ﺑﺮرﺳﯽ ﺗﺄﺛﯿﺮﺳﺎﻟﯿﺴﯿﻠﯿﮏ اﺳﯿﺪ ﺑﺮ ﻣﯿﺰان ﻣﺘﺎﺑﻮﻟﯿﺖ ﻫﺎي ﺛﺎﻧﻮﯾﻪ رﯾﺸﻪ ﻫﺎي ﻧﺎﺑﺠﺎ و ﮐﺎﻟﻮس ﮔﯿﺎه ﺳﺮﺧﺎرﮔﻞ Echinacea purpurea L

The study of micropropagation and salicylic acid treatment effect on Echinacea purpurea L callus and root secondary metabolites in tissue culture

در اﯾﻦ ﺗﺤﻘﯿﻖ ﺑﻪ ﺑﺮرﺳﯽ ﺗﺎﺛﯿﺮ ﺳﺎﻟﯿﺴﯿﻠﯿﮏ اﺳﯿﺪ در ﻣﺤﯿﻂ ﮐﺸﺖ ﺑﺎﻓﺖ ﺑﺮ ﻣﯿﺰان ﻣﺘﺎﺑﻮﻟﯿﺖﻫﺎي ﺛﺎﻧﻮﯾﻪ ﮐﺎﻟﻮس و رﯾﺸﻪﻫﺎي ﻧﺎﺑﺠﺎي ﮔﯿﺎه ﺳﺮﺧﺎرﮔﻞ )Echinacea purpurea L( ﭘﺮداﺧﺘﻪ ﺷﺪ. ﻟﭙﻪ ﺟﻬﺖ ﺗﺸﮑﯿﻞ ﮐﺎﻟﻮس در ﻏﺎﻟﺐ آزﻣﺎﯾﺶ ﻓﺎﮐﺘﻮرﯾﻞ ﺑﺎ ﻫﻮرﻣﻮن ﺑﻨﺰﯾﻞ آدﻧﯿﻦ )0/5 و 1 ﻣﯿﻠﯽ ﮔﺮم ﺑﺮ ﻟﯿﺘﺮ( و ﺗﻮﻓﻮردي و ﻧﻔﺘﺎﻟﯿﻦ اﺳﺘﯿﮏ اﺳﯿﺪ )0/5 و 1 ﻣﯿﻠﯽ ﮔﺮم ﺑﺮ ﻟﯿﺘﺮ( ﮐﺸﺖ ﺷﺪﻧﺪ. ﺟﻬﺖ ﺗﻬﯿﻪ رﯾﺸﻪ ﻧﺎﺑﺠﺎ ﻧﯿﺰ ﻟﭙﻪ در ﻣﺤﯿﻂ ﺣﺎوي ﻧﻔﺘﺎﻟﯿﻦ اﺳﺘﯿﮏ اﺳﯿﺪ و اﯾﻨﺪول ﺑﻮﺗﯿﺮﯾﮏ اﺳﯿﺪ )0، 0/2، 1، 1/5 و 2 ﻣﯿﻠﯽ ﮔﺮم ﺑﺮ ﻟﯿﺘﺮ( ﺑﺮ ﭘﺎﯾﻪ ﻃﺮح ﮐﺎﻣﻼ ﺗﺼﺎدﻓﯽ ﮐﺸﺖ ﺷﺪﻧﺪ. ﺗﯿﻤﺎرﻫﺎي اﺳﯿﺪﺳﺎﻟﯿﺴﯿﻠﯿﮏ در ﻏﻠﻈﺖ ﻫﺎي 0، 50، 100 و 200 ﻣﯿﻠﯽ ﮔﺮم ﺑﺮ ﻟﯿﺘﺮ در ﻣﺤﯿﻂ ﮐﺸﺖ MS ﻣﺎﯾﻊ اﻋﻤﺎل ﺷﺪ و ﺳﭙﺲ ﺗﺮﮐﯿﺒﺎﺗﯽ ﺷﺎﻣﻞ ﻣﯿﺰان ﮐﻠﺮوﻓﯿﻞ ﮐﻞ، ﻓﻨﻞ، ﻓﻼوﻧﻮﺋﯿﺪ، ﻓﻌﺎﻟﯿﺖ آﻧﺘﯽ اﮐﺴﯿﺪاﻧﯽ، اﺳﯿﺪﮐﺎﻓﺌﯿﮏ و اﺳﯿﺪﮔﺎﻟﯿﮏ اﻧﺪازهﮔﯿﺮي ﺷﺪ. ﺑﻬﺘﺮﯾﻦ ﺗﯿﻤﺎر از ﻧﻈﺮ ﺗﻮﻟﯿﺪ ﮐﺎﻟﻮس ﻣﺤﯿﻂ ﮐﺸﺖ MS ﺣﺎوي ﻧﯿﻢ ﻣﯿﻠﯽ ﮔﺮم ﺑﺮ ﻟﯿﺘﺮ ﻫﻮرﻣﻮن ﺑﻨﺰﯾﻞ آدﻧﯿﻦ و ﯾﮏ ﻣﯿﻠﯽ ﮔﺮم ﺑﺮ ﻟﯿﺘﺮ ﻫﻮرﻣﻮن ﺗﻮﻓﻮردي و ﻫﻤﭽﻨﯿﻦ ﺑﻬﺘﺮﯾﻦ ﺗﯿﻤﺎر ﺟﻬﺖ ﺗﻮﻟﯿﺪ رﯾﺸﻪﻫﺎي ﻧﺎﺑﺠﺎ از ﻣﺤﯿﻂ ﮐﺸﺖ1/2MS ﺣﺎوي ﻧﯿﻢ ﻣﯿﻠﯽ ﮔﺮم ﺑﺮ ﻟﯿﺘﺮ ﻫﻮرﻣﻮن اﯾﻨﺪول ﺑﻮﺗﯿﺮﯾﮏ اﺳﯿﺪ ﺑﺪﺳﺖ آﻣﺪ. ﺑﯿﺸﺘﺮﯾﻦ ﻣﯿﺰان ﻓﻌﺎﻟﯿﺖ آﻧﺘﯽ اﮐﺴﯿﺪاﻧﯽ ﺑﺘﺮﺗﯿﺐ در ﺗﯿﻤﺎر 100 و 200 ﻣﯿﻠﯽ ﮔﺮم ﺑﺮ ﻟﯿﺘﺮ اﺳﯿﺪﺳﺎﻟﯿﺴﯿﻠﯿﮏ در ﮐﺎﻟﻮس و رﯾﺸﻪﻫﺎي ﻧﺎﺑﺠﺎي ﻣﺸﺎﻫﺪه ﺷﺪ. ﺑﯿﺸﺘﺮﯾﻦ ﻣﻘﺪار اﺳﯿﺪﮔﺎﻟﯿﮏ ﺑﺎ 2/11 ﻣﯿﻠﯽ ﮔﺮم در ﮔﺮم وزن ﺗﺮ در رﯾﺰﻧﻤﻮﻧﻪ رﯾﺸﻪ در ﻏﻠﻈﺖ100 ﻣﯿﻠﯽ ﮔﺮم ﺑﺮ ﻟﯿﺘﺮ اﺳﯿﺪﺳﺎﻟﯿﺴﯿﻠﯿﮏ در روز ﺳﻮم ﻧﻤﻮﻧﻪ ﺑﺮداري ﺛﺒﺖ ﺷﺪ. ﺑﺎﻻﺗﺮﯾﻦ ﻣﻘﺪار اﺳﯿﺪﮐﺎﻓﺌﯿﮏ ﺑﺎ 2/58 ﻣﯿﻠﯽ ﮔﺮم ﺑﺮ ﮔﺮم وزن ﺗﺮ در رﯾﺰﻧﻤﻮﻧﻪ رﯾﺸﻪ در ﻏﻠﻈﺖ 200 ﻣﯿﻠﯽ ﮔﺮم در ﻟﯿﺘﺮ اﺳﯿﺪ ﺳﺎﻟﯿﺴﯿﻠﯿﮏ در روز ﭘﻨﺠﻢ ﻧﻤﻮﻧﻪﺑﺮداري ﻣﺸﺎﻫﺪه ﺷﺪ.

Echinacea purpurea L. is very important in the pharmaceutical industry due to its valuable secondary metabolites. In this experiment, the investigation of salicylic acid in tissue culture medium on callus and adventitious root secondary metabolites of Echinacea was conducted. The best treatment for callus production was MS medium contain 0.5 mg/l BA and 1 mg/l 2,4-D. The best treatment for adventitious root was obtained in 1/2 MS medium contain 0.5 mg/l IBA in dark condition. Salicylic acid at concentrations of 0, 50, 100 and 200 mg/l in liquid MS medium, and then fresh weight, phenol content, flavonoids and antioxidant activities were measured. The highest antioxidant activity was observed in 100 and 200 mg/l of salicylic acid in callus and adventitious root respectively. Salicylic acid promoted phenol and flavonoid content rather than control medium (without hormone). The highest gallic acid content with 2.11 mg/g root fresh weight was recorded in 100 mg/l 3 days after inoculation. Also the best medium for cafeic acid content was observed in 200 mg/g root fresh weight 5 days after inoculation.