القای ریشه مویین در گیاه کتان سفید بواسطه آگروباکتریوم رایزوژنز
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عنوان دوره: سیزدهمین کنگره زراعت و اصلاح نباتات ایران
نویسندگان
1دانشجوی کارشناسی ارشد، گروه اصلاح و بیوتکنولوژی گیاهی، دانشکده کشاورزی، دانشگاه ارومیه
2استادیار، گروه اصلاح و بیوتکنولوژی کشاورزی، دانشکده کشاورزی، دانشگاه ارومیه
چکیده
رﻳﺸـﻪ ﻫـﺎی ﻣـﻮﻳﻴﻦ ﻣﻨﺒـﻊ ارزﺷـﻤﻨﺪی ﺑـﺮای ﺗﻮﻟﻴـﺪ ﺗﺮﻛﻴﺒـﺎت ﻣﻬـﻢ داروﻳـﻲ ﮔﻴﺎﻫـﺎن ﻫﺴـﺘﻨﺪ. بسیاری از ﮔﻮﻧـﻪ ﻫـﺎ ی ﮔﻴـ ﺎﻫﻲ آگروباکتریوم رایزوژنز ﺣﺴﺎس ﻣﻲ ﺑﺎﺷﻨﺪ و ﺗﻠﻘـﻴﺢ آﻧﻬـﺎ ﺑـﺎ اﻳـﻦ ﺑـﺎﻛﺘﺮی ﻣﻨﺠﺮﺑـﻪ اﻟﻘـﺎ ی رﻳﺸـﻪ ﻫـﺎی ﻣـﻮﻳﻴﻦ ﻣـﻲ ﮔـﺮدد. اﻳـﻦ رﻳﺸـﻪ ﻫﺎ در اﺛﺮ ﺗﻠﻔﻴﻖ ﺑﺨﺸﻲ از DNA ﭘﻼﺳـﻤﻴﺪ اﻟﻘﺎﻛﻨﻨـﺪه رﻳﺸـﻪ ﺑـﻪ درون ژﻧـﻮم ﮔﻴـﺎه ﺑـﻪ وﺟـﻮد ﻣـﻲ آﻳﻨـﺪ ﮔﻴـﺎه ﻛﺘـﺎن ﺳـﻔﻴﺪ Linum (
)album دارای ﺗﺮﻛﻴﺒﺎت ﻟﻴﮕﻨﺎﻧﻲ ﻣﺎﻧﻨﺪ ﭘﻮدوﻓﻴﻠﻮﺗﻮﻛﺴـﻴﻦ ﻣـﻲ ﺑﺎﺷـﺪ ﻛـﻪ ﺑـﻪ دﻟﻴـﻞ ﺧـﻮاص ﺿـﺪ ﺳـﺮﻃﺎﻧﻲ اﻫﻤﻴـﺖ داروﻳـﻲ زﻳـﺎد ی دارد. در اﻳﻦ ﺗﺤﻘﻴﻖ رﻳﺰﻧﻤﻮﻧـﻪ ﻫـﺎی ﻣﻴـﺎﻧﮕﺮه، ﻛﻮﺗﻴﻠـﺪون و ﺑـﺮگ ﻛﺘـﺎن ﺳـﻔﻴﺪ، ﺑـﻪ ﻣﻨﻈـﻮر اﻟﻘـﺎی رﻳﺸـﻪ ﻫـﺎی ﻣـﻮﻳﻴﻦ ﺑﻮاﺳـﻄﻪ ﺳـﻮﻳﻪ A13 آﮔﺮوﺑﺎﻛﺘﺮﻳﻮم راﻳﺰوژﻧﺰ اﺳﺘﻔﺎده ﺷﺪ. آﻧـﺎﻟﻴﺰ آﻣـﺎری ﺗﻌـﺪاد رﻳﺸـﻪ ﻫـﺎی اﻟﻘـﺎﻳﻲ اﻳـﻦ آزﻣـﺎﻳﺶ درﻗﺎﻟـﺐ ﻃـﺮح ﻛـﺎﻣﻼ ﺗﺼـﺎدﻓﻲ ﺑـﺎ 3 ﺗﻜــﺮار اﻧﺠــﺎم ﺷــﺪ و ﺑــﺮای ﺗﺄﻳﻴــﺪ درج ﻗﻄﻌــﻪ T-DNA در ژﻧــﻮم رﻳﺸــﻪﻫــﺎی ﻣــﻮﻳﻴﻦ از واﻛــﻨﺶ زﻧﺠﻴــﺮهای ﭘﻠﻴﻤــﺮاز (PCR) ﺑــﺎ اﺳـﺘﻔﺎده از آﻏـﺎزﮔﺮ اﺧﺘﺼﺎﺻـﻲ ﺑـﺮای ﺗﻜﺜﻴـﺮ ﻫﻤﺰﻣـﺎن ﺑﺨـﺶ ﻫـﺎﻳﻲ از ژن ﻫـﺎی rolA و rolB آﻧـﺎﻟﻴﺰ ﻣﻠﻜـﻮﻟﻲ اﻧﺠـﺎم ﮔﺮﻓـﺖ ﻧﺘـﺎﻳﺞ ﺣﺎﺻــﻞ از آﻧــﺎﻟﻴﺰ آﻣــﺎری ﻧﺸــﺎن داد ﻛــﻪ رﻳــﺰﻧﻤﻮﻧــﻪﻫــﺎی ﻛﻮﺗﻴﻠــﺪون ﺑﻴﺸــﺘﺮﻳﻦ ﺗﻌــﺪاد رﻳﺸــﻪ)7-6 رﻳﺸــﻪ در ﻫــﺮ رﻳﺰﻧﻤﻮﻧــﻪ( و ﺑــﺮگ ﻛﻤﺘــﺮﻳﻦ ﺗﻌــﺪاد رﻳﺸــﻪ)3-2 رﻳﺸــﻪ در ﻫــﺮ رﻳﺰﻧﻤﻮﻧــﻪ( را ﺗﻮﻟﻴــﺪ ﻛﺮدﻧــﺪ و ﻣﻴــﺎﻧﮕﺮهﻫــﺎ ﻫــﻴﭻ رﻳﺸــﻪای ﺗﻮﻟﻴــﺪ ﻧﻜﺮدﻧــﺪ. آﻧــﺎﻟﻴﺰ ﻫﻤﺮاه ﺑﺎ رﺷﺪ ﺳﺮﻳﻊ رﻳﺸﻪﻫﺎ در ﻳﻚ ﻣﺤﻴﻂ ﻛﺸﺖ ﺑﺪون ﻫﺮ ﮔﻮﻧﻪ ﺗﻨﻄﻴﻢ ﻛﻨﻨﺪه رﺷﺪ ﻣﻮﻳﻴﻦ ﺑﻮدن رﻳﺸﻪﻫﺎ را ﺗﺎﻳﻴﺪ ﻧﻤﻮد
)album دارای ﺗﺮﻛﻴﺒﺎت ﻟﻴﮕﻨﺎﻧﻲ ﻣﺎﻧﻨﺪ ﭘﻮدوﻓﻴﻠﻮﺗﻮﻛﺴـﻴﻦ ﻣـﻲ ﺑﺎﺷـﺪ ﻛـﻪ ﺑـﻪ دﻟﻴـﻞ ﺧـﻮاص ﺿـﺪ ﺳـﺮﻃﺎﻧﻲ اﻫﻤﻴـﺖ داروﻳـﻲ زﻳـﺎد ی دارد. در اﻳﻦ ﺗﺤﻘﻴﻖ رﻳﺰﻧﻤﻮﻧـﻪ ﻫـﺎی ﻣﻴـﺎﻧﮕﺮه، ﻛﻮﺗﻴﻠـﺪون و ﺑـﺮگ ﻛﺘـﺎن ﺳـﻔﻴﺪ، ﺑـﻪ ﻣﻨﻈـﻮر اﻟﻘـﺎی رﻳﺸـﻪ ﻫـﺎی ﻣـﻮﻳﻴﻦ ﺑﻮاﺳـﻄﻪ ﺳـﻮﻳﻪ A13 آﮔﺮوﺑﺎﻛﺘﺮﻳﻮم راﻳﺰوژﻧﺰ اﺳﺘﻔﺎده ﺷﺪ. آﻧـﺎﻟﻴﺰ آﻣـﺎری ﺗﻌـﺪاد رﻳﺸـﻪ ﻫـﺎی اﻟﻘـﺎﻳﻲ اﻳـﻦ آزﻣـﺎﻳﺶ درﻗﺎﻟـﺐ ﻃـﺮح ﻛـﺎﻣﻼ ﺗﺼـﺎدﻓﻲ ﺑـﺎ 3 ﺗﻜــﺮار اﻧﺠــﺎم ﺷــﺪ و ﺑــﺮای ﺗﺄﻳﻴــﺪ درج ﻗﻄﻌــﻪ T-DNA در ژﻧــﻮم رﻳﺸــﻪﻫــﺎی ﻣــﻮﻳﻴﻦ از واﻛــﻨﺶ زﻧﺠﻴــﺮهای ﭘﻠﻴﻤــﺮاز (PCR) ﺑــﺎ اﺳـﺘﻔﺎده از آﻏـﺎزﮔﺮ اﺧﺘﺼﺎﺻـﻲ ﺑـﺮای ﺗﻜﺜﻴـﺮ ﻫﻤﺰﻣـﺎن ﺑﺨـﺶ ﻫـﺎﻳﻲ از ژن ﻫـﺎی rolA و rolB آﻧـﺎﻟﻴﺰ ﻣﻠﻜـﻮﻟﻲ اﻧﺠـﺎم ﮔﺮﻓـﺖ ﻧﺘـﺎﻳﺞ ﺣﺎﺻــﻞ از آﻧــﺎﻟﻴﺰ آﻣــﺎری ﻧﺸــﺎن داد ﻛــﻪ رﻳــﺰﻧﻤﻮﻧــﻪﻫــﺎی ﻛﻮﺗﻴﻠــﺪون ﺑﻴﺸــﺘﺮﻳﻦ ﺗﻌــﺪاد رﻳﺸــﻪ)7-6 رﻳﺸــﻪ در ﻫــﺮ رﻳﺰﻧﻤﻮﻧــﻪ( و ﺑــﺮگ ﻛﻤﺘــﺮﻳﻦ ﺗﻌــﺪاد رﻳﺸــﻪ)3-2 رﻳﺸــﻪ در ﻫــﺮ رﻳﺰﻧﻤﻮﻧــﻪ( را ﺗﻮﻟﻴــﺪ ﻛﺮدﻧــﺪ و ﻣﻴــﺎﻧﮕﺮهﻫــﺎ ﻫــﻴﭻ رﻳﺸــﻪای ﺗﻮﻟﻴــﺪ ﻧﻜﺮدﻧــﺪ. آﻧــﺎﻟﻴﺰ ﻫﻤﺮاه ﺑﺎ رﺷﺪ ﺳﺮﻳﻊ رﻳﺸﻪﻫﺎ در ﻳﻚ ﻣﺤﻴﻂ ﻛﺸﺖ ﺑﺪون ﻫﺮ ﮔﻮﻧﻪ ﺗﻨﻄﻴﻢ ﻛﻨﻨﺪه رﺷﺪ ﻣﻮﻳﻴﻦ ﺑﻮدن رﻳﺸﻪﻫﺎ را ﺗﺎﻳﻴﺪ ﻧﻤﻮد
کلیدواژه ها
Title
Induction of hairy roots of Linum album by Agrobacterium rhizogenes
Authors
Abstract
Hairy roots of medicinal plants are a valuable source for the production of important compounds. Many
plant species are susceptible to Agrobacterium rhizogenes and inoculated with these bacteria may lead to the
induction of hairy roots. The roots of the root inducing plasmid DNA incorporated into the genome of plants
to come. White flax plant (Linum album) compounds such as Podophyllotoxin Lignan is important because
many drug has anticancer properties. In this study internode explants, cotyledon and leaf of white linen, in
order to induce hairy roots by strain A13 of Agrobacterium rhizogenes was used. Statistical analysis of the
origins of the inducible form of completely randomized design with 3 replications was used to test and
validate the inserted T-DNA fragment in the genome of hairy roots of polymerase chain reaction (PCR)
using specific primers for simultaneous amplification of part of the gene Molecular analysis was performed
by rolA and rolB. PCR analysis with the rapid growth in a medium without any root regulated growth of
hairy roots was confirmed.
plant species are susceptible to Agrobacterium rhizogenes and inoculated with these bacteria may lead to the
induction of hairy roots. The roots of the root inducing plasmid DNA incorporated into the genome of plants
to come. White flax plant (Linum album) compounds such as Podophyllotoxin Lignan is important because
many drug has anticancer properties. In this study internode explants, cotyledon and leaf of white linen, in
order to induce hairy roots by strain A13 of Agrobacterium rhizogenes was used. Statistical analysis of the
origins of the inducible form of completely randomized design with 3 replications was used to test and
validate the inserted T-DNA fragment in the genome of hairy roots of polymerase chain reaction (PCR)
using specific primers for simultaneous amplification of part of the gene Molecular analysis was performed
by rolA and rolB. PCR analysis with the rapid growth in a medium without any root regulated growth of
hairy roots was confirmed.
Keywords
Agrobacterium rhizogenes, explants, hairy root, Linum album