بهينه سازي كالوس زايي و بازايي مستقيم گلرنگ با استفاده از قطعات ريشه، لپه و محور زير لپه

Callugenesis and direct regeneration optimization for root, cotyledon and hypocotyle explants of safflower

ﮔﻠﺮﻧﮓ ﯾﮑﯽ از ﮔﯿﺎﻫﺎن روﻏﻨﯽ و ﺳﺎزﮔﺎرﯾﺎﻓﺘﻪ ﺑﻪ ﺷﺮاﯾﻂ ﺳﺨﺖ اﯾﺮان اﺳﺖ. ارﻗﺎم زراﻋﯽ زﯾﺎد و ﻣﺘﻨﻮﻋﯽ از اﯾﻦ ﮔﯿﺎه در ﮐﺸﻮر وﺟﻮد دارد. ارﻗﺎم ﮔﻠﺮﻧﮓ ﻣﺤﺘﻮاي اﺳﯿﺪ ﭼﺮب اﻣﮕﺎ 6 ﺑﺎﻻﯾﯽ دارﻧﺪ ﮐﻪ ﻧﺸﺎندﻫﻨﺪه وﺟﻮد ﻣﺴﯿﺮﻫﺎي ﭘﯿﺸﺮﻓﺘﻪ ﺳﺎﺧﺖ اﺳﯿﺪﻫﺎي ﭼﺮب ﻏﯿﺮاﺷﺒﺎع ﺑﺎ زﻧﺠﯿﺮه ﻧﺴﺒﺘﺎ ﺑﻠﻨﺪ در اﯾﻦ ﮔﯿﺎه اﺳﺖ. ﺑﻪ ﻫﻤﯿﻦ دﻟﯿﻞ اﺳﺖ ﮐﻪ اﯾﻦ ﮔﯿﺎه ﮔﺰﯾﻨﻪ ﻣﻨﺎﺳﺒﯽ ﺑﺮاي ﻣﻬﻨﺪﺳﯽ ﻣﺴﯿﺮﻫﺎي ﻣﺨﺘﻠﻒ ﺳﻨﺘﺰ اﺳﯿﺪﻫﺎي ﭼﺮب زﻧﺠﯿﺮه ﺑﻠﻨﺪ ﺷﻨﺎﺧﺘﻪ ﻣﯽﺷﻮد. ﮐﺸﺖ ﺑﺎﻓﺖ ﭘﯿﺶ ﻧﯿﺎز اﻧﺠﺎم ﻣﻬﻨﺪﺳﯽ ژﻧﺘﯿﮏ )ﻣﺘﺎﺑﻮﻟﯿﺖ( در ﮔﯿﺎﻫﺎن اﺳﺖ. ﺑﺪﯾﻦ ﻣﻨﻈﻮر در اﯾﻦ ﺗﺤﻘﯿﻖ ﺷﺮاﯾﻂ ﺑﻬﯿﻨﻪ ﮐﺸﺖ ﺑﺎﻓﺖ رﻗﻢ ﮔﻠﻤﻬﺮ ﮔﻠﺮﻧﮓ اﻧﺠﺎم ﺷﺪ. ﺑﺮاي اﯾﻦ ﻣﻨﻈﻮر ﭘﺎﺳﺦ ﺑﻪ ﺳﺎﻗﻪزاﯾﯽ ﺳﻪ رﯾﺰﻧﻤﻮﻧﻪ رﯾﺸﻪ، ﻣﺤﻮر زﯾﺮﻟﭙﻪ و ﻟﭙﻪ ﺑﺮ روي ﻣﺤﯿﻂ MS ﻏﻨﯽﺷﺪه ﺑﺎ ﻏﻠﻈﺖﻫﺎي ﻣﺨﺘﻠﻒ ﺗﯿﺪﯾﺎزورون/ ﻧﻔﺘﺎﻟﯿﻦ اﺳﺘﯿﮏ اﺳﯿﺪ و /Zeatin ﻧﻔﺘﺎﻟﯿﻦ اﺳﺘﯿﮏ اﺳﯿﺪ ﺑﺮرﺳﯽ ﺷﺪ. ﺳﭙﺲ ﺗﯿﻤﺎرﻫﺎي ﭘﺮزاﯾﯽ )Multiplication ( و ﻃﻮﯾﻞﺳﺎزي ﺳﺎﻗﻪﻫﺎي ﺑﺎزاﺷﺪه اﻋﻤﺎل و در ﻧﻬﺎﯾﺖ ﺗﯿﻤﺎرﻫﺎي ﻣﺨﺘﻠﻒ رﯾﺸﻪزاﯾﯽ ﻧﻮﺳﺎﻗﻪﻫﺎ ﺑﺮرﺳﯽ ﺷﺪ. ﺑﯿﺸﺘﺮﯾﻦ ﺑﺎززاﯾﯽﻫﺎي ﻣﺸﺎﻫﺪه ﺷﺪه 25/15 ،%28/8 و 24/53% ﺑﻪ ﺗﺮﺗﯿﺐ ﺑﺮاي ﺗﯿﻤﺎرﻫﺎي ﻣﺤﻮر زﯾﺮ ﻟﭙﻪ )ﺗﯿﺪﯾﺎزورون,0.5 ﻧﻔﺘﺎﻟﯿﻦ اﺳﺘﯿﮏ اﺳﯿﺪ0 (، رﯾﺸﻪ )ﺗﯿﺪﯾﺎزورون,0.5 ﻧﻔﺘﺎﻟﯿﻦ اﺳﺘﯿﮏ اﺳﯿﺪ2 ( و ﻟﭙﻪ )ﺗﯿﺪﯾﺎزورون,0.5 ﻧﻔﺘﺎﻟﯿﻦ اﺳﺘﯿﮏ اﺳﯿﺪ2 ( ﺑﺪﺳﺖ آﻣﺪ. ﻫﻤﭽﻨﯿﻦ ﺗﯿﻤﺎر ﻫﻮرﻣﻮﻧﯽ BA(0.5mg/L)+Thiamin-HCl (4mg/L) ﺗﻮاﻧﺴﺖ ﺑﺎﻋﺚ ﭘﺮزاﯾﯽ در رﯾﺰﻧﻤﻮﻧﻪ-ﻫﺎي ﺑﺎززا ﺷﺪه ﺷﻮد. ﺑﻪ ﻣﻨﻈﻮر اﻓﺰاﯾﺶ ﺑﺎززاﯾﯽ رﯾﺰﻧﻤﻮﻧﻪﻫﺎﯾﯽ ﮐﻪ ﮐﺎﻟﻮس داده ﺑﻮدﻧﺪ، ﺗﯿﻤﺎر ﻫﻮرﻣﻮﻧﯽ + BA(1mg/L) ﻧﻔﺘﺎﻟﯿﻦ اﺳﺘﯿﮏ اﺳﯿﺪ)1mg/L اﻋﻤﺎل ﺷﺪ ﮐﻪ اﯾﻦ ﺗﯿﻤﺎر ﺗﺎﺛﯿﺮ ﻣﻌﻨﯽداري در ﺑﺎززاﯾﯽ رﯾﺰﻧﻤﻮﻧﻪﻫﺎي ﮐﺎﻟﻮس داده ﻧﺪاﺷﺖ. در اﯾﻦ ﺗﺤﻘﯿﻖ ﻧﯿﺰ رﯾﺸﻪدﻫﯽ ﻧﻮﺳﺎﻗﻪﻫﺎ ﺑﺎ ﺗﯿﻤﺎرﻫﺎي ﻫﻮرﻣﻮﻧﯽ ﻣﺨﺘﻠﻒ ﺑﺎ ﻓﺮاواﻧﯽ ﮐﻤﺘﺮ از ﭼﻬﺎر درﺻﺪ ﺑﺪﺳﺖ آﻣﺪ.

Safflower (Carthamus tinctorius) is one of the oil crops which well adapted to Iran's semi-hard land. High content of omega-6 fatty acids in safflower oils representing an advanced semi-long chain poly unsaturated fatty acid (SLC-PUFA) pathway in the plant, and make this plant as a suitable candidate for SLC-PUFA pathway engeeniring. Tissue culture optimization and plant regeneration is a prerequisite for any genetic engineering procedure. In this research, the optimisation of tissue culture of Golmehr cultivar was done. For this, regeneration of three explants include root, hypocotyle and cotyledon on MS riched by different concentration of TDZ/NAA and Zeatin/NAA was investigated. Shoot propagation, shoot elongation and root induction studied in the present work. Shoot regenerations were obtained in TDZ=0.5mg/L with 28.8%, TDZ=0.5mg/L+NAA=2mg/L with 25.15% and TDZ=0.5mg/L+NAA=2mg/L with 24.53% was obtained in hypocotyle, root and cotyledon explant respectively. The highest propagation was observed in BA=0.5mg/L+Thiamin-HCl=4mg/L treatment for regenerated explants. However, BA=1mg/L+NAA=1mg/L treatment have no significant effects on the regeneration of callus inducing explants. In this research, we obtain less than 4% rooting in shoots by hormonal treatment.