ﻣﺠﻠﻪ ﺑﻪﻧﮋادي ﻧﻬﺎل و ﺑﺬر ﺟﻠﺪ ، 30-1ﺷﻤﺎره ،3ﺳﺎل 1393 ﺷﻨﺎﺳﺎﻳﻲ آﻟﻞﻫﺎي ﺧﻮد ﻧﺎﺳﺎزﮔﺎري در ارﻗﺎم ﺑﺎدام اﻳﺮاﻧﻲ ﺑﻪ روش ﺗﻮاﻟﻲﻳﺎﺑﻲ Identification of Self-Incompatibility Alleles in Iranian Almond Cultivars Using Sequencing ﺳﻴﺪاﺻﻐﺮ ﻣﻮﺳﻮي ،1ﻣﺤﻤﺪرﺿﺎ ﻓﺘﺎﺣﻲﻣﻘﺪم ،2ذﺑﻴﺢاﻟﻪ زﻣﺎﻧﻲ ،3ﻣﺮﻳﻢ ﺗﺎﺗﺎري،4 6 D I ﻋﻠﻲ اﻳﻤﺎﻧﻲ 5و اﻧﻜﺎرﻧﺎ اورﺗﮕﺎ ﭘﺎﺳﺘﻮر -1اﺳﺘﺎدﻳﺎر ،ﻣﺮﻛﺰ ﺗﺤﻘﻴﻘﺎت ﻛﺸﺎورزي و ﻣﻨﺎﺑﻊ ﻃﺒﻴﻌﻲ ﭼﻬﺎرﻣﺤﺎل و ﺑﺨﺘﻴﺎري ،ﺷﻬﺮﻛﺮد S f 2و -3ﺑﻪ ﺗﺮﺗﻴﺐ داﻧﺸﻴﺎر و اﺳﺘﺎد ،ﭘﺮدﻳﺲ ﻛﺸﺎورزي و ﻣﻨﺎﺑﻊ ﻃﺒﻴﻌﻲ داﻧﺸﮕﺎه ﺗﻬﺮان ،ﻛﺮج 4و -5ﺑﻪ ﺗﺮﺗﻴﺐ ﻛﺎرﺷﻨﺎس ﺗﺤﻘﻴﻘﺎﺗﻲ و اﺳﺘﺎدﻳﺎر ،ﻣﻮﺳﺴﻪ ﺗﺤﻘﻴﻘﺎت اﺻﻼح و ﺗﻬﻴﻪ ﻧﻬﺎل و ﺑﺬر ،ﻛﺮج -6داﻧﺸﻴﺎر ،ﮔﺮوه اﺻﻼح ﻧﺒﺎﺗﺎت ،ﻣﻮﺳﺴﻪ ﺗﺤﻘﻴﻘﺎت ،CEBAS-SCICﻣﻮرﺳﻴﺎ ،اﺳﭙﺎﻧﻴﺎ o e ﺗﺎرﻳﺦ ﭘﺬﻳﺮش1393/3/8 : ﺗﺎرﻳﺦ درﻳﺎﻓﺖ1392/9/13 : ﭼﻜﻴﺪه v i h ﻣﻮﺳﻮي ،س .ا ،.ﻓﺘﺎﺣﻲﻣﻘﺪم ،م .ر ،.زﻣﺎﻧﻲ ،ذ ،.ﺗﺎﺗﺎري ،م ،.اﻳﻤﺎﻧﻲ ،ع .و اورﺗﮕﺎ ﭘﺎﺳﺘﻮر ،ا .1393.ﺷﻨﺎﺳﺎﻳﻲ آﻟﻞﻫﺎي ﺧـﻮد ﻧﺎﺳـﺎزﮔﺎري در ارﻗـﺎم ﺑـﺎدام اﻳﺮاﻧﻲ ﺑﻪ روش ﺗﻮاﻟﻲﻳﺎﺑﻲ .ﻣﺠﻠﻪ ﺑﻪﻧﮋادي ﻧﻬﺎل و ﺑﺬر .651 - 668 :30-1 c r ﺷﻨﺎﺳﺎﻳﻲ آﻟﻞﻫﺎي ﺧﻮدﻧﺎﺳﺎزﮔﺎري در ارﻗﺎم ﺑﺎدام ) (Prunus dulcisاﻫﻤﻴﺖ زﻳﺎدي در اﺣﺪاث ﺑﺎغﻫـﺎي ﺟﺪﻳـﺪ و اﻧﺘﺨﺎب واﻟﺪﻳﻦ در ﺑﺮﻧﺎﻣﻪﻫﺎي ﺑﻪﻧﮋادي ﺑﺎدام دارد .ﺑﻪ ﻣﻨﻈﻮر ﺷﻨﺎﺳﺎﻳﻲ آﻟﻞﻫﺎي ﺟﺪﻳﺪ ﻧﺎﺳـﺎزﮔﺎري در ﭼﻨـﺪ رﻗـﻢ ﺑﺎدام اﻳﺮاﻧﻲ ،ﻗﻄﻌﺎت ﻣﺮﺑﻮط ﺑﻪ ﻫﺮ آﻟﻞ ﺗﻜﺜﻴﺮ و ﻣﺤﺼﻮل PCRآنﻫﺎ ﺧﺎﻟﺺﺳﺎزي ﺷﺪ .ﺑﺮاي ﻫﻤﺴﺎﻧﻪﺳﺎزي از ﻧﺎﻗـﻞ PCR-Blunt-II-TOPOاﺳﺘﻔﺎده ﺷﺪ .ﻧﺎﻗﻞ ﺑﻪ ﺑﺎﻛﺘﺮيﻫﺎي ﻣﺴﺘﻌﺪ Escherichia coli TOP 10ﻣﻨﺘﻘﻞ و ﻛﻠﻮﻧﻲﻫـﺎي ﺗﺮارﻳﺰش ﺷﺪه ﺷﻨﺎﺳﺎﻳﻲ ﺷﺪﻧﺪ DNA .ﭘﻼﺳﻤﻴﺪي اﺳﺘﺨﺮاج و ﺗﻮاﻟﻲﻳﺎﺑﻲ ﺑـﺎ اﺳـﺘﻔﺎده از آﻏﺎزﮔﺮﻫـﺎي M13ﺗﻮﺳـﻂ دﺳﺘﮕﺎه SECUGENاﻧﺠﺎم ﺷﺪ .ﻧﻬﺎﻳﺘﺎً ﺗﻮاﻟﻲ ﺣﺎﺻﻞ از اﻳﻦ ﭘﻼﺳﻤﻴﺪﻫﺎ ﻣﻘﺎﻳﺴﻪ و ﺗﻮاﻟﻲ ﺛﺎﺑﺖ ﻣﺮﺑﻮط ﺑﻪ ﻫـﺮ آﻟـﻞ ﺑـﺎ ﻧﺮماﻓﺰار SEQMAN IIﺗﻌﻴﻴﻴﻦ ﺷﺪ .ﺑﺮاﺳﺎس ﻧﺘﺎﻳﺞ اﻳﻦ ﺗﺤﻘﻴﻖ ،ﺗﻌﺪاد ﻫﺸﺖ آﻟﻞ در ارﻗﺎم ﺑﺎدام اﻳﺮاﻧﻲ ﺷﺎﻣﻞ ارﻗﺎم ﺗﺠﺎري ) ،(S36ﻫﻠﻮﻳﻲ ) ،(S37ﺳﻔﻴﺪ ) ،(S38ﻳﺰد S39) 17-و ،(S40ﻣـﺸﻬﺪ ،(S41) 40ﺟـﻲ-آر (S42) 16-و ﻣﺎﻣـﺎﻳﻲ )(S43 ﺷﻨﺎﺳﺎﻳﻲ ﺷﺪﻧﺪ .ﻫﻔﺖ آﻟﻞ از ﻣﺠﻤﻮع ﻫﺸﺖ آﻟﻞ ﺷﻨﺎﺳﺎﻳﻲ ﺷﺪه ،ﻗﺒﻼ ﮔﺰارش ﻧﺸﺪه اﺳﺖ ،وﻟﻲ ﻳﻜﻲ از آﻟﻞﻫﺎ )(S40 ﺷﺒﺎﻫﺖ زﻳـﺎدي ﺑـﺎ آﻟـﻞ ﻧﺎﺳـﺎزﮔﺎري ) (S5ﮔـﺰارش ﺷـﺪه در ﮔﻮﻧـﻪ P. webbiiداﺷـﺖ .ﺷـﺒﺎﻫﺖ ﺑـﻴﻦ آﻟـﻞﻫـﺎي ﺧﻮدﻧﺎﺳﺎزﮔﺎري در ﺑﺎدام اﻫﻠﻲ ﺑﺎ ﮔﻮﻧﻪ وﺣﺸﻲ P. webbiiﺑﻪ وﺟﻮد ﻳﻚ ﺟﺪ ﻣﺸﺘﺮك ﺑﺎ ﮔﻮﻧﻪﻫﺎي ﺟﻨﺲ ﭘﺮوﻧﻮس و ﻧﻘﺶ اﻳﻦ ﮔﻮﻧﻪ در ﺗﻜﺎﻣﻞ ﺑﺎدامﻫﺎي اﻫﻠﻲ ﺑﻪ ﻋﻨﻮان ﻳﻜﻲ از اﺟﺪاد اوﻟﻴﻪ اﺷﺎره دارد. A واژهﻫﺎي ﻛﻠﻴﺪي :ﺑﺎدام ،رﻗﻢ ،آﻟﻞ ﻧﺎﺳﺎزﮔﺎري ،ﻫﻤﺴﺎﻧﻪﺳﺎزي ،ﺗﻮاﻟﻲﻳﺎﺑﻲ. ﻧﻮﻳﺴﻨﺪه ﻣﺴﺌﻮل[email protected] : www.SID.ir ﺗﻠﻔـــﻦ03813335640 : 651 ”ﻣﺠﻠﻪ ﺑﻪﻧﮋادي ﻧﻬﺎل و ﺑﺬر” ﺟﻠﺪ ،30-1ﺷﻤﺎره ،3ﺳﺎل 1393 ﻣﻘﺪﻣﻪ ) .(Sassa et al., 1996ﻣﻄﺎﻟﻌــﺎت ﻧــﺸﺎن داده ﺑﺎدام ) (Prunus dulcisاز ﻣﻨـﺎﻃﻖ ﻣﺮﻛـﺰي و اﺳﺖ ﻛـﻪ اﻳـﻦ ژن در ﺟـﻨﺲ ﭘﺮوﻧـﻮس داراي دو ﺟﻨﻮب ﻏﺮﺑﻲ آﺳﻴﺎ ﻣﻨﺸﺎء ﮔﺮﻓﺘﻪ اﺳﺖ و اﮔـﺮ ﭼـﻪ اﻳﻨﺘﺮون ﺑﺎ ﻃﻮل ﻣﺘﻐﻴﺮ اﺳﺖ ﻛـﻪ اﻳﻨﺘـﺮون اول ﺑـﻴﻦ ﮔﺰارش ﺷﺪه ﻛﻪ ﺑﺎدام اﻫﻠﻲ از ﺗﻼﻗﻲ ﮔﻮﻧـﻪﻫـﺎي ﺗــﻮاﻟﻲ اﺑﺘــﺪاﻳﻲ ) (Single Peptideو ﻧﺎﺣﻴــﻪ C1 وﺣــﺸﻲ ﺧﻮﻳــﺸﺎوﻧﺪ ﺑــﻪ دﺳــﺖ آﻣــﺪه اﺳــﺖ، اﻳـــﻦ ژن اﺳـــﺖ و اﻳﻨﺘـــﺮون دوم ﺑـــﻴﻦ ﻧـــﻮاﺣﻲ وﻟـــﻲ اﺟـــﺪاد و واﻟـــﺪﻳﻦ وﺣـــﺸﻲ آن ﻫﻤﭽﻨـــﺎن ﺣﻔﺎﻇـــــــﺖ ﺷـــــــﺪه C2و C3ﻗـــــــﺮار دارد ﻧﺎﺷـــﻨﺎﺧﺘﻪ اﺳـــﺖ )Bortiri et al., 2001؛ ).(Ushijima et al., 1998 .(Ladizinsky,ﺑــــــﺎدام ﮔﻮﻧــــــﻪاي ﻗﺎﺑﻠﻴﺖ دﺳﺘﺮﺳـﻲ ﺑـﻪ ﺗﻌـﺪاد زﻳـﺎدي از ﺗـﻮاﻟﻲ ﺧﻮدﻧﺎﺳــﺎزﮔﺎر از ﻧــﻮع ﮔﺎﻣﺘﻮﻓﻴﺘﻴــﻚ اﺳــﺖ ﻛــﻪ آﻟــﻞﻫــﺎي ﻧﺎﺳــﺎزﮔﺎري در ﺟــﻨﺲ ﭘﺮوﻧــﻮس در ﺗﻮﺳﻂ ﻳﻚ ﻣﻜﺎن ژﻧﻲ ﺑـﻪ ﻧـﺎم Sﺑـﺎ ﭼﻨـﺪﻳﻦ آﻟـﻞ ﭘﺎﻳﮕــﺎه اﻃﻼﻋــﺎﺗﻲ اﻣﻜــﺎن ﻃﺮاﺣــﻲ آﻏﺎزﮔﺮﻫــﺎي ﻛﻨﺘــﺮل ﻣــﻲﺷــﻮد ) .(Nettancourt, 2001در ﻋﻤــﻮﻣﻲ و اﺧﺘــﺼﺎﺻﻲ ﺑــﺮ اﺳــﺎس ﺗــﻮاﻟﻲ ﻧــﻮاﺣﻲ ﻣﺎدﮔﻲ ﻣﺤﺼﻮل ژن Sﺑﺎ ﻓﻌﺎﻟﻴﺖ رﻳﺒﻮﻧﻮﻛﻠﺌﺎزي ﺑﻪ ﺣﻔﺎﻇــﺖ ﺷــﺪه آنﻫــﺎ را ﻓــﺮاﻫﻢ آورده و ﻗﺎﺑﻠﻴــﺖ ﻧــﺎم S-RNaseاﺳــﺖ ﻛــﻪ از رﺷــﺪ ﻟﻮﻟــﻪ ﮔــﺮده ﺗﻜﺜﻴﺮ داﻣﻨﻪ وﺳﻴﻌﻲ از آﻟﻞﻫﺎي ﻧﺎﺳﺎزﮔﺎري را در 1999 D I S f o e ﻧﺎﺳـــﺎزﮔﺎر در ﺧﺎﻣـــﻪ ﺟﻠـــﻮﮔﻴﺮي ﻣـــﻲﻛﻨــــﺪ ) .(McClure et al., 1989آﻟـﻞﻫـﺎي ﻧﺎﺳـﺎزﮔﺎر زﻳــــﺎدي از ﮔﻮﻧــــﻪﻫــــﺎي ﺧــــﺎﻧﻮاده رزاﺳــــﻪ، ﻫﻤﺴﺎﻧﻪﺳﺎزي ) (Cloningو ﺗﻮاﻟﻲﻳـﺎﺑﻲ ﺷـﺪهاﻧـﺪ ﮔﻮﻧﻪ ﻫﺎي ﻣﺨﺘﻠﻒ ﺟﻨﺲ ﭘﺮوﻧﻮس ﺑﺮ اﺳـﺎس اﻳـﻦ آﻏﺎزﮔﺮﻫــــﺎ را اﻣﻜــــﺎنﭘــــﺬﻳﺮ ﻛــــﺮده اﺳــــﺖ iv )Sutherland et al., 2004؛ .(Sonneveld et al., 2003 ﺑــــــﺮ اﺳــــــﺎس آﻏﺎزﮔﺮﻫــــــﺎ در ﻣﺠﻤــــــﻮع h c )Sanzol et al., 2009؛ Tao et al., 2007؛ 30آﻟﻞ )ﺷﺎﻣﻞ 29آﻟـﻞ ﺧﻮدﻧﺎﺳـﺎزﮔﺎري وﻳـﻚ Ushijima et al., 1998؛ .(Sassa et al.,1996 آﻟﻞ ﺧﻮدﺳﺎزﮔﺎري( در ﺑﺎدام ﺷﻨﺎﺳﺎﻳﻲ ﺷﺪه اﺳﺖ ﻣﻄﺎﻟﻌــﺎت ﻣﻮﻟﻜــﻮﻟﻲ و ﻣﻄﺎﺑﻘــﺖ ﺗــﻮاﻟﻲﻫــﺎي ﻛﻪ ﭘﻨﺞ آﻟﻞ از آنﻫﺎ ﻗـﺒﻼ ﺑـﻪ اﺷـﺘﺒﺎه ﻧـﺎمﮔـﺬاري اﺳــﻴﺪﻫﺎي آﻣﻴﻨــﻪ ﺣﺎﺻــﻞ از ژن ﻧﺎﺳــﺎزﮔﺎري در ﺷﺪه ﺑﻮدﻧﺪ و ﺷﺶ آﻟﻞ دﻳﮕﺮ ﺟﺪﻳﺪ ﺑـﻮده و ﻗـﺒﻼً ﻣﺎدﮔﻲ ﻧـﺸﺎن داده اﺳـﺖ ﻛـﻪ ﺳـﺎﺧﺘﻤﺎن اﻳـﻦ ژن ﮔﺰارش ﻧﺸﺪه ﺑﻮدﻧﺪ ).(Ortega et al., 2005 r A ﺷـﺎﻣﻞ ﭘـﻨﺞ ﻧﺎﺣﻴـﻪ ﺣﻔﺎﻇـﺖ ﺷـﺪه ،C3 ،C2 ،C1 آﻏﺎزﮔﺮﻫﺎي اﺧﺘـﺼﺎﺻﻲ و ﻋﻤـﻮﻣﻲ ﻣﺨﺘﻠـﻒ RC4و C5و ﻳــﻚ ﻧﺎﺣﻴــﻪ ﺑــﺎ ﭼﻨــﺪ ﺷــﻜﻠﻲ ﺑــﺎﻻ ﺑــﺮ اﺳــﺎس ﺗــﻮاﻟﻲ ﻣﻜــﺎن ژﻧــﻲ ﻧﺎﺳــﺎزﮔﺎري در )ﺗــﻮاﻟﻲ ﻣﺘﻐﻴــﺮ( ﺑــﻪ ﻧــﺎم RHVاﺳــﺖ ﻛــﻪ در ﺣــﺪ ﺑﺎدام و دﻳﮕﺮ ﮔﻮﻧﻪﻫﺎي ﺟـﻨﺲ Prunusﻃﺮاﺣـﻲ ﻓﺎﺻـــﻞ ﻧـــﻮاﺣﻲ C2و C3ﻗـــﺮار ﮔﺮﻓﺘـــﻪ اﺳـــﺖ ﺷــــﺪه اﺳــــﺖ )Tamura et al., 2000؛ ) .(Igic and Kohn, 2001ﻧﺎﺣﻴـﻪ RHVﻧﻘـﺶ Channuntapipat et al., 2001, 2002؛ ﻣﻬﻤـــﻲ در ﺷﻨﺎﺳـــﺎﻳﻲ RNAداﻧـــﻪ ﮔـــﺮده ﺑـــﺎ 2001 Ma؛ ژﻧﻮﺗﻴــــﭗ ﻣــــﺸﺎﺑﻪ و ﺗﻌﻴــــﻴﻦ ﻧﺎﺳــــﺎزﮔﺎري دارد (Sutherland et al., 2004ﻛــﻪ ﺑــﻪ ﻃــﻮر www.SID.ir 652 Oliveira, and ﺷﻨﺎﺳﺎﻳﻲ آﻟﻞﻫﺎي ﺧﻮد ﻧﺎﺳﺎزﮔﺎري در ارﻗﺎم ﺑﺎدام اﻳﺮاﻧﻲ ... ﻣﻮﻓﻘﻴــﺖآﻣﻴــﺰي ﺑﺎﻋــﺚ ﺗﻜﺜﻴــﺮ آﻟــﻞﻫــﺎي ﺧﻮدﻧﺎﺳــــﺎزﮔﺎري در ارﻗــــﺎم ﻣــــﻲﺷــــﻮﻧﺪ )2005 ) .(Ortega et al., 2006اﮔﺮ ﭼﻪ ﻧﺘـﺎﻳﺞ ﺣﺎﺻـﻞ از اﻳــﻦ آﻏﺎزﮔﺮﻫــﺎ ﻧــﺸﺎن داد ﻛــﻪ ﻗﺎﺑﻠﻴــﺖ ﺑــﺴﻴﺎر ﺑــــﺎدام Ortega؛ ﺑﺎﻻﻳﻲ در ﺷﻨﺎﺳﺎﻳﻲ ژﻧﻮﺗﻴـﭗ ﺧﻮدﻧﺎﺳـﺎزﮔﺎري در .(Kodad et al., 2008ﺷﻨﺎﺳــﺎﻳﻲ آﻟــﻞﻫــﺎي ﺑــﺎدام دارﻧــﺪ ) .(Ortega et al., 2005ﺑــﺎ اﻳــﻦ ﻧﺎﺳﺎزﮔﺎري در ارﻗﺎم ﺑـﺎدام اﻳﺮاﻧـﻲ ﺑـﻪ روشﻫـﺎي وﺟﻮد ،در ﺑﺮﺧﻲ ﻣﻮارد ﺧﺎص ﺑـﻪ ﻧﻈـﺮ ﻣـﻲرﺳـﺪ ﻣﻮﻟﻜــــﻮﻟﻲ ﻣﺒﺘﻨــــﻲ ﺑــــﺮ PCRﺑــــﺎ اﺳــــﺘﻔﺎده ﻛﻪ ﺗﻮاﻟﻲﻳﺎﺑﻲ ﺑﻪ ﻣﻨﻈـﻮر ﺗـﺸﺨﻴﺺ دﻗﻴـﻖ آﻟـﻞﻫـﺎ از آﻏﺎزﮔﺮﻫــــﺎي اﺧﺘــــﺼﺎﺻﻲ و آﻏﺎزﮔﺮﻫــــﺎي ﺿﺮوري اﺳﺖ ).(Halasz et al., 2008 al., et ﻋﻤـــﻮﻣﻲ )Sheikh-Alian et al., 2010؛ ﻛﺪاد و ﻫﻤﻜﺎران ) (Kodad et al., 2008در Rahemi et al., 2010؛ Ebadi et al., 2011؛ ﺑﻴﻦ ﭼﻬـﺎرده رﻗـﻢ ﺑـﻮﻣﻲ اﺳـﭙﺎﻧﻴﺎ ﭘـﻨﺞ ﻋـﺪد آﻟـﻞ (Mousavi et al., 2011ﮔﺰارش ﺷﺪه اﺳﺖ. ﺟﺪﻳـﺪ را از ﻃﺮﻳــﻖ ﻫﻤــﺴﺎﻧﻪﺳـﺎزي و ﺗــﻮاﻟﻲﻳــﺎﺑﻲ D I S f ارﺗﮕﺎ و ﻫﻤﻜﺎران ) (Ortega et al., 2006ﺑﻪ ﺷﻨﺎﺳﺎﻳﻲ ﻛﺮده و ﺑﻪ ﻧﺎم آﻟﻞﻫﺎي S31ﺗـﺎ S35ﻧـﺎم- ﻣﻨﻈﻮر ﺗﺎﻳﻴﺪ آﻟﻞﻫﺎي ﺟﺪﻳﺪ ﺷﻨﺎﺳﺎﻳﻲ ﺷﺪه ،ﻛﻠﻴـﻪ ﮔﺬاري ﻛﺮدﻧﺪ .ﺗﺎﻛﻨﻮن 36آﻟـﻞ ﻧﺎﺳـﺎزﮔﺎري در ﻧﻮارﻫﺎي ﺣﺎﺻﻞ از ﺗﻜﺜﻴﺮ ﺑﺎ آﻏﺎزﮔﺮﻫﺎي اﻳﻨﺘـﺮون ﺑﺎدام ) 35آﻟـﻞ ﻧﺎﺳـﺎزﮔﺎري S1-S35و ﻳـﻚ آﻟـﻞ o e اول و دوم را ﻫﻤــﺴﺎﻧﻪﺳــﺎزي و ﺗــﻮاﻟﻲﻳــﺎﺑﻲ و ﮔﺰارش ﻛﺮدﻧـﺪ ﻛـﻪ ﻣﻮﺗﺎﻧـﺖ آﻟـﻞ S10ﮔـﺰارش ﺷــــﺪه ﺗﻮﺳــــﻂ ﭼﺎﻧﻮﻧﺘﺎﭘﻴﭙــــﺎت و ﻫﻤﻜــــﺎران ) (Channuntapipat et al., 2002در واﻗﻊ ﻳﻚ ﺳﺎزﮔﺎري (Sfﺷﻨﺎﺳﺎﻳﻲ و ﺗـﻮاﻟﻲ آنﻫـﺎ ﮔـﺰارش ﺷـــﺪه اﺳـــﺖ )Ushijima et al., 1998؛ iv Tamura et al., 2000؛ Ma and Oliveira, 2001؛ Channuntapipat et al., 2001, 2002؛ h c آﻟــﻞ ﺟﺪﻳــﺪ اﺳــﺖ و آن را ﺑــﻪ ﻧــﺎم آﻟــﻞ S24 .(Barckley et al., 2006ﺑــﻪ دﻟﻴــﻞ اﻫﻤﻴــﺖ ﻧﺎﻣﮕﺬاري ﻧﻤﻮدﻧﺪ .اﻳﻦ ﻣﺤﻘﻘﺎن آﻟﻞ ﻫـﺎي ﺟﺪﻳـﺪ آﻟﻞﻫﺎي ﺧﻮدﻧﺎﺳﺎزﮔﺎري اﻳﻦ ﭘـﮋوﻫﺶ ﺑـﺎ ﻫـﺪف ﺷﻨﺎﺳــــﺎﻳﻲ ﺷــــﺪه در ﺗﺤﻘﻴــــﻖ ﻗﺒﻠــــﻲ ﺧــــﻮد ﺷﻨﺎﺳﺎﻳﻲ و ﺗﻌﻴﻴﻦ آﻟﻞ ﺧﻮدﻧﺎﺳﺎزﮔﺎري ﺟﺪﻳـﺪ در ) (Ortega et al., 2005را ﺑﻪ ﻧﺎم آﻟﻞﻫﺎي S25ﺗﺎ ارﻗــﺎم ﺑــﺎدام اﻳﺮاﻧــﻲ ﺑــﺎ روش ﻫﻤــﺴﺎﻧﻪﺳــﺎزي و S29ﻧﺎم ﮔﺬاري ﻛﺮدﻧﺪ .آنﻫـﺎ ﻫﻤﭽﻨـﻴﻦ ﮔـﺰارش ﺗﻮاﻟﻲﻳﺎﺑﻲ اﻧﺠﺎم ﺷﺪ. r A ﻛﺮدﻧﺪ ﻛﻪ ﺗﻮاﻟﻲ ﺑﺮﺧﻲ از آﻟﻞﻫﺎي ﻗﺒﻠﻲ ﮔﺰارش ﺷﺪه در ﺑـﺎدام ﻣـﺸﺎﺑﻪ ﻫـﻢدﻳﮕـﺮ ﺑـﻮده و در واﻗـﻊ ﻣﻮاد و روشﻫﺎ آﻟﻞﻫﺎي S16 ،S1و S17ﺑﺎ ﻫﻢ و آﻟـﻞﻫـﺎي S4ﺑـﺎ ﻣﻮاد ﮔﻴﺎﻫﻲ S20و آﻟﻞﻫﺎي S13ﺑﺎ S19و ﻫﻤﭽﻨﻴﻦ آﻟﻞﻫﺎي S5 ﻧﻤﻮﻧﻪ ﻫﺎي ﺑﺮﮔﻲ از ارﻗﺎم ﺑﺎدام اﻳﺮاﻧـﻲ ﺳـﻔﻴﺪ، ﺑﺎ S15ﺗﻮاﻟﻲ ﻳﻜﺴﺎﻧﻲ داﺷﺘﻪ و ﻣﺸﺎﺑﻪ ﻫﻢدﻳﮕﺮﻧﺪ و ﻣﺎﻣﺎﻳﻲ ،ﺗﺠﺎري ،ﻫﻠﻮﻳﻲ ،ﻳﺰد ،17-ﺟـﻲ آر 16-و ﻧﻬﺎﻳﺘﺎ آﻟﻞﻫﺎي ﺧﻮدﻧﺎﺳﺎزﮔﺎري در ﺑـﺎدام را از S1 ﻣﺸﻬﺪ 40-در ﻓﺼﻞ رﺷﺪ از ﻛﻠﻜﺴﻴﻮنﻫﺎي ﺑـﺎدام ﺗـــﺎ S29و Sfﺗﺎﻳﻴـــﺪ و ﻧـــﺎمﮔـــﺬاري ﻛﺮدﻧـــﺪ در ﺷــﻬﺮﻛﺮد و ﻛﻤــﺎل آﺑــﺎد ﻛــﺮج ﺟﻤــﻊآوري www.SID.ir 653 ”ﻣﺠﻠﻪ ﺑﻪﻧﮋادي ﻧﻬﺎل و ﺑﺬر” ﺟﻠﺪ ،30-1ﺷﻤﺎره ،3ﺳﺎل 1393 اﭘﻨﺪورف )آﻟﻤﺎن( اﻧﺠﺎم ﺷﺪ .ﺷﺮاﻳﻂ دﻣﺎﻳﻲ ﺷﺎﻣﻞ ﺷﺪﻧﺪ. واﺳﺮﺷﺖﺳﺎزي اوﻟﻴـﻪ در 94درﺟـﻪ ﺳـﺎﻧﺘﻲﮔـﺮاد اﺳﺘﺨﺮاج DNA ﺑﺮاي دو دﻗﻴﻘﻪ 35 ،ﭼﺮﺧﻪ ﺑﺎ واﺳﺮﺷـﺖﺳـﺎزي در DNAژﻧﻮﻣﻲ ﺑـﻪ روش اﺳـﺘﺨﺮاج در ﻣﻘﻴـﺎس 94درﺟــﻪ ﺳــﺎﻧﺘﻲﮔــﺮاد ﺑــﺮاي 10ﺛﺎﻧﻴــﻪ ،اﺗــﺼﺎل ﻛﻮﭼـــــﻚ ) (Miniprepﺑـــــﺮ اﺳـــــﺎس روش آﻏﺎزﮔﺮ در 50درﺟﻪ ﺳﺎﻧﺘﻲﮔﺮاد ﺑﺮاي دو دﻗﻴﻘﻪ، دوﻳـﻞ و دوﻳـﻞ ) (Doyle and Doyle, 1987و ﻣﺮﺣﻠﻪ ﺑﺴﻂ در دﻣﺎي 68درﺟﻪ ﺳﺎﻧﺘﻲﮔﺮاد ﺑـﺮاي ﺗﻐﻴﻴـــﺮ ﻳﺎﻓﺘــــﻪ ﺗﻮﺳــــﻂ ﺳــــﻮﻧﻮﻟﺪ و ﻫﻤﻜــــﺎران ﭼﻬﺎر دﻗﻴﻘﻪ )ﺑﺎ اﻓﺰاﻳﺶ 10ﺛﺎﻧﻴﻪ ﺑﻪ ازاي ﻫﺮ ﭼﺮﺧﻪ ) (Sonneveld et al., 2001اﻧﺠـﺎم ﺷـﺪ .ﺗﻌﻴـﻴﻦ ﺑﻌﺪ از اﺗﻤﺎم ﭼﺮﺧﻪ دﻫﻢ( و ﺑﺴﻂ ﻧﻬﺎﻳﻲ در دﻣـﺎي ﻛﻤﻴـــﺖ و ﻛﻴﻔﻴـــﺖ DNAروي ژل آﮔـــﺎرز و 68درﺟﻪ ﺳﺎﻧﺘﻲﮔﺮاد ﺑﺮاي 10دﻗﻴﻘـﻪ اﻧﺠـﺎم ﺷـﺪ. ﻣﻘﺎﻳﺴﻪ وﺿﻌﻴﺖ ﻧﻮارﻫﺎي DNAﻧﻤﻮﻧﻪ ﻫﺎ از ﻧﻈـﺮ اﻟﻜﺘﺮوﻓﻮرز روي ژل آﮔﺎرز 1/5درﺻﺪ ﺑﻪ ﻣﺪت ﻛﻤﻴﺖ و ﻛﻴﻔﻴﺖ در ﻣﻘﺎﻳﺴﻪ ﺑـﺎ ﻧـﺸﺎﻧﮕﺮ ﺑـﺎ DNA 5ﺳﺎﻋﺖ در 70وﻟﺖ و رﻧﮓآﻣﻴـﺰي ﺑـﺎ اﺗﻴـﺪﻳﻮم ﻻﻣﺒﺪا اﺳﺘﺎﻧﺪارد ﺑﺮرﺳﻲ ﺷﺪ. ﺑﺮوﻣﺎﻳﺪ ﺑﻪ ﻣﺪت ﻳﻚ ﺳﺎﻋﺖ اﻧﺠﺎم ﺷـﺪ .در زﻳـﺮ D I S f ﺗﻜﺜﻴﺮ آﻟﻞﻫﺎي ﺧﻮدﻧﺎﺳﺎزﮔﺎري ﻧﻮر ﻣﺎوراي ﺑﻨﻔﺶ ) (UVﻋﻜﺲﺑﺮداري اﻧﺠـﺎم و o e ﺑــــﻪ ﻣﻨﻈــــﻮر ﺗﻜﺜﻴــــﺮ آﻟــــﻞﻫــــﺎي ﺟﺪﻳــــﺪ ﺧﻮدﻧﺎﺳﺎزﮔﺎري در اﻳﻦ ارﻗﺎم ،از آﻏﺎزﮔﺮ ﻣﺴﺘﻘﻴﻢ (Sonneveld et al., 2003) PaConsI-Fو اﻧﺪازه ﻧﻮارﻫﺎي ﺗﻜﺜﻴﺮ ﺷﺪه ﺑـﺎ اﺳـﺘﻔﺎده از ﻧـﺸﺎﻧﮕﺮ اﻧﺪازه DNAﻳﻚ ﻛﻴﻠﻮ ﺟﻔـﺖ ﺑـﺎزي ﺗﻌﻴـﻴﻦ ﺷـﺪ iv ).(Ortega et al., 2006 h c آﻏـــــــﺎزﮔﺮ ﻣﻌﻜـــــــﻮس EM-PC5ConsRD ﻫﻤﺴﺎﻧﻪﺳﺎزي و ﺗﻮاﻟﻲﻳﺎﺑﻲ ) (Sutherland et al., 2004ﺑﺮاي ﺗﻜﺜﻴـﺮ ﻧﺎﺣﻴـﻪ -1ﺧﺎﻟﺺﺳﺎزي ﻣﺤﺼﻮل PCR ﺑﻴﻦ SPو C5ﺑﺎ اﺳﺘﻔﺎده از DNAژﻧﻮﻣﻲ اﺳﺘﻔﺎده ﺑﻪ ﻣﻨﻈﻮر اﻧﺠﺎم ﻫﻤﺴﺎﻧﻪﺳﺎزي ،ﻣﺤﺼﻮل PCR ﺷــﺪ .واﻛــﻨﺶ زﻧﺠﻴــﺮهاي ﭘﻠــﻲ ﻣــﺮاز در ﺣﺠــﻢ ﺑﺎ اﺳﺘﻔﺎده از ﻛﻴﺖ QIA quick spinاز ﺷـﺮﻛﺖ 30ﻣﻴﻜﺮوﻟﻴﺘﺮ ﺷﺎﻣﻞ 75ﻧـﺎﻧﻮﮔﺮم DNAژﻧـﻮﻣﻲ، ﻛﻴــﺎژن ) (Qiageneآﻣﺮﻳﻜـــﺎ ﺧـــﺎﻟﺺﺳـــﺎزي و ﺑـﺎﻓﺮ 1X PCRاز ﺷـﺮﻛﺖ ﻧـﻮاژن )(Novagene ﻛﻴﻔﻴﺖ آن روي ژل آﮔﺎرز ﻳﻚ درﺻـﺪ در ﺑـﺎﻓﺮ آﻟﻤﺎن 0/2 ،ﻣﻴﻠﻲﻣﻮﻻر ،dNTPsﻳﻚ ﻣﻴﻠﻲﻣﻮﻻر TAEﺗﻌﻴﻴﻦ ﺷﺪ. r A ﺳﻮﻟﻔﺎت ﻣﻨﻴﺰﻳﻮم ) 0/5 ،(MgSO4ﻣﻴﻜـﺮو ﻣـﻮﻻر -2ﻫﻤﺴﺎﻧﻪﺳﺎزي از ﻫــــﺮ آﻏــــﺎزﮔﺮ ،ﻳــــﻚ واﺣــــﺪ از آﻧــــﺰﻳﻢ در اﻳــــــــــﻦ ﻣﺮﺣﻠــــــــــﻪ از ﻧﺎﻗــــــــــﻞ ﭘﻠــــــﻲ ﻣــــــﺮاز ﻛــــــﺪ ﻫــــــﺎت اﺳــــــﺘﺎرت PCR-Blunt-II-TOPOاز ﺷﺮﻛﺖ اﻳﻨـﻮﻳﺘﺮوژن )(KOD Hot Start DNA Polymerase ) (Invitrogeneآﻣﺮﻳﻜــﺎ ﺟﻬــﺖ ﻫﻤــﺴﺎﻧﻪﺳــﺎزي ازﺷـــﺮﻛﺖ ﻧـــﻮاژن در دﺳـــﺘﮕﺎه ﺗﺮﻣﻮﺳـــﺎﻳﻜﻠﺮ DNAﻫــﺪف و اﻧﺘﻘــﺎل ﺑــﻪ ﺑــﺎﻛﺘﺮيﻫــﺎي ﻣــﺴﺘﻌﺪ www.SID.ir 654 ﺷﻨﺎﺳﺎﻳﻲ آﻟﻞﻫﺎي ﺧﻮد ﻧﺎﺳﺎزﮔﺎري در ارﻗﺎم ﺑﺎدام اﻳﺮاﻧﻲ ... ) E.ﺷـــــﺮﻛﺖ ﺷـــــﺪ و ﺗـــــﻮاﻟﻲ ﻧﻬـــــﺎﻳﻲ آنﻫـــــﺎ ﺗﻮﺳـــــﻂ اﻳﻨــﻮﻳﺘﺮوژن( اﺳــﺘﻔﺎده ﺷــﺪ .ﺑــﺎﻛﺘﺮيﻫــﺎي ﺣﺎﻣــﻞ ﻧـــــــــــــﺮماﻓـــــــــــــﺰار ژن داك )(GenDoc DNAﻫﺪف روي ﻣﺤﻴﻂ ﻛﺸﺖ LBدر ﺣـﻀﻮر ) (www.PSC.edu/biomed/gendocﺗﻌﻴﻴﻦ ﺷﺪ. )coli TOP10 (Competent آﻧﺘــﻲ ﺑﻴﻮﺗﻴــﻚ ﻛﺎﻧﺎﻣﺎﻳــﺴﻴﻦ ) 50ﻣﻴﻜﺮوﮔــﺮم در ﻣﻴﻠــﻲﻟﻴﺘــﺮ( اﻧﺘﺨــﺎب ﺷــﺪﻧﺪ و اﺳــﺘﺨﺮاج DNA ﻧﺘﺎﻳﺞ و ﺑﺤﺚ ﭘﻼﺳـــــــﻤﻴﺪي ﺑـــــــﺎ اﺳـــــــﺘﻔﺎده از ﻛﻴـــــــﺖ ﻧﺘﺎﻳﺞ ﺗﻜﺜﻴﺮ ﺑﺎ اﺳﺘﻔﺎده از آﻏﺎزﮔﺮﻫـﺎي ﺟﻔـﺖ Pure Link Quick Plasmid Miniprep Kit اﻳﻨﺘﺮون و ﺗﻮﺳﻂ آﻧﺰﻳﻢ ﻛﺪ ﻫﺎت اﺳﺘﺎرت ،وﺟﻮد )ﺷﺮﻛﺖ ﻛﻴﺎژن( اﻧﺠﺎم ﺷﺪ .ﺗﻌﻴﻴﻦ ﻛﻴﻔﻴـﺖ DNA ﻗﻄﻌﺎت ﻣﺮﺑﻮط ﺑﻪ آﻟﻞﻫﺎي ﺟﺪﻳﺪ در ارﻗﺎم ﺳﻔﻴﺪ، ﭘﻼﺳﻤﻴﺪي ،روي ژل آﮔﺎرز 1/5درﺻـﺪ در ﺑـﺎﻓﺮ ﻣﺎﻣــﺎﻳﻲ ،ﻣــﺸﻬﺪ ،40-ﻳــﺰد ،17-ﺟــﻲ آر 16-و TAEاﻧﺠﺎم ﺷﺪ .ﻫﻤﭽﻨـﻴﻦ ﺑـﻪ ﻣﻨﻈـﻮر اﻃﻤﻴﻨـﺎن از ﻫﻠﻮﻳﻲ را ﻧﺸﺎن داد )ﺷﻜﻞ .(1ﺑﺮاي اﻃﻤﻴﻨﺎن ﺑﻴﺸﺘﺮ ﻫﻤﺴﺎﻧﻪ ﺳـﺎزي DNA ،ﭘﻼﺳـﻤﻴﺪي ﺑـﺎ اﺳـﺘﻔﺎده از از ﻫﻤــﺴﺎﻧﻪﺳــﺎزي DNA ،ﭘﻼﺳــﻤﻴﺪي ﺣﺎﺻــﻞ از آﻏﺎزﮔﺮﻫــــــــﺎي ) PaconsI-Fﻣــــــــﺴﺘﻘﻴﻢ( و ﻫﻤــﺴﺎﻧﻪﺳــﺎزي در 16ﻛﻠــﻮﻧﻲ ﻣﺮﺑــﻮط ﺑــﻪ رﻗــﻢ ) M-PC5consRDﻣﻌﻜــﻮس( ﺑــﺮ اﺳــﺎس روش ﻳﺰد 17-ﺑﺎ اﺳﺘﻔﺎده از آﻏﺎزﮔﺮﻫﺎي ﭘﻮﺷـﺶدﻫﻨـﺪه D I S f o e ارﺗﮕﺎ و ﻫﻤﻜﺎران ) (Ortega et al., 2006ﺗﻜﺜﻴﺮ ﺷﺪ. -3ﺗﻮاﻟﻲﻳﺎﺑﻲ ﺗـــﻮاﻟﻲﻳـــﺎﺑﻲ ﺳـــﻪ ﭘﻼﺳـــﻤﻴﺪ ﺑـــﺎ اﺳـــﺘﻔﺎده از ﺟﻔﺖ اﻳﻨﺘﺮون ﺗﻜﺜﻴﺮ و اﻧﺪازه ﻧﻮارﻫﺎي ﺗﻜﺜﻴﺮ ﺷﺪه ﻣﺮﺑﻮط ﺑﻪ آﻟﻞﻫﺎي ﺟﺪﻳﺪ در اﻳﻦ رﻗﻢ ﺗﻌﻴﻴﻦ ﺷـﺪ. iv ﻧﺘﺎﻳﺞ ﻧﺸﺎن داد ﻛﻪ ﻗﻄﻌﺎت ﻣﺮﺑﻮط ﺑﻪ آﻟـﻞ ﺟﺪﻳـﺪ S39در 5ﻛﻠﻮﻧﻲ و ﻗﻄﻌﺎت ﻣﺮﺑﻮط ﺑﻪ آﻟﻞ ﺟﺪﻳـﺪ h c آﻏﺎزﮔﺮﻫﺎي M13ﺗﻮﺳﻂ دﺳـﺘﮕﺎه SECUGEN S40در 8ﻛﻠـﻮﻧﻲ ،ﻧﻮارﻫــﺎي ﻣـﺸﺎﺑﻪ و ﻫــﻢ اﻧــﺪازه )ﻣﺎدرﻳﺪ ،اﺳﭙﺎﻧﻴﺎ( ﺑﺮاي ﻫﺮ آﻟﻞ اﻧﺠﺎم ﺷـﺪ ،ﺳـﭙﺲ داﺷﺘﻨﺪ )ﺷﻜﻞ .(2 r A ﺗﻮاﻟﻲ ﺣﺎﺻﻞ از اﻳﻦ ﭘﻼﺳﻤﻴﺪﻫﺎ ﻣﻘﺎﻳـﺴﻪ و ﺗـﻮاﻟﻲ ﻧﺘﺎﻳﺞ ﺗﻜﺜﻴﺮ DNAﺣﺎﺻـﻞ از ﻫﻤـﺴﺎﻧﻪﺳـﺎزي ﺛﺎﺑﺖ ﻣﺮﺑﻮط ﺑﻪ ﻫﺮ آﻟـﻞ ﺑـﺎ ﻧـﺮماﻓـﺰار ﺳـﻜﻤﻦ دو ﺑـــــﺎ آﻏﺎزﮔﺮﻫـــــﺎي ) PaconsI-Fﻣـــــﺴﺘﻘﻴﻢ( و ) (SEQMAN IIﺗﻌﻴﻴـﻴﻦ ﺷـﺪ .در ﻧﻬﺎﻳـﺖ ﺗـﻮاﻟﻲ ) EM-PC5cons RDﻣﻌﻜﻮس( وﺟﻮد ﻧﻮارﻫـﺎي اﺳﻴﺪ آﻣﻴﻨﻪ ﻫـﺮ آﻟـﻞ ﺑـﺎ ﺗـﻮاﻟﻲ اﺳـﻴﺪ آﻣﻴﻨـﻪ ﺳـﺎﻳﺮ ﺗﻜﺜﻴﺮ ﺷﺪه ﺟﺪﻳﺪ ﻣﺮﺑﻮط ﺑﻪ آﻟـﻞﻫـﺎي ) S36رﻗـﻢ آﻟـــﻞﻫـــﺎي ﻣﻮﺟـــﻮد در ﺟـــﻨﺲ ﭘﺮوﻧـــﻮس در ﺗﺠﺎري() S37 ،رﻗﻢ ﻫﻠـﻮﻳﻲ() S38 ،رﻗـﻢ ﺳـﻔﻴﺪ(، ﭘﺎﻳﮕـــﺎه اﻃﻼﻋـــﺎﺗﻲ ﻣﻮﺳـــﺴﻪ ﺑﻴﻮاﻧﻔﻮرﻣﺎﺗﻴـــﻚ ) S39ارﻗــﺎم ﻣــﺸﻬﺪ 40-و ﻳــﺰد) S40 ،(17-رﻗــﻢ اروﭘـــﺎ ) (www.ebi.ac.ukﺑـــﻪ روش BLAST ﻳـــــﺰد) S41 ،(17-ﻣـــــﺸﻬﺪ) S42 ،(40-رﻗـــــﻢ ﺗﻮﺳـــــــﻂ ﺑﺮﻧﺎﻣـــــــﻪ ﻛﻼﺳـــــــﺘﺎل اﻳﻜـــــــﺲ ﺟــﻲ آر (16-و ) S43رﻗــﻢ ﻣﺎﻣــﺎﻳﻲ( را ﻧــﺸﺎن داد ) (Thompson et al., 1997) (Clustal Xﺑـﺎ )ﺷﻜﻞ .(3ﺑـﺪﻳﻦ ﺗﺮﺗﻴـﺐ ﻫـﺸﺖ آﻟـﻞ ﺟﺪﻳـﺪ در ﻧــﺮماﻓــﺰار ﻣــﮓ اﻻﻳــﻦ ) (MegAlignﻣﻘﺎﻳــﺴﻪ ﻫﻔﺖ رﻗـﻢ ﺑـﺎدام اﻳﺮاﻧـﻲ ﺗﺠـﺎري ) ،(S36ﻫﻠـﻮﻳﻲ www.SID.ir 655 ”ﻣﺠﻠﻪ ﺑﻪﻧﮋادي ﻧﻬﺎل و ﺑﺬر” ﺟﻠﺪ ،30-1ﺷﻤﺎره ،3ﺳﺎل 1393 2 M 5 6 1 8 M 7 3 4 bp 4000300020001650- 1000- D I 850650- S f o e ﺷﻜﻞ -1ﻣﺤﺼﻮل PCRﺑﺎ اﻧﺘﻬﺎي ﺻﺎف ﺗﻮﺳﻂ آﻧﺰﻳﻢ ﻛﺪﻫﺎت اﺳﺘﺎرت ﺑﺎ آﻏﺎزﮔﺮﻫﺎي Pacons I-Fو EM-PC5consRDاز ﭼﭗ ﺑﻪ راﺳﺖ ﺷﺎﻣﻞ (Mﻧﺸﺎﻧﮕﺮ اﻧﺪازه DNAﻳﻚ ﻛﻴﻠﻮ ﺟﻔﺖ ﺑﺎزي ) ﺷﺮﻛﺖ ﻛﻴﺎژن((1 ،رﻗﻢ ﺳﻔﻴﺪ (2 ،رﻗﻢ ﻣﺎﻣﺎﻳﻲ (3 ،رﻗﻢ ﻣﺸﻬﺪ (4 ،40-رﻗﻢ ﻳﺰد (5 ،17-رﻗﻢ ﺟﻲ آر (6 ،16-آب ﻣﻘﻄﺮ )ﻛﻨﺘﺮل ﻣﻨﻔﻲ( (7 ،رﻗﻢ ﻫﻠﻮﻳﻲ و (8آب ﻣﻘﻄﺮ )ﻛﻨﺘﺮل ﻣﻨﻔﻲ( Fig. 1. PCR products with Blunt-end by KOD Hot Start DNA polymerase obtained with consensus )primers PaConsI-F and EMPC5consRD from left to right including: M) DNA Ladder 1Kb (Qiagene), 1 )Sefied, 2) Mamaei, 3) Mashhad-40, 4) Yazd-17, 5) Gr-16, 6) Distillated water (negative control), 7 )Holouei and 8) Distillated water (negative control v i h ) ،(S37ﺳﻔﻴﺪ ) ،(S38ﻳﺰد S39) 17-و ،(S40ﻣﺸﻬﺪ ﻧﺎمﮔﺬاري ﺷـﺪ )ﺷـﻜﻞ .(3آﻟـﻞ S25ﻧﻴـﺰ ﻗـﺒﻼً ﺑـﺎ ،(S41) 40ﺟـــﻲ-آر (S42) 16-و ﻣﺎﻣـــﺎﻳﻲ )(S43 ﺟﻔــﺖ آﻏــﺎزﮔﺮ اﻳﻨﺘــﺮون دوم ﺗﻜﺜﻴــﺮ و ﺷﻨﺎﺳــﺎﻳﻲ ﺷﻨﺎﺳــﺎﻳﻲ ﺷــﺪ )ﺷــﻜﻞ .(3در رﻗــﻢ ﻫﻠــﻮﻳﻲ ﻳــﻚ ﺷﺪه ﺑﻮد ).(Mousavi et al., 2011 c r A ﻗﻄﻌﻪ ﺑـﻪ اﻧـﺪازه 950ﺟﻔـﺖ ﺑـﺎز ﺗﻜﺜﻴـﺮ ﺷـﺪ ﻛـﻪ ﺗـــــــــﺎﻛﻨﻮن آﻟـــــــــﻞﻫـــــــــﺎي S1و S3 ﻣﺮﺑﻮط ﺑﻪ آﻟﻞ ﺟﺪﻳﺪ S37ﺑﻮد و ﻗﻄﻌـﻪ ﻣﺮﺑـﻮط ﺑـﻪ ) ،(Ma and Oliveira, 2001آﻟﻞﻫـﺎي ،S7 ،S1 آﻟــﻞ S13ﻛــﻪ ﻗــﺒﻼ ﺑــﺎ آﻏﺎزﮔﺮﻫــﺎي اﻳﻨﺘــﺮون دوم S8و ،(Channuntapipat et al., 2001) Sf ﺗﻜﺜﻴﺮ ﺷﺪه ﺑﻮد ) (Mousavi et al., 2011ﺑﺎ اﻳﻦ آﻟـــــــــــــﻞﻫــــــــــــــﺎي S10 ،S9 ،S2و S23 ﺟﻔﺖ آﻏﺎزﮔﺮ ﺗﻜﺜﻴﺮ ﻧﺸﺪ .ﻋﺪم ﺗﻜﺜﻴﺮ آﻟﻞ S13ﺑـﺎ ) ،(Channuntapipat et al., 2002آﻟـﻞ S5و اﻳﻦ ﺟﻔﺖ آﻏﺎزﮔﺮ در ﻣﻄﺎﻟﻌﺎت ﻗﺒﻠﻲ ﻧﻴﺰ ﮔﺰارش ،(Lopez et al., 2004) S25آﻟﻞﻫﺎي S1ﺗﺎ S29 ﺷﺪه اﺳﺖ ) .(Ortega et al., 2006ﻫﻤﭽﻨـﻴﻦ در )2006 ،(Ortegaآﻟـــــــﻞ ﺑﺎدام ﻣﺎﻣﺎﻳﻲ ﻳﻚ ﻧﻮار ﺑﻪ ﻃـﻮل 1400ﺟﻔـﺖ ﺑـﺎز ،(Halasz et al., 2008) S30آﻟـﻞﻫـﺎي S31ﺗـﺎ ﺗﻜﺜﻴﺮ ﺷﺪ ﻛﻪ ﺑﻌﺪ از ﺗﻮاﻟﻲﻳـﺎﺑﻲ ﺑـﻪ ﻧـﺎم آﻟـﻞ S43 (Kodad etاز ﻃﺮﻳــــﻖ www.SID.ir 2008) S35 656 al., al., et ... ﺷﻨﺎﺳﺎﻳﻲ آﻟﻞﻫﺎي ﺧﻮد ﻧﺎﺳﺎزﮔﺎري در ارﻗﺎم ﺑﺎدام اﻳﺮاﻧﻲ ( ازS39S40) 17- در رﻗﻢ ﻳﺰدE. coli ﭘﻼﺳﻤﻴﺪي از ﻛﻠﻮﻧﻲ ﻫﺎي ﺑﺎﻛﺘﺮيDNA ﻫﻤﺴﺎﻧﻪ ﺳﺎزيPCR ﻣﺤﺼﻮل-2 ﺷﻜﻞ ( ﺷﺎﻣﻞ8 ﺗﺎ3 ،(S39) 1 ( ﻛﻠﻮﻧﻲ2 ،( ﻳﻚ ﻛﻴﻠﻮ ﺟﻔﺖ ﺑﺎزي) ﺷﺮﻛﺖ ﻛﻴﺎژنDNA ( ﻧﺸﺎﻧﮕﺮ اﻧﺪازه1 :ﭼﭗ ﺑﻪ راﺳﺖ ﺷﺎﻣﻞ ،(S40) 11 ( ﻛﻠﻮﻧﻲ12 ،(S40) 10 ( ﻛﻠﻮﻧﻲ11 ،(S39) 9 ( ﻛﻠﻮﻧﻲ10 ،( )اﺷﺘﺒﺎه8 ( ﻛﻠﻮﻧﻲ9 ،(S40) 7و6 ،5 ،4 ،3 ،2 ﻛﻠﻮﻧﻲﻫﺎي (18 ( وS39) (16 ( ﻛﻠﻮﻧﻲ17 ،(S39) 15 ( ﻛﻠﻮﻧﻲ16 ،( )اﺷﺘﺒﺎه14 ( ﻛﻠﻮﻧﻲ15 ،(S39) 13 ( ﻛﻠﻮﻧﻲ14 ،( )اﺷﺘﺒﺎه12 ( ﻛﻠﻮﻧﻲ13 ﻳﻚ ﻛﻴﻠﻮ ﺟﻔﺖ ﺑﺎزيDNA ﻧﺸﺎﻧﮕﺮ اﻧﺪازه D I Fig. 2. PCR products obtained from cloning of DNA Plasmid from E. coli colonies in Yazd-17 cultivar (S39S40) from left to right including: 1) DNA Ladder 1kb (Qiagene), 2) colony 1(S39), 3-8) colonies 2, 3, 4, 5, 6 and 7(S40), 9) colony 8 ( mistake), 10) colony 9 (S39), 11) colony 10 (S40), 12) colony 11 (S40), 13) colony 12 ( mistake), 14) colony 13 (S39), 15) colony 14 ( mistake), 16) colony 15 (S39), 17) colony 16 (S39) and 18) DNA Ladder 1Kb S f o e v i h c r A از ارﻗﺎمEM-PC5cons RD وPacons I-F ﺧﺎﻟﺺ ﺳﺎزي ﺷﺪه از ﻫﻤﺴﺎﻧﻪ ﺳﺎزي ﺑﺎ آﻏﺎزﮔﺮﻫﺎيPCR ﻣﺤﺼﻮل-3 ﺷﻜﻞ 40- ﻣﺸﻬﺪ،(S39S40) 17- ﻳﺰد،(S7S38) ﺳﻔﻴﺪ،(S13S37) ﻫﻠﻮﻳﻲ،(S24S36) ﺑﺎدام اﻳﺮاﻧﻲ از ﭼﭗ ﺑﻪ راﺳﺖ ﺷﺎﻣﻞ ارﻗﺎم ﺗﺠﺎري (S25S43) ( و ﻣﺎﻣﺎﻳﻲS2S42) 16- ﺟﻲ آر،(S39S41) Fig. 3. Purified PCR products from cloning that obtained with consensus primers PaConsI-F and EMPC5consRD for Iranian almond cultivars from left to right including Tejari (S24S36), Holouei (S13S37), Sefied (S7S38), Yazd-17 (S39S40), Mashhad-40 (S39S41), Gr-16 (S2S42), and Mamaei (S25S43) cultivars 657 www.SID.ir ”ﻣﺠﻠﻪ ﺑﻪﻧﮋادي ﻧﻬﺎل و ﺑﺬر” ﺟﻠﺪ ،30-1ﺷﻤﺎره ،3ﺳﺎل 1393 ﻫﻤﺴﺎﻧﻪﺳﺎزي و ﺗـﻮاﻟﻲﻳـﺎﺑﻲ در ﺑـﺎدام ﺷﻨﺎﺳـﺎﻳﻲ و S5ﺷﻨﺎﺳــﺎﻳﻲ ﺷــﺪه در ﻳــﻚ ژﻧﻮﺗﻴــﭗ وﺣــﺸﻲ از ﺗﻌﻴﻴﻦ ﺷﺪه اﻧﺪ .ﺑﻨﺎﺑﺮاﻳﻦ آﻟﻞ ﻫﺎي ﺟﺪﻳـﺪ در ارﻗـﺎم ﮔﻮﻧـــﻪ P. webbiiاز ﻛـــﺸﻮر ﻣـــﻮﻧﺘﻨﮕﺮو دارد ﻣﻮرد ﺑﺮرﺳﻲ ﻛﻪ ﻫﻤﺴﺎﻧﻪﺳﺎزي و ﺗﻮاﻟﻲﻳﺎﺑﻲ ﺷﺪﻧﺪ )ﺟﺪول ،(1اﻣﺎ ﺗﻮاﻟﻲ ﻧﻮﻛﻠﺌﻮﺗﻴﺪي اﻳﻦ آﻟـﻞﻫـﺎ از از آﻟﻞ S36ﺗﺎ S43ﻧﺎمﮔﺬاري و ﺑﻪ ﻋﻨﻮان آﻟﻞﻫﺎي ﻧﻈﺮ ﺗﻮاﻟﻲ در ﻧﻮاﺣﻲ اﻳﻨﺘﺮون اول و دوم ﻣﺘﻔـﺎوت ﺟﺪﻳﺪ در ارﻗﺎم اﻳﺮاﻧﻲ ﺑﺮاي اوﻟﻴﻦ ﺑﺎر ﺷﻨﺎﺳـﺎﻳﻲ و ﺑــﻮد .ﺗــﻮاﻟﻲ اﺳــﻴﺪﻫﺎي آﻣﻴﻨــﻪ آﻟــﻞ S43از رﻗــﻢ ﺗــﻮاﻟﻲ اﺳــﻴﺪﻫﺎي آﻣﻴﻨــﻪ ﻣﺮﺑــﻮط ﺑــﻪ اﻳــﻦ آﻟــﻞﻫــﺎ ﻣﺎﻣﺎﻳﻲ ﺑﺎ ﺗﻮاﻟﻲ اﺳﻴﺪﻫﺎي آﻣﻴﻨﻪ آﻟـﻞ S25از رﻗـﻢ ﮔﺰارش ﮔﺮدﻳﺪ )ﺷﻜﻞ .(4 ﻻﻣﻮﻧـﺎ ) (Ortega et al., 2006ﺷـﺒﺎﻫﺖ زﻳـﺎدي ﻧﺘــﺎﻳﺞ ﺣﺎﺻــﻞ از ﻣﻄﺎﺑﻘــﺖ ﺗــﻮاﻟﻲ اﺳــﻴﺪﻫﺎي ﻧﺸﺎن داد و ﻫﻤﭽﻨﻴﻦ ﺷﺒﺎﻫﺖ ﺑﺎﻻﻳﻲ ﺑﺎ آﻟﻞ S15در آﻣﻴﻨﻪ آﻟﻞﻫﺎي ﺟﺪﻳﺪ ﺑﺎ آﻟﻞﻫﺎي ﮔﺰارش ﺷﺪه در ﮔﻮﻧــﻪ (Vieira et al., 2007) P. spinosa ﺟﻨﺲ ﭘﺮوﻧﻮس ﻧﺸﺎن داد ﻛـﻪ آﻟـﻞ S43ﻣـﺸﺎﺑﻬﺖ داﺷــﺖ )ﺟــﺪول .(1آﻟــﻞ ﺟﺪﻳــﺪ S37در رﻗــﻢ ﺑــﺎﻻﻳﻲ ) 99درﺻــﺪ( ﺑــﺎ آﻟــﻞ S25در ﺑــﺎدام رﻗــﻢ ﻫﻠــﻮﻳﻲ از ﻧﻈــﺮ ﺗــﻮاﻟﻲ اﺳــﻴﺪﻫﺎي آﻣﻴﻨــﻪ ﺷــﺒﺎﻫﺖ ﻻﻣﻮﻧﺎ ) (Ortega et al., 2006داﺷﺖ و ﺗﻔـﺎوت زﻳــﺎدي ﺑــﺎ آﻟــﻞ S8از ﮔﻮﻧــﻪ P. cerasifera اﻳﻦ دو آﻟﻞ در ﻳﻚ اﺳﻴﺪ آﻣﻴﻨﻪ اﺳﺖ .اﻳﻦ ﺗﻔﺎوت ) (Sutherland et al., 2009و آﻟﻞ S7از ﮔﻮﻧـﻪ D I S f o e ﻣﻲﺗﻮاﻧﺪ ﻧﺎﺷﻲ از ﻣﻮﺗﺎﺳﻴﻮن ﻧﻘﻄﻪاي در ﺗﻮاﻟﻲ اﻳﻦ دو آﻟـــﻞ ﺑﺎﺷـــﺪ .وﺟـــﻮد ﺷـــﺒﺎﻫﺖ در ﺗـــﻮاﻟﻲ ﻧﻮﻛﻠﺌﻮﺗﻴﺪﻫﺎ و ﺗـﻮاﻟﻲ اﺳـﻴﺪﻫﺎي آﻣﻴﻨـﻪ ﻣـﻲﺗﻮاﻧـﺪ ﺑﻴﺎﻧﮕﺮ اﻧﺸﻘﺎق اﻳﻦ دو آﻟﻞ از ﻳﻚ آﻟـﻞ اﺟـﺪادي (Vieira et al., 2007) P. spinosaﻧـﺸﺎن داد. ﺳــﺎﻳﺮ آﻟــﻞﻫــﺎي ﺟﺪﻳــﺪ ﺷﻨﺎﺳــﺎﻳﻲ ﺷــﺪه ﻫــﻢ ﺑــﺎ iv آﻟـــﻞﻫـــﺎي ﻣﻮﺟـــﻮد در ﮔﻮﻧـــﻪﻫـــﺎي ﮔـــﻴﻼس ) ،(P. aviumزردآﻟــﻮ ) ،(P. armeniacaآﻟــﻮ h c ﻣﺸﺎﺑﻪ و ﻳﺎ در اﺛﺮ ﻣﻮﺗﺎﺳﻴﻮن از ﺳﺎﻳﺮ آﻟﻞﻫـﺎ ﺑﺎﺷـﺪ ) P. spinosa ،P. domesticaو (P. salicina ) .(Wunsch and Hormaza, 2004ﺗــﻮاﻟﻲ از ﻧﻈﺮ ﺗﻮاﻟﻲ اﺳﻴﺪﻫﺎي آﻣﻴﻨﻪ ﺷﺒﺎﻫﺖ زﻳﺎدي ﻧﺸﺎن اﺳﻴﺪﻫﺎي آﻣﻴﻨﻪ آﻟﻞﻫـﺎي ﺟﺪﻳـﺪ در اﻳـﻦ ﺗﺤﻘﻴـﻖ دادﻧﺪ )ﺟﺪول .(1 r A ﻧﺸﺎن داد ﻛﻪ اﻳﻦ آﻟﻞﻫﺎ از ﻧﻈﺮ اﻧﺪازه و ﺗـﻮاﻟﻲ ﺑـﺎ ﺑﻪ ﻫﺮ ﺣﺎل وﺟـﻮد ﻣـﺸﺎﺑﻬﺖ زﻳـﺎد و ﻳـﺎ ﺗـﺸﺎﺑﻪ ﻫﻢ ﻣﺘﻔﺎوت ﺑﻮده و ﺑﺎ آﻟﻞﻫﺎي ﻗﺒﻠﻲ ﮔﺰارش ﺷﺪه ﻛﺎﻣـــﻞ دو آﻟـــﻞ در ﺗـــﻮاﻟﻲ اﺳـــﻴﺪﻫﺎي آﻣﻴﻨـــﻪ ﻧﻴﺰ ﻣﺘﻔﺎوت ﻫﺴﺘﻨﺪ و اﻳﻦ ﺗﻔﺎوت در ﺗﻮاﻟﻲ ،ﻋﻤﺪﺗﺎً ﻧﻤــﻲﺗﻮاﻧــﺪ ﺑﻴــﺎﻧﮕﺮ ﻋﻤــﻞ ﻣــﺸﺎﺑﻪ دو آﻟــﻞ ﺑﺎﺷــﺪ در ﻣﻮﻗﻌﻴﺖﻫﺎي ﺑـﻴﻦ ﻧـﻮاﺣﻲ ﺣﻔﺎﻇـﺖ ﺷـﺪه ﺑـﻮد ) .(Ortega et al., 2006ﻟــﻮﭘﺰ و ﻫﻤﻜــﺎران )ﺷﻜﻞ .(4ﺗﻮاﻟﻲ اﺳﻴﺪﻫﺎي آﻣﻴﻨﻪ در اﻳﻦ آﻟﻞﻫﺎي ) (Lopez et al., 2004ﮔــﺰارش ﻛﺮدﻧــﺪ ﻛــﻪ ﺟﺪﻳﺪ ﻣـﺸﺎﺑﻬﺖ زﻳـﺎدي )ﺑـﻴﺶ از 80درﺻـﺪ( ﺑـﺎ ﻣﻮﺗﺎﺳﻴﻮنﻫﺎي ﻧﻘﻄﻪاي و ﺣﺬف ﻗﺴﻤﺘﻲ از ﺗـﻮاﻟﻲ آﻟﻞ ﻫﺎي ﻧﺎﺳﺎزﮔﺎري ﮔﺰارش ﺷﺪه در ﮔﻮﻧﻪﻫـﺎي اﻳﻨﺘﺮون آﻟﻞﻫﺎي S5و S25ﻣﻨﺠﺮ ﺑـﻪ ﺗﻔـﺎوت ﺑـﻴﻦ دﻳﮕــﺮ ﭘﺮوﻧــﻮس دارد )ﺟــﺪول .(1ﺗــﻮاﻟﻲ اﺳــﻴﺪ اﻳﻦ آﻟﻞﻫﺎ ﻣـﻲﺷـﻮد ،ﺑﻨـﺎﺑﺮاﻳﻦ ﺗﻔـﺎوت در ﺗـﻮاﻟﻲ آﻣﻴﻨﻪ آﻟﻞ S40در ﻳﺰد 17-ﺷﺒﺎﻫﺖ ﻛﺎﻣﻞ ﺑـﺎ آﻟـﻞ آﻟﻞﻫـﺎ ﻣﻨﺠـﺮ ﺑـﻪ اﻳﺠـﺎد ﭼﻨـﺪ ﺷـﻜﻠﻲ ﺑـﻴﻦ آنﻫـﺎ www.SID.ir 658 ... ﺷﻨﺎﺳﺎﻳﻲ آﻟﻞﻫﺎي ﺧﻮد ﻧﺎﺳﺎزﮔﺎري در ارﻗﺎم ﺑﺎدام اﻳﺮاﻧﻲ fS eo 659 iv ch ... ﺷﻨﺎﺳﺎﻳﻲ آﻟﻞﻫﺎي ﺧﻮد ﻧﺎﺳﺎزﮔﺎري در ارﻗﺎم ﺑﺎدام اﻳﺮاﻧﻲ Ar ID )آﻣﻴﻨﻮClustal X و ﻣﺎﻣﺎﻳﻲ ﺑﻪ روش16- ﺟﻲ آر،40- ﻣﺸﻬﺪ،17- ﻳﺰد، ﺳﻔﻴﺪ، ﻫﻠﻮﻳﻲ، ﺑﻪ ﺗﺮﺗﻴﺐ در ارﻗﺎم ﺑﺎدام اﻳﺮاﻧﻲ ﺗﺠﺎريS43 ﺗﺎS36 ﻫﻢﭘﻮﺷﺎﻧﻲ ﺗﻮاﻟﻲ اﺳﻴﺪﻫﺎي آﻣﻴﻨﻪ آﻟﻞﻫﺎي ﺟﺪﻳﺪ-4 ﺷﻜﻞ (S-RNases) و ﻣﻮﻗﻌﻴﺖ اﻳﻨﺘﺮونﻫﺎي اول و دوم در ﺳﺎﺧﺘﻤﺎن ژن ﺧﻮدﻧﺎﺳﺎزﮔﺎريSP ﻧﺎﺣﻴﻪ،(C1-C5) ﻧﻮاﺣﻲ ﺣﻔﺎﻇﺖ ﺷﺪه.اﺳﻴﺪﻫﺎي ﻣﺘﻔﺎوت در ﺗﻮاﻟﻲ اﻳﻦ آﻟﻞﻫﺎ ﺑﺎ رﻧﮓ ﺳﻴﺎه ﻣﺸﺨﺺ ﺷﺪهاﻧﺪ . ﻫﻴﺴﺘﻴﺪﻳﻦ ﺣﻔﺎﻇﺖ ﺷﺪه ﺑﺎ داﻳﺮهﻫﺎي ﺳﻔﻴﺪ )ﺗﻮﺧﺎﻟﻲ( و ﺳﻴﺴﺘﻴﻦ ﺣﻔﺎﻇﺖ ﺷﺪه ﺑﺎ داﻳﺮهﻫﺎي ﺳﻴﺎه )ﺗﻮﭘﺮ( ﻣﺸﺨﺺ ﺷﺪهاﻧﺪ.( در ﺗﻮاﻟﻲ اﺳﻴﺪﻫﺎي آﻣﻴﻨﻪ ﻣﺸﺨﺺ ﺷﺪه اﺳﺖUshijima et al., 1998) Fig. 4. Alignment by ClustalX method of amino acid sequences of S-RNases S36-S43 from almond cultivars Tejari, Holouei, Sefied, Yazd-17, Mashhad-40, Gr-16, and Mamaei, respectively. Different amino acids from a consensus sequence are indicated with black shading. Conserved regions (C1-C5) and hypervariable region (RHV) of Rosaceae S-RNases described by Ushijima et al. (1998), and the signal peptide (SP) are underlined. Position of the two introns is indicated by arrows. Conserved histidine residues are indicated with white circles, and conserved cysteine residues with black circles www.SID.ir 659 ”ﻣﺠﻠﻪ ﺑﻪﻧﮋادي ﻧﻬﺎل و ﺑﺬر” ﺟﻠﺪ ،30-1ﺷﻤﺎره ،3ﺳﺎل 1393 ﺟﺪول -1درﺻﺪ ﺷﺒﺎﻫﺖ ﺗﻮاﻟﻲ اﺳﻴﺪﻫﺎي آﻣﻴﻨﻪ آﻟﻞﻫﺎي ﺟﺪﻳﺪ S36ﺗﺎ S43در ارﻗﺎم ﺑﺎدام اﻳﺮاﻧﻲ ﺑﺎ آﻟﻞﻫﺎي Sدر ﮔﻮﻧﻪﻫﺎي ﭘﺮوﻧﻮس Table 1. Percentage of identity of amino acid sequences of S36-S43 almond RNases from Iranian almond cultivars with S-RNases of Prunus species ﻣﻨﺒﻊ Reference )Yaegaki et al. (2001 Surbanovski et al. )(2007 )Sutherland et al. (2009 آﻟﻞ در ﮔﻮﻧﻪ ﻣﺘﻌﻠﻖ ﺑﻪ ﺟﻨﺲ ﭘﺮوﻧﻮس آﻟﻞ در ارﻗﺎم اﻳﺮاﻧﻲ Allele in species belong to Prunus آﻟﻞ ﮔﻮﻧﻪ Allele in Iranian cultivars درﺻﺪ ﺷﺒﺎﻫﺖ رﻗﻢ اﻳﺮاﻧﻲ Species Prunus mume Allele MSRN-2 P. tenella S7 98 P. cerasifera S8 98 P. lannesiana S02 97 P. tenella S9 98 ﺳﻔﻴﺪ P. webbii Sk 97 ﻳﺰد17 - )Halasz et al. (2007 P. armeniaca S9 96 )Banovic et al. (2009 P. webbii S5 100 )Sutherland et al. (2007 P. domestica S9 98 )Kato et al. (2007 P. lannesiana SX-MK13 97 )Vaughan et al. (2008 P. avium S14 )Wunsch et al. (2004 P. avium S23 )Beppu et al. (2002 P. salicina Si )Boskovic et al. (2007 P. webbii Sn3 D I )Kato et al. (2007 Surbanovski et al. )(2007 )Boskovic et al. (2007 S f o e آﻟﻞ Allele Iranian cultivar )Identity (% ﺗﺠﺎري S36 Tejari 85 iv 97 ﻫﻠﻮﻳﻲ Holouei S37 Sefid S38 Yazd- 17 S39 S40 ﻣﺸﻬﺪ40 - Mashhad- 40 S41 96 h c 96 83 ﺟﻲآر16 - G.R- 16 S42 )Ortega et al. (2006 P. dulcis S25 99 ﻣﺎﻣﺎﻳﻲ Mamaei S43 )Vieira et al. (2007 P. spinosa S15 98 r A ﻣﻲﺷﻮد و اﻳﻦ ﻣﻮﺗﺎﺳﻴﻮنﻫـﺎ ﻣـﻲﺗﻮاﻧﻨـﺪ در ﺗـﻮاﻟﻲ ﻣﺘﻔﺎوت وﻟﻲ ﺑﺎ ﺗـﻮاﻟﻲ ﻣـﺸﺎﺑﻪ در ﻧﺎﺣﻴـﻪ RHVدر اﺳــﻴﺪﻫﺎي آﻣﻴﻨـــﻪ ﻣـــﻮﺛﺮ ﺑـــﻮده و ﺑﺎﻋـــﺚ ﻧﻘـــﺶ ﮔﻼﺑــﻲ اروﭘــﺎﻳﻲ )(Zisovich et al., 2008 اﺧﺘﺼﺎﺻﻲ آﻟﻞ ﻫﺎ و ﺗﻔﺎوت ﺑﻴﻦ آنﻫـﺎ ﺷـﻮﻧﺪ .در ﮔﺰارش ﺷﺪه اﺳﺖ ﻛﻪ ﺑﻴﺎﻧﮕﺮ اﻳﻦ اﺳﺖ ﻛﻪ ﻓﻘـﻂ واﻗﻊ اﻳﺠﺎد آﻟﻞ ﺟﺪﻳـﺪ در اﺛـﺮ ﺗﺠﻤـﻊ ﻣﻮﺗﺎﺳـﻴﻮن ﻧﺎﺣﻴﻪ RHVﺑﻪ ﺗﻨﻬﺎﻳﻲ ﻧﻤﻲﺗﻮاﻧﺪ ﻧﻘـﺶ اﺻـﻠﻲ در ﻫﺎي ﻧﻘﻄﻪاي در ﻧﻬﺎﻳﺖ ﻣﻨﺠﺮ ﺑﻪ ﺗﻔﺎوت در ﺗـﻮاﻟﻲ ﺷﻨﺎﺳﺎﻳﻲ داﻧﻪ ﮔﺮده ﺧـﻮدي و ﺗﻮﻗـﻒ رﺷـﺪ ﻟﻮﻟـﻪ اﺳﻴﺪﻫﺎي آﻣﻴﻨﻪ ﺷـﺪه و ﺑﺎﻋـﺚ اﻳﺠـﺎد ﺗﻔـﺎوت در ﮔﺮده را در ﺣﺎﻟﺖ ﻧﺎﺳﺎزﮔﺎري ﻧـﺸﺎن دﻫـﺪ .ﺗﻨـﻮع ﻧﻘــﺶ اﺧﺘــﺼﺎﺻﻲ آﻟــﻞﻫــﺎ و اﻳﺠــﺎد ﺗﻨــﻮع آﻟﻠــﻲ آﻟﻞﻫﺎ ﺗﻨﻬﺎ ﻧﺎﺷﻲ از ﻣﻮﺗﺎﺳﻴﻮن ﻧﻘﻄﻪاي ﻧﻴﺴﺖ ﺑﻠﻜﻪ ﻣﻲﺷﻮد ) .(Kodad et al., 2008دو آﻟﻞ ﺑﺎ ﻧﻘﺶ وﺟـــﻮد ﻛﺮاﺳـــﻴﻨﮓآور ) (Crossing overو www.SID.ir 660 ﺷﻨﺎﺳﺎﻳﻲ آﻟﻞﻫﺎي ﺧﻮد ﻧﺎﺳﺎزﮔﺎري در ارﻗﺎم ﺑﺎدام اﻳﺮاﻧﻲ ... ﻧﻮﺗﺮﻛﻴﺒﻲ در ﺗـﻮاﻟﻲ ﻧﺰدﻳـﻚ ﺑـﻪ اﻳﻨﺘـﺮونﻫـﺎ ﻫـﻢ آﻟﻞ S1از ﻓﺮاﻧﻴﺲ و آﻟﻞ S6از ﮔﻮﻧﻪ P. webbii ﻣﻲﺗﻮاﻧﻨﺪ ﻧﻘﺶ ﺗﻌﻴﻴﻦ ﻛﻨﻨـﺪهاي در ﺗﻨـﻮع آﻟﻠـﻲ و از ﻳــﻚ ﺟــﺪ ﻣــﺸﺘﺮك ﻣﻨــﺘﺞ ﺷــﺪهاﻧــﺪ .ﻫﻤﭽﻨــﻴﻦ وﺟــﻮد ﭼﻨــﺪ ﺷــﻜﻠﻲ ﺑــﻴﻦ آﻟــﻞﻫــﺎ داﺷــﺘﻪ ﺑﺎﺷــﺪ ﺗﺸﺎﺑﻪ ﺑﺎﻻ ) 99/8درﺻﺪ( ﺑﻴﻦ آﻟـﻞ S21از ﺑـﺎدام و ) .(Ortega et al., 2006وﺟﻮد ﺗﺸﺎﺑﻪ ﺑﻴﻦ ﺗـﻮاﻟﻲ آﻟﻞ S2از ﮔﻮﻧﻪ P. webbiiﮔﺰارش ﺷـﺪه اﺳـﺖ اﺳـــﻴﺪﻫﺎي آﻣﻴﻨـــﻪ در آﻟـــﻞﻫـــﺎي ﻧﺎﺳـــﺎزﮔﺎري ) .(Sanchez and Oliveira, 2005اﻳـﻦ ﺗـﺸﺎﺑﻪ ﺑــﺎدام ﺑــﺎ ﺳــﺎﻳﺮ ﮔﻮﻧــﻪﻫــﺎي ﺟــﻨﺲ ﭘﺮوﻧــﻮس ﻧﻘﺶ ﮔﻮﻧﻪ وﺣﺸﻲ P. webbiiرا در ﺗﻼﻗﻲﻫـﺎي ﺗﻮﺳــﻂ ﺳــﺎﻳﺮ ﻣﺤﻘﻘــﻴﻦ ﻧﻴــﺰ ﮔــﺰارش ﺷــﺪه اﺳــﺖ ﺑﻴﻦ ﮔﻮﻧﻪاي ﺑﻪ ﻋﻨﻮان ﻳﻜﻲ از ﮔﻮﻧﻪﻫﺎي ﻣـﻮﺛﺮ در )Kodad et al., 2008؛ Ortega et al., 2006؛ اﻫﻠــــﻲ ﺷــــﺪن ﺑــــﺎدام ﻧــــﺸﺎن ﻣــــﻲدﻫــــﺪ .(Surbanovski et al., 2007 ) .(Socias i Company, 2004وﺟﻮد آﻟﻞﻫـﺎي D I وﺟــﻮد ﺷــﺒﺎﻫﺖ ﺗــﻮاﻟﻲ آﻟــﻞ ﻧﺎﺳــﺎزﮔﺎري در ﺑﺎ ﺗﺸﺎﺑﻪ ﺑﺎﻻ در ﺗﻮاﻟﻲﻫﺎي اﺳﻴﺪﻫﺎي آﻣﻴﻨﻪ در ﺑـﻴﻦ ارﻗـﺎم اﻫﻠـﻲ ﺑــﺎدام ﺑـﺎ ﮔﻮﻧــﻪ وﺣـﺸﻲ P. webbii ﮔﻮﻧـﻪﻫـﺎي ﺟـﻨﺲ ﭘﺮوﻧـﻮس ﺑﻴـﺎﻧﮕﺮ وﺟـﻮد ﻣﻨــﺸﺎ اﺷــﺎره ﺑــﻪ اﻳــﻦ دارد ﻛــﻪ اﻳــﻦ ﮔﻮﻧــﻪ در ﺗﻜﺎﻣــﻞ ﻳﻜﺴﺎن اﻳﻦ ﮔﻮﻧﻪﻫﺎ در ﻓﺮآﻳﻨﺪ ﺗﻜﺎﻣﻞ اﻳـﻦ ﺟـﻨﺲ ﺑﺎدامﻫﺎي اﻫﻠـﻲ ﺑـﻪ ﻋﻨـﻮان ﻳﻜـﻲ از اﺟـﺪاد اوﻟﻴـﻪ اﺳﺖ. S f o e ﻣﻄﺮح اﺳﺖ .در ﻣﻄﺎﻟﻌﺎت ﻗﺒﻠـﻲ از اﻳـﻦ ﮔﻮﻧـﻪ ﺑـﻪ ﻋﻨﻮان ﻳﻜﻲ از ﮔﻮﻧﻪﻫﺎي ﻣﻮﺛﺮ در اﻳﺠﺎد و ﺗﻜﺎﻣـﻞ ﺑــــﺎدامﻫــــﺎي اﻫﻠــــﻲ اﺷــــﺎره ﺷــــﺪه اﺳــــﺖ )Socias i Company, 1990 and 2004؛ ارزﻳــــﺎﺑﻲ آﻟــــﻞﻫــــﺎي ﺧﻮدﻧﺎﺳــــﺎزﮔﺎري ) (S-alellesدر اﻳــﻦ ﺗﺤﻘﻴــﻖ و ﺗﺤﻘﻴﻘــﺎت ﻗﺒﻠــﻲ iv )2012 al., 2009 Ershadi Zenolabedini؛ h c et and Valizadeh؛ .(Ladizinsky, 1999وﺟــــﻮد ﺗــــﺸﺎﺑﻪ ﺑــــﺎﻻ (Mousavi et al., 2011ﻧﺸﺎن داد ﻛـﻪ اﺣﺘﻤـﺎل ) 100درﺻﺪ( ﺑﻴﻦ آﻟﻞ S1از ﺑﺎدام رﻗﻢ ﻓﺮاﻧﻴﺲ از وﺟﻮد آﻟـﻞﻫـﺎي ﺟﺪﻳـﺪ در ارﻗـﺎم ،ژﻧﻮﺗﻴـﭗﻫـﺎ و ﻓﺮاﻧـــﺴﻪ ﺑـــﺎ آﻟـــﻞ S6ﮔـــﺰارش ﺷـــﺪه از ﮔﻮﻧـ ـﻪ ﮔﻮﻧﻪﻫﺎي وﺣـﺸﻲ ﺑـﺎدام اﻳﺮاﻧـﻲ وﺟـﻮد دارد ﻛـﻪ P.از ﻣﻨﻄﻘــــﻪ ﭘﻮﮔﻠﻴــــﺎ در اﻳﺘﺎﻟﻴــــﺎ ﺑﻴــﺎﻧﮕﺮ وﺟــﻮد ﭼﻨــﺪ ﺷــﻜﻠﻲ ﺑــﺎﻻ در ژن ﺧــﻮد ﺗﻮﺳـــــــــــﻂ ﺳـــــــــــﺎﻧﭽﺰ و اﻟﻴـــــــــــﻮﻳﺮا ﻧﺎﺳــﺎزﮔﺎري در ﺟﻤﻌﻴــﺖﻫــﺎي ﺑــﺎدام اﺳــﺖ ﻛــﻪ ) (Sanchez and Oliveira, 2005ﻧﻴـﺰ ﮔـﺰارش r A ﺑﻴـــﺎﻧﮕﺮ آن اﺳـــﺖ ﻛـــﻪ اﻳـــﺮان ﺑﺎﻳـــﺪ ﻳﻜـــﻲ از ﺷﺪه اﺳﺖ .از آنﺟﺎﻳﻲ ﻛﻪ ﻓﺮاﻧﻴﺲ از ﺗﻼﻗﻲ رﻗـﻢ ﻣﺮاﻛـــﺰ اﺻـــﻠﻲ ﺗﻨـــﻮع ﺑـــﺎدام ﺑﺎﺷـــﺪ .وﺟـــﻮد ﻓﺮاﻧــــــﺴﻮي آي ) (S3S4و رﻗــــــﻢ اﻳﺘﺎﻟﻴــــــﺎﻳﻲ آﻟــﻞﻫــﺎي ﺟﺪﻳــﺪ در ژرم ﭘﻼﺳــﻢ ﺑــﺎدامﻫــﺎي ﻛﺮﻳﺴﺘﻮﻣﻮرﺗﻮ ) (S1S2در ﻣﻨﻄﻘﻪ ﭘﻮﮔﻠﻴﺎ در اﻳﺘﺎﻟﻴﺎ، ﻣﺤﻠـــﻲ اﺳـــﭙﺎﻧﻴﺎﻳﻲ ﺗﻮﺳـــﻂ ارﺗﮕـــﺎ و ﻫﻤﻜـــﺎران ﻣﻨﻄﻘــﻪاي ﻛــﻪ ﺑــﺎدامﻫــﺎي اﻫﻠــﻲ و ﮔﻮﻧـﻪ وﺣــﺸﻲ ) (Ortega et al., 2005 and 2006و ﻛـﺪاد و P. webbiiدر ﻛﻨﺎر ﻫﻢ وﺟﻮد دارﻧـﺪ ،ﺑـﻪ دﺳـﺖ ﻫﻤﻜـﺎران )(Kodad et al., 2008 and 2010 آﻣﺪه اﺳﺖ ،ﻣﻲﺗﻮان ﭼﻨﻴﻦ ﻧﺘﻴﺠﻪﮔﻴﺮي ﻛـﺮد ﻛـﻪ ﻧﻴﺰ ﮔﺰارش ﺷﺪه اﺳـﺖ ﻛـﻪ ﺑﻴـﺎﻧﮕﺮ وﺟـﻮد ﺗﻨـﻮع webbii www.SID.ir 661 ”ﻣﺠﻠﻪ ﺑﻪﻧﮋادي ﻧﻬﺎل و ﺑﺬر” ﺟﻠﺪ ،30-1ﺷﻤﺎره ،3ﺳﺎل 1393 )2008 al., ﻧﺘﺎﻳﺞ اﻳﻦ ﺗﺤﻘﻴﻖ ﻧﺸﺎن داد ﻫﺸﺖ آﻟـﻞ ﺟﺪﻳـﺪ 2004 Hormaza, در ارﻗﺎم ﺑﺎدام اﻳﺮاﻧـﻲ ﻣـﻮرد ﻣﻄﺎﻟﻌـﻪ وﺟـﻮد دارد. 2001 اﻳـــﻦ ارﻗـــﺎم ﻗـــﺒﻼً در ﺷﻨﺎﺳـــﺎﻳﻲ آﻟـــﻞﻫـــﺎي )2011 ﺧﻮدﻧﺎﺳــﺎزﮔﺎري ارﻗــﺎم ﺑــﺎدام اﻳﺮاﻧــﻲ ﺑــﻪ روش (Zhang et al., 2008ﮔﺰارش ﺷﺪه اﺳﺖ. آﻟﻠﻲ ﺑﺎﻻ در ژن ﺧﻮدﻧﺎﺳﺎزﮔﺎري اﺳﺖ. al. et and Wunsch؛ (Sonneveldو زردآﻟــــﻮ et al., Vaughan؛ Gharesheikhbayat؛ et واﻛــﻨﺶ زﻧﺠﻴــﺮهاي ﭘﻠــﻲﻣــﺮاز ) (PCRﻗﻄﻌــﺎت ژﻧﻮﺗﻴﭗ ﻧﺎﺳﺎزﮔﺎري ارﻗﺎم ﺑﺎدام ﻣﻮرد ﺑﺮرﺳـﻲ ﺟﺪﻳﺪ و ﻣﺘﻔﺎوت از اﻧـﺪازه ﻣﺮﺑـﻮط ﺑـﻪ آﻟـﻞﻫـﺎي در اﻳﻦ ﺗﺤﻘﻴﻖ در ﺟﺪول 2اراﺋﻪ ﺷـﺪه اﺳـﺖ .ﺑـﻪ ﻧﺎﺳﺎزﮔﺎري ﺷﻨﺎﺧﺘﻪ ﺷﺪه در ارﻗﺎم اﺳﺘﺎﻧﺪارد ﻧﺸﺎن دﻟﻴــﻞ اﻳــﻦﻛــﻪ ارﻗــﺎم ﻣــﻮرد ﺑﺮرﺳــﻲ ژﻧﻮﺗﻴــﭗ داده ﺑﻮدﻧﺪ ﻛﻪ ﻣﺮﺑﻮط ﺑﻪ آﻟﻞﻫﺎي ﺟﺪﻳـﺪ در ﻧﻈـﺮ ﺧﻮدﻧﺎﺳــﺎزﮔﺎري ﻣﺘﻔــﺎوﺗﻲ از ﺳــﺎﻳﺮ ارﻗــﺎم اﻳﺮاﻧــﻲ ﮔﺮﻓﺘـﻪ ﺷـﺪه ﺑﻮدﻧـﺪ ).(Mousavi et al., 2011 )2009 Ershadi and Valizadeh؛ ﻧﺘـﺎﻳﺞ روش ﺗـﻮاﻟﻲﻳــﺎﺑﻲ در اﻳـﻦ ﺗﺤﻘﻴـﻖ ،وﺟــﻮد 2011 al., et Mousavi؛ آﻟﻞﻫﺎي ﺟﺪﻳـﺪ ﺧﻮدﻧﺎﺳـﺎزﮔﺎري در ارﻗـﺎم ﺑـﺎدام (Zenolabedini et al., 2012و ﺧـــﺎرﺟﻲ اﻳﺮاﻧــﻲ را ﻣــﻮرد ﺗﺎﻳﻴــﺪ ﻗــﺮار داد .ﺑﻨــﺎﺑﺮاﻳﻦ روش ) (Ortega et al., 2005, 2006ﮔـﺰارش ﺷـﺪه D I S f o e دارﻧﺪ ،از ﻧﻈﺮ ﮔﺮوهﻫﺎي دﮔﺮﻧﺎﺳـﺎزﮔﺎر در ﮔـﺮوه ﺗــــﻮاﻟﻲﻳـــــﺎﺑﻲ ﻧﻮﻛﻠﺌﻮﺗﻴــــﺪﻫﺎي ﻣﻮﺟـــــﻮد در ﺳــﺎﺧﺘﻤﺎن آﻟــﻞﻫــﺎي ﺧﻮدﻧﺎﺳــﺎزﮔﺎري ،ﻳﻜــﻲ از روشﻫــﺎي دﻗﻴــﻖ و ﻛﺎرآﻣــﺪ ﺟﻬــﺖ ﺷﻨﺎﺳــﺎﻳﻲ آﻟــﻞﻫــﺎي ﺧﻮدﻧﺎﺳـــﺎزﮔﺎري اﺳــﺖ .در ﻫﻤـــﻴﻦ ﻋﻤﻮﻣﻲ ) (O Groupﻗﺮار ﻣﻲﮔﻴﺮﻧﺪ و ﺑـﻪ ﻋﻨـﻮان iv ﻳــﻚ دﻫﻨــﺪه داﻧــﻪ ﮔــﺮده ﻋﻤــﻮﻣﻲ و ﺳــﺎزﮔﺎر ﺑــــﺎ ﺳــــﺎﻳﺮ ارﻗــــﺎم ﻣﺤــــﺴﻮب ﻣــــﻲﺷــــﻮﻧﺪ h c ارﺗﺒــــﺎط ﻗــــﺮه ﺷــــﻴﺦ ﺑﻴــــﺎت و ﻫﻤﻜــــﺎران ) .(Mousavi et al., 2011ﮔﺮوهدﻫﻨﺪه ﻋﻤﻮﻣﻲ ) (Gharesheikhbayat et al., 2011آﻟـﻞﻫـﺎي ﺷﺎﻣﻞ رﻗﻢﻫﺎﻳﻲ اﺳﺖ ﻛـﻪ ﻣـﻲﺗﻮاﻧﻨـﺪ رﻗـﻢﻫـﺎي ﺧﻮدﻧﺎﺳﺎزﮔﺎري را در 24رﻗﻢ زردآﻟـﻮ از ﻃﺮﻳـﻖ ﻣﻮﺟـﻮد در ﺳـﺎﻳﺮ ﮔـﺮوهﻫـﺎي دﮔﺮﻧﺎﺳـﺎزﮔﺎري روشﻫــﺎي ﻛﻼﺳــﻴﻚ و ﻣﻮﻟﻜــﻮﻟﻲ ﺷﻨﺎﺳــﺎﻳﻲ و را ﺑــﻪ ﻃــﻮر ﻣﻮﻓﻘﻴــﺖآﻣﻴــﺰي ﮔــﺮدهاﻓــﺸﺎﻧﻲ و ﮔــــﺰارش ﻛﺮدﻧــــﺪ ﻛــــﻪ ﺗﻌﻴــــﻴﻦ ژﻧﻮﺗﻴــــﭗ ﺗﻠﻘﻴﺢ ﻛﻨﻨﺪ و ﺑﻪ ﺻﻮرت ﻣﺘﻘﺎﺑﻞ ﺗﻮﺳﻂ رﻗـﻢﻫـﺎي ﺧﻮدﻧﺎﺳــﺎزﮔﺎري ﺑــﻪ ﻛﻤــﻚ روش ﺗــﻮاﻟﻲﻳــﺎﺑﻲ آن ﮔـــﺮوهﻫـــﺎ ﮔـــﺮدهاﻓـــﺸﺎﻧﻲ و ﺗﻠﻘـــﻴﺢ ﺷـــﻮﻧﺪ اﺳﻴﺪﻫﺎي ﻧﻮﻛﻠﺌﻴﻚ داراي اﻃﻤﻴﻨﺎن و دﻗﺖ ﺑﺎﻻﻳﻲ ) .(Valizadeh and Ershadi, 2009ﺷﻨﺎﺳـﺎﻳﻲ در ﻣﻘﺎﻳﺴﻪ ﺑﺎ ﺳﺎﻳﺮ روشﻫﺎ اﺳﺖ .اﺳﺘﻔﺎده از روش آﻟــﻞﻫــﺎي ﺧﻮدﻧﺎﺳــﺎزﮔﺎري و ﺗﻌﻴــﻴﻦ ژﻧﻮﺗﻴــﭗ ﺗﻮاﻟﻲﻳﺎﺑﻲ اﺳﻴﺪﻫﺎي ﻧﻮﻛﻠﺌﻴﻚ ،در ﺗﻌﻴﻴﻦ آﻟﻞﻫﺎي ﺧﻮدﻧﺎﺳﺎزﮔﺎري ارﻗﺎم ﺑﺎدام ﻣﻮﺟﻮد در ﻛـﺸﻮر در ﺟﺪﻳــﺪ و ﻳــﺎ ﺗﺎﻳﻴــﺪ آﻟــﻞﻫــﺎي ﺷﻨﺎﺳــﺎﻳﻲ ﺷــﺪه اﻳـــــﻦ ﭘـــــﮋوﻫﺶ و ﭘـــــﮋوﻫﺶﻫـــــﺎي ﻗﺒﻠـــــﻲ در ارﻗـــﺎم ﺑـــﺎدام )Ortega et al., 2006؛ Zenolabedini؛ 2001 www.SID.ir Oliveira, and r A )2012 ،(Maﮔــــﻴﻼس 662 al., et ... ﺷﻨﺎﺳﺎﻳﻲ آﻟﻞﻫﺎي ﺧﻮد ﻧﺎﺳﺎزﮔﺎري در ارﻗﺎم ﺑﺎدام اﻳﺮاﻧﻲ ژﻧﻮﺗﻴﭗ ﺧﻮدﻧﺎﺳﺎزﮔﺎري ارﻗﺎم ﺑﺎدام اﻳﺮاﻧﻲ در اﻳﻦ ﺑﺮرﺳﻲ-2 ﺟﺪول Table 2. Genotype of self-incompatibility in Iranian almond cultivars in this study رﻗﻢ ﻣﻨﺸﺎ ﺷﺠﺮه ژﻧﻮﺗﻴﭗ ﻧﺎﺳﺎزﮔﺎري ﮔﺮوه دﮔﺮﻧﺎﺳﺎزﮔﺎري Origin Pedigree Incompatibility genotype Tejari ﺗﺠﺎري Iran Unknown S24S36 Cross incompatibility group O Holuei ﻫﻠﻮﻳﻲ Iran Unknown S13S37 O ﺳﻔﻴﺪ Iran Unknown S7S38 O 17-ﺳﺰد Iran Unknown S39S40 O Mashhad040 40-ﻣﺸﻬﺪ Iran Unknown S39S41 O G.R-16 16-ﺟﻲآر Iran Unknown S2S42 Mamaei ﻣﺎﻣﺎﻳﻲ Iran Unknown S25S43 Cultivar Sefid Yaszd-17 D I O O S f . در اﻳﻦ ارﻗﺎم ﺑﻪ روش ﻫﻤﺴﺎﻧﻪﺳﺎزي و ﺗﻮاﻟﻲﻳﺎﺑﻲ ﺷﻨﺎﺳﺎﻳﻲ ﺷﺪهاﻧﺪS43 اﻟﻲS36 آﻟﻞﻫﺎي ﺟﺪﻳﺪ New alleles from S36 to S43 in these cultivars were identified by cloning and sequencing method. o e ﻫــﻢﭘﻮﺷــﺎﻧﻲ دوره ﮔــﻞدﻫــﻲ ﺑــﻴﻦ رﻗــﻢ اﺻــﻠﻲ و ؛Ebadi et al., 2011 ؛Mousavi et al., 2011 ﮔــﺮدهزا( و ﺗﺮﻛﻴــﺐ ﻣﻨﺎﺳــﺐ ﻛﺎﺷــﺖ ارﻗــﺎم در اﺣﺪاث ﺑﺎغﻫﺎي ﺟﺪﻳﺪ ﺗﺠﺎري ﺑﻪ ﻣﻨﻈﻮر ﻣﻮﻓﻘﻴـﺖ ( اﻧﺠـﺎم ﺷـﺪهValizadeh and Ershadi 2009 v i h ﮔﺮدهاﻓﺸﺎﻧﻲ و ﺗﺸﻜﻴﻞ ﻣﻴﻮه و در ﻧﻬﺎﻳـﺖ اﻓـﺰاﻳﺶ ﺑﺮاي ﺗﻮﻟﻴﺪﻛﻨﻨﺪﮔﺎن ﺑﺴﻴﺎر،ﻋﻤﻠﻜﺮد ﺑﺎغﻫﺎي ﺑﺎدام .ﻣﻔﻴﺪ و داراي اﻫﻤﻴﺖ اﺳﺖ c r References اﻳــﻦ ارﻗــﺎم ﻣــﻲﺗﻮاﻧﻨــﺪ ﺑــﻪ ﻋﻨــﻮان واﻟــﺪﻳﻦ.اﺳــﺖ ﻣﻨﺎﺳﺐ در ﺑﺮﻧﺎﻣﻪﻫﺎي ﺑﻪﻧﮋادي ﺟﻬـﺖ اﻃﻤﻴﻨـﺎن از ﻣﻮﻓﻘﻴــﺖ در ﺗﻼﻗــﻲﻫــﺎي ﻛﻨﺘــﺮل ﺷــﺪه ﺑــﺮاي ﺑــﻪ ﻫﻤﭽﻨﻴﻦ.ﻧﮋادﮔﺮان ﺑﺎدام ﻣﻮرداﺳﺘﻔﺎده ﻗﺮار ﮔﻴﺮﻧﺪ اﻧﺘﺨـــﺎب ارﻗـــﺎم ﻣﻨﺎﺳـــﺐ و ﺳـــﺎزﮔﺎر از ﻧﻈـــﺮ ﮔﺮدهاﻓﺸﺎﻧﻲ ﺑﻪ ﻋﻨﻮن درﺧﺘﺎن ﮔﺮدهزا )ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ A Banovic, B., Surbanovski, N., Konstantinovic, M., and Maksimovic, V. 2009. Basic RNases of wild almond (Prunus webbii): Cloning and characterization of six new SRNase and one “non-S RNase” genes. Journal of Plant Physiology 166: 395-402. Barckley, K. K., Uratsu, S. L., Gradziel, T. M., and Dandekar, A. M. 2006. Multidimensional analysis of S-alleles from cross-incompatible groups of California almond cultivars. Journal of the American Society for Horticultural Sciences 131: 632-636. 663 www.SID.ir 1393 ﺳﺎل،3 ﺷﻤﺎره،30-1 ”ﻣﺠﻠﻪ ﺑﻪﻧﮋادي ﻧﻬﺎل و ﺑﺬر” ﺟﻠﺪ Beppu, K., Yamane, H., Yaegaki, H., Yamaguchi, M., Kataoka, I., and Tao, R. 2002. Diversity of S-RNase genes and S-haplotypes in Japanese plum (Prunus salicina L.). Journal of Horticultural Science and Biotechnology 77: 658-664. Bortiri, E., Oh, S. H., Jiang, J., Baggett, S., Granger, A., Weeks, C., Buckingham, M., Potter, D., and Parfitt, D. E. 2001. Phylogeny and systematics of Prunus (Rosaceae) as determined by sequence analysis of ITS and chloroplast trnL-trnF spacer DNA. Systematic Botany 26: 797-807. Boskovic, R. I., Tobutt, K. R., Ortega, E., Sutherland, B. G., and Godini, A. 2007. Self-(in) compatibility of the almonds P. dulcis and P. webbii: detection and cloning D I of ‘wild-type Sf‘ and new self-compatibility alleles encoding inactive S-RNases. Molecular Genetics and Genomics 278: 665-676. Channuntapipat, C., Sedgley, M., Batlle, I., Arus, P., and Collins, G. 2002. S f Sequences of the genomic DNAs encoding the S2, S9, S10, and S23 alleles from almond, Prunus dulcis. Journal of Horticultural Science and Biotechnology 77: 387- o e 392. Channuntapipat, C., Sedgley, M., and Collins, G. 2001. Sequences of the cDNAs and genomic DNAs encoding the S1, S7, S8, and Sf alleles from almond, Prunus dulcis. v i h Theoretical and Applied Genetics 103: 1115-1122. Doyle, J. J., and Doyle, J. L. 1987. A rapid DNA isolation procedure for small quantities of fresh tissue. Phytochemical Bulletin 19: 11-15. c r Ebadi, A., Kamali, K., Fatahi-Moghadam, M. R., Naghavi, M. R., Imani, A., and Ofoghi, H. 2011. Determination of self-compatible almond genotypes obtained from A controlled crosses and identification of S-alleles in some foreign cultivars and genotypes by PCR. Seed and Plant Improvment Journal 27-1(1): 57-67 (in Persian). Gharesheikhbayat, R., Dendini, L., and Sansavini, S. 2011. Identification of selfincompatibility allels in apricot (Prunus armeniaca L.) using multi-level approaches. Seed and Plant Improvment Journal 27-1: 411-426 (in Persian). Halasz, J., Fodor, A., Hegedus, A., and Pedryc, A. 2008. Identification of a new selfincompatibility allele (S31) in a Hungarian almond cultivar and its reliable detection. Scientia Horticulturae 116: 448-451. Halasz, J., Pedryc, A., and Hegedus, A. 2007. Origin and dissemination of the pollenpart mutated SC haplotype which confers self-compatibility in apricot (Prunus 664 www.SID.ir ... ﺷﻨﺎﺳﺎﻳﻲ آﻟﻞﻫﺎي ﺧﻮد ﻧﺎﺳﺎزﮔﺎري در ارﻗﺎم ﺑﺎدام اﻳﺮاﻧﻲ armeniaca). New Phytologist 176: 792-803. Igic, B., and Kohn, J. R. 2001. Evolutionary relationships among self-incompatibility RNases. Proceedings of the National Academy of Sciences of the United States of America 98: 13167-13171. Kato, S., Iwata, H., Tsumura, Y., and Mukai, Y. 2007. Distribution of S-alleles in island populations of flowering cherry, Prunus lannesiana var. speciosa. Genes and Genetics Systems 82: 65-75. Kodad, O., Alonso, J. M., Fernandez I., Marti, A., Oliveira, M. M., and Socias i Company, R. 2010. Molecular and physiological identification of new S-alleles D I associated with self-(in)compatibility in local Spanish almond cultivars. Scientia Horticulturae 123: 308-311. Kodad, O., Sanchez, A. M., Saibo, N., Oliveira, M., and Socias i Company, R. 2008. S f Identification and characterization of new S-alleles associated with selfincompatibility in almond. Plant Breeding 127: 632-638. o e Ladizinsky, G. 1999. On the origin of almond. Genetic Resources and Crop Evoluation 46: 143-147. Lopez, M., Mnejja, M., Rovira, M., Collins, G., Vargas, F. J., Arus, P., and Batlle, v i h I. 2004. Self-incompatibility genotypes in almond re-evaluated by PCR,stylar ribonucleases, sequencing analysis and controlled pollinations. Theoretical and Applied Genetics 109: 954-964. c r Ma, R. C., and Oliveira, M. M. 2001. Molecular cloning of the self-incompatibility genes S1 and S3 from almond (Prunus dulcis cv. Ferragnes). Sexual Plant Reprod. A 14: 163-167. McClure, B. A., Haring, V., Ebert, P. R., Anderson, M. A., Simpson, R. J., Sakiyama, F., and Clarke, A. E. 1989. Style self-incompatibility gene products of Nicotiana alata are ribonucleases. Nature 342: 955-957. Mousavi, S. A., Fatahi Moghadam, M. R., Zamani, Z., Imani, A., Ortega, E., and Dicenta, F. 2011. Identification of self-incompatibility alleles in Iranian almond cultivars and genotypes using PCR. Iranian Journal of Horticultural Sciences 42(2): 169-183 (in Persian). Nettancourt de, D. 2001. Incompatibility and Incongruity of Wild and Cultivated Plants. Springer-Verlag, Berlin, Heidelberg, Germany. 322 pp. 665 www.SID.ir 1393 ﺳﺎل،3 ﺷﻤﺎره،30-1 ”ﻣﺠﻠﻪ ﺑﻪﻧﮋادي ﻧﻬﺎل و ﺑﺬر” ﺟﻠﺪ Ortega, E., Boskovic, R. I., Sargent, D. J., and Tobutt, K. R. 2006. Analysis of SRNase alleles of almond (Prunus dulcis): characterization of new sequences, resolution of synonyms and evidence of intragenic recombination. Molecular Genetics and Genomics 276: 413-426. Ortega, E., Sutherland, B. G., Dicenta, F., Boskovic, R. I., and Tobutt, K. R. 2005. Determination of incompatibility genotypes in almond using first and second intron consensus primers: detection of new S alleles and correction of reported S genotypes. Plant Breeding 124: 188-196. Rahemi, A. R., Fatahi, R., Ebadi, A., Taghavi, T., Hassani, D., Gradziel, T., and D I Chaparro, J. 2010. Genetic variation of S-alleles in wild almonds and their related Prunus species. Australian Journal of Crop Science 4(8): 648-659. Sanchez, A. M., and Oliveira, M. M. 2005. S-alleles in self-compatible Prunus webbii. S f Options Mediterranian 63: 147-152. Sanzol, J. 2009. Genomic characterization of self-incompatibility ribonucleases (S- o e RNases) in European pear cultivars and development of PCR detection for 20 alleles. Tree Genetics and Genomes 5: 393-405. Sassa, H., Nishio, T., Kowyama, Y., Hirano, H., Koba, T., and Ikehashi, H. 1996. v i h Self-incompatibility (S) alleles of the Rosaceae encode members of a distinct class of the T2/S ribonuclease superfamily. Molecular Genetics and Genomics 250: 547-557. Sheikh-Alian, A., Vezvaei, A., Ebadi, A., Fatahi-Moghadam, M. R., and Sarkhosh, c r A. 2010. Determination and identification of self-incompatibility alleles in selective Iranian and foreign almond (Prunus dulcis M.) cultivars by PCR Method. Iranian A Journal of Horticultural Sciences 41 (3): 247-252 (in Persian). Socias i Company, R. 1990. Breeding self-compatible almonds. Plant Breeding Reviews 8: 313-338. Socias i Company, R. 2004. The contribution of Prunus webbii to almond evolution. Plant Genetic Resources Newsletter 140: 9-13. Sonneveld, T., Robbins, T. P., Boskovic, R., and Tobutt, K. R. 2001. Cloning of six cherry self-incompatibility alleles and development of allele-specific PCR detection. Theoretical and Applied Genetics 102: 1046-1055. Sonneveld, T., Tobutt, K. R., and Robbins, T. P. 2003. Allele-specific PCR detection of sweet cherry self-incompatibility (S) alleles S1 to S16 using consensus and allele- 666 www.SID.ir ... ﺷﻨﺎﺳﺎﻳﻲ آﻟﻞﻫﺎي ﺧﻮد ﻧﺎﺳﺎزﮔﺎري در ارﻗﺎم ﺑﺎدام اﻳﺮاﻧﻲ specific primers. Theoretical and Applied Genetics 107: 1059-1070. Surbanovski, N., Tobutt, K. R., Konstantinovic, M., Maksimovic, V., Sargent, D. J., Stevanovic, V., Ortega, E., and Boskovic, R. I. 2007. Self-incompatibility of Prunus tenella and evidence that reproductively isolated species of Prunus have different SFB alleles coupled with an identical S-RNase allele. The Plant Journal 50: 723-734. Sutherland, B. G., Cerovic, R., Robbins, T. P., and Tobutt, K. R. 2009. The myrobalan (Prunus cerasifera L.): a useful diploid model for studying the molecular genetics of self-incompatibility in plums. Euphytica 166: 385-398. D I Sutherland, B. G., Tobutt, K. R., and Robbins, T. P. 2004. Primers amplifying a range of Prunus S alleles. Plant Breeding 123: 582-584. Sutherland, B. G., Tobutt, K. R., and Robbins, T. P. 2007. Trans-specific S-RNase S f and SFB alleles in Prunus self-incompatibility haplotypes. Molecular Genetics and Genomics 279: 95-106. o e Tamura, M., Ushijima, K., Sassa, H., Hirano, H., Tao, R., Gradziel, T. M., and Dandekar, A. M. 2000. Identification of self-incompatibility genotypes of almond by allele-specific PCR analysis. Theoretical and Applied Genetics 101: 334-349. v i h Tao, R., Watari, A., Hanada, T., Habu, T., Yaegaki, H., Yamaguchi, M., and Yamane, H. 2007. Self-compatible peach (Prunus persica) has mutant versions of the S haplotypes found in self-incompatible Prunus species. Plant Molecular Biology c r 63: 109-123. Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F., and Higgins, D. G. A 1997. The CLUSTAL X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25: 4876-4882. Ushijima, K., Sassa, H., Tao, R., Yamane, H., Dandekar, A. M., Gradziel, T., and Hirano, H. 1998. Cloning and characterization of cDNAs encoding S-RNases from almond (Prunus dulcis): primary structural features and sequence diversity of the SRNases in Rosaceae. Molecular Genetics and Genomics 260: 261-268. Valizadeh, B., and Ershadi A. 2009. Identification of self-incompatibility alleles in Iranian almond cultivars by PCR using consensus and allele-specific primers. Journal of Horticultural Science and Biotechnology 84 (3): 285-290. Vaughan, S. P., Boskovic, R. I., Gisbert-Climent, A., Russell, K., and Tobutt, K. R. 667 www.SID.ir 1393 ﺳﺎل،3 ﺷﻤﺎره،30-1 ”ﻣﺠﻠﻪ ﺑﻪﻧﮋادي ﻧﻬﺎل و ﺑﺬر” ﺟﻠﺪ 2008. Characterisation of novel S-alleles from cherry (Prunus avium L.). Tree Genetics and Genomes 4: 531-541. Vieira, C. P., Santos, R. A. M., Morales-Hojas, R., and Vieira, J. 2007. Different positively selected sites at the gametophytic self-incompatibility pistil S-RNase gene in the Solanaceae and Rosaceae (Prunus, Pyrus, and Malus). Journal of Molecular Evoluation 65: 175-185. Wunsch, A., and Hormaza, J. I. 2004. Cloning and characterization of genomic DNA sequences of four self-incompatibility alleles in sweet cherry (Prunus avium L.) Theoretical and Applied Genetics 108: 299-305. D I Yaegaki, H., Shimada, T., Moriguchi, T., Hayama, H., Hajime, T., and Yamaguchi, M. 2001. Molecular characterization of S-RNase genes and S-genotypes in the Japanese apricot (Prunus mume Sieb. et Zucc.). Sexual Plant Report. 13: 251-257. S f Zeinolabedini, M., Khayam-Nekoui, M., Imani, A., and Majidian, P. 2012. Identification of self-compatibility and self-incompatibility genotypes in almond and o e some Prunus species using molecular markers. Seed and Plant Improvment Journal 28-1(2): 227-238 (in Persian). Zhang, L., Chen, X., Chen, X., Zhang, C., Liu, X., Ci, Z., Zhang, H., Wu, C., and v i h Liu, C. 2008. Identification of self-incompatibility (S-) genotypes of Chinese apricot cultivars. Euphytica 160: 241-248. Zisovich, A. H., Stern, R. A., Shafir, S., and Goldway, M. 2008. Identification of c r seven S-alleles from the European pear (Pyrus communis) and the determination of compatibility among cultivars. Journal of Horticultural Science and Biotechnology A 79: 101-106. 668 www.SID.ir
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