تاثير نسبت هاي مختلف نيترات به آمونيوم بر ميزان زيست توده و برخي ويژگي هاي بيوشيميايي ريشه هاي موئين گياه بذرالبنج مشبك (L. Hyoscyamus reticulatus)

Effect of different nitrate/ammonium ratios on biomass content and some biochemical characteristics of lattice henbane plant (Hyoscyamus reticulatus L.) hairy roots

بذرالبنج مشبك گياهي علفي، دوساله و متعلق به تيره سيب زميني سانان مي باشد و به دليل توليد آلكالوييدهاي تروپاني هيوسيامين و آسكوپولامين به طور گسترده اي در پزشكي مورد استفاده قرار مي گيرد. در پژوهش حاضر، به منظور افزايش توليد آلكالوييدهاي تروپاني، كشت ريشه مويين مشتق شده از ريزنمونه برگ هاي لپه اي تراريخت شده با سويه ي A7 Agrobacterium rhizogenes، توسط نسبت هاي مختلف نيترات به آمونيوم تحريك گرديدند. تاثير نسبت هاي مختلف نيترات به آمونيوم ... ، 10/25 : 19/75، )ﺷﺎﻫﺪ( 20/5 : 39/5، 41 : 79 و 82 : 158 ﻣﻴﻠﻲ ﻣﺘﺮ( در دو ﺗﻴﻤﺎر زﻣﺎﻧﻲ 24 و 48 ﺳﺎﻋﺖ( ﻣﻮرد ﺑﺮرﺳﻲ ﻗﺮارﮔﺮﻓﺖ. ﺑﺮاﺳﺎس ﻣﻘﺎﻳﺴﻪ ﻣﻴﺎﻧﮕﻴﻦ، ﺑﻴﺸﺘﺮﻳﻦ ﻣﺤﺘﻮاي ﻫﻴﻮﺳﻴﺎﻣﻴﻦ و آﺳﻜﻮﭘﻮﻻﻣﻴﻦ 1/5 و 1/17 ﺑﺮاﺑﺮ ﺑﻴﺸﺘﺮ از ﺷﺎﻫﺪ(، ﺑﻪ ﺗﺮﺗﻴﺐ در ﺗﻴﻤﺎر ﻧﺴﺒﺖ 5/1 : 9/9 ﻣﻴﻠﻲﻣﻮﻻر ﻧﻴﺘﺮات ﺑﻪ آﻣﻮﻧﻴﻮم ﺑﻪﻣﺪت 24 ﺳﺎﻋﺖ ﻣﺸﺎﻫﺪه ﮔﺮدﻳﺪ. ﻫﻤﭽﻨﻴﻦ، ﺑﻴﺸﺘﺮﻳﻦ ﻓﻌﺎﻟﻴﺖ آﻧﺰﻳﻢﻫﺎي آﻧﺘﻲ اﻛﺴﻴﺪاﻧﻲ ﻛﺎﺗﺎﻻز 35/612 ﻣﻴﻜﺮوﻣﻮل ﺑﺮ دﻗﻴﻘﻪ ﺑﺮ ﻣﻴﻠﻲﮔﺮم ﭘﺮوﺗﺌﻴﻦ(، 24 ﺳﺎﻋﺖ ﭘﺲ از ﺗﻴﻤﺎر ﺑﺎ ﻧﺴﺒﺖ 82 : 158 ﻣﻴﻠﻲﻣﻮﻻر ﻧﻴﺘﺮات ﺑﻪ آﻣﻮﻧﻴﻮم، ﭘﺮاﻛﺴﻴﺪاز 64/373 ﻣﻴﻜﺮوﻣﻮل ﺑﺮ دﻗﻴﻘﻪ ﺑﺮ ﻣﻴﻠﻲﮔﺮم ﭘﺮوﺗﺌﻴﻦ(، 48 ﺳﺎﻋﺖ ﭘﺲ از ﺗﻴﻤﺎر ﺑﺎ ﻧﺴﺒﺖ ... ﻣﻴﻠﻲﻣﻮﻻر ﻧﻴﺘﺮات ﺑﻪ آﻣﻮﻧﻴﻮم و آﺳﻜﻮرﺑﺎت ﭘﺮاﻛﺴﻴﺪاز 156/16 ﻣﻴﻜﺮوﻣﻮل ﺑﺮ دﻗﻴﻘﻪ ﺑﺮ ﻣﻴﻠﻲﮔﺮم ﭘﺮوﺗﺌﻴﻦ(، 48 ﺳﺎﻋﺖ ﭘﺲ از ﺗﻴﻤﺎر ﺑﺎ ﻧﺴﺒﺖ 10/25 : 19/75 ﻣﻴﻠﻲﻣﻮﻻر ﻧﻴﺘﺮات ﺑﻪ آﻣﻮﻧﻴﻮم ﺑﻪ دﺳﺖ آﻣﺪ. ﺑﺎﺗﻮﺟﻪ ﺑﻪ اﻳﻦ ﻧﺘﺎﻳﺞ، ﻣﺤﺮك ﻧﻴﺘﺮات ﺑﻪ آﻣﻮﻧﻴﻮم ﻣﻲﺗﻮاﻧﺪ در ﺑﻴﻮﺗﻜﻨﻮﻟﻮژي ﮔﻴﺎﻫﻲ ﺑﻪ ﻋﻨﻮان ﻣﺤﺮك ﻣﺆﺛﺮ ﺟﻬﺖ ﺗﻮﻟﻴﺪ ﻣﺘﺎﺑﻮﻟﻴﺖﻫﺎي ﺛﺎﻧﻮي ﮔﻴﺎﻫﻲ ازﺟﻤﻠﻪ آﻟﻜﺎﻟﻮﺋﻴﺪﻫﺎي ﺗﺮوﭘﺎﻧﻲ ﻣﻮرد اﺳﺘﻔﺎده ﻗﺮار ﮔﻴﺮد.

Hyoscyamus reticulatus L. is an herbaceous, biennial plant, belonging to Solanaceae family and Due to production of tropane alkaloids such as hyoscyamine and scopolamine are widely used in medicine. In the present study, in order to increase production of tropane alkaloids, cotyledon-derived hairy root cultures transformed with Agrobacterium rhizogenes strain A7, elicited by various ratio of nitrate to ammonium. The effect of different ratio of nitrate to ammonium (9.9:5.1, 19.75:10.25, 39.5:20.5 (control), 79:41 and 158:82 mM) in two exposure times (24 and 48 h) were investigated. According to the ANOVA results, the highest hyoscyamine and scopolamine production (about 1.5-fold and 1.17-fold increase over the control) was observed in treatment 9.9: 5.1 mM nitrate to ammonium at 24 hours of exposure time, respectively. Therefore, the highest catalase antioxidant enzymes activity (23/40 µmol min-1 mg-1 pro) and peroxidase (145/32 µmol min-1 mg-1 pro) and ascorbate peroxidase (24/22 µmol min-1 mg-1 pro), was obtained in nitrate to ammonium ratio (158: 82 mM) with 24 hours exposure time. According to these results, nitrate to ammonium ratio as elicitor can be used as effective strategies in plant biotechnology in order toenhance production of plant secondary metabolites such as tropane alkaloids.