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Isolation and Identification of Maple Tar Spot Pathogen in Acer Velutinum Trees in Dr

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Isolation and Identification of Maple Tar Spot Pathogen in Acer Velutinum Trees in Dr ﺑﻮمﺷﻨﺎﺳﯽ ﺟﻨﮕﻞﻫﺎي اﯾﺮان ﺳﺎل دوم/ ﺷﻤﺎره ﺳﻮم/ ﺑﻬﺎر و ﺗﺎﺑﺴﺘﺎن 1393 ................................................................................................... 26 داﻧﺸﮕﺎه ﻋﻠﻮم ﮐﺸﺎورزي و ﻣﻨﺎﺑﻊ ﻃﺒﯿﻌﯽ ﺳﺎري ﺑﻮمﺷﻨﺎﺳﯽ ﺟﻨﮕﻞﻫﺎي اﯾﺮان ﺟﺪاﺳﺎزي و ﺷﻨﺎﺳﺎﯾﯽ ﻋﺎﻣﻞ ﺑﯿﻤﺎري ﻟﮑﻪ ﻗﯿﺮي درﺧﺘﺎن اﻓﺮاﭘﻠﺖ در ﺟﻨﮕﻞ آﻣﻮزﺷﯽ و ﭘﮋوﻫﺸﯽ دﮐﺘﺮ ﺑﻬﺮامﻧﯿﺎ ﺷﻬﺮام ﻣﻬﺪي ﮐﺮﻣﯽ1، ﻣﺤﻤﺪرﺿﺎ ﮐﺎوﺳﯽ2 و اﮐﺮم اﺣﻤﺪي3 1- داﻧﺶآﻣﻮﺧﺘﻪ ﮐﺎرﺷﻨﺎﺳﯽ ارﺷﺪ، داﻧﺸﮕﺎه ﻋﻠﻮم ﮐﺸﺎورزي و ﻣﻨﺎﺑﻊ ﻃﺒﯿﻌﯽ ﮔﺮﮔﺎن، (ﻧﻮﯾﺴﻨﺪه ﻣﺴﺌﻮل: [email protected]) 2 و 3- داﻧﺸﯿﺎر و داﻧﺸﺠﻮي دﮐﺘﺮي، داﻧﺸﮕﺎه ﻋﻠﻮم ﮐﺸﺎورزي و ﻣﻨﺎﺑﻊ ﻃﺒﯿﻌﯽ ﮔﺮﮔﺎن ﺗﺎرﯾﺦ درﯾﺎﻓﺖ: 2/9/1392 ﺗﺎرﯾﺦ ﭘﺬﯾﺮش: 1393/2/10 ﭼﮑﯿﺪه ﻟﮑﻪ ﻗﯿﺮي از ﺟﻤﻠﻪ ﺑﯿﻤﺎريﻫﺎي ﻗﺎرﭼﯽ اﺳﺖ ﮐﻪ درﺧﺘﺎن اﻓﺮا را ﻣﺒﺘﻼ ﻧﻤﻮده اﺳﺖ. اﯾﻦ ﺑﯿﻤﺎري ﺑﺎﻋﺚ ﺧﺰان زودرس، از ﺑﯿﻦ رﻓﺘﻦ ﺑﺮگ، ﺳﺒﺐ ﮐﺎﻫﺶ رﺷﺪ، ﮐﺎﻫﺶ ﺗﻮﻟﯿﺪ ﭼﻮب و ﺑﺬر درﺧﺘﺎن اﻓﺮا ﻣﯽﮔﺮدد. ﺑﻪ ﻫﻤﯿﻦ ﻣﻨﻈﻮر ﺟﺪاﺳﺎزي و ﺷﻨﺎﺳﺎﯾﯽ ﻋﺎﻣﻞ ﺑﯿﻤﺎري روي ﺑﺮگ درﺧﺘﺎن اﻓﺮاﭘﻠﺖ در ﺟﻨﮕﻞ آﻣﻮزﺷﯽ و ﭘﮋوﻫﺸﯽ دﮐﺘﺮ ﺑﻬﺮامﻧﯿﺎ (ﺷﺼﺖﮐﻼﺗﻪ ﮔﺮﮔﺎن) اﻧﺠﺎم ﮔﺮﻓﺖ. ﭘﺲ از ﺗﻬﯿﻪ ﻧﻤﻮﻧﻪ و ﮐﺸﺖ آن روي ﻣﺤﯿﻂ PDA، ﭘﺮﮔﻨﻪ ﻗﺎرچ ﻋﺎﻣﻞ ﺑﯿﻤﺎري ﮐﻪ ﺳﻔﯿﺪ رﻧﮓ و ﮐﺮﮐﯽ ﺷﮑﻞ ﺑﻮد، ﺑﺪﺳﺖ آﻣﺪ. ﻋﺎﻣﻞ ﺑﯿﻤﺎريزاﯾﯽ ﭘﺲ از ﺧﺎﻟﺺﺳﺎزي، ﺟﺪاﺳﺎزي و ﺑﺮرﺳﯽ ﺧﺼﻮﺻﯿﺎت رﻧﮓ و ﺷﮑﻞ اﺳﭙﻮر، ﻋﻼﯾﻢ ﺑﯿﻤﺎري ﺷﻨﺎﺳﺎﯾﯽ ﮔﺮدﯾﺪ. در ﻣﺤﯿﻂ ﮐﺸﺖ ﻣﺎﯾﻊ PDA، آﺳﮏﻫﺎي ﻗﺎرچ ﺳﻔﯿﺪ رﻧﮓ و ﭼﻤﺎﻗﯽ ﺷﮑﻞ ﺑﻮدﻧﺪ و آﺳﮑﻮﺳﭙﻮرﻫﺎي ﺗﮏ ﺳﻠﻮﻟﯽ ﻧﺨﯽ ﺷﮑﻞ و ﮐﺸﯿﺪه در درون آﻧﻬﺎ ﻣﺸﺎﻫﺪه ﺷﺪ. ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ ﺑﺮرﺳﯽ ﺑﻪﻋﻤﻞ آﻣﺪه، ﻋﺎﻣﻞ ﺑﯿﻤﺎري ﻟﮑﻪ ﻗﯿﺮي در درﺧﺘﺎن اﻓﺮا در ﻣﻨﻄﻘﻪ ﻣﻮرد ﺗﺤﻘﯿﻖ، ﻗﺎرچ Rhytisma acerinum ﺗﺸﺨﯿﺺ داده ﺷﺪ. Downloaded from ifej.sanru.ac.ir at 23:41 +0330 on Tuesday September 28th 2021 واژهﻫﺎي ﮐﻠﯿﺪي: اﻓﺮاﭘﻠﺖ، ﻟﮑﻪ ﻗﯿﺮي، Rhytisma acerinum ﻣﻘﺪﻣﻪ ﻣﯽﺑﺎﺷﻨﺪ (13). ﯾﮑﯽ از ﻣﻬﻢﺗﺮﯾﻦ ﺑﯿﻤﺎريﻫﺎي 1 درﺧﺖ اﻓﺮاﭘﻠﺖ (Acer velutinum) ﺟﺰء ﺗﯿﺮه درﺧﺖ اﻓﺮا، ﺑﯿﻤﺎري ﻟﮑﻪ ﻗﯿﺮي ﻣﯽﺑﺎﺷﺪ ﮐﻪ ﺑﻪ Aceraceae، راﺳﺘﻪ Sapindales و در ﺷﺎﺧﻪ اﺳﺎﻣﯽ ﻟﮑﻪ ﺳﯿﺎه ﺑﺮگ اﻓﺮا و ﻣﺮض ﺳﯿﺎه ﭘﻮﺳﺖ Magnoliophyta ﻗﺮار دارد. درﺧﺘﺎن اﻓﺮا از اﻓﺮا ﻧﺎﻣﯿﺪه ﻣﯽﺷﻮد. از ﻋﻮاﻣﻞ ﺑﯿﻤﺎري ﻟﮑﻪ ﻗﯿﺮي ﻣﻬﻢﺗﺮﯾﻦ ﮔﻮﻧﻪﻫﺎي ﺻﻨﻌﺘﯽ ﮐﺸﻮر ﺑﻪ ﺣﺴﺎب ﺑـﺮگ درﺧﺘـﺎن اﻓـﺮا، ﻗـﺎرچﻫـﺎي Rhytisma ﻣﯽآﯾﻨﺪ ﮐﻪ داراي ﺣﺠﻤﯽ ﺗﺎ ﺣﺪود 5/7 درﺻﺪ ﺑﺎ R. Punctatum ،acerinum و R. Salicium 2 ﻣﺘﻮﺳﻂ ﻣﻮﺟﻮدي 210 ﻣﺘﺮﻣﮑﻌﺐ در ﻫﮑﺘﺎر ﻣﯽﺑﺎﺷﻨﺪ ﮐﻪ در رده آﺳﮑﻮﻣﯿﺴﺖﻫﺎ ﻗﺮار دارﻧﺪ 1- Rhytisma acerinum 2- Ascomycetes ﺑﻮمﺷﻨﺎﺳﯽ ﺟﻨﮕﻞﻫﺎي اﯾﺮان ﺳﺎل دوم/ ﺷﻤﺎره ﺳﻮم/ ﺑﻬﺎر و ﺗﺎﺑﺴﺘﺎن 1393 ................................................................................................................................. 27 (15،8،2،1). اﯾﻦ ﺑﯿﻤﺎري ﺑﺮاي اوﻟﯿﻦ ﺑﺎر در اﯾﺮان ﻟﮑﻪﻫﺎي ﺳﯿﺎه رﻧﮓ روي ﺑﺮگ درﺧﺘﺎن اﻓﺮا در ﺳﺎل 1318 ﺗﻮﺳﻂ ﭘﺘﺮاك در ﻣﻨﺎﻃﻖ ﺷﻤﺎل و ﻣﯽﺷﻮد. اﻃﺮاف ﺗﻬﺮان ﻣﺸﺎﻫﺪه ﺷﺪ (2). ﻣﻄﺎﻟﻌﺎﺗﯽ در ﻣﻮرد ﺑﯿﻤﺎريزاﯾﯽ، ﭼﺮﺧﻪ ﻧﺸﺎﻧﻪﻫﺎي ﺑﯿﻤﺎري در ﺗﯿﺮﻣﺎه روي ﺑﺮگ زﻧﺪﮔﯽ و ﺳﯿﺘﻮﻟﻮژي اﯾﻦ ﻗﺎرچ اﻧﺠﺎم ﺷﺪه اﺳﺖ. درﺧﺘﺎن اﻓﺮا ﺑﻪﺻﻮرت ﻟﮑﻪﻫﺎي ﮔﺮدي ﺑﻪ ﻗﻄﺮ روﻧﺎﻟﺪ و ﻫﻤﮑﺎران (14) و ﭘﺎرﺗﺮﯾﺪﯾﮓ و وو (16)، 8 -7 ﻣﯿﻠﯽﻣﺘﺮ ﺗﺸﮑﯿﻞ ﻣﯽﮔﺮدد. ﺗﻌﺪاد اﯾﻦ ﻟﮑﻪﻫﺎ ﭼــﺮﺧــﻪ زﻧــﺪﮔﯽ و ﺳﯿــﺘﻮﻟﻮژي ﻗـــﺎرچ زﯾﺎد و ﮔﺎﻫﯽ ﺗﺎ 50 ﻋﺪد ﻧﯿﺰ در ﺳﻄﺢ ﺑﺮگ R. punctaphylum را ﺑﺮرﺳﯽ ﻧﻤﻮدﻧﺪ (16). ﻣﯽرﺳﺪ. رﻧﮓ ﻟﮑﻪﻫﺎ اﺑﺘﺪا زرد رﻧﮓ، ﺳﭙﺲ ﻫﻤﭽﻨﯿﻦ دوراوﺗﺰ و ﻣﻮرﮔﺎن (6) ﻣﺮاﺣﻞ رﺷﺪ و ﻗﻬﻮهاي و در ﻧﻬﺎﯾﺖ ﺳﯿﺎه رﻧﮓ ﺷﺪه ﮐﻪ ﺧﺰان ﻧﻤﻮ آﺳﮑﻮﮐﺎرپ در ﻗﺎرچﻫﺎي ﻋﺎﻣﻞ ﻟﮑﻪ ﻗﯿﺮي اﻓﺮا زودرس ﺑﺮگﻫﺎ را ﺑﺎﻋﺚ ﻣﯽﮔﺮدد. اﯾﻦ ﺑﯿﻤﺎري را ﻣﻮرد ﻣﻄﺎﻟﻌﻪ ﻗﺮار دادﻧﺪ. ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ ﻧﺘﺎﯾﺞ، ﻋﻼوه ﺑﺮ درﺧﺘﺎن ﻣﺴﻦ، در ﺧﺰاﻧﻪﻫﺎ روي ﺷﮑﻞ اﺳﭙﻮرﻫﺎي ﻗﺎرچ R. acerinum ﺑﻪ ﺻﻮرت ﻧﻬﺎلﻫﺎي ﺟﻮان درﺧﺘﺎن اﻓﺮا ﻧﯿﺰ ﮔﺴﺘﺮش دارد و ﻧﺨﯽ ﺷﮑﻞ و ﮐﺸﯿﺪه و ﺑﻪ ﺻﻮرت ﺑﯽرﻧﮓ ﻣﯿﺰان ﺧﺴﺎرت آن در ﻧﻬﺎﻟﺴﺘﺎن ﺑﯿﺸﺘﺮ اﺳﺖ. ﻣﯽﺑﺎﺷﻨﺪ (9،7،4). ﻧﺤﻮهي اﻧﺘﺸﺎر اﯾﻦ ﻗﺎرچ ﺑﻪ اﯾﻦ ﺻﻮرت ﻣﯽﺑﺎﺷﺪ از ﺟﻤﻠﻪ ﺧﺴﺎرات اوﻟﯿﻪ اﯾﻦ ﺑﯿﻤﺎري ﺧﺰان ﮐﻪ اﺳﺘﺮوﻣﺎي اﯾﻦ ﻗﺎرچ در ﭘﻬﻨﮏ ﺑﺮگ در زﯾﺮ زودرس ﺑﺮگ درﺧﺘﺎن ﻣﯽﺑﺎﺷﺪ ﮐﻪ ﺑﺮ روي ﻧﺤﻮه ﻻﯾﻪ اﭘﯿﺪرﻣﯽ ﮔﺴﺘﺮش ﻣﯽﯾﺎﺑﺪ و ﺑﻪ ﺗﺪرﯾﺞ رﺷﺪ درﺧﺖ ﺗﺎﺛﯿﺮ ﻣﯽﮔﺬارد (1). اﯾﻦ ﺗﺤﻘﯿﻖ ﺟﺎﻧﺸﯿﻦ ﺑﺎﻓﺖﻫﺎي ﻣﯿﺰﺑﺎن ﻣﯽﺷﻮد (7،4،2). ﺑﻪﻣﻨﻈﻮر ﺷﻨﺎﺳﺎﯾﯽ ﻋﺎﻣﻞ اﯾﻦ ﺑﯿﻤﺎري روي ﺑﺮگ اﻓﺮاﭘﻠﺖ، ﮐﺮب، ﺷﯿﺮدار، ﺳﻔﯿﺪﮐﺮﮐﻮ، ﮐﺮﮐﻮ، اﻓﺮاي درﺧﺘﺎن اﻓﺮا ﭘﻠﺖ در ﺟﻨﮕﻞ آﻣﻮزﺷﯽ و ﭘﮋوﻫﺸﯽ ﺳﯿﺎه، ﮐﺮﮐﻒ و اﻓﺮاي ﺷﺒﻪﭼﻨﺎري از ﮔﻮﻧﻪﻫﺎي دﮐﺘﺮ ﺑﻬﺮامﻧﯿﺎ (ﺷﺼﺖﮐﻼﺗﻪ ﮔﺮﮔﺎن) ﺑﺮاي ﻣﺪﯾﺮﯾﺖ ﻣﻬﻢ ﺟﻨﺲ اﻓﺮا (Acer) ﻫﺴﺘﻨﺪ ﮐﻪ در اﯾﺮان ﻣﻮرد و ﺣﻔﺎﻇﺖ ﺑﻬﺘﺮ درﺧﺘﺎن اﻓﺮا اﻧﺠﺎم ﮔﺮﻓﺘﻪ اﺳﺖ. ﺣﻤﻠﻪ ﻗﺎرچﻫﺎي ﻋﺎﻣﻞ ﺑﯿﻤﺎري ﻟﮑﻪ ﻗﯿﺮي ﻗﺮار Downloaded from ifej.sanru.ac.ir at 23:41 +0330 on Tuesday September 28th 2021 ﮔﺮﻓﺘﻪاﻧﺪ. ﻣﺤﻞ اﻧﺘﺸﺎر اﯾﻦ ﺑﯿﻤﺎري در ﺳﻮاﺣﻞ ﻣﻮاد و روشﻫﺎ درﯾﺎي ﺧﺰر، اﻃﺮاف ﺗﻬﺮان، ﮐﺮج و اﺣﺘﻤﺎﻻً ﺳﺎﯾﺮ ﻧﻤﻮﻧﻪﺑﺮداري ﻧﻘﺎط ﮐﺸﻮر ﻣﯽﺑﺎﺷﺪ (10،7،2،1). ﮐﺎﻧﻮن و ﺑﻪﻣﻨﻈﻮر ﮐﺸﺖ و ﺗﺸﺨﯿﺺ ﻋﺎﻣﻞ ﺑﯿﻤﺎري ﻫﻤﮑﺎران (5) در ﺗﺤﻘﯿﻘﺎت ﺧﻮد ﭘﯽ ﺑﺮدﻧﺪ ﮐﻪ ﻟﮑﻪﻗﯿﺮي، ﻧﻤﻮﻧﻪ ﺑﺮگ آﻟﻮده درﺧﺘﺎن اﻓﺮا ﭘﻠﺖ ﺑﻪ R. acerinum روي ﺑﺴﯿﺎري از ﮔﻮﻧﻪﻫﺎي اﻓﺮا در ﻟﮑﻪ ﻗﯿﺮي از ﺟﻨﮕﻞ آﻣﻮزﺷﯽ و ﭘﮋوﻫﺸﯽ دﮐﺘﺮ اروﭘﺎ و آﻣﺮﯾﮑﺎ ﭘﺮاﮐﻨﺶ دارﻧﺪ ﻫﻢﭼﻨﯿﻦ ﻻﭘﻮﯾﻨﺖ ﺑﻬﺮامﻧﯿﺎ (ﺷﺼﺖﮐﻼﺗﻪ ﮔﺮﮔﺎن) واﻗﻊ در ﺟﻨﻮب و ﺑﺮﯾﺴﻮن (12) در ﮐﺸﻮر آﻣﺮﯾﮑﺎ ﺑﻪ ﺑﺮرﺳﯽ ﻗﺎرچ ﻏﺮﺑﯽ ﺷﻬﺮ ﮔﺮﮔﺎن ﺑﻪ ﺻﻮرت ﺗﺼﺎدﻓﯽ ﺟﻤﻊآوري R. acerinum ﭘﺮداﺧﺘﻨﺪ ﮐﻪ ﺳﺒﺐ ﺑﻪوﺟﻮد آﻣﺪن ﮔﺮدﯾﺪ. ﺟﺪاﺳﺎزي و ﺷﻨﺎﺳﺎﯾﯽ ﻋﺎﻣﻞ ﺑﯿﻤﺎري ﻟﮑﻪ ﻗﯿﺮي درﺧﺘﺎن اﻓﺮاﭘﻠﺖ ..................................................................................................................................................... 28 ﺟﺪاﺳﺎزي و ﺷﻨﺎﺳﺎﯾﯽ ﻗﺎرچ ﻋﺎﻣﻞ ﺑﯿﻤﺎري ﻟﮑﻪ ﻧﺘﺎﯾﺞ و ﺑﺤﺚ ﻗﯿﺮي اﻓﺮاﭘﻠﺖ ﻋﻼﺋﻤﯽ ﮐﻪ ﺗﻮﺳﻂ اﯾﻦ ﺑﯿﻤﺎري روي درﺧﺘﺎن ﺑﺮاي ﺟﺪاﺳﺎزي ﻗﺎرچ ﻋﺎﻣﻞ ﺑﯿﻤﺎري، اﻓﺮا ﻣﺸﺎﻫﺪه ﮔﺮدﯾﺪ، اﯾﺠﺎد ﻟﮑﻪﻫﺎي ﺳﺒﺰ ﻣﺎﯾﻞ ﺑﻪ ﻧﻤﻮﻧﻪﻫﺎﯾﯽ ﺑﻪ اﺑﻌﺎد 5×5 ﻣﯿﻠﯽﻣﺘﺮ از روي زرد رﻧﮓ ﺑﻪ ﺻﻮرت ﮔﺮوﻫﯽ روي ﺑﺮگﻫﺎ، ﺗﺒﺪﯾﻞ ﺑﺮگﻫﺎي ﻣﺒﺘﻼ ﺑﻪ ﻗﺎرچ ﺟﺪا ﺷﺪﻧﺪ (ﺷﮑﻞ 1). ﺷﺪن ﻟﮑﻪﻫﺎ ﺑﻪ ﻟﮑﻪﻫﺎي ﻗﯿﺮي ﺷﮑﻞ ﺑﺎ ﺣﺎﺷﯿﻪ زرد ﺑﺮاي ﺿﺪﻋﻔﻮﻧﯽ ﻧﻤﻮﻧﻪﻫﺎ 3-2 دﻗﯿﻘﻪ در ﻣﺤﻠﻮل رﻧﮓ ﺑﻮد اﺳﺘﺮوﻣﺎي ﻗﺎرچ ﺑﻪ ﺧﻮﺑﯽ در ﺳﻄﺢ ﺑﺮگ ﻫﯿﭙﻮﮐﻠﺮﯾﺖ ﺳﺪﯾﻢ 10 درﺻﺪ ﻗﺮار داده ﺷﺪﻧﺪ و ﻗﺎﺑﻞ ﻣﺸﺎﻫﺪه ﺑﻮد (ﺷﮑﻞ 2). روي ﻫﺮ ﻧﺎﺣﯿﻪ ﺳﭙﺲ ﺑﺎ اﻟﮑﻞ 96% ﺑﺮاي ﻣﺪت زﻣﺎن 1 دﻗﯿﻘﻪ آﺳﯿﺐ دﯾﺪه دواﯾﺮي ﺑﻮﺟﻮد ﻣﯽآﯾﺪ ﮐﻪ ﮐﻨﯿﺪي و اﺳﺘﺮﯾﻞ ﺷﺪﻧﺪ. ﭘﺲ از اﺳﺘﺮﯾﻞ ﻧﻤﻮﻧﻪﻫﺎ، در ﺷﺮاﯾﻂ در داﺧﻞ ﻫﺮ ﮐﺪام ﮐﻨﯿﺪﯾﻮﻓﻮرﻫﺎ ﺗﺸﮑﯿﻞ ﻣﯽﺷﻮﻧﺪ. آزﻣﺎﯾﺸﮕﺎﻫﯽ در زﯾﺮ ﻻﻣﯿﻨﺎر اﯾﺮﻓﻠﻮ در ﻣﺤﯿﻂ ﻫﻤﭽﻨﯿﻦ، ﻧﺘﺎﯾﺞ ﺗﺤﻘﯿﻖ ﺣﺎﺻﻞ از ﮐﺸﺖ ﺣﺎﺷﯿﻪ 1 ﮐﺸﺖ PDA ﺑﺎ اﺳﯿﺪﯾﺘﻪ 5/4 ﮐﺸﺖ ﺷﺪﻧﺪ و در ﻟﮑﻪﻫﺎي روي ﺑﺮگ ﻧﺸﺎن داد ﮐﻪ ﺷﮑﻞ ﭘﺮﮔﻨﻪاي دﺳﺘﮕﺎه اﻧﮑﻮﺑﺎﺗﻮر در دﻣﺎي 1±25 درﺟﻪ ﮐﻪ در اﺛﺮ ﻗﺎرچ ﻋﺎﻣﻞ ﺑﯿﻤﺎري روي ﻟﮑﻪ ﻗﯿﺮي اﻓﺮا ﺳﺎﻧﺘﯽﮔﺮاد و در ﺷﺮاﯾﻂ ﺗﺎرﯾﮑﯽ ﻗﺮار داده ﺷﺪﻧﺪ. در ﻣﺤﯿﻂ ﮐﺸﺖ ﺑﻮﺟﻮد آﻣﺪ، ﺳﻔﯿﺪ رﻧﮓ، ﮐﺮﮐﯽ در ﻧﻬﺎﯾﺖ ﺳﻪ ﻫﻔﺘﻪ ﺑﻌﺪ از ﮐﺸﺖ ﻗﺎرچ، ﻣﺠﺪد ﺷﮑﻞ و داراي رﺷﺪي ﺑﻪ ﺻﻮرت ﺷﻌﺎﻋﯽ ﺑﻮد ﻋﻤﻞ ﺧﺎﻟﺺﺳﺎزي ﻗﺎرچ اﻧﺠﺎم ﮔﺮﻓﺖ. (ﺷﮑﻞ 3) ﮐﻪ ﺑﻌﺪ از 2 ﺗﺎ 3 ﻫﻔﺘﻪ رﺷﺪ ﻗﺎرچ ﮐﺎﻣﻞ ﻫﻤﭽﻨﯿﻦ، ﺑﻌﺪ از اﻧﺠﺎم ﻣﺮاﺣﻞ ﺧﺎﻟﺺﺳﺎزي و و در ﻣﺪت زﻣﺎن ﯾﮏ ﻫﻔﺘﻪ داراي ﻣﯿﺎﻧﮕﯿﻦ رﺷﺪي ﺑﺪﺳﺖ آوردن ﭘﺮﮔﻨﻪ ﻗﺎرچ، ﻣﻘﺪاري از ﭘﺮﮔﻨﻪ ﻗﺎرچ 8/41 ﻣﯿﻠﯽﻣﺘﺮ ﺑﺎ اﻧﺤﺮاف ﻣﻌﯿﺎر 78/6± ﺑﻮد. ﺗﻮده در ﻣﺤﯿﻂ ﮐﺸﺖ (ﻣﺎﯾﻊ ﺳﯿﺐزﻣﯿﻨﯽ و آب ﻣﻘﻄﺮ) ﻣﯿﺴﯿﻠﯿﻮﻣﯽ ﺗﺸﮑﯿﻞ ﺷﺪه ﺑﻪ ﺷﮑﻞ ﮐﺮوي و از ﻫﻢ روي دﺳﺘﮕﺎه ﺷﯿﮑﺮ ﺑﻪ ﻣﺪت ﯾﮏ ﻣﺎه ﻗﺮار داده ﺟﺪا ﺑﻮد (ﺷﮑﻞ 4). در اﺛﺮ اﺳﺘﻘﺮار اﺳﺘﺮوﻣﺎي ﻗﺎرچ ﺷﺪ. ﺟﻬﺖ ﺟﺪاﺳﺎزي اﺳﭙﻮرﻫﺎ از ﺗﻮده ﻣﯿﺴﯿﻠﯿﻮﻣﯽ ﻋﺎﻣﻞ ﻟﮑﻪ ﻗﯿﺮي اﻓﺮا روي ﺑﺮگ درﺧﺖ اﻓﺮا، در ﺗﺸﮑﯿﻞ ﺷﺪه، اﺳﺘﻔﺎده ﮔﺮدﯾﺪ. ﺳﻮﺳﭙﺎﻧﺴﯿﻮن ﺗﻬﯿﻪ درون اﺳﺘﺮوﻣﺎ، ﭘﯿﮑﻨﯿﺪﻫﺎ و ﺑﻌﺪ ﭘﺮﯾﺘﺴﯿﻮم ﮐﻪ Downloaded from ifej.sanru.ac.ir at 23:41 +0330 on Tuesday September 28th 2021 ﺷﺪه، از داﺧﻞ دﺳﺘﮕﺎه ﺧﻼء ﻋﺒﻮر داده ﺷﺪه و ﭼﻤﺎﻗﯽ ﺷﮑﻞ، ﺷﻔﺎف و ﺑﯽرﻧﮓ ﺑﻮدﻧﺪ، ﻣﺸﺎﻫﺪه ﺳﭙﺲ ﻋﺼﺎره ﺑﺪﺳﺖ آﻣﺪه روي دﺳﺘﮕﺎه ﮔﺮدﯾﺪ (اﺷﮑﺎل 5 و 6). در داﺧﻞ ﭘﯿﮑﻨﯿﺪ ﻋﺪﺳﯽ ﺳﺎﻧﺘﺮﯾﻔﯿﻮژ ﺑﺎ 3000 دور در دﻗﯿﻘﻪ ﺑﺮاي ﺳﻪ ﺷﮑﻞ، ﮐﻨﯿﺪيﻫﺎي ﮐﻮﭼﮏ ﮐﺸﯿﺪه و ﺑﯿﻀﻮي ﻣﺪت زﻣﺎن 20، 40 و 60 ﻗﺮار داده ﺷﺪ. ﺑﺎ ﺗﻮﺟﻪ ﺷﮑﻞ ﻣﺸﺎﻫﺪه ﮔﺮدﯾﺪ. آﺳﮏﻫﺎي ﻗﺎرچ ﺑﻪ رﻧﮓ ﺑﻪ ﻧﺘﺎﯾﺞ ﺑﺪﺳﺖ آﻣﺪه اﺳﭙﻮرﻫﺎي ﺑﺪﺳﺖ آﻣﺪه ﺑﺎ ﺳﻔﯿﺪ و ﭼﻤﺎﻗﯽ ﺷﮑﻞ ﻣﺸﺎﻫﺪه ﺷﺪﻧﺪ. در داﺧﻞ دور 3000 دور در دﻗﯿﻘﻪ ﺑﺮاي ﻣﺪت زﻣﺎن 40 ﻫﺮ آﺳﮏ، دﺳﺘﻪاي از آﺳﮑﻮﺳﭙﻮرﻫﺎي وﺟﻮد دﻗﯿﻘﻪ ﻧﺘﺎﯾﺞ ﺑﻬﺘﺮي را ﺑﺮاي ﺷﻨﺎﺳﺎﯾﯽ ﻗﺎرچ ﻧﺸﺎن داﺷﺖ ﮐﻪ ﻧﺨﯽ ﺷﮑﻞ، ﺳﺎده، ﺷﻔﺎف و ﺑﯽرﻧﮓ داد. ﺑﻌﺪ از ﺟﻮاﻧﻪزﻧﯽ اﺳﭙﻮرﻫﺎ، ﻣﻄﺎﻟﻌﺎت ﺑﻌﺪي ﺑﺎ ﺑﻮدﻧﺪ (4). آﺳﮑﻮﺳﭙﻮرﻫﺎ ﺗﮏ ﺳﻠﻮﻟﯽ و ﺳﻮزﻧﯽ اﺳﺘﻔﺎده از ﻣﯿﮑﺮوﺳﮑﻮپ ﻧﻮري ﺻﻮرت ﮔﺮﻓﺖ. ﺷﮑﻞ ﺑﻮدﻧﺪ (14). آﺳﮑﻮﺳﭙﻮرﻫﺎ ﭼﺴﺒﻨﺎك ﻫﺴﺘﻨﺪ 1- Potato Dextrose Agar ﺑﻮمﺷﻨﺎﺳﯽ ﺟﻨﮕﻞﻫﺎي اﯾﺮان ﺳﺎل دوم/ ﺷﻤﺎره ﺳﻮم/ ﺑﻬﺎر و ﺗﺎﺑﺴﺘﺎن 1393 ................................................................................................................................. 29 و روي ﺳﻄﺢ ﺑﺮگ ﺟﻮاﻧﻪ ﻣﯽزﻧﻨﺪ و از راه روزﻧﻪﻫﺎ ﻣﺮﺣﻠﻪ ﺗﻮﻟﯿﺪﻣﺜﻞ ﺟﻨﺴﯽ ﻗﺎرچ اﺳﺖ ﮐﻪ در ﻓﺼﻞ وارد ﺑﺮگﻫﺎ ﻣﯽﺷﻮﻧﺪ (6) و ﻣﻮﺟﺐ آﻟﻮدﮔﯽ زﻣﺴﺘﺎن ﯾﺎ اواﯾﻞ ﺑﻬﺎر روي ﺑﺮگﻫﺎي اﻓﺘﺎده ﺳﻠﻮلﻫﺎي اﭘﯿﺪرم و ﻣﺰوﻓﯿﻞ ﻣﯽﺷﻮﻧﺪ. ﺑﺎ رﺷﺪ ﺗﺸﮑﯿﻞ ﻣﯽﮔﺮدد و ﺑﺎ ﮐﻤﮏ ﺑﺎد ﺑﺎﻋﺚ اﻧﺘﺸﺎر و ﻗﺎرچ، ﺳﻠﻮلﻫﺎ ﭘﺎره ﺷﺪه و اﺳﺘﺮوﻣﺎﯾﯽ ﺑﺎ ﭘﻮﺷﺶ اﻧﺘﻘﺎل ﺑﯿﻤﺎري ﻣﯽﺷﻮﻧﺪ (8،2). ﺳﯿﺎه رﻧﮓ ﺗﺸﮑﯿﻞ ﻣﯽدﻫﻨﺪ. ﻣﺮﺣﻠﻪ ﭘﺮﯾﺘﺴﯿﻮم، ﺷﮑﻞ 1- ﺑﯿﻤﺎري ﻟﮑﻪ ﻗﯿﺮي روي ﺑﺮگ درﺧﺖ اﻓﺮا ﺷﮑﻞ 2- اﺳﺘﺮوﻣﺎي ﻗﺎرچ R. acerinum روي ﺑﺮگ اﻓﺮاﭘﻠﺖ ﺷﮑﻞ 3- ﭘﺮﮔﻨﻪ ﻗﺎرچ R. acerinum در ﻣﺤﯿﻂ ﺷﮑﻞ 4- ﺗﻮدهﻫﺎي ﻣﯿﺴﯿﻠﯿﻮﻣﯽ ﺣﺎﺻﻞ از ﮐﺸﺖ ﻗﺎرچ ﮐﺸﺖ R. acerinum PDA اﻟﻒ ب Downloaded from ifej.sanru.ac.ir at 23:41 +0330 on Tuesday September 28th 2021 ﺷﮑﻞ 5- اﻟﻒ: آﺳﮑﻮﺳﭙﻮر، ب: آﺳﮏ ﻗﺎرچ ﺷﮑﻞ 6- آﺳﮏ ﻗﺎرچ R. acerinum R. acerinum ﺑﺮ اﺳﺎس ﻣﺸﺨﺼﺎت ﻗﺎرچ و ﻋﻼﺋﻢ ﻇﺎﻫﺮي ﺗﺸﺨﯿﺺ داده ﺷﺪ ﮐﻪ ﺑﺎ ﻧﺘﺎﯾﺞ ﺗﺤﻘﯿﻖ ﮐﺎﻧﻮن و ﻟﮑﻪ ﺑﺮﮔﯽ، ﺷﮑﻞ و ﺳﺎﯾﺮ ﻣﺸﺨﺼﺎت ﭘﺮﮔﻨﻪ، ﺷﮑﻞ ﻫﻤﮑﺎران (5) و ﻻﭘﻮﯾﻨﺖ و ﺑﺮﯾﺴﻮن (12) در و ﻣﺸﺨﺼﺎت آﺳﮑﻮﺳﭙﻮر، ﻗﺎرچ ﻋﺎﻣﻞ ﺑﯿﻤﺎري روي ﮐﺸﻮر آﻣﺮﯾﮑﺎ ﻣﻄﺎﺑﻘﺖ دارد. ﻗﺎرچﻫﺎي Rhytisma ﺑــﺮگ درﺧﺘــﺎن اﻓــﺮاﭘـﻠـﺖ از ﻧـﻮع Rhytisma ﭘﺎرازﯾﺖ ﻫﺴﺘﻨﺪ ﮐﻪ روي ﺑﺮگﻫﺎي ﻧﻬﺎﻧﺪاﻧﮕﺎن acerinum از رده آﺳﮑﻮﻣﯿﺴﺖ، راﺳﺘــﻪ ﻣﺸﺎﻫﺪه ﻣﯽﺷﻮﻧﺪ (11) و ﺳﺒﺐ ﺑﯿﻤﺎري در Rhytismatales و ﺧــﺎﻧـﻮاده Rhytismataceae ﮔﻮﻧﻪﻫﺎي ﻣﺨﺘﻠﻒ اﻓﺮا ﻣﯽﮔﺮدد (12). ﻣﻌﻤﻮﻻً اﯾﻦ ﺟﺪاﺳﺎزي و ﺷﻨﺎﺳﺎﯾﯽ ﻋﺎﻣﻞ ﺑﯿﻤﺎري ﻟﮑﻪ ﻗﯿﺮي درﺧﺘﺎن اﻓﺮاﭘﻠﺖ ..................................................................................................................................................... 30 ﺑﯿﻤﺎري در اواﺧﺮ زﻣﺴﺘﺎن روي ﻣﯿﺰﺑﺎن و ﺑﺮگﻫﺎي ﺟﻨﮕﻞﻫﺎ ﺑﻪﻋﻨﻮان واﺣﺪ ﺗﻮﻟﯿﺪ ﻧﺎﺧﺎﻟﺺ ﻣﻠﯽ اﻓﺘﺎده، ﮐﯿﺴﻪﻫﺎي آﺳﮏ را ﺗﺸﮑﯿﻞ ﻣﯽدﻫﻨﺪ ﮐﻪ در ﺑﺴﯿﺎري از ﮐﺸﻮرﻫﺎي دﻧﯿﺎ ﻣﻄﺮح ﻫﺴﺘﻨﺪ و از ﺳﺒﺐ ﻣﯽﺷﻮﻧﺪ، اﺳﭙﻮر ﻗﺎرچ روي ﺑﺮگﻫﺎي اﯾﻦ ﻧﻈﺮ ﺟﺎﯾﮕﺎه ﺧﺎﺻﯽ را در ﻋﺮﺻﻪ اﻗﺘﺼﺎد ﺑﻪ درﺧﺘﺎن اﻓﺮا در ﻓﺼﻞ روﯾﺶ ﺑﻌﺪي ﻧﻔﻮذ ﮐﺮده و ﺧﻮد اﺧﺘﺼﺎص دادهاﻧﺪ. ﺟﻨﮕﻞﻫﺎ ﻋﻼوه ﺑﺮ ﺗﻮﻟﯿﺪ ﻟﮑﻪﻫﺎي ﺳﯿﺎه اﺳﺘﺮوﻣﺎﯾﯽ روي ﺑﺮگ درﺧﺘﺎن ﭼﻮب داراي ﻧﻘﺶ زﯾﺴﺖ ﻣﺤﯿﻄﯽ ﺑﺴﯿﺎر ﻣﻬﻤﯽ اﯾﺠﺎد ﮐﻨﻨﺪ، اﯾﻦ ﻟﮑﻪﻫﺎ در ﺻﻮرت ﺗﻮﺳﻌﻪ ﺑﺎﻋﺚ ﻫﺴﺘﻨﺪ ﮐﻪ ﺿﺎﻣﻦ ﺑﻘﺎي ﭼﺮﺧﻪ آﻟﯽ، ﺧﺎﮐﯽ و آب و ﺧﺰان زودرس، از ﺑﯿﻦ رﻓﺘﻦ ﺑﺮگ درﺧﺖ اﻓﺮا، ﻫﻮاﯾﯽ ﻣﻨﺎﻃﻖ ﺗﺤﺖ ﭘﻮﺷﺶ ﺧﻮد ﻣﯽﺑﺎﺷﻨﺪ.
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