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Archive of SID ﺑﻴﻤﺎﺭﻱﻫﺎﻱ ﮔﻴﺎﻫﻲ / ﺟﻠﺪ ۴۷ / ﺷﻤﺎﺭﻩ ۴ / ﺳﺎﻝ ۱۳۹۰: ۴۰۵-۴۱۸ ﻣﻘﺎﻳﺴﺔ ﺟﻤﻌﻴ ﺖﻫﺎﻱ ﻣﺨﺘﻠﻒ Heterodera schachtii ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ ﺍﺯ ﻣﺸﺨﺼﺎﺕ * ﻣﺮﻓﻮﻟﻮﮊﻳﮑﻲ ﻭ ﻣﻮﻟﮑﻮﻟﻲ COMPARISON OF MORPHOLOGICAL AND MOLECULAR CHARACTERS OF DIFFERENT POPULATIONS OF Heterodera schachtii ** ﻋﺒﺎﺱ ﻣﮑﺮﻡ ﺣﺼﺎﺭ۱، ﻋﺼﻤﺖ ﻣﻬﺪﻳﺨﺎﻧﻲ ﻣﻘﺪﻡ۱ ﻭ ﺯﻫﺮﺍ ﺗﻨﻬﺎ ﻣﻌﺎﻓﻲ۲ (ﺗﺎﺭﻳﺦ ﺩﺭﻳﺎﻓﺖ: ۱۷/۹/۱۳۸۹؛ ﺗﺎﺭﻳﺦ ﭘﺬﻳﺮﺵ: ۱۳۹۰/۴/۲۹) ﭼﻜﻴﺪﻩ ﮔﻮﻧﻪ Heterodera schachtii، ﻳﮑﻲ ﺍﺯ ﻣﻬﻢﺗﺮﻳﻦ ﻋﻮﺍﻣﻞ ﺑﻴﻤﺎﺭﻱﺯﺍﻱ ﭼﻐﻨﺪﺭﻗﻨﺪ ﺩﺭ ﺩﻧﻴﺎﺳﺖ. ﺍﻳﻦ ﻧﻤﺎﺗﻮﺩ ﺩﺭ ﺍﮐﺜﺮ ﻣﻨﺎﻃﻖ ﺗﺤﺖ ﮐﺸﺖ ﺍﻳﻦ ﮔﻴـ ﺎﻩ ﺩﺭ ﺍﻳﺮﺍﻥ ﮔﺴﺘﺮﺵ ﺩﺍﺭﺩ ﻭ ﻣﻮﺟﺐ ﮐﺎﻫﺶ ﻋﻤﻠﮑﺮﺩ ﻭ ﻣﻴﺰﺍﻥ ﻗﻨﺪ ﻣﻮﺟﻮﺩ ﭼﻐﻨﺪﺭ ﺗﻮﻟ ﻴﺪ ﺷﺪﻩ ﺩ ﺭ ﻣﺰﺍﺭﻉ ﺁﻟـﻮﺩﻩ ﻣـﻲ ﺷـﻮﺩ . ﺟﻤﻌﻴـﺖ ﻫـﺎﻱ ﻣﺨﺘﻠـﻒ H. schachtii ﺍﺯ ﻟﺤﺎﻅ ﺧﺼﻮﺻﻴﺎﺕ ﻣﺮﻓﻮﻟﻮﮊﻳﮑﻲ ﺑﺴﻴﺎﺭ ﻣﺘﻔﺎﻭﺕ ﻫﺴﺘﻨﺪ ﻭ ﺷﻨﺎﺳﺎﻳﻲ ﺁﻧﻬﺎ ﺑﺮﺍﺳﺎﺱ ﺧـﺼﻮﺻﻴﺎﺕ ﻣﺮﻓﻮﻟـﻮﮊ ﻳﮑﻲ ﻭ ﻣﺮﻓـﻮﻣﺘﺮﻳﮑﻲ ﺳﻴﺴﺖﻫﺎﻭﻻﺭﻭﻫﺎﻱ ﺳﻦ ﺩﻭﻡ ﺑﺴﻴﺎﺭ ﻭﻗﺖﮔﻴﺮ ﺑﻮﺩﻩ ﻭ ﻧﻴﺎﺯﻣﻨﺪ ﻣﻬﺎﺭﺕ ﻭ ﺩﻗﺖ ﺯﻳﺎﺩ ﻣﻲ ﺑﺎﺷﺪ. ﺩﺭ ﺳﺎﻝ ﻫـﺎ ﻱ ﺍﺧ ﻴـ ﺮ ﺍﻣﮑـﺎﻥ ﺍﺳـﺘﻔ ﺎﺩﻩ ﺍﺯ ﺭﻭﺵ -ITS PCR-RFLP ﺑﺮﺍﻱ ﺗﻔﮑﻴﮏ ﺍﻳﻦ ﮔﻮﻧﻪ ﺍﺯ ﺳﺎﻳﺮ ﮔﻮﻧﻪﻫﺎﻱ ﻣﻮﺟﻮﺩ ﺑﺮﺭﺳﻲ ﺷﺪﻩ ﺍﺳﺖ . ﺩﺭ ﺍ ﻳﻦ ﺑﺮﺭﺳ ﻲ ۱۲۰ ﺟﻤﻌﻴﺖ ﺍﺯ ﻧﻤﺎﺗﻮﺩ ﺳ ﻴﺴﺘﻲ ﭼﻐﻨﺪﺭﻗﻨـﺪ ﺍﺯ ﻣﺰﺍﺭﻉ ﻣﺨﺘﻠﻒ ﺩﺭ ﺍﺳﺘﺎﻥ ﺧﺮﺍﺳﺎﻥ ﺭﺿﻮﻱ ﺟﻤﻊ ﺁﻭﺭﻱ ﺷﺪ ﻭ ﺳ ﻴ ﺴﺖﻫﺎ ﺍﺯ ﻟﺤﺎﻅ ﺧﺼﻮﺻ ﻴﺎﺕ ﻣﺮﻓﻮﻟﻮﮊ ﻳﮑﻲ ﻭ ﻣﺮﻓﻮﻣﺘﺮ ﻳﮑﻲ ﻣﻮﺭﺩ ﺑﺮﺭﺳـ ﻲ ﻗـﺮﺍﺭ ﮔﺮﻓﺘﻨﺪ. ﺩﺭ ﻣﻴﺎﻥ ﺍﻳﻦ ﺟﻤﻌﻴﺖ ﻫﺎ، ۸۸ ﺟﻤﻌﻴﺖ ﮐﻪ ﺗﻔﺎﻭﺕ ﻫﺎﻱ ﻣﺮﻓﻮﻟﻮﮊﻳﮑﻲ ﻧﺴﺒﺘﺎ ﺯﻳﺎﺩﻱ ﺑﺎ ﻫﻢ ﺩﺍﺷﺘﻨﺪ، ﺍﻧﺘﺨﺎﺏ ﺷﺪﻧﺪ . ﭘـﺲ ﺍﺯ ﺍﺳـﺘﺨﺮﺍﺝ DNA، ﺗﮑﺜﻴﺮ ﻧﺎﺣﻴﻪ rDNA-ITS ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ ﺍﺯ ﺁﻏﺎﺯﮔﺮﻫﺎﻱ ﻋﻤﻮﻣ ﻲ ﺍ ﻳﻦ ﻧﺎﺣ ﻴﻪ ﺍﻧﺠﺎﻡ ﺷﺪ . ﻣﺤﺼﻮﻝ PCR ﻫﺮ ﻧﻤﻮﻧﻪ ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ ﺍﺯ ﺁﻧـﺰﻳﻢ ﺑﺮﺷـﻲ MvaI ﻫﻀﻢ ﺷﺪ . ﺗﻤﺎﻣﻲ ۸۸ ﻧﻤﻮﻧﻪ ﺍﻟﮕﻮﻱ ﺑﺎﻧﺪ ﻱ ﻣﺸﺎﺑﻬ ﻲ ﺭﺍ ﺗﻮﻟﻴﺪ ﮐﺮﺩﻧﺪ ﮐﻪ ﺑﺎ ﺍﻟﮕﻮﻱ ﺍﺭﺍ ﺋﻪ ﺷﺪﻩ ﺑﺮﺍ ﻱ ﺍ ﻳﻦ ﮔﻮﻧﻪ ﺩﺭ ﻣﻨﺎﺑﻊ ﻣ ﻌﺘﺒﺮ ﻫﻤﺨﻮﺍﻧ ﻲ ﺩﺍﺭﺩ . ﭘﺲ ﺍﺯ ﺗﺎﻳﻴﺪ ﮔﻮﻧﻪ، ﮔﺮﻭﻩ ﺑﻨﺪﻱ ﺟﻤﻌ ﻴ ﺖﻫﺎﻱ ﺫﮐﺮ ﺷﺪﻩ ﺑﺮ ﺍﺳﺎﺱ ﺧﺼﻮﺻ ﻴﺎﺕ ﻣﺮﻓﻮﻣﺘﺮ ﻳﮑﻲ ﻧﺸﺎﻥ ﺩﺍﺩ ﮐﻪ ﺟﻤﻌ ﻴ ﺖﻫﺎﻱ ﻣﻨﺎﻃﻖ ﺷﻤﺎﻟ ﻲ ﺍﺳﺘﺎﻥ ﺍﺯﺟﻤﻌ ﻴ ﺖﻫﺎﻱ ﻣﻨﺎﻃﻖ ﺟﻨﻮﺑ ﻲ ﻗﺎﺑﻞ ﺗﻔﮑ ﻴﮏ ﺍﺳﺖ، ﻫﺮ ﭼﻨﺪ ﺑﺮﺍﺳﺎﺱ ﻣﻨﺎﻃﻖ ﮐﻮﭼﮏ ﺗﺮ ﺟﻐﺮﺍﻓ ﻴﺎﻳﻲ ﺗﻔﮑ ﻴﮏ ﻗﺎﺑﻞ ﻗﺒﻮﻟ ﻲ ﺻﻮﺭﺕ ﻧﮕﺮﻓﺖ . ﻫﻢﭼﻨﻴﻦ ﺑﺮ ﺍﺳـﺎﺱ ﻃـﻮﻝ ﺑﺎﻧﺪﻫﺎﻱ ﮐﻮﺗ ﻴﮑﻮﻟﻲ ﻧﺎﺣﻴﻪ ﻭﺍﮊﻥ ﻭ ﺑﺮﺟﺴﺘﮕ ﻲﻫﺎﻱ ﮔﺮﻩ ﻣﺎﻧﻨﺪ ﻣﻮﺟﻮﺩ ﺩﺭ ﻣﺨﺮﻭﻁ ﺍﻧﺘﻬﺎ ﻱ ﺑﺪﻥ ﺳ ﻴ ﺴﺖﻫﺎ، ﭼﻬﺎﺭ ﺗ ﻴﭗ ﻣﺮﻓﻮﻟـﻮﮊ ﻳﮑﻲ ﻧـﺸﺨ ﻴﺺ ﺩﺍﺩﻩ ﺷﺪ. ﻧﺘﺎﻳﺞ ﺍﻳﻦ ﺗﺤﻘﻴﻖ ﻧﺸﺎﻥ ﺩﺍﺩ ﮐﻪ ﺭﻭﺵ ITS-PCR-RFLPof SID ﻣﻲﺗﻮﺍﻧﺪ ﺩﺭ ﺷﻨﺎﺳﺎﺋﻲ ﺟﻤﻌﻴ ﺖﻫـﺎ ﻱ ﻣﺨﺘﻠـﻒ H. schachtiiArchive ﺑـﺎ ﺩﺍﻣﻨـﻪ ﻭﺳـ ﻴﻊ ﺗﻔﺎﻭﺕﻫﺎﻱ ﻣﺮﻓﻮﻟﻮﮊﻳﮑﻲ ﺑﺴﻴﺎﺭ ﮐﺎﺭﺁﻣﺪﺗﺮ ﻭ ﺳﺮﻳ ﻊﺗﺮ ﺍﺯ ﺭﻭﺵﻫﺎﻱ ﻣﺮﻓﻮﻟﻮﮊﻳﮑﻲ ﺑﺎﺷﺪ. ﻭﺍﮊﻩﻫﺎ ﻱ ﻛﻠﻴﺪ ﻱ: ﮔﺮﻭﻩﺑﻨﺪﻱ، ﻧﻤﺎﺗﻮﺩ ﺳﻴﺴﺘﻲ ﭼﻐﻨﺪﺭﻗﻨﺪ، ﺧﺮﺍﺳﺎﻥ ﺭﺿﻮﻱ، ﻣﺮﻓﻮﻟﻮﮊﻱ،Heterodera schachtii ،ITS-PCR-RFLP. *: ﺑﺨﺸﻲ ﺍﺯ ﭘﺎﻳﺎﻥ ﻧﺎﻣﻪ ﮐﺎﺭﺷﻨﺎﺳﻲ ﺍﺭﺷﺪ ﻧﮕﺎﺭﻧﺪﻩ ﺍﻭﻝ ﺍﺭﺍﺋﻪ ﺷﺪﻩ ﺑﻪ ﺩﺍﻧﺸﮑﺪﻩ ﮐﺸﺎﻭﺭﺯﻱ، ﺩﺍﻧﺸﮕﺎﻩ ﻓﺮﺩﻭﺳﻲ ﻣﺸﻬﺪ **: ﻣﺴﺌﻮﻝ ﻣﻜﺎﺗﺒﺎﺕ، ﭘﺴﺖ ﺍﻟﻜﺘﺮﻭﻧﻴﻜﻲ: [email protected] ١. ﺑﻪ ﺗﺮﺗﻴﺐ ﺩﺍﻧﺸﺠﻮﻱ ﺳﺎﺑﻖ ﮐﺎﺭﺷﻨﺎﺳ ﻲﺍﺭﺷﺪ ﻭ ﺩﺍﻧﺸﻴﺎﺭ ﺑﻴﻤﺎﺭ ﻱﺷﻨﺎﺳﻲ ﮔﻴﺎﻫﻲ، ﺩﺍﻧﺸﮑﺪﻩ ﮐﺸﺎﻭﺭﺯﻱ، ﺩﺍﻧﺸﮕﺎﻩ ﻓﺮﺩﻭﺳﻲ ﻣﺸﻬﺪ ۲. ﺩﺍﻧﺸﻴﺎﺭ ﺑﺨﺶ ﺗﺤﻘﻴﻘﺎﺕ ﻧﻤﺎﺗﻮﺩﺷﻨﺎﺳﻲ، ﻣﺆﺳﺴﻪ ﺗﺤﻘﻴﻘﺎﺕ ﮔﻴ ﺎﻩﭘﺰﺷﮑﻲ ﮐﺸﻮﺭ ۴۰۵ www.SID.ir ﺑﻴﻤﺎﺭﻱﻫﺎﻱ ﮔﻴﺎﻫﻲ / ﺟﻠﺪ ۴۷ / ﺷﻤﺎﺭﻩ ۴ / ﺳﺎﻝ ۱۳۹۰: ۴۰۵-۴۱۸ ﻣﻘﺪﻣﻪ Moghadam & Kheiri 1995) ﺍﺯ ﺧـــﺎﮎ ﻣـــﺰﺍﺭﻉ ﻧﻤـﺎﺗﻮﺩ ﺳ ﻴـ ﺴﺘﻲ ﭼﻐﻨﺪﺭﻗﻨـﺪ ﺑـﺎ ﻧـﺎﻡ ﻋﻠﻤـ ﻲ Heterodera ﭼﻐﻨﺪﺭﻗﻨﺪ ﺍﺳﺘﺎﻥ ﺧﺮﺍﺳـﺎﻥ ﺭﺿـﻮ ﻱ ﺟﺪﺍﺳـﺎﺯ ﻱ ﻭ ﺷﻨﺎﺳـﺎ ﻳﻲ schachtii Schmidt 1871 ، ﻳﮑـ ﻲ ﺍﺯ ﻧﻤﺎﺗﻮﺩﻫـﺎ ﻱ ﻣﻬـﻢ ﺷﺪﻩﺍﻧﺪ. ﻻﺯﻡ ﺑﻪ ﺫﮐﺮ ﺍﺳﺖ ﮐﻪ ﺷﻨﺎﺳﺎﻳﻲ ﺍﻳﻦ ﮔﻮﻧﻪ ﻫـﺎ ﺗﻨﻬـﺎ ﺍﻧﮕﻞ ﮔ ﻴﺎﻫﻲ ﻭ ﺧﺴﺎﺭﺕ ﺯﺍ ﺩﺭ ﻧﻘﺎﻁ ﻣﺨﺘﻠـﻒ ﺩﻧ ﻴـ ﺎ ﻣـ ﻲﺑﺎﺷـﺪ ﺑﺮﺍﺳﺎﺱ ﺧﺼﻮﺻﻴﺎﺕ ﻣﺮﻓﻮﻟﻮﮊﻳﮑﻲ ﻭ ﻣﺮﻓﻮﻣﺘﺮﻳﮑﻲ ﺍﻧﺠـﺎﻡ ( & Baldwin & Mundo-Ocampo 1991; Evans ﺷﺪﻩ ﺍﺳﺖ . ﮔﻮﻧﻪ H. schachtii ﺩﺭ ﮔـﺮﻭﻩ schachtii ﻭ ﺩﺭ Rowe 1998; Plantard & Porte 2003). ﺍﻳـ ﻦ ﻧﻤـﺎﺗﻮﺩ ﮐﻨﺎﺭ ﮔﻮﻧﻪﻫﺎﻳﻲ ﻣﺜﻞ H. daverti ،H. ciceri ،H. betae، ﺑﻪ ۲۰۰ ﮔﻮﻧﻪ ﺍﺯ ۹۵ ﺟﻨﺲ ﻭ ۲۳ ﺧـﺎﻧﻮﺍﺩﻩ ﻣﺨﺘﻠـﻒ ﮔ ﻴـ ﺎﻫﻲ H. lespedezae ، H. glycines ، H. galeopsidis ، ﺣﻤﻠــﻪ ﻣ ــ ﻲﮐﻨــﺪ. ﺍﮐﺜــﺮ ﻣﻴ ﺰﺑــﺎﻥﻫــﺎﻱ ﺁﻥ ﺩﺭ ﺧــﺎﻧﻮﺍﺩﻩﻫــﺎﻱ H. rosii ، H. medicaginis ﻭ H. trifolii ﻗــــﺮﺍﺭ ﺩﺍﺭﺩ Chenopodiaceae ﻭ Cruciferae ﻳﺎﻓـــﺖ ﻣـــ ﻲﺷـــﻮﻧﺪ (Subbotin et al. 2001; Tanha Maafi et al. 2003). (Steele 1965). ﺍﻳﻦ ﻧﻤﺎﺗﻮﺩ ﺳﺒﺐ ﮐﺎﻫﺶ ﻋﻤﻠﮑﺮﺩ ﻭ ﻣﻴﺰﺍﻥ ﺗﻔﺎﻭﺕﻫﺎﻱ ﻣﺮﻓﻮﻟﻮﮊ ﻳﮑﻲ ﺑ ﻴﻦ ﺍ ﻳﻦ ﮔﻮﻧﻪ ﻫﺎ، ﺑـﻪ ﺧـﺼﻮﺹ ﺩﻭ ﺗﻮﻟﻴــﺪ ﻗﻨــﺪ ﺩﺭ ﭼﻐﻨــﺪﺭﮐﺎﺭ ﻱﻫــﺎﻱ ﺁﻟــﻮﺩﻩ ﺑــﻪ ﺁﻥ ﻣــ ﻲﺷــﻮﺩ، ﮔﻮﻧﻪ H. schachtii ﻭ H. trifolii ﺑﺴﻴﺎﺭ ﺟﺰﺋﻲ ﻣ ﻲﺑﺎﺷـﻨﺪ . ﺑﻪﻃﻮﺭﻳﮑﻪ ﺩﺭ ﮐﺸﻮﺭﻫﺎ ﻱ ﺍﺭﻭﭘﺎ ﻳﻲ ﻣ ﻴـ ﺰﺍﻥ ﺧـﺴﺎﺭﺕ ﻧﺎﺷـ ﻲ ﺍﺯ ﮔﻮﻧــﻪ ﺩﻭﻡ ﺩﺭ ﻣــﺰﺍﺭﻉ ﭼﻐﻨﺪﺭﻗﻨــﺪ ﺧﺮﺍﺳــﺎﻥ ﺩﺍﺭﺍﻱ ﭘــﺮﺍﮐﻨﺶ ﺍﻳﻦ ﻧﻤﺎﺗﻮﺩ ﺩﺭ ﺣﺪﻭﺩ ۹۰ ﻣ ﻴﻠﻴﻮﻥ ﻳﻮﺭﻭ ﺩﺭ ﺳﺎﻝ ﺑﺮﺁﻭﺭﺩ ﺷـﺪﻩ ﺯﻳﺎﺩﻱ ﻣ ﻲﺑﺎﺷﺪ ﻭ ﻋﻼﻭﻩ ﺑﺮ ﺧﺎﮎ ﺍ ﺯ ﺭ ﻳ ﺸﻪﻫﺎﻱ ﭼﻐﻨﺪﺭﻗﻨﺪ ﻧﻴﺰ ﺍﺳﺖ (Müller 1999). ﺟﺪﺍﺳﺎﺯﻱ ﺷﺪﻩ ﺍﺳﺖ . ﺑﻪ ﻫﻤ ﻴﻦ ﺩﻟ ﻴﻞ ﺍﺣﺘﻤﺎﻝ ﺑﺮﻭﺯ ﺧﻄـﺎ ﺩﺭ ﮔﻮﻧــﻪ H. schachtii ﺩﺭ ﺍ ﻳ ــﺮﺍﻥ ﺑــﺮﺍﻱ ﺍﻭﻟــﻴﻦ ﺑــﺎﺭ ﺍﺯ ﺷﻨﺎﺳـﺎ ﻳﻲ ﻭﺟـﻮﺩ ﺩﺍﺭﺩ ( .Mahdikhani Moghadam et al ﭼﻐﻨــﺪﺭﮐﺎﺭ ﻱﻫــﺎﻱ ﺗﺮﺑــﺖ ﺣﻴﺪﺭﻳــﻪ ﺩﺭ ﺍﺳــﺘﺎﻥ ﺧﺮﺍﺳــﺎﻥ Mahdikhani Moghadam & Jafarpour 2008 ;1996). ﮔﺰﺍﺭﺵﺷﺪ (Esmailpour & Schaffer 1970). ﺩﺭ ﺑﺮﺭﺳـ ﻲ ﺍﺯ ﺳﻮ ﻱ ﺩ ﻳﮕﺮ ﺷﻨﺎﺳـﺎ ﻳﻲ ﺍ ﻳـ ﻦ ﻧﻤﺎﺗﻮﺩﻫـﺎ ﺑ ﻴـ ﺸﺘﺮ ﺑـﺮ ﺍﺳـﺎﺱ ﻧﻤﺎﺗﻮﺩﻫـــﺎﻱ ﺳﻴـــﺴﺘﻲ ﺍﻳـــﺮﺍﻥ، ﻫﻔـــﺖ ﮔﻮﻧـــﻪ ﻧﻤـــﺎﺗﻮﺩ ﻣﺸﺨﺼﺎﺕ ﻣﺮﻓﻮﻟـﻮﮊ ﻳﮑﻲ ﻭ ﻣﺮﻓـﻮﻣﺘﺮ ﻳﮑﻲ ﻣﺨـﺮﻭﻁ ﺍﻧﺘﻬـﺎ ﻱ ﺳﻴـــﺴﺘﻲ ،H. rosii ، H. mothi، H. iri ، H. schachtii ﺳﻴﺴﺖ ﻧﻤﺎﺗﻮﺩ ﺻﻮﺭﺕ ﻣ ﻲﮔﻴﺮﺩ. ﺗﻬﻴﻪ ﺑﺮﺵ ﺍﺯ ﺍ ﻳـ ﻦ ﻗـﺴﻤﺖ ،H. latipons ،H. trifolii ، H. galeopsidis ﻭ H. avenae ﺑﺪﻥ ﺩﺭ ﻣﻮﺍﺭﺩ ﻱ ﮐﻪ ﻧ ﻴﺎﺯ ﺑـﻪ ﺑﺮﺭﺳـ ﻲ ﺟﻤﻌ ﻴـ ﺖﻫـﺎ ﻱ ﺯ ﻳـ ﺎﺩﻱ group ﺍﺯ ﭼﻐﻨــﺪﺭﮐﺎﺭ ﻱﻫــﺎﻱ ﮐــﺸﻮﺭ ﺷﻨﺎﺳــﺎ ﻳﻲ ﺷــﺪﻩ ﻭ ﻣ ﻲﺑﺎﺷﺪ، ﮐﺎﺭ ﻱ ﻭﻗﺖ ﮔﻴﺮ ﺑـﻮﺩﻩ ﻭ ﺩﺭ ﺑﻌـﻀ ﻲ ﻣـﻮﺍﺭﺩ ﻣﻤﮑـﻦ ﺍﺳﺘﺎﻥﻫﺎﻱ ﺧﺮﺍﺳﺎﻥ، ﺁﺫﺭﺑﺎ ﻳﺠـﺎﻥ ﻏﺮﺑـ ﻲ ﻭ ﻓـﺎﺭﺱ ﺑـﻪ ﻋﻨـﻮﺍﻥ ﺍﺳﺖ ﺳ ﻴ ﺴﺖﻫﺎﻱ ﻣﻨﺎﺳـﺒ ﻲ ﺑـﺮﺍ ﻱ ﺗﻬ ﻴـ ﻪ ﺑـﺮﺵ ﺩﺭ ﺩﺳـﺘﺮﺱ ﺁﻟﻮﺩﻩﺗﺮﻳﻦ ﻣﻨﺎﻃﻖ ﺑﻪ ﻧﻤـﺎﺗﻮﺩ ﺳ ﻴـ ﺴﺘﻲ ﭼﻐﻨﺪﺭﻗﻨـﺪ ﺗـﺸﺨ ﻴﺺ ﻧﺒﺎﺷﺪ (Karegar 2006 )، ﻣﺨﺼﻮﺻﺎ ﺯﻣﺎﻧﻲ ﮐﻪ ﺁﺯﻣﻮﻥ ﺧﺎﮎ ﺩﺍﺩﻩ ﺷﺪﻩﺍﻧﺪ (TalachianSID et al. 1976). ﺩﺭ ﻃﻲ of ﺳﺎﻝﻫـﺎ ﻱ ﺑﻪ ﻣﻨﻈﻮﺭ ﺗﻌ ﻴﻴﻦ ﺁﻟﻮﺩﮔ ﻲ ﺑﻪ ﺍ ﻳﻦ Archiveﻧﻤﺎﺗﻮﺩ ﻗﺒﻞ ﺍﺯ ﮐﺎﺷـﺖ ﺯﻣـ ﻴﻦ ﺑﻌﺪ، ﭘﺮﺍﮐﻨﺶ ﺍ ﻳﻦ ﮔﻮﻧﻪ ﺩﺭ ﻧﻘﺎﻁ ﻣﺨﺘﻠﻒ ﺍﻳﺮﺍﻥ ﻣﻮﺭﺩ ﺑﺮﺭﺳـ ﻲ ﺍﻧﺠﺎﻡ ﻣ ﻲﺷﻮﺩ ﮐﻪ ﺩﺭ ﺍﻳﻦ ﻣﻮﺍﻗـﻊ ﻣ ﻌ ﻤ ـ ﻮ ﻻﹰ ﺳﻴـ ﺴﺖﻫـﺎ ﺷـﮑﻞ ﻗـﺮﺍﺭ ﮔﺮﻓـﺖ ( Mahdikhani Moghadam & Kheiri ﻣﻨﺎﺳﺒﻲ ﻧﺪﺍﺭﻧﺪ ﻭ ﺍﻣﮑﺎﻥ ﺷﻨﺎﺳﺎ ﻳﻲ ﺑﺎ ﺍﺳﺘﻔﺎﺩﻩ ﺍﺯ ﻻﺭﻭﻫﺎ ﻱ ﺳﻦ .Akhiani et al. 1996; Khezrinejad et al ;1995 ﺩﻭﻡ ﻧ ﻴﺰ ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ ﻣﺸﺎﺑﻬﺖ ﺯ ﻳﺎﺩ ﺑ ﻴﻦ ﮔﻮﻧـﻪ ﻫـﺎ ﺍﻣﮑـﺎﻥ ﭘـﺬ ﻳﺮ 2006; Karegar 2006; Mahdikhani Moghdam & Jafarpour 2008). ﻫﻢﭼﻨـ ﻴﻦ ﮔﻮﻧـﻪ ﻫـﺎ ﻱ H. carotae ﻭ ﻧﻴﺴﺖ. ﺍﺯ ﺍ ﻳﻦ ﺭﻭ ﺷﻨﺎﺳﺎ ﻳﻲ ﻣﺮﻓﻮﻟـﻮﮊ ﻳﮑﻲ ﺍ ﻳـ ﻦ ﻧﻤﺎﺗﻮﺩﻫـﺎ ﺩﺭ Shiqi et al. Mahdikhani Moghadam & ) H. cruciferae ﻣﺠﻤﻮﻉ ﮐـﺎﺭ ﻱ ﺩﺷـﻮﺍ ﺭ ﻭ ﻭﻗـﺖ ﮔﻴـ ﺮ ﺍﺳـﺖ ( 2008; Amiri et al. 2003 Jafarpour, 2008) ﻭ ﻧﻴﺰ ﮔﻮﻧـﻪ Mahdikhani ) H. mani ). ﺷﻨﺎﺳــﺎﻳﻲ ﻣﻮﻟﮑــﻮﻟﻲ ۴۰۶ www.SID.ir ﻣﮑﺮﻡ ﻭ ﻫﻤﮑﺎﺭﺍﻥ: ﻣﻘﺎﻳﺴﺔ ﺟﻤﻌﻴﺖﻫﺎﻱ ﻣﺨﺘﻠﻒ Heterodera schachtii ... ﻧﻤﺎﺗﻮﺩﻫﺎﻱ ﺍﻧﮕﻞ ﮔ ﻴﺎﻫﻲ، ﺑﻪ ﺧـﺼﻮﺹ ﻧﻤﺎﺗﻮﺩﻫـﺎ ﻱ ﺳ ﻴـ ﺴﺘﻲ ﻧﻤﺎﺗﻮﺩ ﻭ ﺗﻌ ﻴﻴﻦ ﺍﺷﮑﺎﻝ ﻣﺨﺘﻠـﻒ ﻣﺮﻓﻮﻟـﻮﮊ ﻳﮑﻲ ﺁﻥ ﺩﺭ ﺍ ﻳـ ﺮﺍﻥ ﺑﺮ ﭘﺎ ﻳﻪ ﺭﻭﺵ ﻫﺎﻱ ﻣﺒﺘﻨ ﻲ ﺑﺮ PCR ﻣ ﻲﺗﻮﺍﻧﺪ ﺑﺴ ﻴﺎﺭ ﺳﺮ ﻳ ﻊﺗـﺮ ﻭ ﺻﻮﺭﺕ ﻧﮕﺮﻓﺘﻪ ﺍﺳﺖ، ﺑﺮﺭﺳـ ﻲ ﺣﺎﺿـﺮ ﺭﻭ ﻱ ﺟﻤﻌ ﻴـ ﺖﻫـﺎ ﻱ ﺭﺍﺣﺖﺗﺮ ﺍﺯ ﺭﻭﺵ ﻫﺎﻱ ﺩﺷﻮﺍﺭ ﻭ ﻭﻗﺖ ﮔﻴﺮ ﻣﺮﻓﻮﻟﻮﮊ ﻳﮑﻲ ﺑﺎﺷﺪ ﻣﺨﺘﻠﻒ ﻧﻤـﺎﺗﻮﺩ ﺳ ﻴـ ﺴﺘﻲ ﭼﻐﻨـﺪﺭ ﻗﻨـﺪ ﺩﺭ ﺍﺳـﺘﺎﻥ ﺧﺮﺍﺳـﺎﻥ ( ;Powers et al. 1997; Subbotin et al. 2001 ﺭﺿﻮﻱ ﺍﻧﺠﺎﻡ ﮔﺮﺩ ﻳﺪ. ﺩﺭ ﺍ ﻳﻦ ﺗﺤﻘ ﻴﻖ ﺟﻤﻌ ﻴ ﺖﻫﺎﻱ ﻣﺨﺘﻠﻒ ﺑﺎ Tanha Maafi et al. 2003). ﺍﺳــﺘﻔﺎﺩﻩ ﺍﺯ ﭼﻨﺪﺷــﮑﻠﻲ ﺍﺳﺘﻔﺎﺩﻩ ﺍﺯ ﺧﺼﻮﺻ ﻴﺎﺕ ﻣﺮﻓﻮﻟﻮﮊ ﻳﮑﻲ ﻭ ﻣﺮﻓﻮﻣﺘﺮ ﻳﮑﻲ ﻣﺨﺮﻭﻁ ﻃﻮﻟﻲ ﻗﻄﻌﺎﺕ ﺑﺮﺷـ ﻲ (RFLPs) ﻧـﻮﺍﺣ ﻲ ITS ﮊﻥ rDNA ﺍﻧﺘﻬﺎﻱ ﺳ ﻴ ﺴﺖﻫﺎ ﻭ ﻻﺭﻭﻫﺎ ﻱ ﺳﻦ ﺩﻭﻡ ﺷﻨﺎﺳﺎ ﻳﻲ ﺷﺪﻩ ﻭ ﺑﺮﺍ ﻱ ﻣ ﻲﺗﻮﺍﻧﺪ ﺑﺴ ﻴﺎﺭ ﻣﻔ ﻴﺪ ﺑﺎﺷﺪ. ﺩﺭ ﺍﻳﻦ ﺭﻭﺵ ﺍﺑﺘﺪﺍ ﻧﺎﺣﻴـ ﻪ ITS1- ﺁﺯﻣــﻮﻥ ﮐ ــ ﺎﺭﺍﻳﻲ ﺭﻭﺵﻫــﺎﻱ ﻣﺮﻓﻮﻟــﻮﮊﻳﮑﻲ ﺍﺯ ﺭﻭﺵ -ITS 5.8S-ITS2 ﺗﻮﺳﻂ ﺁﻏﺎﺯﮔﺮﻫﺎ ﻱ ﻋﻤﻮﻣ ﻲ ﺍ ﻳﻦ ﻧﺎﺣ ﻴـ ﻪ ﺗﮑﺜ ﻴـ ﺮ PCR-RFLP ﺍﺳــﺘﻔﺎﺩﻩ ﮔﺮﺩ ﻳ ــﺪ. ﺩﺭ ﻧﻬﺎ ﻳ ــﺖ ﺑــﺎ ﺗﻮﺟــﻪ ﺑــﻪ ﻣ ﻲﺷﻮﺩ ﻭ ﺳﭙﺲ ﻗﻄﻌﻪ ﺗﮑﺜ ﻴﺮ ﺷﺪﻩ ﺗﻮﺳﻂ ﺁﻧﺰ ﻳ ﻢﻫـﺎ ﻱ ﺑﺮﺷـ ﻲ ﺍﺧﺘﻼﻓﺎﺕ ﻣﺮﻓﻮﻟﻮﮊ ﻳﮑﻲ ﺯ ﻳﺎﺩﻱ ﮐﻪ ﺑ ﻴﻦ ﺟﻤﻌ ﻴ ﺖﻫﺎﻱ ﻣﺨﺘﻠﻒ ﻣﺨﺘﻠﻒ ﻣﻮﺭﺩ ﻫﻀﻢ ﻗﺮﺍﺭ ﻣ ﻲﮔﻴﺮﺩ. ﺍﻳﻦ ﺭﻭﺵ ﺩﺭ ﮐﻨـﺎﺭ ﺭﻭﺵ ﺍﻳﻦ ﮔﻮﻧﻪ ﻗﺎﺑﻞ ﻣﺸﺎﻫﺪﻩ ﺑـﻮﺩ، ﺗـﻼﺵ ﺷـﺪ ﺗـﺎ ﺑـﺎ ﺍﺳـﺘﻔﺎﺩﻩ ﺍﺯ ﺗﻮﺍﻟ ﻲﻳﺎﺑﻲ ﻧﺎﺣ ﻴﻪ ITS، ﺑﺮﺍ ﻱ ﺷﻨﺎﺳـﺎ ﻳﻲ ﻧﻤﺎﺗﻮﺩﻫـﺎ ﻱ ﺳ ﻴـ ﺴﺘﻲ ﮔﺮﻭﻩﺑﻨﺪﻱ ﻣﺮﻓﻮﻣﺘﺮ ﻳﮑﻲ ﺟﻤﻌﻴ ﺖﻫﺎﻱ ﻣﺨﺘﻠـﻒ، ﮔـﺮﻭﻩ ﺑﻨـﺪ ﻱ ﺍﻳــﺮﺍﻥ ﻣــﻮﺭﺩ ﺍﺳــﺘﻔﺎﺩﻩ ﻗﺮﺍﺭﮔﺮﻓﺘــﻪ ﻭ ﺗﻌــﺪﺍﺩ ﻣﺤــﺪﻭﺩ ﻱ ﻣﺮﻓﻮﻟﻮﮊﻳﮑﻲ ﻣﻨﺎﺳﺒ ﻲ ﺑﺮﺍ ﻱ ﺟﻤﻌ ﻴ ﺖﻫﺎﻱ ﻣﺨﺘﻠﻒ ﺍ ﻳـ ﻦ ﮔﻮﻧـﻪ ﺍﺯﺟﻤﻌﻴ ﺖﻫﺎﻱ ﮔﻮﻧـﻪ H. schachtii ﻧﻴـ ﺰ ﺷﻨﺎﺳـﺎ ﻳﻲ ﺷـﺪﻩ ﺗﻌﻴﻴﻦ ﺷﻮﺩ. Tanha Maafi et al. 2003 ﺍﺳﺖ ( ). ﻫﻢﭼﻨـ ﻴﻦ ﺍﺯ ﺍ ﻳـ ﻦ ﺭﻭﺵ ﺑـﺮﺍ ﻱ ﺷﻨﺎﺳـﺎ ﻳﻲ ﺟﻤﻌﻴـ ﺖﻫـﺎ ﻱ ﻣﻮﺍﺩ ﻭ ﺭﻭﺵﻫﺎ ﻣﺨﺘﻠﻒ ﮔﺮﻭﻩ avenae ﺩﺭ ﻣﺰﺍﺭﻉ ﭼﻐﻨﺪﺭﻗﻨﺪ ﺍﺳﺘﺎﻥ ﺧﺮﺍﺳـﺎﻥ ﺩﺭ ﻃﻮﻝ ﺳﺎﻝ ﻫﺎﻱ ۱۳۸۹-۱۳۸۷ ﺗﻌﺪﺍﺩ ۱۲۰ ﻧﻤﻮﻧـﻪ ﺧـﺎﮎ ﻭ ﺭﺿﻮﻱ ﺍﺳﺘﻔﺎﺩﻩ ﺷﺪﻩ ﺍﺳـﺖ (Mokaram Hesar et al. 2010). ﺭﻳﺸﻪ ﭼﻐﻨﺪﺭﻗﻨﺪ ﺍﺯ ﻣﺰﺍﺭﻉ ﺁﻟﻮﺩﻩ ﺑﻪ ﻧﻤﺎﺗﻮﺩ ﺳ ﻴﺴﺘﻲ ﭼﻐﻨﺪﺭﻗﻨﺪ ﺩﺭ ﻣﻮﺭﺩ ﮔﻮﻧـﻪ H. schachtii ﻧـﺸﺎﻥ ﺩﺍﺩﻩ ﺷـﺪﻩ ﺍﺳـﺖ ﮐـﻪ ﺩﺭ ﺍﺳﺘﺎﻥ ﺧﺮﺍﺳﺎﻥ ﺭﺿﻮ ﻱ ﺟﻤﻊ ﺁﻭﺭﻱ ﮔﺮﺩ ﻳﺪ ﮐﻪ ﺍﺯ ﻣ ﻴﺎﻥ ﺁﻧﻬﺎ ﻫﻀﻢ ﻗﻄﻌﻪ ﺗﮑﺜ ﻴﺮﻱ ﻣﻮﺭﺩ ﻧﻈﺮ ﺑـﺎ ﺍﺳـﺘﻔﺎﺩﻩ ﺍﺯ ﺁﻧـﺰ ﻳﻢ ﺑﺮﺷـ ﻲ ۸۸ ﺟﻤﻌ ﻴﺖ ﮐﻪ ﻣﺸﺨـﺼﺎﺕ ﺁﻧﻬـﺎ ﺩﺭ ﺟـﺪﻭﻝ ۱ ﺁﻭﺭﺩﻩ ﺷـﺪﻩ MvaI ﻣ ﻲﺗﻮﺍﻧﺪ ﺍﻟﮕﻮ ﻱ ﺑﺮﺷ ﻲ ﻣﻨﺤـﺼﺮ ﺑـﻪ ﻓـﺮﺩ ﻱ ﺭﺍ ﺑـﺮﺍ ﻱ ﺍﺳﺖ، ﺍﻧﺘﺨﺎﺏ ﺷ ﺪﻧﺪ. ﺟﻬﺖ ﺍﺳﺘﺨﺮﺍﺝ ﻧﻤﺎﺗﻮﺩﻫﺎ ﻱ ﺳ ﻴﺴﺘﻲ ﺍﺯ ﺟﻤﻌﻴــ ﺖﻫــﺎﻱ ﻣﺨﺘﻠــﻒ ﺍﻳــﻦ ﮔﻮﻧــﻪ ﺍﻳﺠــﺎﺩ ﮐﻨــﺪ ﻭ ﺁﻥ ﺭﺍ ﺍﺯ ﺧــــﺎﮎ، ﺗﺮﮐﻴﺒــــﻲ ﺍﺯ ﺭﻭﺵﻫــــﺎﻱ (Dunn, (1969 ﮔﻮﻧﻪﻫﺎﻱ ﻧﺰﺩﻳﮏ ﺑﻪ ﺁﻥ ﺩﺭ ﮔﺮﻭﻩ ﻣﺘﻤﺎﻳﺰ ﻧﻤﺎﻳـ ﺪ ( Amiri et ﻭ (Seinhorst, (1964 ﺍﺳﺘﻔﺎﺩﻩ ﺷﺪ. ﺑﺮﺵﻫﺎﻱ ﻻﺯﻡ ﺍﺯ ﻣﺨـﺮﻭﻁ .(al. 2001, 2002; Tanha Maafi et al. 2003 ﺍﻧﺘﻬﺎﻱ ﺳﻴﺴﺖ ﻫﺎ ﺗﻬﻴﻪ ﮔﺮﺩﻳﺪ . ﻫﻢﭼﻨﻴﻦ ﺑﺮﺍﻱ ﺗﺜﺒﻴﺖ ﻻﺭﻭﻫﺎﻱ ﺳـﻦ ﻧﺨﺴﺘﻴﻦ ﮔـﺎﻡ ﺑـﺮﺍ ﻱ ﺍﻋﻤـﺎﻝ ﺭﻭﺵ ﻫـﺎ SIDﻱ ﻣﺨﺘﻠـﻒ ofﻣـﺪ ﻳﺮﻳ ﺘﻲ Archive ﺩﻭﻡ ﺩﺍﺧـﻞ ﺳﻴـﺴﺖ ﻫـﺎ ﺍﺯ ﺭﻭﺵ (De Grisse (1969 ﺍﺳـﺘﻔﺎﺩﻩ ﺷﻨﺎﺳﺎﻳﻲ ﺻﺤﻴﺢ ﮔﻮﻧﻪ ﻧﻤﺎﺗﻮﺩ ﻣ ﻲﺑﺎﺷﺪ. ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ ﺍﺧﺘﻼﻓﺎﺕ ﺷﺪ.
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