جنين زايي سوماتيكي مستقيم و توليد بذر مصنوعي در بلوط ايراني (Quercus brantii L.) با استفاده از هورمون 2,4-D

Direct somatic embryogenesis and synthetic seed production of Persian oak (Quercus brantii L.) using 2,4-D hormone

ﺑﻠﻮط اﻳﺮاﻧﻲ )Quercus brantii L.( از اﺻﻠﻲ ﺗﺮﻳﻦ ﮔﻮﻧﻪ ﻫﺎي درﺧﺘﻲ ﺟﻨﮕﻞ ﻫﺎي زاﮔﺮس در اﻳﺮان اﺳﺖ. ﺑﺮاي ﺣﻔﻆ ﺑﻘﺎي اﻳﻦ ﮔﻮﻧﻪ اﺳﺘﻔﺎده از روش ﻫﺎي آزﻣﺎﻳﺸﮕﺎﻫﻲ ﻣﺜﻞ ﻛﺸﺖ ﺑﺎﻓﺖ و ﺟﻨﻴﻦ زاﻳﻲ روﻳﺸﻲ ﻣﻨﺎﺳﺐ اﺳﺖ، ﺑﻪ ﻫﻤﻴﻦ ﻣﻨﻈﻮر دو آزﻣﺎﻳﺶ ﻣﺠﺰا ﺑﺼﻮرت ﺟﻨﻴﻦ زاﻳﻲ ﺳﻮﻣﺎﺗﻴﻜﻲ و ﭘﻮﺷﺶ دار ﻛﺮدن اﻳﻦ ﺟﻨﻴﻦ ﻫﺎ ﻃﺮاﺣﻲ و اﺟﺮا ﺷﺪ. آزﻣﺎﻳﺶ ﺟﻨﻴﻦ زاﻳﻲ ﺳﻮﻣﺎﺗﻴﻜﻲ در ﻗﺎﻟﺐ ﻃﺮح ﻛﺎﻣﻼً ﺗﺼﺎدﻓﻲ ﺑﺎ ﭼﻬﺎر ﺗﻜﺮار اﺟﺮا ﺷﺪ، دراﻳﻦ آزﻣﺎﻳﺶ ﺟﻨﻴﻦ زﻳﮕﻮﺗﻲ ﻧﺎﺑﺎﻟﻎ ﺑﻌﻨﻮان ﻣﻨﺒﻊ رﻳﺰ ﻧﻤﻮﻧﻪ و ﻫﻮرﻣﻮن D-2,4 در ﭼﻬﺎر ﺳﻄﺢ 0،0/25، 0/5 و 0/75 ﻣﻴﻠﻲ ﮔﺮم ﺑﺮ ﻟﻴﺘﺮ( اﺳﺘﻔﺎده ﺷﺪ. آزﻣﺎﻳﺶ ﻛﭙﺴﻮﻟﻪ ﻛﺮدن ﺟﻨﻴﻦ ﻫﺎي ﺳﻮﻣﺎﺗﻴﻜﻲ ﺑﺼﻮرت ﻓﺎﻛﺘﻮرﻳﻞ در ﻗﺎﻟﺐ ﻃﺮح ﻛﺎﻣﻼً ﺗﺼﺎدﻓﻲ ﺑﺎ ﭼﻬﺎر ﺗﻜﺮار اﺟﺮا ﺷﺪ، ﻓﺎﻛﺘﻮر اول ﺷﺎﻣﻞ ﺳﻪ ﻏﻠﻈﺖ آﻟﮋﻳﻨﺎت ﺳﺪﻳﻢ 3/5، 4 و 4/5 درﺻﺪ( و ﻓﺎﻛﺘﻮر دوم دو ﻏﻠﻈﺖ ﻛﻠﺮﻳﺪ ﻛﻠﺴﻴﻢ 50 و 100 ﻣﻴﻠﻲ ﻣﻮﻻر( ﺑﻮد. ﻧﺘﺎﻳﺞ ﻧﺸﺎن داد ﻛﻪ در ﻏﻠﻈﺖ 0/25 ﻣﻴﻠﻲ ﮔﺮم ﺑﺮ ﻟﻴﺘﺮ D-2,4 ﺑﻴﺸﺘﺮﻳﻦ ﺗﻌﺪاد و درﺻﺪ ﺟﻨﻴﻦ زاﻳﻲ 66/6 درﺻﺪ( ﻣﺸﺎﻫﺪه ﺷﺪ. ﻧﺘﺎﻳﺞ آزﻣﺎﻳﺶ ﭘﻮﺷﺶ دار ﻛﺮدن ﻧﺸﺎن داد ﻛﻪ ﺑﺮﻫﻢ ﻛﻨﺶ آﻟﮋﻳﻨﺎت ﺳﺪﻳﻢ و ﻛﻠﺮﻳﺪ ﻛﻠﺴﻴﻢ ﺑﺮاي ﺗﻤﺎم ﺗﺮﻛﻴﺒﺎت در ﺳﻄﺢ اﺣﺘﻤﺎل 1 درﺻﺪ ﻣﻌﻨﻲ دار ﺷﺪ، ﻏﻠﻈﺖ 3/5 درﺻﺪ آﻟﮋﻳﻨﺎت ﺳﺪﻳﻢ ﻫﻤﺮاه ﺑﺎ 50 ﻣﻴﻠﻲ ﻣﻮﻻر ﻛﻠﺮﻳﺪ ﻛﻠﺴﻴﻢ ﺑﻬﺘﺮﻳﻦ ﺗﻴﻤﺎر ﺑﺮاي ﭘﻮﺷﺶ دار ﻛﺮدن ﺑﻮد ﻛﻪ ﺑﻴﺸﺘﺮﻳﻦ درﺻﺪ) 100 ( ﺟﻮاﻧﻪ زﻧﻲ در اﻳﻦ ﺗﺮﻛﻴﺐ ﻣﺸﺎﻫﺪه ﺷﺪ.

Persian oak (Quercus brantii L.) is the most important Zagros forest tree in Iran. Therefore, in order to maintain the survival of this specie, use the laboratory methods such as tissue culture and somatic embryogenesis is suitable. So, two separate experiments were designed includes somatic embryogenesis and encapsulation of somatic embryos. Somatic embryogenesis experiment was done in CRD with 4 replications. In this experiment immature zygotic embryos as explant source and 2,4-D in 4 level (0, 0.25, 0.5 and 0.75 mg l-1) were used. Encapsulation test was done in factorial based on CRD with 4 replications; factors include sodium alginate in 3 concentrations (3.5, 4 and 4.5 %) and CaCl2 in two concentrations (50 and 100 Mm). Results of somatic embryogenesis showed that the maximum number of somatic embryos and the highest somatic embryogenesis percentage (66.6%) were observed in 0.25 mg l-1 2,4-D. Results of encapsulation showed that, sodium alginate 3.5% with CaCl2 50 mM was the best treatment for encapsulation of somatic embryos and had highest germination percentage (100%).