تأثير پيش تيمار دمايي و نور بر القاء جنين و كالوس در كشت بساك خيار

Effect of Temperature Pre-Treatment and Light on Embryogenesis and Callus Induction in Anther Culture of Cucumber

ﺑﺎﻫﺪف ﺑﺮرﺳ ﻲ ﺗﺄﺛﻴﺮ ﺗﺮﻛﻴﺐﻫﺎ و ﻏﻠﻈﺖﻫﺎي ﻣﺨﺘﻠﻒ ﻫﻮرﻣﻮﻧﻲ و ﻫﻤﭽﻨﻴﻦ ﺗﺄﺛﻴﺮ ﭘﻴﺶﺗﻴﻤﺎر دﻣﺎﻳﻲ و ﻧﻮر ﺑﺮ ﺗﻐﻴﻴﺮ ﻣﺴﻴﺮ ﺑﻴﻮﻟﻮژﻳﻜﻲ و ﺣﺮﻛﺖ ﺑﻪ ﺳﻤﺖ ﻛﺎﻟﻮسزاﻳﻲ، ﺟﻨﻴﻦزاﻳﻲ و ﺑﺎززاﻳﻲ در رﻳ ﺰﻧﻤﻮﻧﻪﻫﺎي ﺑﺴ ﺎك ﺧ ﻴﺎر، ﭘﮋوﻫﺸﻲ ﺑﻪ ﺻ ﻮرت دو آزﻣﺎﻳﺶ ﻓﺎﻛ ﺘﻮرﻳ ﻞ ﺟﺪاﮔﺎﻧﻪ در ﻗﺎﻟ ﺐ ﻃﺮح ﻛ ﺎﻣ ﻼً ﺗﺼ ﺎدﻓ ﻲ ﺑﺎ ﺳ ﻪ ﺗﻜ ﺮار ﺑﺮ روي رﻗﻢ ﺑﻮﻣﻲ ﻓﻀﺎي ﺑﺎز ﺧﻴﺎر اﺻﻔﻬﺎﻧﻲ و ﻻﻳﻦ اﺻﻼﺣﻲ ﺧﻴﺎر ﮔﻠﺨﺎﻧﻪاي اﺟ ﺮا ﮔ ﺮدﻳ ﺪ. ﻓﺎﻛﺘﻮرﻫﺎ ﺷﺎﻣﻞ ﭘﻴﺶ ﺗﻴﻤﺎر دﻣﺎﻳﻲ )ﺳﺮﻣﺎي 4 درﺟﻪ ﺳﺎﻧﺘﻲ ﮔﺮاد و ﺑﺪون ﭘﻴﺶﺗﻴﻤﺎر ﺳﺮﻣﺎﻳﻲ )ﮔﻞ ﺗﺎزه((، ﺗﻴﻤﺎر ﻧﻮري ﭘﺲ از ﻛﺸﺖ )ﻧﻮر و ﺗﺎرﻳﻜﻲ( و ﻏﻠﻈﺖﻫﺎي ﻣﺨﺘﻠﻒ ﻫﻮرﻣﻮﻧﻲ ﺷﺎﻣﻞ KIN )1 ﺗﺎ 2/5 ﻣﻴﻠﻲﮔﺮم/ﻟﻴﺘﺮ(، 2,4-D )1 ﺗﺎ 3/5 ﻣﻴﻠﻲ- ﮔﺮم/ﻟﻴﺘﺮ( و BAP )0/1 ﺗﺎ 3/5 ﻣﻴﻠﻲﮔﺮم/ﻟﻴﺘﺮ( ﺑﻮدﻧﺪ. ﻧ ﺘﺎﻳ ﺞ ﺣ ﺎﻛ ﻲ از ﺑﺮﺗ ﺮي ﭘ ﻴﺶ ﺗ ﻴﻤ ﺎر ﺑﺪون ﺳﺮﻣﺎ ﮔ ﻞ ﺗﺎزه( ﻧﺴﺒﺖ ﺑﻪ ﭘﻴﺶﺗﻴﻤﺎر ﺳﺮﻣﺎ در ﺟ ﻨ ﻴﻦ زاﻳ ﻲ و ﻛ ﺎﻟ ﻮس زاﻳ ﻲ در ﻻﻳﻦ اﺻﻼﺣﻲ و ﻛ ﺎﻟ ﻮس زاﻳ ﻲ در رﻗ ﻢ ﺑﻮﻣ ﻲ ﺑﻮد. ﺗﻴﻤﺎر ﺗﺎرﻳﻜﻲ ﺑﺎﻋﺚ اﻓﺰاﻳﺶ ﻣﻌﻨﻲدار درﺻﺪ ﻛﺎﻟﻮسزاﻳﻲ و ﻗﻄﺮ ﻛﺎﻟﻮس ﻻﻳﻦ اﺻﻼﺣﻲ ﺷﺪ، اﻣﺎ ﺗﺄﺛﻴﺮ ﻣﻌﻨﻲداري ﺑﺮ روي ﺻﻔﺎت ﻣﻮرد ﺑﺮرﺳﻲ در رﻗﻢ ﺑﻮﻣﻲ ﻧﺪاﺷﺖ. ﺗﺮﻛ ﻴﺐ ﻫ ﻮرﻣ ﻮﻧ ﻲ BAP و 2,4-D )1+3/5 ﻣﻴﻠﻲﮔﺮم/ﻟﻴﺘﺮ(در ﻫ ﺮ دو ژﻧﻮﺗﻴﭗ ﺑﺎﻋ ﺚ ﺑ ﻴﺸ ﺘﺮﻳ ﻦ اﻟ ﻘ ﺎي ﺟﻨﻴﻦزاﻳﻲ و ﻛﺎﻟﻮسزاﻳﻲ ﺷ ﺪ. ﺟﻨﻴﻦزاﻳﻲ در ﻻﻳﻦ اﺻﻼﺣﻲ ﺗﺤﺖ ﺗﺄﺛﻴﺮ ﻓﺎﻛﺘﻮرﻫﺎي ﻣﻮرد ﻣﻄﺎﻟﻌﻪ ﻗﺮارﮔﺮﻓﺖ اﻣﺎ اﻳﻦ ﻓﺎﻛﺘﻮرﻫﺎ ﺗﺄﺛﻴﺮ ﻣﻌﻨﻲداري ﺑﺮ ﺟﻨﻴﻦزاﻳﻲ رﻗﻢ ﺑﻮﻣﻲ ﻧﺪاﺷﺘﻨﺪ. اﺛﺮﻣﺘﻘﺎﺑﻞ ﺳﻪﮔﺎﻧﻪ ﭘﻴﺶﺗﻴﻤﺎر دﻣﺎﻳﻲ، ﺗﻴﻤﺎر ﻧﻮري و ﻫﻮرﻣﻮﻧﻲ ﻣﺤﻴﻂ ﻛﺸﺖ ﻧﺸﺎن داد ﻛﻪ ﺑﻪﻣﻨﻈﻮر ﺑﻴﺸﺘﺮﻳﻦ ﺟﻨﻴﻦزاﻳﻲ، ﻛﺎﻟﻮسزاﻳﻲ و ﻗﻄﺮ ﻛﺎﻟﻮس در ﻛﺸﺖ ﺑﺴﺎك ﺧﻴﺎر، اﺳﺘﻔﺎده از ﮔﻞ ﺗﺎزه و ﻣﺤﻴﻂ ﻛﺸﺖ D+BAP-2,4 ) 1+3/5 ﻣﻴﻠﻲﮔﺮم/ﻟﻴﺘﺮ( ﺑﺎﻋﺚ ﺣﺎﺻﻞ ﺷﺪن ﺑﻬﺘﺮﻳﻦ ﻧﺘﻴﺠﻪ ﺧﻮاﻫﺪ ﺷﺪ و ﺷﺮاﻳﻂ ﻧﻮر و ﺗﺎرﻳﻜﻲ ﻧﻘﺶ ﭼﻨﺪاﻧﻲ در ﺟﻨﻴﻦزاﻳﻲ ﻧﺪارد.

The aim of this study was to investigate the effects of hormonal compositions and concentrations, temperature and light pre-treatments in changing the biological pathways and moving toward callugenesis, embryogenesis and regeneration in cucumber anther explants. Two seperate factorial experiments were conducted in completely randomized design with three replications on open pollination Isfahan native cucumber and greenhouse inbred line. Factors consisted of tempreture (4°C cold and without cold pretreatment (fresh flower)), Light treatment (brightness and darkness) and different hormone compositions and concentrations including KIN (1 to 2.5 mg/L), BAP (0.1 to 3.5 mg/L) and 2.4-D (1 to 3.5 mg/L) at five levels. The results showed the superiority of the fresh flower in cold pre-treatment for embrygenesis and callugenesis in inbred line, and for callugenesis in native cultivar. Darkness treatment caused a significant increasing in callugenesis and callus diameter of inbred line, however did not significant effects on native cultivar. The combination of BAP and 2,4-D hormones at a rate of 3.5+1mg/L (in both genotypes) caused the highest induction of embryogenesis and callugenesis. Embryogenesis was affected by studied factors in inbred line, however these factors did not have significant effects on embryogenesis in native cucumber. Triple interaction of temperature, light and hormonal compositions in culture media showed that in order to obtain the highest embryogenesis, callus induction and callus diameter, use of fresh flowers and 2,4-D+BAP hormones (3.5+1 mg/L) are suitable in anther culture of cucumber. The conditions of lightness and darkness did not play important role in embryogenesis.