باززايي درون شيشه اي ماميران ايراني (Chelidonium majus L.)

In vitro regeneration of Iranian celandine (Chelidonium majus L

ﻣﺎﻣﻴﺮان Chelidonium majus L.)( ﮔﻴﺎﻫﻲ داروﻳﻲ از ﺧﺎﻧﻮاده ﺧﺸﺨﺎش و ﻏﻨﻲ از آﻟﻜﺎﻟﻮﺋﻴﺪﻫﺎي اﻳﺰوﻛﻮﻳﻨﻮﻟﻴﻦ اﺳﺖ. از آﻧﺠﺎﻳﻲ ﻛﻪ ﺳﻨﺘﺰ ﺷﻴﻤﻴﺎﻳﻲ اﻳﻦ آﻟﻜﺎﻟﻮﺋﻴﺪﻫﺎ ﻣﻘﺪور ﻧﻴﺴﺖ، ﺻﻨﻌﺖ داروﺳﺎزي ﺑﻪ ﻣﻘﺎدﻳﺮ زﻳﺎدي ﮔﻴﺎه ﺑﺮاي اﺳﺘﺨﺮاج اﻳﻦ ﺗﺮﻛﻴﺒﺎت ﻧﻴﺎزﻣﻨﺪ اﺳﺖ. ﻫﺪف اﻳﻦ ﭘﮋوﻫﺶ ﺑﻬﻴﻨﻪﺳﺎزي ﻳﻚ روش ﺗﻜﺮارﭘﺬﻳﺮ ﺑﺮاي ﺑﺎز زاﻳﻲ ﻣﺎﻣﻴﺮان ﺑﻮاﺳﻄﻪ ﻛﺎﻟﻮسزاﻳﻲ ﺑﻮد. ﺑﺪﻳﻦ ﻣﻨﻈﻮر، ﺑﺬور ﻣﺎﻣﻴﺮان ﺑﻌﺪ از ﺿﺪﻋﻔﻮﻧﻲ ﺳﻄﺤﻲ ﺑﺮاي ﺟﻮاﻧﻪزﻧﻲ در ﻣﺤﻴﻂ 1/2 MSﻛﺸﺖ ﺷﺪﻧﺪ. رﻳﺰ ﻧﻤﻮﻧﻪ ﻫﺎي ﻛﻮﺗﻴﻠﺪون و ﻫﻴﭙﻮﻛﻮﺗﻴﻞ از ﮔﻴﺎﻫﭽﻪﻫﺎي 20 روزه و رﻳﺰ ﻧﻤﻮﻧﻪ ﻫﺎي ﺑﺮگ از ﮔﻴﺎﻫﺎن 3 ﻣﺎﻫﻪ ﺗﻬﻴﻪ و ﺑﺮاي ﻛﺎﻟﻮسزاﻳﻲ در ﻣﺤﻴﻂ ﻛﺸﺖ ﻣﻮراﺷﻴﮓ و اﺳﻜﻮگ (MS) ﺗﻜﻤﻴﻞ ﺷﺪه ﺑﺎ ﺗﻨﻈﻴﻢ ﻛﻨﻨﺪهﻫﺎي رﺷﺪ ﮔﻴﺎﻫﻲ ﻣﺨﺘﻠﻒ ﻛﺸﺖ ﺷﺪﻧﺪ. ﺑﻴﺸﺘﺮﻳﻦ وزن ﺗﺮ ﻛﺎﻟﻮس از رﻳﺰ ﻧﻤﻮﻧﻪ ﻫﺎي ﻣﺴﺘﻘﺮ در ﻣﺤﻴﻂ MS داراي 1 ﻣﻴﻠﻲ ﮔﺮم ﺑﺮ ﻟﻴﺘﺮ2و4 دي ﻛﻠﺮو ﻓﻨﻮﻛﺴﻲ اﺳﺘﻴﻚ اﺳﻴﺪ )2,4-D در ﺗﺮﻛﻴﺐ ﺑﺎ 1 ﻳﺎ 0/5 ﻣﻴﻠﻲ ﮔﺮم ﺑﺮ ﻟﻴﺘﺮ از ﻳﻜﻲ از ﺳﻴﺘﻮﻛﻨﻴﻦﻫﺎي ﺑﻨﺰﻳﻞ آﻣﻴﻨﻮ ﭘﻮرﻳﻦ )(BAP ﻳﺎ ﺗﻴﺪﻳﺎزورون (TDZ) ﺑﻪ دﺳﺖ آﻣﺪ. در ﭼﻨﺪ ﺗﻴﻤﺎر ﺑﻌﺪ از ﺑﺎز ﻛﺸﺖﻫﺎي ﻣﻜﺮر اﻧﺪامزاﻳﻲ ﻧﻴﺰ ﻣﺸﺎﻫﺪه ﺷﺪ. ﺑﺎز زاﻳﻲ دو ﻣﺎه ﭘﺲ از ﺑﺎز ﻛﺸﺖ ﻛﺎﻟﻮسﻫﺎي ﺣﺎﺻﻞ از رﻳﺰ ﻧﻤﻮﻧﻪ ﻫﺎي ﻛﻮﺗﻴﻠﺪون در ﻣﺤﻴﻂ ﻛﺸﺖ ﻓﺎﻗﺪ ﺗﻨﻈﻴﻢ ﻛﻨﻨﺪه ﻫﺎي رﺷﺪ ﮔﻴﺎﻫﻲ ﺑﺎ ﻓﺮاواﻧﻲ 11/11 درﺻﺪ و ﻳﻚ ﺷﺎﺧﺴﺎره ﺑﻪ ازاي ﻫﺮ رﻳﺰ ﻧﻤﻮﻧﻪ ﻣﺸﺎﻫﺪه ﺷﺪ. ﺷﺎﺧﺴﺎرهﻫﺎ در ﻣﺤﻴﻂ MS ﻓﺎﻗﺪ ﺗﻨﻈﻴﻢ ﻛﻨﻨﺪه-ﻫﺎي رﺷﺪ ﮔﻴﺎﻫﻲ ﺑﺎ ﻓﺮاواﻧﻲ 77/78 درﺻﺪ و 2/36 رﻳﺸﻪ ﺑﻪ ازاي ﻫﺮ ﺷﺎﺧﺴﺎره رﻳﺸﻪدار ﺷﺪﻧﺪ.

Celandine (Chelidonium majus L.) is a medicinal plant belonging to the Papaveraceae and rich in isoquinolin alkaloids. Chemical synthesis of these alkaloids is not possible. So, the pharmaceutical industry requires large quantities of plants to extract these compounds. The purpose of this study was optimization of a repeatable method for regeneration of celandine by callus induction. Celandine seeds after surface sterilizing were placed on 1/2MS medium for germination. Cotyledon and hypocotyl explants were prepared from 20-day-old sterile seedlings and leaf explants from 3-month-old plants. Explants were put on Murashige and Skoog (MS) medium supplemented with different plant growth regulators (PGRs) to callus induction. The highest fresh weight was for calli in mediums containing 1 mg/l 2,4-Dichlorophenoxyacetic acid (2,4-D) in combination with 0.5 or 1 mg/l Amino Purine (BAP) or Tidiazuron (TDZ). Organogenesis was observed in few treatments after repetitive subculture. Plant regeneration only happened in PGRs free MS medium, from cotyledon calli after 2 months with frequency of 11.11 % by one shoot per explant. Shoots were rooted in PGRs free MS medium with frequency of 77.78 % by 2.36 roots per explant.