 3.90 mg.gm-1) respectively.
When vegetative growths were treated with different concentrations of tryptophan, the concentration of 15 mg.L-1 achieved the highest average concentration of Trigonelline and Diosgenine compounds, which reached (277.23 and 212.73) μg.g-1, respectively. While the irradiation treatment at minute 20 gave the highest rate of the same compounds (286.45 and 210.60) µg.g-1, respectively.
The shoot tip significantly outperformed the leaves in achieving a response rate of 10% in all combinations of 2,4-D with BA, while the leaf achieved a response rate of 30% at the 1 mg.l-1 2,4-D concentration and overlapping with all BA concentrations.
The concentration of 2 mg.l-1 2,4-D achieved the highest average fresh and dry weight of the induced callus from the growing top, which amounted to (172.75 and 15.31) mg, respectively, while concentration 0.2 mg.l-1 BA achieved the highest weight rate for the same traits that reached (141.60 and 12.22) mg, respectively.
When the callus induced from the growing top was treated with different concentrations of tryptophan, the concentration of 15 mg.l-1 achieved the highest average of the fresh and dry weight of the callus reached (299.49 and 27.23) mg, respectively, and the irradiation treatment at minute 20 achieved the highest rate for the same characteristics amounted to (265.14 and 24.08) mg, respectively.
The concentration of 20 mg.l-1 of tryptophan gave the highest concentration of Trigonelline compound which was 239.89 µg. g-1, while the highest concentration of Diosgenine compound was in the prepared medium with a concentration of 15 mg.l-1 of tryptophan which was 190.34 μg. g-1.
Regarding the effect of UV rays on the rate of concentration of metabolic compounds in the callus of the fenugreek plant, the 20th minute excelled in giving the highest average concentration of Trigonelline and Diosgenine, which reached (248.06 and 192.21) μg. g-1 respectively.