ﻑﺮﻌﻣ ﻥﺍﺭﺎﮑﻤﻫ ﻭ ﻩﺩﺍﺯ :
ﻲﺧﺮﺑ ﻲﻳﺁﺭﺎﻛ ﻲﺳﺭﺮﺑ ﺵﻭﺭ
ﺎﻫ ﺮﺑ ﻲﻨﺘﺒﻣ ﻭ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ ﻱ ﺭﺩ PCR
...
۵۵
ﻪﻠﺠﻣ ﻩﺎﻴﮔ ﺶﻧﺍﺩ ﻲﻜﺷﺰﭘ ﻥﺍﺮﻳﺍ
ﺩ ﻩﺭﻭ ۴۰ ﻩﺭﺎﻤﺷ ، ۱ ، ۱۳۸۸ ) ۶۴
‐ ۵۵ (
ﺳﺭﺮﺑ ﻲ ﺁﺭﺎﻛ ﻲﻳ ﺧﺮﺑ ﻲ ﺵﻭﺭ ﺎﻫ ﻱ ﮊﻮﻟﻭﺮﺳ ﻳ
ﻜ ﻲ ﻨﺘﺒﻣ ﻭ ﻲ ﺮﺑ ﺭﺩ PCR
ﺨﺸﺗ ﻴ ﺩﺭ ﻭ ﺺ ﻳ
ﺑﺎ ﻲ ﺮﺘﻛﺎﺑ ﻱ ﺑ ﻞﻣﺎﻋ ﻴ ﺭﺎﻤ ﻱ ﻣ ﻥﺎﺘﺧﺭﺩ ﻚﺸﺗﺁ ﻴ
ﻪﻧﺍﺩ ﻩﻮ ﺭﺍﺩ
ﻫﺎﻧ ﻴ ﻑﺮﻌﻣ ﺪ
۱ﻩﺩﺍﺯ ﺒﺘﺠﻣ ،* ﻲ ﺪﻤﺤﻣ
۲ﻱ ﺮﺷ ﺱﺎﺒﻋ ، ﻳ
ﻔ ﻲ ﻧﺍﺮﻬﺗ
۳ﻲ ﻭ ﺮﻛﺍﺫ ﺍﺮﻫﺯ
۴ﻱ
۱ ، ۲ ، ۳ ، ۴
، ﺪﺷﺭﺍ ﻲﺳﺎﻨﺷﺭﺎﮐ ﻖﺑﺎﺳ ﻱﻮﺠﺸﻧﺍﺩ ،
،ﻲﻤﻠﻋ ﺖﺌﻴﻫ ﻮﻀﻋ ﻭ ﺩﺎﺘﺳﺍ ،ﻲﻤﻠﻋ ﺖﺌﻴﻫ ﻮﻀﻋ
ﺩﺮﭘ ﻳ ﺯﺭﻭﺎﺸﻛ ﺲ ﻱ
ﺒﻃ ﻊﺑﺎﻨﻣ ﻭ ﻴ ﻌ ﻲ ﻩﺎﮕﺸﻧﺍﺩ ﻥﺍﺮﻬﺗ
) ﺖﻓﺎﻳﺭﺩ ﺦﻳﺭﺎﺗ :
۷ / ۱۱ / ۸۵
‐ ﺐﻳﻮﺼﺗ ﺦﻳﺭﺎﺗ :
۲۶ / ۱ / ۸۸ (
ﻩﺪﻴﮑﭼ
ﺑﻴ ﺭﺎﻤ ﻱ ﺮﺘﻛﺎﺑ ﻞﻣﺎﻋ ﺎﺑ ﻚﺸﺗﺁ ﻳ
ﻲﻳﺎ
Erwinia amylovora
ﺯﺍ ﻱﺭﺎﻤﻴﺑ ﺎﻫ ﻩﻮﻴﻣ ﻢﻬﻣ ﻱ ﻱﺎﻫ
ﻪﻧﺍﺩ ﺭﺍﺩ ﺏﻮﺴﺤﻣ
ﻲﻣ ﺩﻮﺷ . ﻃ ﻱﺭﺎﻤﻴﺑ ﻦﻳﺍ ﻲ
ﻝﺎﺳ ﺎﻫ ﻱ ﻪﺘﺷﺬﮔ ﺕﺭﺎﺴﺧ ﺎﻫ ﻱ ﺘﻔﮕﻨﻫ ﻲ ﻪﺑ ﻍﺎﺑ ﺎﻫ ﻱ ﻣ ﻴ ﻩﻮ ﻪﻧﺍﺩ ﻤﺤﺗ ﻥﺍﺮﻳﺍ ﺭﺍﺩ ﻴ
ﻩﺩﻮﻤﻧ ﻞ
ﺖﺳﺍ . ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺵﻭﺭ
ﺎﻫ ﻱ ﻗﺩ ﻴ ﻤﻃﺍ ﻞﺑﺎﻗ ﻭ ﻖ ﻴ
ﺎﺳﺎﻨﺷ ﺖﻬﺟ ﻥﺎﻨ ﻲﻳ
ﺩﺭ ﻭ ﻳ ﺑﺎ ﻲ ﺑ ﻊﻗﻮﻣ ﻪﺑ ﻴ ﺩﻭﺯ ﻝﺮﺘﻨﻛ ﻪﺑ ،ﺮﮔﺭﺎﻤ
ﮔﻮﻠﺟ ﻭ ﻡﺎﮕﻨﻫ ﻴ
ﺮ ﻱ ﺑ ﺵﺮﺘﺴﮔ ﺯﺍ ﻴ
ﺭﺎﻤ ﻱ ﻣ ﻚﻤﻛ ﻩﺩﻮﻟﺁ ﻥﺎﺘﺧﺭﺩ ﻑﺬﺣ ﻭ ﻲ
ﺎﻤﻧ ﻳ ﺪ . ﺭﺩ ﻭﺩ ﻖﻴﻘﺤﺗ ﻦﻳﺍ ﻲﺘﻧﺁ
ﻡﺮﺳ
ﻠﭘ ﻲ ﺻﺎﺼﺘﺧﺍ ﻝﺎﻧﻮﻠﻛ ﻲ
ﻬﺗ ﻴ ﺍﺮﺑ ﻭ ﻩﺪﺷ ﻪ ﻱ
ﺩﺭ ﻳ ﺑﺎ ﻲ ﻝﻮﻠﺳ ﺎﻫ ﻱ ﺮﺘﻛﺎﺑ ﻱ
E. amylovora (Ea)
ﺭﺩ ﻥﻮﻣﺯﺁ ﺎﻫ ﻱ
ﻭ ﺭﺎﮔﺁ ﺭﺩ ﺖﺸﻧ DIBA
ﺪﻨﺘﻓﺮﮔ ﺭﺍﺮﻗ ﻩﺩﺎﻔﺘﺳﺍ ﺩﺭﻮﻣ .
ﺎﺒﻳﺩ ﻥﻮﻣﺯﺁ ﺩﺭ ﻪﺑ ﺭﺩﺎﻗ
ﻳ ﺑﺎ
٧ ﻲ ١٠
× ١ ﺮﺘﻛﺎﺑ ﻝﻮﻠﺳ ﻱ
ﺮﻫ ﺭﺩ
ﻣ ﻴﻠ ﻲ ﻟ ﻴ ﻧ ﻭ ﺺﻟﺎﺧ ﺖﺸﻛ ﺮﺘ ﻴ
ﺩﻮﺑ ﻩﺩﻮﻟﺁ ﻩﺭﺎﺼﻋ ﺰ .
ﻨﻏ ﺎﺑ ﻲ ﺯﺎﺳ ﻱ ﺤﻣ ﺭﺩ ﻴ ﺎﻣ ﻂ ﻳ ﻛ ﻊ ﻴ ﺳﺎﺴﺣ ﺢﻄﺳ ،ﻲﺑ ﮓﻨ ﻴ
ﺖ
ﻪﺑ ﺎﺒﻳﺩ ١٠ ﺍﺰﻓﺍ ﻝﻮﻠﺳ ﻳ
ﺶ ﻳ ﺖﻓﺎ . ﺍ ﺭﺩ ﻳ ﺳﺭﺮﺑ ﻦ ﻲ ﺍﺭﺎﮐ ﻲﻳ ﺵﻭﺭ ﺎﻫ ﻱ ﮊﻮﻟﻭﺮﺳ ﻳ ﮑ ﻲ ﺧﺮﺑ ﺎﺑ ﻲ ﺯﺍ ﺵﻭﺭ ﺎﻫ ﻱ ﻨﺘﺒﻣ ﻲ
PCR ﺮﺑ ﺎﺳﺎﻨﺷ ﺖﻬﺟ ﻪﻛ ﻲﻳ
ﺍ ﻳ ﺮﺘﮐﺎﺑ ﻦ ﻱ ﻩﺪﺷ ﺮﺸﺘﻨﻣ ﺎﻘﻣ ﺰﻴﻧ ﺪﻧﺍ
ﻳ ﺩﺮﮔ ﻪﺴ ﻳ ﺪ . ﺍ ﻪﺑ ﻳ ﻪﻌﻄﻗ ﺭﻮﻈﻨﻣ ﻦ ﺍ
ﻱ ﻳ ﻛ ﻚ ﻴ ﻮﻠ
ﺯﺎﺑ ﻱ ﻣﺯﻼﭘ ﺯﺍ ﻴ
pEA29 ﺪ ﻡﺎﻨﺑ
pstI
ﺎﻫﺮﮔﺯﺎﻏﺁ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ
Aﻱ
Bﻭ ﺵﻭﺭ ﺭﺩ ﺪﻳﺩﺮﮔ ﺮﻴﺜﻜﺗ PCR
. ﺡﻮﻄﺳ
ﺳﺎﺴﺣ ﻴ ﺍ ﺖ ﻳ ﮔ ﻩﺭﺎﺼﻋ ﻭ ﺏﺁ ﺭﺩ ﺵﻭﺭ ﻦ ﻴ
ﻫﺎ ﻲ ﺗﺮﺗ ﻪﺑ ﻴ ﺐ ١٠٤
× ٢ ﻭ ١٠٦
× ٢ ﺩﻮﺑ ﺶﻨﻛﺍﻭ ﻁﻮﻠﺨﻣ ﺭﺩ ﻝﻮﻠﺳ .
ﺎﺳﺎﻨﺷ ﻪﺑ ﺭﺩﺎﻗ ﻩﺪﺷ ﺮﮐﺫ ﺵﻭﺭ ﻪﺳ ﻲﻳ
ﻦﻳﺮﺘﺳﺍ ﺎﻫ ﻱ ﻪﺑ ﻖﻠﻌﺘﻣ ﻥﺎﺑﺰﻴﻣ ﺎﻫ ﺖﻬﺟ ﻭ ﻩﺩﻮﺑ ﺭﻮﺸﮐ ﻒﻠﺘﺨﻣ ﻖﻃﺎﻨﻣ ﻭ
ﺎﺳﺎﻨﺷ
Ea ﻲﻳ ﺎﻘﻣ ﺭﺩ ﻳ ﺑ ﻪﺴ ﺮﻳﺎﺳ ﺎ ﺲﻨﺟ ﺎﻫ ﻱ ﺮﺘﮐﺎﺑ ﻳﺎ ﻲﻳ ﺻﺎﺼﺘﺧﺍ ﻲ ﺪﻧﺩﻮﻤﻧ ﻞﻤﻋ .
Nested-PCR ﺭﺩ ﺎﺑ
ﺎﻫﺮﮔﺯﺎﻏﺁ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﻱ
AJ75
AJ76 ﻭ ﻳ ﻪﻌﻄﻗ ﻚ ٨٠٠ ﺯﺎﺑ ﺖﻔﺟ ﻱ ﺜﻜﺗ ﻴ ﺩﺮﮔ ﺮ ﻳ ﺪ . ﺍﻳ ﺴﺑ ﺵﻭﺭ ﻦ ﻴ
ﺱﺎﺴﺣ ﺭﺎ
ﺎﺗ ﺶﻨﮐﺍﻭ ﻁﻮﻠﺨﻣ ﺭﺩ ﺖﺴﻧﺍﻮﺗ ﻳ
ﺮﺘﻛﺎﺑ ﻝﻮﻠﺳ ﻚ ﻳﺎ
ﻲﻳ ﺎﺗ ﻭ ﺏﺁ ﺭﺩ ﺍﺭ ١٠٠
ﮔ ﻩﺭﺎﺼﻋ ﺭﺩ ﺍﺭ ﻝﻮﻠﺳ ﻴ
ﻫﺎ ﻲ
ﺩﺭ ﻳ ﺑﺎ ﻲ ﺎﻤﻧ ﻳ ﺪ .
ﻩﮊﺍﻭ ﺎﻫ ﻱ ﻠﮐ ﻴ ﺪ ﻱ ،ﺎﺒﻳﺩ ،ﻲﺑﺎﻳﺩﺭ ،ﻱﮊﻮﻟﻭﺮﺳ :
Erwinia amylovora Nested-PCR ،
.
ﻪﻣﺪﻘﻣ
ﻣ ﻴ ﻩﻮ ﺎﻫ ﻱ ﻪﻧﺍﺩ ﺳ ﺹﻮﺼﺧ ﻪﺑ ﺭﺍﺩ ﻴ
ﺑﻼﮔ ،ﺐ ﻲ ﺯﺍ ﻪﺑ ﻭ
ﻏﺎﺑ ﻢﻬﻣ ﺕﻻﻮﺼﺤﻣ ﻲ
ﺍ ﻳ ﻥﺍﺮ ﺬﻐﺗ ﻅﺎﺤﻟ ﺯﺍ ﻭ ﻩﺩﻮﺑ ﻳ
ﻪ ﺍ ﻱ ،
ﺎﻨﺻ ﻳ ﺪﺒﺗ ﻊ ﻳﻠ ﻲ ﻝﺎﻐﺘﺷﺍ ، ﺍﺯ ﻲﻳ ﺎﻫﺪﻣﺁﺭﺩ ﺐﺴﻛ ﻭ ﺕﺍﺭﺩﺎﺻ ، ﻱ
ﺯﺭﺍ ﻱ ﺎﺟ ﻳ ﻤﻬﻣ ﻩﺎﮕ ﻲ ﻣ ﺍﺭﺍﺩ ﺭﻮﺸﻛ ﺩﺎﺼﺘﻗﺍ ﺭﺩ ﺍﺭ ﻲ
ﺪﻨﺷﺎﺑ .
ﺑﻴ ﺭﺎﻤ ﻱ ﻚﺸﺗﺁ ﻞﻣﺎﻋ ﺎﺑ
Erwinia amylovora (Burr.)
Winslow et al.
(1920)
ﻃ ﺭﺩ ﻲ ﻝﺎﺳ ﺎﻫ ﻱ ﻪﺘﺷﺬﮔ
ﺘﻔﮕﻨﻫ ﺕﺍﺭﺎﺴﺧ ﻲ
ﻪﺑ ﻍﺎﺑ ﺎﻫ ﻱ ﻣ ﻴ ﻪﻧﺍﺩ ﺓﻮ ﺭﻮﺸﻛ ﺭﺍﺩ ﻩﺩﺭﻭﺁ ﺩﺭﺍﻭ
ﺖﺳﺍ
. ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺵﻭﺭ
ﺎﻫ ﻱ ﺮﺳ ﻳ ﭘ ،ﻊ ﻴ ﻤﻃﺍ ﻞﺑﺎﻗ ﻭ ﻪﺘﻓﺮﺸ ﻴ
ﻥﺎﻨ
ﻣ ﻲ ﺪﻧﺍﻮﺗ ﻪﺑ ﮏﻤﮐ ﺎﺑ ﺨﺸﺗ
ﻴ ﺩﺭ ﻭ ﺺ ﻳ
ﺑﺎ ﻲ ﻊﻗﻮﻣ ﻪﺑ ﻞﻣﺎﻋ
ﺑﻴ ﺎﻤ ﺭ ﻱ
، ﻄﻧﺮﻗ ﺕﺎﻣﺍﺪﻗﺍ ﺭﺩ ﻴ
ﻪﻨ ﺍ ﻱ ﻧ ﻭ ﻴ ﺰ ﭘ ﻴ ﻫﺎﮔﺁ ﺶ ﻲ ﻭ
ﺪﻣ ﻳ ﺮ ﻳ ﺑ ﺖ ﻴ ﺭﺎﻤ ﻱ ﻔﻣ ﻴ ﺩﻮﺷ ﻊﻗﺍﻭ ﺪ .
ﻟﻭﺍ ﻴ ﻦ ﺵﻭﺭ ﺎﻫ ﻱ ﺑ ﻪ ﺍﺮﺑ ﻪﺘﻓﺭ ﺭﺎﻛ ﻱ
ﺎﺳﺎﻨﺷ ﻲﻳ
E. amylovora
ﺎﺳ ﺯﺍ ﻳ ﻣ ﺮ ﻴ ﻧﺎﮔﺭﺍﻭﺮﻜ ﻴ ﻡﺰ ﻪﻧﻮﻤﻧ ﺭﺩ ﺎﻫ ﺎﻫ
ﻱ
ﮔ ﻙﻮﻜﺸﻣ ﻴ
ﻫﺎ ﻲ ﺤﻣ ، ﻴ ﻂ ﺎﻫ ﻱ ﻧ ﻴ ﺑﺎﺨﺘﻧﺍ ﻪﻤ ﻲ ﻩﺩﻮﺑ ﺭﺍﺩﺭﺎﮔﺁ ﺪﻧﺍ
(Miller & Schroth, 1972; Falkenstein et al., 1988;
Bereswill et al., 1998)
.
ﻝﻮﺌﺴﻣ ﻩﺪﻨﺴﻳﻮﻧ *
: ﻑﺮﻌﻣ ﺪﻴﻫﺎﻧ ﻩﺩﺍﺯ
ﻦﻔﻠﺗ : ۱۵۴۸۱۵۱
‐ ۰۹۱۱
Email: nmoarref@gmail.com
۵۶ ﻪﻠﺠﻣ
ﻩﺎﻴﮔ ﺶﻧﺍﺩ ﻲﻜﺷﺰﭘ
ﺩ ،ﻥﺍﺮﻳﺍ ﻩﺭﻭ ۴۰ ، ﻩﺭﺎﻤﺷ ۱ ، ۱۳۸۸
ﻘﻘﺤﻣ ﻴ ﺴﺑ ﻦ ﻴ ﺭﺎ ﻱ ﺯﺍ ﺵﻭﺭ ﺎﻫ ﻱ ﮊﻮﻟﻭﺮﺳ ﻳ ﮑ ﻲ ﺍﺮﺑ ﻱ ﺩﺭ ﻳ ﺑﺎ ﻲ
E. amylovora
ﻩﺩﺮﮐ ﻩﺩﺎﻔﺘﺳﺍ ﺪﻧﺍ
.
Thampson & Cherath
(1976)
ﺘﻧﺁ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﻲ
ﻡﺮﺳ ﺎﻫ ﻱ ﺯﺍ ،ﻒﻠﺘﺨﻣ ﺵﻭﺭ
ﺍﻳ ﻤ ﻴ ﺲﻨﺳﺭﻮﻠﻓﻮﻧﻮ ﺎﺳﺎﻨﺷ ﺖﻬﺟ١
ﻲﻳ
E. amylovora
ﻩﺩﺎﻔﺘﺳﺍ
ﮐ ﺪﻧﺩﺮ
(Vanneste, 2000)
.
Lin et al.
(1987)
ﺘﻧﺁ ﺯﺍ ﻲ -
ﺩﺎﺑ ﻳ ﺎﻬ ﻱ ﻝﺎﻧﻮﻠﻛﻮﻧﻮﻣ ﺵﻭﺭ ﺭﺩ
ﺍ ﻳ ﻤ ﻴ ﺲﻨﺳﺭﻮﻠﻓﻮﻧﻮ ﺖﻬﺟ
ﺩﺭ ﻳ ﺑﺎ ﻲ ﻩﺩﺎﻔﺘﺳﺍ ﻚﺸﺗﺁ ﺪﻧﺩﺮﮐ
(Lelliot & Stead, 1987;
McLaughlin et al., 1989; Vanneste, 2000)
.
Zielke
(1993)
ﮊﻮﻟﻭﺮﺳ ﺵﻭﺭ ﻪﺳ ﻳ
ﻜ ﻲ ﺍﺮﺑ ﺍﺭ ﻒﻠﺘﺨﻣ ﻱ
ﺩﺭ ﻳ ﺑﺎ ﻲ
E. amylovora
ﺴﻧﺎﭙﺳﻮﺳ ﺭﺩ ﻴ
ﺤﻣ ﻥﻮ ﻴ ﻭ ﺖﺸﻛ ﻂ ﺖﻓﺎﺑ
ﺎﻫ ﻱ
ﺎﻘﻣ ﻩﺩﻮﻟﺁ ﻳ
ﺪﻧﺩﻮﻤﻧ ﻪﺴ .
Gorris et al.
(1996)
ﺵﻭﺭ
ﻻﺍ ـ ﺱﺍﺩ ﻳ
ﻨﻏ ﺔﻠﺣﺮﻣ ﻩﺍﺮﻤﻫ ﻪﺑ ﺍﺭ ﺍﺰ ﻲ
ﺯﺎﺳ
٢ﻱ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﻭ
ﺘﻧﺁ ﻲ ﺩﺎﺑ ﻳ ﺎﻬ ﻱ ﭘ ﻝﺎﻧﻮﻠﻛﻮﻧﻮﻣ ﻴ
ﺪﻧﺩﺮﻛ ﺩﺎﻬﻨﺸ
(Gorris et al.,
1996; Vanneste 2,000)
.
Falkenstein et al.
(1988)
ﻣﺯﻼﭘ ﺱﺎﺳﺍ ﺮﺑ ﻴ
ﺪ
pEA29
ﺭﺩ ﻪﮐ ﺍﺪﺟ ﻳ ﻪ ﺎﻫ ﻱ ﻣ ﺩﻮﺟﻮﻣ Ea
ﻲ ﺑ ﺪﺷﺎ ﺎﻫﺮﮕﺷﻭﺎﻛ ، ﻱ
DNA
ﺎﺑ ﻩﺪﺷ ﺭﺍﺪﻧﺎﺸﻧ
P32
ﻬﺗ ﺍﺭ ﻴ ﺍﺭﺎﻛ ﻭ ﻩﺩﺮﻛ ﻪ ﻲﻳ
ﻥﺁ ﺎﻫ ﺵﻭﺭ ﺭﺩ ﺍﺭ
ﻫ ﻴ ﺮﺒ ﻳ ﺍﺪ ﻳ ﺰ ﻳ
٣ﻦﺸ ﻨﻠﻛ ﻲ ﺩﻮﻤﻧ ﺖﺑﺎﺛ
(Zhang & Geider,
1997; Zhang et al., 2000)
. ﻟﻭﺍ ﻴ ﺭﺎﺑ ﻦ
Bereswill et al.
(1992)
ﺵﻭﺭ ﺯﺍ ﺩﺭ ﺖﻬﺟ ،PCR
ﻳ ﺑﺎ ﻲ
E. amylovora
ﺪﻧﺩﺮﻛ ﻩﺩﺎﻔﺘﺳﺍ
(Bereswill et al., 1992; Bereswill et al.,
1992; Zhang & Geider, 1997; Vanneste, 2000;
Zhang et al., 2000)
.
McManus & Jones (1995)
ﺳﺎﺴﺣ ﻴ ﺻﺎﺼﺘﺧﺍ ﻭ ﺖ ﻲ
ﺵﻭﺭ ﺭﺎﻬﭼ ﻥﺩﻮﺑ ﻨﺘﺒﻣ
ﻲ ﺮﺑ
ﻞﻣﺎﺷ PCR
،PCR Nested-PCR
،
PCR-Dot-Blot
ybridization
ﻭ
Reverse-Blot Hybridization
ﺖﻬﺟ ﺍﺭ
ﺩﺭ ﻳ ﺑﺎ ﻲ
E. amylovora
ﺑ ﺎﻘﻣ ﻢﻫ ﺎ ﻳ ﺪﻧﺩﻮﻤﻧ ﻪﺴ .
ﻞﻴﻟﺩ ﻪﺑ ﻤﻫﺍ ﻴ ﺨﺸﺗ ﺖ ﻴ ﺩﺭ ﻭ ﺺ ﻳ ﺑﺎ ﻲ ﺤﺻ ﻴ ﻊﻗﻮﻣ ﻪﺑ ﻭ ﺢ
ﺍﻳ ﺑ ﻦ ﻴ ﺭﺎﻤ ﻱ ﻭ ﺍ ﻪﺑ ﻪﺟﻮﺗ ﺎﺑ ﻳ
ﺍ ﺭﺩ ﻪﮑﻨ ﻳ
ﻥﻮﻨﮐﺎﺗ ﺩﺭﻮﻣ ﻦ
ﻘﺤﺗ ﻴ ﻘ ﻲ ،ﺩﻮﺑ ﻩﺪﺸﻧ ﻡﺎﺠﻧﺍ ﺭﻮﺸﮐ ﻞﺧﺍﺩ ﺭﺩ ﻘﺤﺗ
ﻴ ﻖ ﺮﺿﺎﺣ ﺎﺑ
ﻑﺪﻫ ﺳﺭﺮﺑ ﻲ ﺍﺭﺎﻛ ﻲﻳ ﺧﺮﺑ ﻲ ﺵﻭﺭ ﺎﻫ ﻱ ﮊﻮﻟﻭﺮﺳ ﻳ ﮑ ﻲ ﻭ ﻨﺘﺒﻣ ﻲ
ﺮﺑ ﺨﺸﺗ ﺭﺩ PCR
ﻴ ﺩﺭ ﻭ ﺺ ﻳ ﺑﺎ ﻲ ﺑ ﻞﻣﺎﻋ ﻴ ﺭﺎﻤ ﻱ ﮏﺸﺗﺁ ﺭﻮﺻ ﺕ
ﺖﻓﺮﮔ .
1. Immunofluorescence
2. Enrichment
3. Colony hybridization
ﻭ ﺩﺍﻮﻣ ﺵﻭﺭ ﺎﻫ
ﻃ ﻲ ﺎﭘ ﻭ ﺭﺎﻬﺑ ﻴﻳ ﺰ ﻝﺎﺳ ﺎﻫ ﻱ ١٣٧٩ ﻭ ١٣٨٠ ﺯﺍ ، ﻍﺎﺑ ﺎﻫ ﻱ
ﺭﻮﺸﻛ ﻒﻠﺘﺨﻣ ﻖﻃﺎﻨﻣ ﻞﻣﺎﺷ
ﺝﺮﻛ ) ﻭ ﺱﻮﻟﺎﭼ ﺓﺩﺎﺟ ،ﻥﺎﻏﺮﺑ
ﻥﺎﻘﻟﺎﻃ ( ﺮﻬﺷ ، ﻳ ﺭﺎ
، ﻭﺰﻗ ﻳ ﻣ ،ﻦ ﻴ ﻣﻭﺭﺍ ،ﻪﻏﺍﺮﻣ ،ﻪﻧﺎ ﻴ
ﺮﺒﺗ ﻭ ﻪ ﻳ ﺰ
ﺩﺯﺎﺑ ﻳ ﺑ ﻭ ﺪﻣﺁ ﻞﻤﻋ ﻪﺑ ﺪ ﻴ
ﺯﺍ ﺶ ٣٠٠ ﮔ ﺔﻧﻮﻤﻧ ﻴ ﻫﺎ ﻲ ﻭ ﻙﻮﻜﺸﻣ
ﺍﺭﺍﺩ ﻱ ﺑ ﻢﺋﻼﻋ ﻴ ﺭﺎﻤ ﻱ ﻣ ،ﮒﺮﺑ ،ﻪﻓﻮﻜﺷ ﺯﺍ ﻴ
ﻮ
،ﻩ ﻭ ﻪﺧﺎﺷﺮﺳ
ﺍﺭﺍﺩ ﺔﺧﺎﺷ ﻱ
ﻣ ﻥﺎﺘﺧﺭﺩ ﺯﺍ ﺮﻜﻧﺎﺷ ﻴ
ﻪﻧﺍﺩ ﺓﻮ ﺳ ﻞﻣﺎﺷ ﺭﺍﺩ ﻴ
،ﺐ
ﺑﻼﮔ ﻲ ﻊﻤﺟ ﻚﻟﺍﺰﻟﺍﺯ ﻭ ﻪﺑ ، ﺭﻭﺁ
ﻱ ﺪﺷ . ﺯﺎﺳﺍﺪﺟ ﺖﻬﺟ ﻱ
ﺮﺘﻛﺎﺑ ﻱ ﺯﺍ ﻪﻧﻮﻤﻧ ﺎﻫ
، ﺵﻭﺭ ﻖﺑﺎﻄﻣ
Lelliot & Stead (1987)
ﺤﻣ ﺯﺍ ﻴ ﺬﻐﻣ ﺭﺎﮔﺁ ﺖﺸﮐ ﻂ ﻱ
(Nutrient agar)
ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ .
ﮏﺘﺸﺗ ﺎﻫ ﻱ ﺮﺘﭘ ﻱ
،ﻩﺪﺷ ﺖﺸﮐ ﻟﺍ ﻭﺩ
ﻲ ﺭﺩ ﺯﻭﺭ ﻪﺳ ﻣﺩ
ﺎ ﻱ
١ ٢٦± ﺘﻧﺎﺳ ﻪﺟﺭﺩ ﻴ
ﺭﺍﺪﻬﮕﻧ ﺩﺍﺮﮕ ﻱ
ﻭ ﻩﺪﺷ ﻨﻠﻛ ﻲ ﺎﻫ ﻱ ،ﻚﭼﻮﻛ
ﺷ ﺎﺗ ﻡﺮﻛ ،ﻕﺍﺮﺑ ﻴ
ﺮ ﻱ ﺷﺎﺣ ﺎﺑ ﮓﻧﺭ ﻴ
ﺏﺪﺤﻣ ﻭ ﺩﺮﮔ ،ﻞﻣﺎﻛ ﺔ
ﺩﺮﮔ ﺏﺎﺨﺘﻧﺍ ﻳ
ﺪﻧﺪ . ﻨﭽﻤﻫ ﻴ ﺩﺍﺪﻌﺗ ﻦ ﻱ ﺍﺪﺟ ﻳ ﺯ ﺡﺮﺷ ﻪﺑ ﻪ ﻳ
ﺯﺍ ﺮ
ﺎﺳ ﻳ ﺭﺩ ﺰﮐﺍﺮﻣ ﺮ ﻳ
ﺪﻧﺪﺷ ﺖﻓﺎ :
ﺶﺷ ﺍﺪﺟ ﻳ ﺮﺘﻛﺎﺑ ﺔ ﻱ ﻖﻃﺎﻨﻣ ﺯﺍ
ﻭﺰﻗ ﻭ ﻥﺎﺳﺍﺮﺧ ،ﻥﺎﻨﻤﺳ ﻳ
ﻦ ﻘﺤﺗ ﻪﺴﺳﺆﻣ ﺯﺍ ﻴ
ﺕﺎﻘ ﻲﮑﺷﺰﭙﻫﺎﻴﮔ
،ﺭﻮﺸﮐ ﻳ ﺍﺪﺟ ﻚ ﻳ ﻪ ﺑﻼﮔ ﻪﺑ ﻁﻮﺑﺮﻣ ﻲ
ﻘﺤﺗ ﺰﻛﺮﻣ ﺯﺍ ﻴ
ﺕﺎﻘ
ﺯﺭﻭﺎﺸﻛ ﻱ ﻭﺰﻗ ﻳ ﻦ ﻭ ﺍﺪﺟ ﻭﺩ ﻳ ﻪ ﺯﺍ ﻥﺂﺘﺳﺍ ﺎﻫ ﻱ ﻥﺍﺮﻬﺗ ﻭ ﻥﺎﻨﻤﺳ
ﻪﮐ ﻳ ﮑ ﻲ ﺯﺍ ﻥﺁ ﺎﻫ ﺷ ﻪﺑ ﻁﻮﺑﺮﻣ ﻴ
ﺖﺸﺧﺮ ﺯﺍ ﺩﻮﺑ ﻭ ﻡﻮﻠﻋ ﺪﺣﺍﻭ
ﻘﺤﺗ ﻴ ﻣﻼﺳﺍ ﺩﺍﺯﺁ ﻩﺎﮕﺸﻧﺍﺩ ﺕﺎﻘ ﻲ
. ﻳ ﺍﺪﺟ ﻚ ﻳ ﻪ ﻊﺟﺮﻣ
E.a
ﻧ 322A
ﻴ ﺰ ﺯﻭﻼﮐ ﺯﺍ ﺎﮔ ﻳ
٤ﺭﺪ ) ﺘﺴﻧﺍ ﻴ ﻚﻧﻼﭘ ﺲﻜﻣ ﻮﺘ ﺭﺩ
ﺭﻮﺸﻛ
ﻥﺎﻤﻟﺁ ( ﺭﺩ ﻳ ﺖﻓﺎ ﮔ ﺩﺮ ﻳ ﺪ ﺍ ﺭﺩ ﻪﮐ ﻳ ﻡﺎﻧ ﺎﺑ ﺎﺠﻨ ﻩﺭﺎﺷﺍ ﻥﺁ ﻪﺑ Ea1
ﺪﺷ ﺪﻫﺍﻮﺧ .
ﺍ ﻳ ﺍﺪﺟ ﻦ ﻳ ،ﻪ ﻳ ﻥﺎﺗﻮﻣ ﻚ ﺍﺪﺟ ﺯﺍ Tn5
ﻳ ﺔ
E.a 322
ﻣ ﻲ ﮕﺸﻧﺍﺩ ﺭﺩ ﻪﻛ ﺪﺷﺎﺑ ﺎ
ﻞﻧﺭﻮﻛ ﻩ ﻧ ﺭﺩ ﻊﻗﺍﻭ ٥
ﻴ ﻮ ﻳ ﻬﺗ ﻙﺭﻮ ﻴ ﻪ
ﺩﺮﮔ ﻳ ﻩﺪ ﻭ ﺑ ﺪﻗﺎﻓ ﻴ ﺭﺎﻤ ﻳ ﺍﺰ ﻲﻳ ﻣ ﻲ ﺪﺷﺎﺑ
(Steinberger & Beer,
1988)
. ﻨﭽﻤﻫ ﻴ ﺩﺍﺪﻌﺗ ﻦ ﻱ
ﺍﺪﺟ ﻳ ﺧﺮﺑ ﻪﺑ ﻖﻠﻌﺘﻣ ﻪ ﻲ
ﺲﻨﺟ ﺎﻫ ﻱ ﺮﺘﮐﺎﺑ ﻳﺎ ﻲﻳ ﺩ ﻳ ﻣﺎﺳﺍ ﻪﮐ ﺮﮕ ﻲ
ﻥﺁ ﺎﻫ ،ﺪﺷ ﺪﻫﺍﻮﺧ ﺮﮐﺫ
ﺎﻣﺯﺁ ﺯﺍ ﻳ ﺮﺘﮐﺎﺑ ﻩﺎﮕﺸ ﻱ
ﺳﺎﻨﺷ ﻲ ﻲﮑﺷﺰﭙﻫﺎﻴﮔ ﻩﻭﺮﮔ ﻩﺎﮕﺸﻧﺍﺩ
ﻬﺗ ﻥﺍﺮﻬﺗ ﻴ ﺪﻧﺪﺷ ﻪ .
ﻲﻳﺍﺰﻳﺭﺎﻤﻴﺑ ﻥﻮﻣﺯﺁ
ﻠﻛ ﻴ ﺍﺪﺟ ﺔ ﻳ ﻪ ﺎﻧﺍﻮﺗ ﺮﻈﻧ ﺯﺍ ﺍﺪﺘﺑﺍ ،ﺎﻫ ﻲﻳ
ﺍ ﻳ ﻕﻮﻓ ﺶﻨﻛﺍﻭ ﺩﺎﺠ
ﺳﺎﺴﺣ ﻴ ﺖ ﻥﻮﺗﻮﺗ ﺭﺩ (HR)
ﻪﺑ ﺵﻭﺭ
Klement et al.
(1990)
ﺳﺭﺮﺑ ﻲ ﺪﻧﺪﺷ . ﺑ ﺕﺎﺒﺛﺍ ﺖﻬﺟ ﻴ
ﺭﺎﻤ ﻳ ﺍﺰ ﻲﻳ ﺯﺍ ﻣ ﻴ ﻩﻮ ﺎﻫ ﻱ
ﺳ ﺱﺭﺎﻧ ﻴ
،ﺐ ﺑﻼﮔ ﻲ ﺵﻭﺭ ﻖﺑﺎﻄﻣ ﺥﺮﺳ ﻞﮔ ﺞﻨﻬﻧ ﻭ
Schaad et al.
(2001)
ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ .
4. Klaus Geider
5. Cornell University
ﻑﺮﻌﻣ ﻥﺍﺭﺎﮑﻤﻫ ﻭ ﻩﺩﺍﺯ :
ﻲﺧﺮﺑ ﻲﻳﺁﺭﺎﻛ ﻲﺳﺭﺮﺑ ﺵﻭﺭ
ﺎﻫ ﺮﺑ ﻲﻨﺘﺒﻣ ﻭ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ ﻱ ﺭﺩ PCR
...
۵۷
ﻪﻳﺍﺪﺟ ﻲﭙﻴﺗﻮﻨﻓ ﺕﺎﻴﺻﻮﺼﺧ ﻲﺳﺭﺮﺑ ﺎﻫ
ﻮﻠﺳﺎﺳ ﺵﻭﺭ ﻪﺑ ﺪﺻﺭﺩ ﻪﺳ ﺱﺎﺘﭘ ﺭﺩ ﻡﺮﮔ ﻥﻮﻣﺯﺁ ﻭ
ﻥﺍﺭﺎﮑﻤﻫ
(Fahy & Hayward, 1983)
ﺪﺷﺭ ﻥﻮﻣﺯﺁ ، ﺯﺍﻮﻫ
ﻱ /
ﺑ ﻲ ﺯﺍﻮﻫ ﻱ ﻫ ﺵﻭﺭ ﻪﺑ ﻴ
ﻮ ﻭ ﻻ ﻳ ﻦﺳ ﻒ
(Schaad et al., 2001)
،
ﺗ ﻟﻮ ﻴ ﺵﻭﺭ ﻪﺑ ﻥﺍﻮﻟ ﺪ
Lelliot & Stead (1987)
ﻟﻮﺗ ، ﻴ ﺪ
ﺤﻣ ﺭﺩ ﺖﻨﺳﺭﻮﻠﻓ ﻪﻧﺍﺪﮕﻧﺭ ﻴ
ﻂ
KB (King’s medium B)
ﻪﺑ
ﺵﻭﺭ
Schaad et al.
(2001)
ﻥﻮﻣﺯﺁ ، ﺴﻛﺍ ﻴ ﺵﻭﺭ ﻪﺑ ﺯﺍﺪ
ﺲﻛﺍﻮﻛ ، ﻟﻮﺗ ﻥﻮﻣﺯﺁ ﻴ
ﻣ ﺪ ﺍﻮ ﺣﺍ ﺩ ﻴ ﻖﺑﺎﻄﻣ ﺯﻭﺮﻛﻮﺳ ﺯﺍ ﻩﺪﻨﻨﻛﺀﺎ
ﺵﻭﺭ
Fahy & Hayward (1983)
ﻭ ﻥﻮﻣﺯﺁ ﺎﻫ ﻱ ،ﺯﻻﺎﺗﺎﻛ
ﺘﻜﭘ ،ﺯﺎﺗﺎﻔﺴﻓ ﻴ
ﺎﻨ ﻫ ،ﺯ ﻴ ﻟﻭﺭﺪ ﻴ ﺗﻻﮊ ﺰ ﻴ ﻚﻤﻧ ﻞﻤﺤﺗ ،ﻦ ﻃ
ﻡﺎﻌ ٥ ﻭ
٦ ﻫ ،ﺪﺻﺭﺩ ﻴ
ﻟﻭﺭﺪ ﻴ ﻟﻮﻜﺳﺍ ﺰ ﻴ ﻫ ،ﻦ ﻴ ﻟﻭﺭﺪ ﻴ ﻟﻮﺗ ،ﻪﺘﺳﺎﺸﻧ ﺰ ﻴ
ﺪ
ﺍﻳ ﻫ ﻭ ﻝﻭﺪﻨ ﻴ
ﻟﻭﺭﺪ ﻴ ﻮﺗ ﺰ ﻴﻳ ﻦ ٨٠ ﺵﻭﺭ ﻪﺑ
Lelliot & Stead
(1987)
ﺪﺷ ﻡﺎﺠﻧﺍ .
ﻨﭽﻤﻫ ﻴ ﻟﻮﺗ ﻥﻮﻣﺯﺁ ﻦ ﻴ
،ﺰﻛﻮﻠﮔ ﺯﺍ ﺯﺎﮔ ﺪ
ﺣﺍ ﻴ ﻧ ﺀﺎ ﻴ ﻩﺭﻭﺍ ،ﺕﺍﺮﺘ ﻟﻮﺗ ،ﺯﺁ
ﻴ ﻭﺭ ﻪﻧﺍﺪﮕﻧﺭ ﺪ ﻱ
ﺤﻣ ﻴ ﻂ
YDC
)
Yeast extract-dextrose- CaCO3
( ، ﻭﺭ ﺪﺷﺭ ﻱ ﺤﻣ ﻴ ﻂ
ﺭﺎﮔﺁ EMB
)
Eusine methylene blue agar
( ﻭ ﺭﺩ ﺪﺷﺭ
ﺎﻣﺩ ﻱ ٣٦ ﺘﻧﺎﺳ ﺔﺟﺭﺩ ﻲ
ﺵﻭﺭ ﻪﺑ ﺩﺍﺮﮔ
Schaad et al.
(2001)
ﻭ ﺳ ﻑﺮﺼﻣ ﻥﻮﻣﺯﺁ ﻴ
ﺤﻣ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﺕﺍﺮﺘ ﻴ
ﺳ ﻂ ﻴ ﻥﻮﻤ
ﺳ ﻴ ﺭﺎﮔﺁ ﺕﺍﺮﺘ ﺖﻓﺮﮔ ﺕﺭﻮﺻ ١
. ﻭﺭ ﺮﺛﺍ ﻥﻮﻣﺯﺁ ﻱ
ﺷ ﻴ ﺮ
ﻟﻴ ﺱﻮﻤﺘ ﻧ ﺭﺍﺩ ﻴ ﺵﻭﺭ ﻪﺑ ﺰ
Schaad et al.
(2001)
ﻭ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ
ﺤﻣ ﺯﺍ ﻴ ﺩ ﺓﺩﺎﻣﺁ ﻂ ﻳ
٢ﻮﻜﻔ ، ﻧﮊﺭﺁ ﻥﻮﻣﺯﺁ ﻴ
ﻫﺩ ﻦ ﻴ ﺵﻭﺭ ﻪﺑ ﺯﻻﻭﺭﺪ
ﻠﻧﺭﻮﺗ
٣ﻲ
، ﻟﻮﺗ ﻴ ﻔﻟﻮﺳ ﺯﺎﮔ ﺪ ﻴ
ﻫ ﺪ ﻴ ﺵﻭﺭ ﻪﺑ ﻪﻧﻮﺘﭙﭘ ﺏﺁ ﺯﺍ ﻥﮊﻭﺭﺪ
Schaad et al.
(2001)
، ﻟﺎﻌﻓ ﻴ ﺔﺘﺴﻫ ﺖ ﻳ
ﻟﻮﺗ ،ﺦ ﻴ ﺪ
٣ ‐ ﻟﻮﺗ ﻭ ﺯﻮﺘﻛﻻﻮﺘﻛ ﻴ
ﺘﻣ ﺪ ﻴ ﺋﻮﺘﺳﺍ ﻭ ﺩﺭ ﻞ ﻴ
٤ﻦ
(MR-VP)
ﻧ ﻴ ﺰ
ﺵﻭﺭ ﻪﺑ
Fahy & Hayward (1983)
ﻡﺎﺠﻧﺍ ﺪﺷ .
ﻣﺯﺁ ﻥﻮ ﺎﻫ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ ﻱ
ﻟﻮﺗ ﺖﻬﺟ ﻴ ﺘﻧﺁ ﺪ ﻲ ،ﻡﺮﺳ ﻖﺑﺎﻄﻣ ﺵﻭﺭ
Jones et al.
(1983)
ﺪﺷ ﻞﻤﻋ . ﻳ ﮏ ﺍﺪﺟ ﻳ ﺮﺘﻛﺎﺑ ﺔ ﻳﺎ ﻲﻳ ﻪﮐ ﻥﺎﻏﺮﺑ ﻪﻘﻄﻨﻣ ﺯﺍ
ﺍ ﺭﺩ ﻳ ﻡﺎﻧ ﺎﺑ ﺎﺠﻨ ﻩﺭﺎﺷﺍ ﻥﺁ ﻪﺑ Ea2
ﻲﻣ ﺩﻮﺷ ﻧ ﻭ ﻴ ﺍﺪﺟ ﺰ ﻳ ﻪ
Ea1
ﺑ ﻪ ﻫﺎﻣ ﻥﻭﺭﺩ ﺕﺭﻮﺻ ﻴ
ﻪﭽ ﺍ ﻱ ﺵﻮﮔﺮﺧ ﻭﺩ ﻪﺑ ،ﺍﺰﺠﻣ ﺭﻮﻃ ﻪﺑ ﻭ
ﻔﺳ ﻴ ﻧ ﺪ ﻴ ﺯﻮ ﻳ ﺪﻨﻠ ﻱ ﺭﺰﺗ ﻳ ﺩﺮﮔ ﻖ ﻳ ﺪ ﺪﻧ . ﺭﺰﺗ ﺞﻨﭘ ﻳ ﻖ ﺪﻌﺑ ﻱ ﻪﺑ
ﻞﺻﺍﻮﻓ ﻳ ﻪﺘﻔﻫ ﻚ ﺍ ﻱ ﻫﺎﻣ ﻥﻭﺭﺩ ﺕﺭﻮﺼﺑ ﻥﺎﻨﭽﻤﻫ ﻭ ﻴ
ﻪﭽ ﺍ ﻱ
ﺪﻧﺪﺷ ﻡﺎﺠﻧﺍ .
ﺮﺧﺁ ﺯﺍ ﺲﭘ ﻪﺘﻔﻫ ﻭﺩ ﻳ
ﺭﺰﺗ ﻦ ﻳ ﻥﻮﺧ ،ﻖ ﮔ ﻴ ﺮ ﻱ
ﺵﻮﮔﺮﺧ ﺯﺍ ﻞﻣﺎﻛ ﻭ ﺖﻓﺮﮔ ﻡﺎﺠﻧﺍ ﺎﻫ
ﻲﺘﻧﺁ ﻡﺮﺳ ﻥﺁ ﺯﺍ
ﮔ ﻝﺎﺼﺤﺘﺳﺍ ﺩﺮ
ﻳ ﺪ . ﻌﺗ ﺖﻬﺟ ﻴﻴ ﺗ ﻦ ﻴ ﺮﺘ ﻲﺘﻧﺁ ﻡﺮﺳ ﻥﻮﻣﺯﺁ ﺯﺍ ،
1. Simmon's citrate agar
2. Difco 3. Thornley
4. Methyl Red and Voges-Proskauer Tests
ﻣ ﺩﺎﻘﻌﻧﺍ ﻴ ﭘﻮﮑﺳﻭﺮﮑ ﻲ ﺵﻭﺭ ﻪﺑ
Hampton et al.
(1993)
ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ .
ﺭﺎﮔﺁ ﺭﺩ ﻪﻓﺮﻃ ﻭﺩ ﺖﺸﻧ ﻥﻮﻣﺯﺁ :
ﻬﺗ ﺖﻬﺟ ﻴ ﮊ ﺔ ﻝ ،ﺭﺎﮔﺁ ﺯﺍ
ﺓﺪﺷ ﺡﻼﺻﺍ ﺵﻭﺭ
Schaad et al.
(2001)
ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ .
ﻲﺘﻧﺁ ﻡﺮﺳ ﺎﻫ ﻱ ﻓﺮﺼﻣ ﻲ ﺰﻛﺮﻣ ﻚﻫﺎﭼ ﺭﺩ ﻱ
ﻩﺩﻮﻣﺁ ﻭ ﺎﻫ ﻱ
ﺘﻧﺁ ﻲ ﻧﮊ ﻲ ﺴﻧﺎﭙﺳﻮﺳ ﺕﺭﻮﺼﺑ ﻒﻠﺘﺨﻣ ﻴ
ﺓﺪﺷ ﻭﻼﻛﻮﺗﺍ ﻥﻮ
ﻝﻮﻠﺳ ﻱﺎﻫ ﺮﺘﻛﺎﺑ ﻞﻣﺎﻛ ﻳﺎ
ﻲﻳ ﻚﻫﺎﭼ ﺭﺩ ﺎﻫ
ﻱ ﺭﺎﻨﻛ ﻱ ﺭ ﻳ ﻪﺘﺨ
ﺪﺷ ﺪﻧ . ﻚﺘﺸﺗ ﺕﺪﻣ ﻪﺑ ﺎﻫ ﻳ
ﻪﺘﻔﻫ ﻚ ﺭﺩ ﻭ ﺭﻮﺗﺎﺑﻮﮑﻧﺍ ﻥﻭﺭﺩ
ﺎﻣﺩ ﻱ ٢٧ ﺘﻧﺎﺳ ﻪﺟﺭﺩ ﻴ
ﺩﺍﺮﮕ ﺭﺍﺪﻬﮕﻧ ﻱ ﺩﺮﮔ ﻳ ﺮﻈﻧ ﺯﺍ ﻭ ﻩﺪ
ﻜﺸﺗ ﻴ ﺎﻫﺭﺍﻮﻧ ﻞ ﻱ ﺑﻮﺳﺭ ﻲ ﺳﺭﺮﺑ ﺩﺭﻮﻣ ﻲ
ﺪﻨﺘﻓﺮﮔ ﺭﺍﺮﻗ .
ﺍ ﻳ ﻦ
ﺯﺁ ﺎﻣ ﻳ ﺍﺮﺑ ﻪﻧﺎﮔﺍﺪﺟ ﺭﻮﻄﺑ ﺶ ﻱ
ﺘﻧﺁ ﻭﺩ ﺮﻫ ﻲ
ﻬﺗ ﻡﺮﺳ ﻴ ﻩﺪﺷ ﻪ
ﺘﻧﺁ ﺮﻫ ﻭ ﻡﺎﺠﻧﺍ ﻲ
ﻧ ﻡﺮﺳ ﻴ ﻠﻛ ﺎﺑ ﺰ ﻴ ﺍﺪﺟ ﺔ ﻳ ﻪ ﺎﻫ ﻱ ﺮﺘﻛﺎﺑ ﻱ ﻖﻠﻌﺘﻣ
ﻪﺑ ﻥﺎﺑﺰﻴﻣ ﺎﻫ ﺣﺍﻮﻧ ﻭ ﻲ ﻓﺍﺮﻐﺟ ﻴ ﺎ ﻲﻳ ﺍ ﻒﻠﺘﺨﻣ ﻳ
ﺎﻣﺯﺁ ،ﻥﺍﺮ ﻳ ﺶ
ﺩﺮﮔ ﻳ ﺪ . ﺳﺭﺮﺑ ﺖﻬﺟ ﻲ
ﺻﺎﺼﺘﺧﺍ ﻲ ﺍ ﻥﺩﻮﺑ ﻳ
،ﺵﻭﺭ ﻦ ﺶﻨﻛﺍﻭ
ﺘﻧﺁ ﻲ ﻡﺮﺳ ﺎﻫ ﻱ ﻪﻧﻮﮔ ﻞﺑﺎﻘﻣ ﺭﺩ ﻞﺻﺎﺣ ﺎﻫ
ﻲﻳ ﺩ ﺯﺍ ﻳ ﺮﮕ
ﺲﻨﺟ ﺎﻫ ﻱ ﺮﺘﻛﺎﺑ ﻳﺎ ﻲﻳ ﻞﻣﺎﺷ
Pectobacterium carotovorum
subsp.carotovorum
،
P.carotovorum subsp.wasabiae
،
Pseudomonas syringae pv. syringae
،
P. fluorescens
،
Rhizobium vitis
،
Bacillus subtilis
ﻭ
Xanthomonas
axonopodis pv. begoniae
ﺳﺭﺮﺑ ﺩﺭﻮﻣ ﻲ
ﺖﻓﺮﮔ ﺭﺍﺮﻗ .
ﺎﺒﻳﺩ ﻥﻮﻣﺯﺁ :٥
ﺩ ﻥﻮﻣﺯﺁ ﻳ ﺵﻭﺭ ﻪﺑ ﺎﺒ
Hawkes et al.
(1982)
ﻭ
Leach et al.
(1987)
ﻛﺪﻧﺍ ﺎﺑ ، ﻲ ﻐﺗ ﻴﻴ ﻡﺎﺠﻧﺍ ﺮ
ﺖﻓﺮﮔ . ﺖﺸﻛ ﺯﺍ ٢٤
ﺍﺪﺟ ﺔﺘﻋﺎﺳ ﻳ
ﻪ ﺎﻫ ﻱ ﺮﺘﻛﺎﺑ ﻒﻠﺘﺨﻣ ﻳﺎ
ﻲﻳ
ﻭﺭ ﻱ ﺤﻣ ﻴ ﺎﮔﺁ ﻂ ﺬﻐﻣ ﺭ ﻱ ﻭﺎﺣ ﻱ ﻳ ،ﺰﻛﻮﻠﮔ ﺪﺻﺭﺩ ﻚ
ﺴﻧﺎﭙﺳﻮﺳ ﻴ ﻧﻮ ﻲ ﺭﺩ ﻳ ﻣ ﻚ ﻴ ﻠ ﻲ ﻟ ﻴ ﺮﺘﺳ ﺮﻄﻘﻣ ﺏﺁ ﺮﺘ ﻥﻭ
ﻬﺗ ﻴ ﻭ ﻪ
ﺴﻧﺍﺩ ﻴ ﺭﻮﻧ ﺔﺘ ﻱ ﻝﻮﻃ ﺭﺩ ،ﺮﺘﻣﻮﺘﻓﻭﺮﺘﻜﭙﺳﺍ ﻩﺎﮕﺘﺳﺩ ﻂﺳﻮﺗ ﻥﺁ
ﺝﻮﻣ ٦٠٠ ﺮﺘﻣﻮﻧﺎﻧ
، ﻝﺩﺎﻌﻣ ﻳ ﻈﻨﺗ ﻚ ﻴ ﺩﺮﮔ ﻢ ﻳ ﺪ . ﻨﭽﻤﻫ ﻴ ﺯﺍ ﻦ
ﺴﻧﺎﭙﺳﻮﺳ ﻴ ﺍﺪﺟ ﻥﻮ ﻳ ﺮﺘﻛﺎﺑ ﺔ ﻱ ﻊﺟﺮﻣ
، ﺮﺳ ﻱ ﺖﻗﺭ ﻱﺎﻫ
ﻔﻠﺘﺨﻣ ﻲ ﺯﺍ ١٠ ﺎﺗ ١٠٩ ﻜﺸﺗ ﺪﺣﺍﻭ ﻴ
ﻞ ﻠﻛ ﺓﺪﻨﻫﺩ ﻨ ﻲ ) (CFU
ﻣ ﺭﺩ ﻴ ﻠ ﻲ ﻟ ﻴ ﻬﺗ ﺮﺘ ﻴ ﺯﺭﺍ ﺖﻬﺟ ﻭ ﻪ ﻳ
ﺑﺎ ﻲ ﺳﺎﺴﺣ ﻴ ﺩ ﺵﻭﺭ ﺖ ﻳ
ﺎﺒ
ﺖﻓﺮﮔ ﺭﺍﺮﻗ ﻩﺩﺎﻔﺘﺳﺍ ﺩﺭﻮﻣ .
ﻌﺗ ﺖﻬﺟ ﻴﻴ ﻜﺸﺗ ﺪﺣﺍﻭ ﻦ ﻴ
ﻞ ﺓﺪﻨﻫﺩ
ﻨﻠﻛ ﻲ ، ١٠٠ ﻣ ﻴ ﻟﻭﺮﻜ ﻴ ﺮﺳ ﺯﺍ ﺮﺘ ﻱ
ﺎﻫ ﻱ ﻭﺭ ﻞﺻﺎﺣ ﺖﻗﺭ ﻱ
ﺤﻣ ﻴ ﺬﻐﻣ ﺭﺎﮔﺁ ﻂ ﻱ
ﻭﺎﺣ ﻱ ﻳ ﺪﺷ ﻩﺩﺍﺩ ﺖﺸﻛ ﺰﻛﻮﻠﮔ ﺪﺻﺭﺩ ﻚ .
ﺳﺭﺮﺑ ﺖﻬﺟ ﻲ
ﺻﺎﺼﺘﺧﺍ ﻲ ﺍ ﻥﺩﻮﺑ ﻳ ﺵﻭﺭ ﻦ ﺴﻧﺎﭙﺳﻮﺳ ﻴ ﻥﻮ ﻲﻳﺎﻫ
ﺩ ﺯﺍ ﻳ ﺮﮕ ﻨﺟ ﺲ ﺎﻫ ﻱ ﺮﺘﻛﺎﺑ ﻳﺎ ﻲﻳ ﺮﮐﺫ ﻩﺪﺷ ﻬﺗ ﻴ ﺩﺮﮔ ﻪ ﻳ ﻭ ﻩﺪ ﺭﺩ
ﺩ ﻥﻮﻣﺯﺁ ﻳ ﺎﺒ ﺪﻧﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ .
ﺩﺭ ﺖﻬﺟ ﻳ ﺑﺎ ﻲ ﺮﺘﻛﺎﺑ ﻱ ﺖﻓﺎﺑ ﺭﺩ
5. Dot Immunobinding Assay
۵۸ ﻪﻠﺠﻣ
ﻩﺎﻴﮔ ﺶﻧﺍﺩ ﻲﻜﺷﺰﭘ
ﺩ ،ﻥﺍﺮﻳﺍ ﻩﺭﻭ ۴۰ ، ﻩﺭﺎﻤﺷ ۱ ، ۱۳۸۸
ﮔ ﻴ ﻫﺎ ﻲ ﻣ ، ﻴ ﻥﺍﺰ ﻳ ﮔﺩﻮﻟﺁ ﻪﺑ ﻙﻮﻜﺸﻣ ﺖﻓﺎﺑ ﺯﺍ ﻡﺮﮔ ﻚ ﻲ
ﺭﺩ
ﻣ ﺭﺎﻬﭼ ﻴ ﻠ ﻲ ﻟ ﻴ ﺮﻄﻘﻣ ﺏﺁ ﺮﺘ ﻥﻭﺮﺘﺳ
ﻥﻭﺭﺩ ﻭ ﻳ ﺮﺘﭘ ﻚ ﻱ ﺩﺮﺧ
ﻓﺎﺻ ﺬﻏﺎﻛ ﺯﺍ ﻥﺩﺍﺩ ﺭﻮﺒﻋ ﺎﺑ ﻞﺻﺎﺣ ﺓﺭﺎﺼﻋ ﻭ ﻩﺪﺷ ﻲ
،
ﻳ ﺩﺮﮔ ﺖﺧﺍﻮﻨﻜ ﻳ
ﺪ .
ﺖﻬﺟ ﻡﺎﺠﻧﺍ ﺩ ﻥﻮﻣﺯﺁ ﻳ ﺎﺒ ، ﻧ ﺬﻏﺎﻛ ﻴ ﻭﺮﺘ ﺕﺪﻣ ﻪﺑ ﺰﻟﻮﻠﺳ ١٥
ﻗﺩ ﻴ ﻥﻭﺭﺩ ﻪﻘ ﺮﻓﺎﺑ ) TBS
٨٤ / ٤ ﺮﺗ ﻡﺮﮔ ﻳ ﺯﺎﺑ ﺲ ﻱ ، ٤ / ٥٨ ﻡﺮﮔ
ﻭﺩ ﺭﺩ ﻢﻳﺪﺳ ﺭﻭﺮﻠﮐ ﻟ
ﻴ
،ﺮﻄﻘﻣ ﺏﺁ ﺮﺘ ٥ pH
/ ٧ ( ﻧ ﺭﺍﺪﻬﮕ ﻱ
ﺩﺮﮔ ﻳ ﺪ . ﻧ ﺬﻏﺎﻛ ﻥﺩﻮﻤﻧ ﻚﺸﺧ ﺯﺍ ﺲﭘ ﻴ
ﻭﺮﺘ ﺰﻟﻮﻠﺳ ﻣ ﻴ ﻩﺩ ﻥﺍﺰ
ﻣ ﻴ ﻟﻭﺮﻜ ﻴ ﺘﻧﺁ ﺮﺘ ﻲ ﻧ ﺬﻏﺎﻛ ﻊﺑﺮﻣ ﺮﻫ ﺰﻛﺮﻣ ﺭﺩ ﻥﮊ ﻴ
ﻭﺮﺘ ﺰﻟﻮﻠﺳ ﻪﺑ
ﺕﺭﻮﺻ ﻪﻄﻘﻧ ﺍ ﻱ ﺭﺍﺮﻗ ﻭ ﺪﺷ ﻩﺩﺍﺩ ﺯﺍ ﺲﭘ
ﻞﻣﺎﻛ ﻥﺪﺷ ﻚﺸﺧ
ﺘﻧﺁ ﻲ ﺕﺪﻣ ﻪﺑ ﻥﮊ ٣٠
ﻗﺩ ﻴ ﻥﻭﺭﺩ ﻪﻘ ﻪﮐﻮﻠﺑ ﻝﻮﻠﺤﻣ ﻩﺪﻨﻨﮐ
) ٥ / ٢
ﺷ ﻡﺮﮔ ﻴ ﺑﺮﭼ ﻥﻭﺪﺑ ﮏﺸﺧ ﺮ ﻲ
ﺭﺩ ٥٠ ﻲﻠﻴﻣ ﺮﺘﻴﻟ ﺮﻓﺎﺑ
(TBS
ﻥﻭﺭﺩ ﻥﺁ ﺯﺍ ﺪﻌﺑ ﻭ ﺮﻓﺎﺑ
ﻭﺎﺣ TBS
ﻱ ﺘﻧﺁ ﻲ ﺩﺎﺑ ﻱ ﻟﺎﺼﺤﺘﺳﺍ ﻲ
ﺵﻮﮔﺮﺧ ﺯﺍ )
ﺖﺒﺴﻧ ﻪﺑ ١:١٠٠٠
( ﻪﺘﺷﺍﺬﮔ ﺪﺷ . ﺲﭙﺳ
ﺎﺑ ﻮﺸﺘﺴﺷ ﺎﻫﺮﻓﺎﺑ
ﻱ ) TTBS
ﻳ ﻟ ﮏ ﻴ ﺮﺘ ﻓﺎﺑ ﺮ ﻭﺎﺣ TBS
ﻱ ﻧ ﻴ ﻢ
ﻲﻠﻴﻣ ﺮﺘﻴﻟ ﻮﺗ ﻴﻳ ﻦ ٢٠ ( ﻭ ﺗﺮﺗ ﻪﺑ TBS
ﻴ ﺮﻫ ﻭ ﺭﺎﺑﻭﺩ ﻡﺍﺪﻛ ﺮﻫ ﺐ
ﻗﺩ ﺞﻨﭘ ﺕﺪﻣ ﻪﺑ ﺭﺎﺑ ﻴ
ﺖﻓﺮﮔ ﻡﺎﺠﻧﺍ ﻪﻘ .
ﻧ ﺬﻏﺎﻛ ﻴ ﻪﺑ ﺰﻟﻮﻠﺳ ﻭﺮﺘ
ﺘﻧﺁ ﻝﻮﻠﺤﻣ ﻞﺧﺍﺩ ﺭﺩ ﺖﻋﺎﺳ ﻭﺩ ﺕﺪﻣ ﻲ
ﺩﺎﺑ ﻱ ﻩﺩﺍﺩ ﺭﺍﺮﻗ ﻡﻭﺩ
ﺪﺷ . ﺍﻳ ﺘﻧﺁ ﻦ ﻲ ﺩﺎﺑ ﻱ ﻉﻮﻧ ﺯﺍ
Goat anti rabbit IgG
ﻞﺼﺘﻣ
ﺰﻧﺁ ﻪﺑ ﻳ ﻢ
horseradish peroxidase (HRP)
ﺖﺒﺴﻧ ﺎﺑ ﻭ ﺩﻮﺑ
١:٤٠٠٠ ﺮﻓﺎﺑ ﺭﺩ ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ TBS
. ﺲﭙﺳ ﻮﺸﺘﺴﺷ ﻱ
ﺩ ﻳ ﺮﮕ ﻱ ﺖﻓﺮﮔ ﺕﺭﻮﺻ ﻞﺒﻗ ﻪﻠﺣﺮﻣ ﺪﻨﻧﺎﻣ .
ﻣ ﻴ ﻥﺍﺰ ٣٠
ﻣ ﻴ ﻠ ﻲ ﺭﺩﻮﭘ ﻡﺮﮔ ٤
‐ ﻣ ﻩﺩ ﺭﺩ ﻝﻮﺘﻔﻧﻭﺮﻠﻛ ﻴ
ﻠ ﻲ ﻟ ﻴ ﺺﻟﺎﺧ ﻝﻮﻧﺎﺘﻣ ﺮﺘ
ﺩﺮﺳ
، ﻞﺣ ﺩﺮﮔ ﻳ ﺪ . ﺧﺍ ﺓﺩﺎﻣ ﻴ ﺖﺒﺴﻧ ﻪﺑ ﺮ ١:٦
) ﻳ ﻣ ﻚ ﻴ ﻠ ﻲ ﻟ ﻴ ﺮﺘ
ﻣ ﺞﻨﭘ ﻩﺍﺮﻤﻫ ﻪﺑ ﺍﺮﺘﺴﺑﻮﺳ ﻴ
ﻠ ﻲ ﻟ ﻴ ﺮﻓﺎﺑ ﺮﺘ (TBS
ﻗﺭ ﻴ ﺪﺷ ﻖ .
ﺲﭙﺳ ﻣ ﺭﺎﻬﭼ ﻴ ﻟﻭﺮﻜ ﻴ ﺮﺘ ﺴﻛﺍ ﺏﺁ ﻴ ﺔﻧﮋ ٣٠ % ﺑ ﻪ ﺭﻮﻧ ﺯﺍ ﺭﻭﺩ ﻪﺑ
ﻥﺁ ﻧ ﺬﻏﺎﻛ ﻭ ﻪﺘﺸﮔ ﻪﻓﺎﺿﺍ ﻴ
ﻭﺮﺘ ﻝﺎﻘﺘﻧﺍ ﻥﺁ ﻥﻭﺭﺩ ﻪﺑ ﺰﻟﻮﻠﺳ
ﻳ ﺭﺍﺪﻬﮕﻧ ﻥﺁ ﻥﻭﺭﺩ ﺶﻔﻨﺑ ﻁﺎﻘﻧ ﺭﻮﻬﻇ ﺎﺗ ﻭ ﺖﻓﺎ ﻱ
ﺪﺷ . ﺖﻬﺟ
ﻧ ﺬﻏﺎﻛ ،ﺶﻨﻛﺍﻭ ﻒﻗﻮﺗ ﻴ
ﻭﺮﺘ ﻗﺩ ﺞﻨﭘ ﺕﺪﻣ ﻪﺑ ﺰﻟﻮﻠﺳ ﻴ
ﻪﻘ ﺭﺩ
ﻝﻮﻠﺤﻣ ﻞﺧﺍﺩ ﻓ
ﻴ ﺗﺎﺴﻜ ﻴ ﻮ ) ٤٨٤ ﻣ ﻴ ﻠ ﻲ ﺮﺗ ﻡﺮﮔ ﻳ ﺯﺎﺑ ﺲ ﻱ ﻭ
٧٤٥ ﻣ ﻴ ﻠ ﻲ ﻡﺮﮔ
Na2-EDTA
ﺭﺩ ٤٠٠ ﻲﻠﻴﻣ ﺮﺘﻴﻟ ﺮﻄﻘﻣ ﺏﺁ
ﻥﻭﺮﺘﺳ
، ٥ / ٧ (pH
ﻩﺩﺍﺩ ﺭﺍﺮﻗ ﺪﺷ .
ﻱﺯﺎﺳ ﻲﻨﻏ :١
ﺖﻗﺭ ﻱﺎﻫ ﻔﻠﺘﺨﻣ ﻲ ﺴﻧﺎﭙﺳﻮﺳ ﺯﺍ ﻴ
ﺍﺪﺟ ﻥﻮ ﻳ ﺔ
ﺮﺘﺳ ﺮﻄﻘﻣ ﺏﺁ ﺭﺩ ﺩﺭﺍﺪﻧﺎﺘﺳﺍ ﻥﻭ
ﻬﺗ ﻴ ﺩﺮﮔ ﻪ ﻳ ﺪ . ﺮﻫ ﻥﻭﺭﺩ ﻪﺑ
ﻪﻟﻮﻟ ﻭﺎﺣ ﻑﺭﻭﺪﻨﭘﺍ ﻱ
١٠٠ ﻣ ﻴ ﻟﻭﺮﻜ ﻴ ﺤﻣ ﺮﺘ ﻴ ﺎﻣ ﻂ ﻳ ﻛ ﻊ ﻴ ﮓﻨ
،ﺏ ١٠٠ ﻣ ﻴ ﻟﻭﺮﮑ ﻴ ﺯﺍ ﺮﺘ ﺖﻗﺭ ﻱﺎﻫ ﺩﺮﮔ ﻪﻓﺎﺿﺍ ﺭﻮﺑﺰﻣ ﻳ
ﺪﻧﺪ .
ﻪﻟﻮﻟ ﺎﻫ ﻱ ﺍﺮﺑ ﻑﺭﺪﻨﭘﺍ ﻱ
ﻩﺭﻭﺩ ﺎﻫ ﻱ ﻧﺎﻣﺯ ﻲ ﻒﻠﺘﺨﻣ ) ،ﺮﻔﺻ ٢٤ ،
1. Enrichment
٤٨ ﻭ ٧٢ ﺖﻋﺎﺳ ( ﺭﻮﺗﺎﺑﻮﻜﻧﺍ ﻥﻭﺭﺩ ﺎﻣﺩ ﺭﺩ
ﻱ ٢٧ ﻪﺟﺭﺩ
ﺘﻧﺎﺳ ﻴ ﺭﺍﺪﻬﮕﻧ ﺩﺍﺮﮕ ﻱ
ﺪﻧﺪﺷ . ﻪﺑ ﻭ ﺪﻫﺎﺷ ﻥﺍﻮﻨﻌﺑ ﺮﻔﺻ ﻥﺎﻣﺯ
ﻡﺎﺠﻧﺍ ﻡﺪﻋ ﺔﻟﺰﻨﻣ ﻲﻨﻏ
ﻱﺯﺎﺳ ﺩﻮﺑ . ﻩﺩﻮﻣﺁ ﺎﻫ ﻱ ﺘﻧﺁ ﻲ ﻧﮊ ﻲ ﺲﭘ ،
ﻃ ﺯﺍ ﻲ ﺩ ﻥﻮﻣﺯﺁ ﺭﺩ ،ﻪﻃﻮﺑﺮﻣ ﻥﺎﻣﺯ ﻳ
ﺪﻨﺘﻓﺭ ﺭﺎﻜﺑ ﺎﺒ .
ﻥﻮﻣﺯﺁ ﺎﻫ ﺮﺑ ﻲﻨﺘﺒﻣ ﻱ PCR
ﺝﺍﺮﺨﺘﺳﺍ ﻱﺪﻴﻣﺯﻼﭘ DNA
ﺯﺎﺳﺍﺪﺟ :
ﻱ ﺝﺍﺮﺨﺘﺳﺍ ﻭ
ﻣﺯﻼﭘ ﻴ ﻟ ﺵﻭﺭ ﻪﺑ ﺪ ﻴ
ﻠﻗ ﺰ ﻴ ﺎ
٢ﻲﻳ ﺪﺷ ﻡﺎﺠﻧﺍ
(Sambrook et al.,
1989)
. ﻣﺯﻼﭘ ﺯﺭﻮﻓﻭﺮﺘﮑﻟﺍ ﺖﻬﺟ ﻴ
ﻝﮊ ﺯﺍ ،ﺪ ﺯﻭﺭﺎﮔﺁ ﻳ ﺪﺻﺭﺩ ﮏ
ﺮﻓﺎﺑ ﺭﺩ ) TBE
٧٥ / ١٠ ﺮﺗ ﻡﺮﮔ ﻳ ﺯﺎﺑ ﺲ ﻱ ، ٥ / ٥ ﺳﺍ ﻡﺮﮔ ﻴ ﺪ
ﺭﻮﺑ ﻳ ﻭ ﮏ ٩٣٠ ﻣ ﻴ ﻠ ﻲ ﻡﺮﮔ
Na2-EDTA
ﺭﺩ ﻳ ﻟ ﮏ ﻴ ﺏﺁ ﺮﺘ
ﻣ ﺮﻄﻘ ٨ (pH
ﺩﺮﮔ ﻩﺩﺎﻔﺘﺳﺍ ﻳ
ﺪ . ﻣ ﻴ ﻥﺍﺰ ١٥ ﻣ ﻴ ﻟﻭﺮﻜ ﻴ ﺮﻫ ﺯﺍ ﺮﺘ
ﻧﻮﻤﻧ ﻪ ﻣ ﺞﻨﭘ ﻩﺍﺮﻤﻫ ﻪﺑ ﻴ
ﻟﻭﺮﻜ ﻴ ﻪﻧﻮﻤﻧ ﺮﻓﺎﺑ ﺮﺘ )
ﻝﻮﻠﺤﻣ ٥٠
ﻠﮔ ﺪﺻﺭﺩ ﻴ ﺮﻓﺎﺑ ﺭﺩ ﻝﻭﺮﺴ ﻭ TBE
١ / ٠ ﻢﺠﺣ ﻪﺑ ﻥﺯﻭ ﺪﺻﺭﺩ
ﻮﻠﺑ ﻞﻨﻓ ﻡﺮﺑ (
ﺩﺮﮔ ﻁﻮﻠﺨﻣ ﻳ
ﻚﻫﺎﭼ ﻥﻭﺭﺩ ﺲﭙﺳ ﻭ ﺪ ﺎﻫ
ﻱ ﻝﮊ
ﺭ ﻳ ﺪﺷ ﻪﺘﺨ . ﻪﻧﻮﻤﻧ ﺖﺑﺎﺛ ﮊﺎﺘﻟﻭ ﺎﺑ ﺎﻫ ٩٠
ﺖﻟﻭ ﻪﺑ ﻭﺩ ﺕﺪﻣ
ﺮﺘﻜﻟﺍ ﺖﻋﺎﺳ ﺪﻧﺪﺷ ﺯﺭﻮﻓﻭ
. ﺕﺪﻣ ﻪﺑ ﻝﮊ ﺲﭙﺳ ﻳ
ﺭﺩ ﺖﻋﺎﺳ ﻚ
ﻭﺎﺣ ﻑﺮﻇ ﻱ
ﺗﺍ ﻴ ﺪ ﻳ ﺎﻣﻭﺮﺑ ﻡﻮ ﻳ
٣ﺪ ) ﺖﻈﻠﻏ ﻪﺑ ﻢﻴﻧ
ﻣ ﻴ ﻠ ﻲ ﺭﺩ ﻡﺮﮔ
ﻳ ﻟ ﮏ ﻴ ﺮﻄﻘﻣ ﺏﺁ ﺮﺘ (
ﮓﻧﺭ ﻣﺁ ﻴ ﺰ ﻱ ﻭ ﺮﻄﻘﻣ ﺏﺁ ﺭﺩ ﺲﭙﺳ
ﺩﺮﮔ ﻮﺸﺘﺴﺷ ﻳ
ﺪ . ﻝﮊ ﺭﺩ ﺍﺭﻭﺎﻣ ﺭﻮﻧ ﻱ
ﻩﺎﮕﺘﺳﺩ ﺶﻔﻨﺑ
White/ultraviolet transilluminator
ﻂﺳﻮﺗ ﻭ ﻩﺪﻫﺎﺸﻣ
ﺑﺭﻭﺩ ﻴ ﭙﻣﺎﮐ ﻪﺑ ﻞﺼﺘﻣ ﻦ ﻴ
ﺍﺭﺍﺩ ﺮﺗﻮ ﻱ ﺮﻧ ﺭﺍﺰﻓﺍ ﻡ
UVP Gel
documentation
ﺲﮑﻋ ﻥﺁ ﺯﺍ ﺭﺍﺩﺮﺑ
ﻱ ﺪﺷ . ﺝﺍﺮﺨﺘﺳﺍ DNA
ﺑ ﻩﺪﺷ ﻪ ﻋ ﻥﺍﻮﻨ ﺭﺩ ﺐﻟﺎﻗDNA
ﺑ PCR
ﻪ ﺖﻓﺭ ﺭﺎﻛ .
ﻭ ﻩﺮﻴﺠﻧﺯ ﺶﻨﻛﺍ ﺯﺍﺮﻤﻴﻠﭘ ﻱﺍ
(PCR)٤
: ﻡﺎﺠﻧﺍ ﺖﻬﺟ ،PCR
ﺵﻭﺭ
McManus & Jones (1995)
ﻛﺪﻧﺍ ﺎﺑ ﻲ ﻐﺗ ﻴﻴ ﺑ ﺮ ﻪ ﺎﻛ ﺭ
ﺖﻓﺭ . ﺳﺭﺮﺑ ﺭﻮﻈﻨﻣ ﻪﺑ ﻲ
ﻥﺎﮑﻣﺍ ﺩﺭ ﻳ ﺑﺎ ﻲ ﻘﺘﺴﻣ ﻴ
٥ﻢ ﺮﺘﮐﺎﺑ ﻱ
ﻂﺳﻮﺗ ﺝﺍﺮﺨﺘﺳﺍ ﻡﺎﺠﻧﺍ ﻥﻭﺪﺑ ﻭPCR
،DNA
ﺴﻧﺎﭙﺳﻮﺳ ﻴ ﻧﻮ ﻲ
ﺯﺍ ﺖﺸﮐ ٢٤ ﻪﺘﻋﺎﺳ ﺍﺪﺟ ﻳ ﻪ ﺎﻫ ﻱ ﺮﺘﻛﺎﺑ ﻳﺎ ﻲﻳ ﺎﻤﻧ ﻳ ﺓﺪﻨ ﻥﺎﺑﺰﻴﻣ ﺎﻫ
ﻓﺍﺮﻐﺟ ﻒﻠﺘﺨﻣ ﻖﻃﺎﻨﻣ ﻭ ﻴ
ﺎ ﻲﻳ
، ﺩ ﺏﺁ ﺭﺩ ﺍﺰﺠﻣ ﺭﻮﻄﺑ ﻳ
ﻧﻮ ﻴ ﺓﺰ
ﺮﺘﺳ
٦ﻥﻭ ﻪﻟﻮﻟ ﻞﺧﺍﺩ ﺭﺩ ﺎﻫ
ﻱ ﺪﻨﭘﺍ ﻭ ﻬﺗ ﻑﺭ ﻴ ﺴﻧﺍﺩ ﻭ ﻩﺪﺷ ﻪ ﻴ
ﺔﺘ
ﺭﻮﻧ ﻱ ﺝﻮﻣ ﻝﻮﻃ ﺭﺩ ﻥﺁ ٦٠٠
ﻝﺩﺎﻌﻣ ،ﺮﺘﻣﻮﻧﺎﻧ ﻳ
ﻈﻨﺗ ﻚ ﻴ ﻭ ﻢ
ﻳ ﻣ ﻚ ﻴ ﻟﻭﺮﻜ ﻴ ﺮﺘ ﻡﺎﺠﻧﺍ ﺖﻬﺟ ﻥﺁ ﺯﺍ ﺑ PCR
ﻪ ﺖﻓﺭ ﺭﺎﻛ .
ﺳﺭﺮﺑ ﺖﻬﺟ ﻲ
ﺳﺎﺴﺣ ﻴ ﺖ ،PCR
ﺴﻧﺎﭙﺳﻮﺳ ﻴ ﻧﻮ ﻲ ﺯﺍ
ﻝﻮﻠﺳ ﻱﺎﻫ ﺮﺘﻛﺎﺑ ﻳﺎ ﻲﻳ ﺍﺪﺟ ﻳ ﻪ ﻊﺟﺮﻣ ﺴﻧﺍﺩ ﺎﺑ ﻴ ﺭﻮﻧ ﺔﺘ ﻱ ﻝﺩﺎﻌﻣ
ﻳ ﺝﻮﻣ ﻝﻮﻃ ﺭﺩ ﻚ ٦٠٠
ﻬﺗ ﺮﺘﻣﻮﻧﺎﻧ ﻴ
ﺪﺷ ﻪ . ﺮﺳ ﻱ ﺎﻫ ﻱ ﻒﻠﺘﺨﻣ
2. Alkaline lysis
3. Ethidium bromide 4. Polymerase chain reaction 5. Direct detection
6. Deionized water
ﻑﺮﻌﻣ ﻥﺍﺭﺎﮑﻤﻫ ﻭ ﻩﺩﺍﺯ :
ﻲﺧﺮﺑ ﻲﻳﺁﺭﺎﻛ ﻲﺳﺭﺮﺑ ﺵﻭﺭ
ﺎﻫ ﺮﺑ ﻲﻨﺘﺒﻣ ﻭ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ ﻱ ﺭﺩ PCR
...
۵۹
ﺮﺘﻛﺎﺑ ﺖﻗﺭ ﻱ
ﺏﺁ ﺭﺩ ﻳ ﺓﺭﺎﺼﻋ ﺎ ﻳ ﻩﺪﺷ ﺖﺧﺍﻮﻨﻜ ﮒﺮﺑ ﺖﻓﺎﺑ
ﺳ ﻴ ﺐ ﻓﺎﺻ ﺬﻏﺎﻛ ﻂﺳﻮﺗ ﻲ
ﻩﺭﺎﻤﺷ ﻦﻤﺗﺍﻭ ﻳ
ﻚ ) ﻳ ﻚ ﺖﻓﺎﺑ ﻡﺮﮔ
ﻣ ﻩﺩ ﺭﺩ ﻢﻟﺎﺳ ﻴ
ﻠ ﻲ ﻟ ﻴ ﺮﻄﻘﻣ ﺏﺁ ﺮﺘ ﺮﺘﺳ
ﻥﻭ ( ﻬﺗ ﻴ ﺩﺮﮔ ﻪ ﻳ ﺪ . ﺖﻬﺟ
ﻌﺗ ﻴﻴ ﻜﺸﺗ ﺪﺣﺍﻭ ﻦ ﻴ
ﻞ ﻨﻠﻛ ﺓﺪﻨﻫﺩ ﻲ
، ١٠٠ ﻣ ﻴ ﻟﻭﺮﻜ ﻴ ﺯﺍ ﺮﺘ
ﺮﺳ ﻳ ﺎﻬ ﻱ ﻭﺭ ﺮﺑ ﻞﺻﺎﺣ ﺖﻗﺭ ﻱ
ﺤﻣ ﻴ ﺬﻐﻣ ﺭﺎﮔﺁ ﻂ ﻱ
ﻭﺎﺣ ﻱ ﻳ ﻚ
ﺪﺷ ﻩﺩﺍﺩ ﺖﺸﻛ ﺰﻛﻮﻠﮔ ﺪﺻﺭﺩ .
ﻨﭽﻤﻫ ﻴ ،ﻦ ﻳ ﻣ ﻚ ﻴ ﻟﻭﺮﻜ ﻴ ﺯﺍ ﺮﺘ
ﻌﺗ ﺖﻬﺟ ﺖﻗﺭ ﺮﻫ ﻴﻴ
ﺳﺎﺴﺣ ﻦ ﻴ
ﺭ ﺖ ﺵﻭ ﻩﺩﺎﻔﺘﺳﺍ PCR
ﺩﺮﮔ ﻳ ﺪ . ﺴﻧﺎﭙﺳﻮﺳ ﻴ ﻥﻮ ﺍﺪﺟ ﻳ ﻪ ﺎﻫ ﻱ ﺮﺘﻛﺎﺑ ﻳﺎ ﻲﻳ ﻪﺑ ﻖﻠﻌﺘﻣ
ﺲﻨﺟ ﺎﻫ ﻱ ﺩ ﻳ ﺮﮕ ﺗ ﻬ ﻴ ﻪ ﺍﺪﺟ ﻩﺍﺮﻤﻫ ﻪﺑ ﻭ ﻩﺪﺷ ﻳ
ﺔ
،ﻊﺟﺮﻣ
ﺳﺭﺮﺑ ﺖﻬﺟ ﻲ
ﺻﺎﺼﺘﺧﺍ ﻲ ﺵﻭﺭ ﻥﺩﻮﺑ ﺩﺭ ﺭﺩ PCR
ﻳ ﺑﺎ ﻲ
E. amylovora
ﺪﻨﺘﻓﺮﮔ ﺭﺍﺮﻗ ﻩﺩﺎﻔﺘﺳﺍ ﺩﺭﻮﻣ .
ﻝﻭﺍ ﺭﻭﺩ ﻡﺎﺠﻧﺍ ﺖﻬﺟ ،PCR
ﺯﺍ ﻭﺩ ﺮﮔﺯﺎﻏﺁ ١٨
ﺗﻮﺌﻠﻛﻮﻧ ﻴ ﺪ
A(5′-CGGTTTTTAACGCTGGG-3′) ﻱ ﻭ
B(5′-GGGCAAATACTCGGATT-3′)
ﻪﮐ ﺱﺎﺳﺍﺮﺑ
ﺔﻌﻄﻗ ﻳ ﮏ ﻛ ﻴ ﺯﺎﺑ ﻮﻠ ﻱ ﻣﺯﻼﭘ ﻴ ﺪ
pEA29
ﺮﺘﮐﺎﺑ ﻱ
E.amylovora
ﻡﺎﻨﺑ ﺣﺍﺮﻃ PstI
ﻲ ﺪﻧﺩﻮﺑ ﻩﺪﺷ )
١٩
، ٥ ( ﻭ
ﻱﺍﺮﺑ ﻡﺎﺠﻧﺍ
Nested-PCR
ﺯﺍ ﻭﺩ ﺮﮔﺯﺎﻏﺁ ٢١ ﺗﻮﺌﻠﻛﻮﻧ ﻴ ﺪ ﻱ
AJ75(5′-CGTATTCACGGCTTCGCAGAT-3′)
ﻭ
AJ76(5′-ACCCGCCAGGATAGTCGCATA-3′)
ﻪﻛ
ﻟﺍﻮﺗ ﺱﺎﺳﺍ ﺮﺑ ﻲ
ﺎﻫ ﻱ ﺎﻣ ﻴﺑ ﺁ ﻭﺩ ﻦ ﺮﮔﺯﺎﻏ ﻭ A
ﺣﺍﺮﻃ B
ﻲ ﻩﺪﺷ
ﺩﺮﮔ ﻩﺩﺎﻔﺘﺳﺍ ،ﺪﻧﺩﻮﺑ ﻳ
ﺪ
(McManus & Jones, 1995)
.
ﺎﻬﻧ ﻢﺠﺣ ﻲﻳ ﺩ ﺏﺁ ﺭﺩ ﻪﻛ ﺶﻨﻛﺍﻭ ﻁﻮﻠﺨﻣ ﻳ
ﻧﻮ ﻴ ﻬﺗ ﻩﺰ ﻴ ﻪ
ﺩﺮﮔ ﻳ ،ﺪ ٥٠ ﻣ ﻴ ﻟﻭﺮﻜ ﻴ ﺪﺷ ﻪﺘﻓﺮﮔ ﺮﻈﻧ ﺭﺩ ﺮﺘ .
ﺍﻳ ﻁﻮﻠﺨﻣ ﻦ
ﻭﺎﺣ ﻱ ٥ ﻣ ﻴ ﻟﻭﺮﻜ ﻴ ﺮﺘ ﺮﻓﺎﺑ
10x PCR
، ٥ / ١ ﻣ ﻴ ﻠ ﻲ ﺭﻻﻮﻣ ﺭﻭﺮﻠﻛ
ﻨﻣ ﻴ ﺰ ﻳ ،ﻢ ٦٠٠ ﻣ ﻴ ﻝﻮﻣﻭﺮﻜ ،dNTP
٥٠ ﭘ ﻴ ﺮﮔﺯﺎﻏﺁ ﻝﻮﻣﻮﻜ ،A
٥٠ ﭘ ﻴ ﻝﻮﻣﻮﻜ ﺮﮔﺯﺎﻏﺁ ،B
١٠ ﻣ ﻴ ﻠ ﻲ ﻝﻮﻣ ٢ ـ ،ﻝﻮﻧﺎﺗﺍﻮﺘﭘﺎﻛﺮﻣ ٨
ﻣ ﻴ ﻡﺮﮔﻭﺮﻜ
١BSA
، ٥ % ﺩ ﻱ ﺘﻣ ﻴ ﺎﺴﻛﻮﻔﻟﻮﺳ ﻞ ﻳ
،ﺪ ١ / ٠ ﺪﺻﺭﺩ
ﻮﺗ ﻴﻳ ﻦ ٢٠ ، ٥ / ٢ ﺪﺣﺍﻭ
Taq DNA polymerase
ﻭ ﺐﺳﺎﻨﺘﻣ
ﺑ ﺶﻨﮐﺍﻭ ﻭ ﻪﻧﻮﻤﻧ ﻉﻮﻧ ﺎﺑ ﻴ
ﻦ ﻳ ﻣ ﻭﺩ ﺎﺗ ﮏ ﻴ
ﻟﻭﺮﮑ ﻴ ﺮﺘ
DNA
٢ﺐﻟﺎﻗ ﺩﻮﺑ . ﺶﻨﻛﺍﻭ ﻁﻮﻠﺨﻣ ﻪﻟﻮﻟ ﻪﺑ
ﺎﻫ ﻱ ﻑﺭﺪﻨﭘﺍ ٢٠٠ ﻣ ﻴ ﻭﺮﮑ ﻟ ﻴ ﺮﺘ ﻱ
ﻥﻭﺮﺘﺳ ﺪﺷ ﻩﺩﺍﺩ ﻝﺎﻘﺘﻧﺍ .
ﻪﻟﻮﻟ ﺜﻜﺗ ﺖﻬﺟ ﺎﻫ ﻴ
ﻩﺎﮕﺘﺳﺩ ﻞﺧﺍﺩ ،ﺮ
ﻣﺮﺗ ﻝﺎ ﺎﺳ ﻳ ﻝﺪﻣ ﺮﻠﻜ
Palm-Cycler
ﺖﻛﺮﺷ ﺖﺧﺎﺳ
Corbett
Research
ﺳ ﻭ ﺪﻨﺘﻓﺮﮔ ﺭﺍﺮﻗ ﻴ
ﻞﻜ ﻂﺳﻮﺗ ﺎﻫ ﻳ
ﺔﻣﺎﻧﺮﺑ ﻚ
ﭙﻣﺎﻛ ﻴ ﺮﺗﻮ ﻱ ﺵﻭﺭ ﻖﺑﺎﻄﻣ
McManus & Jones (1995)
ﻈﻨﺗ ﻴ ﺪﻧﺪﺷ ﻢ . ﺖﺷﺮﺳﺍﻭ ﺯﺎﺳ ﻱ ﻟﻭﺍ ﻴ ﺎﻣﺩ ﺭﺩ ،ﻪ ﻱ
٩٤ ﻪﺟﺭﺩ
ﺘﻧﺎﺳ ﻴ ﺑ ﺩﺍﺮﮕ ﻪ ﺕﺪﻣ ٥ ﻗﺩ ﻴ ﺳ ﺭﺩ ﻪﻘ ﻴ ﺲﭙﺳ ﺪﺷ ﻡﺎﺠﻧﺍ ﻝﻭﺍ ﻞﻜ
٣٧ ﺳ ﻴ ﺑ ﻞﻜ ﻦﻳﺍ ﻪ ﺖﻓﺮﮔ ﺕﺭﻮﺻ ﺡﺮﺷ :
ﺖﺷﺮﺳﺍﻭ ﺔﻠﺣﺮﻣ
1. Bovine serumalbumin
2. DNA template
ﻪﺘﺷﺭ ﻭﺩ ﻥﺪﺷ ﺭﺩ
٩٤ ﺘﻧﺎﺳ ﻪﺟﺭﺩ ﻴ
ﺕﺪﻤﺑ ﺩﺍﺮﮕ ﻳ
ﻗﺩ ﻚ ﻴ ﻪﻘ ،
ﺒﺴﭼ ﻴ ﺎﻫﺮﮔﺯﺎﻏﺁ ﻥﺪ ﻪﺑ
ﻑﺪﻫ DNA
ﺭﺩ ٥٢ ﺘﻧﺎﺳ ﺔﺟﺭﺩ ﻴ
ﺩﺍﺮﮕ
ﺑ ﻪ ﻗﺩ ﻭﺩ ﺕﺪﻣ ﻴ
ﻪﻘ ﻭ ﺜﻜﺗ ﻴ ﺮ ﻑﺪﻫ DNA
ﺭﺩ ٧٢ ﺔﺟﺭﺩ
ﺘﻧﺎﺳ ﻴ ﺑ ﺩﺍﺮﮕ ﻪ ﻗﺩ ﻭﺩ ﺕﺪﻣ ﻴ
ﻪﻘ . ﻂﺴﺑ ﺎﻬﻧ ﻲﻳ ﺪﻫ DNA
ﻑ ﺰﻴﻧ
ﺑ ﻪ ﺕﺪﻣ ١٥ ﻗﺩ ﻴ ﻪﻘ ﺭﺩ ٧٢ ﺘﻧﺎﺳ ﺔﺟﺭﺩ ﻴ
ﺩﺍﺮﮕ ﺪﺷ ﻡﺎﺠﻧﺍ .
ﻡﺎﺠﻧﺍ ﺖﻬﺟ
Nested-PCR
ﺭﺍﺪﻘﻣ ﻳ ﻣ ﻚ ﻴ ﻟﻭﺮﻜ ﻴ ﺯﺍ ﺮﺘ
ﺭﻭﺩ ﺯﺍ ﻞﺻﺎﺣ ﺓﺩﺭﻭﺁﺮﻓ ﻝﻭﺍ
ﻪﺑPCR
٤٩ ﻣ ﻴ ﻟﻭﺮﻜ ﻴ ﻁﻮﻠﺨﻣ ﺮﺘ
ﻭﺎﺣ ﻪﻛ ﺶﻨﻛﺍﻭ ﻱ
ﺎﻫﺮﮔﺯﺎﻏﺁ ﻱ ﻭ AJ75
) AJ76
٥٠ ﭘ ﻴ ﻝﻮﻣﻮﻜ
ﺮﮔﺯﺎﻏﺁ ﺮﻫ ﺯﺍ (
ﺩﺮﮔ ﻪﻓﺎﺿﺍ ،ﺩﻮﺑ ﻳ
ﺪ . ﺍ ﺭﺩ ﻳ ﻪﻠﺣﺮﻣ ﻦ ﻧ ﻴ ﻁﻮﻠﺨﻣ ﺰ
ﻣ ﺶﻨﻛﺍﻭ ﺭﻭﺩ ﺪﻨﻧﺎ ﻝﻭﺍ ﺩﻮﺑ PCR
ﻭ ﻭ ﺎﻫﺮﮔﺯﺎﻏﺁ ﻂﻘﻓ
DNA
ﻮﮕﻟﺍ ﺪﻨﺘﺷﺍﺩ ﺕﻭﺎﻔﺗ .
ﻠﻛ ﻴ ﻡﺎﺠﻧﺍ ﺭﺩ ﻩﺩﺎﻔﺘﺳﺍ ﺩﺭﻮﻣ ﺩﺍﻮﻣ ﺔ
ﺶﻨﻛﺍﻭ ﺎﻫ ﻱ ﻭ PCR Nested-PCR
ﺳ ﺖﻛﺮﺷ ﺯﺍ ﻴ
ﻥﮊﺎﻨ ﻬﺗ ﻴ ﻪ
ﺪﻧﺪﺷ .
ﺮﻓ ﺁ ﻩﺩﺭﻭ ﺎﻫ ﻱ ﺮﻫ ﻳ ﺯﺍ ﮏ ﻥﻮﻣﺯﺁ ﺎﻫ ﻱ ﻨﺘﺒﻣ ﻲ ﺮﺑ ﻭ PCR
Nested-PCR
ﻩﺍﺮﻤﻫ ﻪﺑ ﻳ
ﻧﺎﺑﺩﺮﻧ ﺮﮐﺭﺎﻣ ﮏ ﻲ
ﻳ ﮐ ﮏ ﻴ ﺖﻔﺟ ﻮﻠ
ﺯﺎﺑ ﻱ )
GeneRulerTM 1Kbp
( ﺭﺩ ﻩﺪﺷ ﺮﮐﺫ ﺵﻭﺭ ﻖﺑﺎﻄﻣ
ﺖﻤﺴﻗ ﻣﺯﻼﭘ DNA
ﻴ ﺪ ﻱ ﺭﺩ ، ﮊ ﻝ ﺯﻭﺭﺎﮔﺁ ﻳ ﺪﺻﺭﺩ ﮏ
ﺩﺮﮔ ﺯﺭﻮﻓﻭﺮﺘﮑﻟﺍ ﻳ
ﺪﻧﺪ .
ﺘﻧ
ﺎﻳ ﺞ
ﺯﺍ ٣٠٠ ﮔ ﺔﻧﻮﻤﻧ ﻴ ﻫﺎ ﻲ ، ٢٠٠ ﺍﺪﺟ ﻳ ﻪ ﻱﺮﺘﮐﺎﺑ ﺮﺑ ﺱﺎﺳﺍ
ﺵﻭﺭ ﺎﻫ ﻱ ﺿﻮﺗ ﻴ ﺯﺎﺳﺍﺪﺟ ﻩﺪﺷ ﻩﺩﺍﺩ ﺢ ﻱ
ﺩﺮﮔ ﻳ ﺪ . ﺱﺎﺳﺍﺮﺑ
ﺎﺘﻧ ﻳ ﺞ ﻥﻮﻣﺯﺁ ﺎﻫ ﻱ ﺗﺎﻣﺪﻘﻣ ﻲ ﻣ ﺯﺍ ، ﻴ ﻥﺎ ﻥﺁ ﺎﻫ ٦٠ ﺍﺪﺟ ﻳ ﻡﺮﮔ ﻪﻛ ﻪ
ﺴﻛﺍ ﻭ ﻴ ﻔﻨﻣ ﺯﺍﺪ ﻲ ﺍﺭﺍﺩ ،ﺖﺒﺜﻣ ﻥﺍﻮﻟ ، ﻱ
ﻠﺑﺎﻗ ﻴ ﺑ ﺪﺷﺭ ﺖ ﻲ ﺯﺍﻮﻫ ﻱ ،
ﺍﻳ ﺳﺎﺴﺣ ﻕﻮﻓ ﺶﻨﻛﺍﻭ ﺩﺎﺠ ﻴ
ﻟﻮﺗ ،ﻥﻮﺗﻮﺗ ﮒﺮﺑ ﺭﺩ ﺖ ﻴ
ﺪ
ﺮﺘﻛﺎﺑ ﺕﺎﺷﻭﺍﺮﺗ ﻳﺎ
ﻲﻳ ﻭﺭ ﻱ ﻣ ﻴ ﺳ ﺱﺭﺎﻧ ﺓﻮ ﻴ
ﺑﻼﮔ ،ﺐ ﻲ ﺞﻨﻬﻧ ﻭ
ﺥﺮﺳ ﻞﮔ ﻟﻮﺗ ﻡﺪﻋ ﻭ ﻴ
ﺤﻣ ﺭﺩ ﺖﻨﺳﺭﻮﻠﻓ ﮓﻧﺭ ﺪ ﻴ
ﻛ ﻂ ﻴ ﺏ ﮓﻨ
ﻥﺍﻮﻨﻌﺑ ،ﺪﻧﺩﻮﺑ
E. amylovora
ﺎﺳﺎﻨﺷ ﻲﻳ ﺪﻧﺪﺷ . ﻨﻠﻛ ﻲ ﺎﻫ ﻱ
ﺍﻳ ﺍﺪﺟ ﻦ ﻳ ﻪ ﻭﺭ ﺎﻫ ﻱ ﺤﻣ ﻴ ﺭﺎﮔﺁ ﻂ ﺬﻐﻣ ﻱ ، ٢ ‐ ١ ﻣ ﻴ ﻠ ﻲ ﺮﻄﻗ ﺮﺘﻣ
ﺍﺭﺍﺩ ،ﻕﺍﺮﺑ ﻭ ﻪﺘﺴﺟﺮﺑ ،ﺭﻭﺪﻣ ﻭ ﻪﺘﺷﺍﺩ ﻱ
ﺷﺎﺣ ﻴ ﻭ ﻞﻣﺎﻛ ﺔ ﻪﺑ
ﺪﻧﺩﻮﺑ ﻦﺷﻭﺭ ﻡﺮﻛ ﮓﻧﺭ .
٣٠ ﺍﺪﺟ ﻳ ﺑ ﺯﺍ ﻪ ﻴ ﻦ ٦٠ ﺍﺪﺟ ﻳ ﻕﻮﻓ ﻪ ﺮﻛﺬﻟﺍ ﻪﺑ ﻓﺩﺎﺼﺗ ﺭﻮﻃ ﻲ
ﻩﺍﺮﻤﻫ ﻪﺑ ،ﻩﺪﺷ ﺏﺎﺨﺘﻧﺍ ٩
ﺍﺪﺟ ﻳ ﺭﺩ ﺔ ﻳ ﺘﻓﺎ ﻲ ﺕﺎﺴﺳﺆﻣ ﺯﺍ
ﻘﺤﺗ ﻴ ﺗﺎﻘ ﻲ ﻞﺒﻗ ﺭﺩ ﻩﺪﺷ ﺮﻛﺫ ﻭ
ﺍﺪﺟ ﻳ ﺔ ﻊﺟﺮﻣ ، ﺭﺩ ﻥﻮﻣﺯﺁ ﺎﻫ ﻱ
ﺪﻌﺑ ﻱ ﺪﻨﺘﻓﺮﮔ ﺭﺍﺮﻗ ﻩﺩﺎﻔﺘﺳﺍ ﺩﺭﻮﻣ .
ﻲﻳﺍﺰﻳﺭﺎﻤﻴﺑ ﺕﺎﺒﺛﺍ
ﺳﺎﺴﺣ ﻕﻮﻓ ﻥﻮﻣﺯﺁ ﺭﺩ ﻴ
ﻭﺭ ﺖ ﻱ ﺍﺪﺟ ،ﻥﻮﺗﻮﺗ ﮒﺮﺑ ﻳ
ﻪ ﺎﻫ ﻱ
ﺎﻨﺜﺘﺳﺍ ﻪﺑ ﺐﺨﺘﻨﻣ ﻱ
ﺍﺪﺟ ﻳ ﺔ
،ﻊﺟﺮﻣ ﻑﺮﻇ ٤٨
‐ ٢٤ ﺖﻋﺎﺳ
ﻌﺿﻮﻣ ﻥﺪﺷ ﺯﻭﺮﻜﻧ ﺐﺟﻮﻣ ﻲ
ﮒﺮﺑ ﺎﻫ ﻱ ﺎﻣ ﻳ ﻪ ﻧﺯ ﻲ ﻩﺪﺷ
ﺩﺮﮔ ﻳ ﺪﻧﺪ . ﺎﻣ ﻳ ﻪ ﻧﺯ ﻲ ﻣ ﻴ ﻩﻮ ﺎﻫ ﻱ ﺳ ﺱﺭﺎﻧ ﻴ ﺑﻼﮔ ﻭ ﺐ ﻲ
ﺞﻨﻬﻧ ﻭ
۶۰ ﻪﻠﺠﻣ
ﻩﺎﻴﮔ ﺶﻧﺍﺩ ﻲﻜﺷﺰﭘ
ﺩ ،ﻥﺍﺮﻳﺍ ﻩﺭﻭ ۴۰ ، ﻩﺭﺎﻤﺷ ۱ ، ۱۳۸۸
ﺍﺪﺟ ﺎﺑ ﺥﺮﺳ ﻞﮔ ﻳ
ﻪ ﺎﻫ ﻱ
E. amylovora
ﺯﺍ ﺲﭘ ٣
‐ ٢ ،ﺯﻭﺭ
ﻜﺸﺗ ﻪﺑ ﺮﺠﻨﻣ ﻴ
ﻪﻜﻟ ﻞ ﺎﻫ ﻱ ﺎﻬﻠﺤﻣ ﻑﺍﺮﻃﺍ ﺭﺩ ﻪﺘﺧﻮﺴﺑﺁ ﻱ
ﻤﺧﺯ ﻲ ﺩﺮﮔ ﻳ ﻪﻜﻟ ،ﻥﺎﻣﺯ ﺖﺷﺬﮔ ﺎﺑ ﻪﻛ ﺪ ﺵﺮﺘﺴﮔ ﺎﻫ
ﻳ ﻭ ﻪﺘﻓﺎ
ﭘ ﻢﻬﺑ ﻴ ﺪﻨﺘﺳﻮ . ﺪﭘ ﻳ ﺮﺘﻛﺎﺑ ﺕﺎﺷﻭﺍﺮﺗ ﻥﺪﺷ ﺭﺍﺪ ﻳﺎ
ﻲﻳ ﻭﺭ ﻱ ﺢﻄﺳ
ﻣ ﻴ ﻩﻮ ﺮﺗﺯﺭﺎﺑ ﺯﺍ ،ﺎﻫ ﻳ
ﺑ ﺕﺎﺼﺨﺸﻣ ﻦ ﻴ
ﺭﺎﻤ ﻳ ﺍﺰ ﻲﻳ ﺍ ﻳ ﺮﺘﻛﺎﺑ ﻦ ﻱ ﺩﻮﺑ .
ﻣ ﺭﺩ ﻴ ﺍﺪﺟ ﻥﺎ ﻳ ﻪ ﺎﻫ ﻱ ﺎﻣﺯﺁ ﺩﺭﻮﻣ ﻳ
ﺍﺪﺟ ،ﺶ ﻳ ﺍ ﺔ ﺩﺭﺍﺪﻧﺎﺘﺳ ﻭ
ﺍﺪﺟ ﻳ ﻪ ﺎﻫ ﻱ
Ea38, Ea39, Ea40
) ﻥﺎﺘﺳﺍ ﺯﺍ ﻪﺳ ﺮﻫ ﻪﮐ
ﺪﻧﺩﻮﺑ ﻥﺎﺳﺍﺮﺧ (
ﻫ ﻴ ﺮﺘﻛﺎﺑ ﺕﺎﺤﺷﺮﺗ ﻪﻧﻮﮕﭽ ﻳﺎ
ﻲﻳ ﻭﺭ ﻱ
ﻣ ﻴ ﻩﻮ ﺎﻫ ﻱ ﻕﻮﻓ ﺱﺭﺎﻧ ﺍ ﺮﻛﺬﻟﺍ
ﻳ ﺪﻧﺩﺮﻜﻧ ﺩﺎﺠ .
ﻪﻳﺍﺪﺟ ﻲﭙﻴﺗﻮﻨﻓ ﺕﺎﻴﺻﻮﺼﺧ ﺎﻫ
ﻠﻛ ﻴ ﺍﺪﺟ ﺔ ﻪﻳ ﺎﻫ ﺴﻛﺍ ﻭ ﻡﺮﮔ ﻥﻮﻣﺯﺁ ﺮﻈﻧ ﺯﺍ ﻴ
ﻔﻨﻣ ،ﺯﺍﺪ ﻲ ﻭ
ﺑ ﻲ ﺯﺍﻮﻫ ﻱ ﺘﺧﺍ ﻴ ﺭﺎ ﻱ ﺪﻧﺩﻮﺑ . ﻨﻠﻛ ﻲ ﻥﺁ ﺎﻫ ﻭﺭ ﻱ ﺤﻣ ﻴ ﻂ ﺭﺎﮔﺁ
ﺬﻐﻣ ﻱ ﻭﺎﺣ ﻱ ٥ % ﺮﻛﻮﺳ ﻭ ﻔﺳ ،ﺯ ﻴ ﻕﺍﺮﺑ ﻭ ﺭﺍﺪﺑﺎﻌﻟ ،ﻪﺘﺴﺟﺮﺑ ،ﺪ
) ﺖﺒﺜﻣ ﻥﺍﻮﻟ (
ﺩﻮﺑ . ﻣﺎﻤﺗ ﻲ ﺍﺪﺟ ﻳ ﻪ ﻭﺭ ﺎﻫ ﻱ ﺤﻣ ﻴ ﻛ ﻂ ﻴ ،ﺏ ﮓﻨ
ﻠﻛ ﻨ ﻴ ﺎﻬ ﻱ ﺷ ﻴ ﺮ ﻱ ﺖﻨﺳﺭﻮﻠﻓ ﺔﻧﺍﺪﮕﻧﺭ ﺪﻗﺎﻓ ﻭ ﺭﺍﺪﺑﺎﻌﻟ ،ﮓﻧﺭ
ﺍﻳ ﺪﻧﺩﻮﻤﻧ ﺩﺎﺠ .
ﻠﻛ ﺭﺩ ﻴ ﺍﺪﺟ ﺔ ﻳ ﻪ ،ﺎﻫ ﺁ ﻟﻮﺗ ﻥﻮﻣﺯ ﻴ
ﺩﺍﻮﻣ ﺪ
ﺣﺍ ﻴ ﺯﻭﺮﻛﻮﺳ ﺯﺍ ﻩﺪﻨﻨﻛﺀﺎ
، ﻟﻮﺗ ﻴ ﺋﻮﺘﺳﺍ ﺪ ﻴ ﻭ ﺖﺒﺜﻣ ﺯﻻﺎﺗﺎﻛ ﻭ ﻦ
ﺁ ﻥﻮﻣﺯ ﺎﻫ ﻱ ﺘﻜﭘ ،ﺯﺎﺗﺎﻔﺴﻓ ﻴ
ﻫ ،ﺯﺎﻨ ﻴ ﻟﻭﺭﺪ ﻴ ﻟﻮﻜﺳﺍﺰ ﻴ ﻫ ،ﻦ ﻴ ﻟﻭﺭﺪ ﻴ ﺰ
ﻟﻮﺗ ،ﻪﺘﺳﺎﺸﻧ ﻴ
ﺍ ﺪ ﻳ ﻝﻭﺪﻨ ، ﻫ ﻴ ﻟﻭﺭﺪ ﻴ ﻮﺗ ﺰ ﻴﻳ ﻦ ٨٠
، ﻧﮊﺭﺁ ﻴ ﺩ ﻦ ﻱ
ﻫ ﻴ ﻟﻮﺗ ،ﺯﻻﻭﺭﺪ ﻴ
ﻔﻟﻮﺳ ﺯﺎﮔ ﺪ ﻴ
ﻫ ﺪ ﻴ ،ﻪﻧﻮﺘﭙﭘ ﺏﺁ ﺯﺍ ﻥﮊﻭﺭﺪ
ﻟﺎﻌﻓ ﻴ ﺔﺘﺴﻫ ﺖ ﻳ
ﻟﻮﺗ ،ﺦ ﻴ ﺪ ٣ ﺘﻣ ﻭ ﺯﻮﺘﻛﻻﻮﺘﻛ –
ﻴ ﻔﻨﻣ ﺩﺭ ﻞ ﻲ
ﺩﻮﺑ . ﺍﺪﺟ ﻳ ﻪ ﺎﻫ ) ﺎﻨﺜﺘﺳﺍ ﻪﺑ ﻱ ﺍﺪﺟ ﻳ ﻪ ﺎﻫ ﻱ
Ea38, Ea39, Ea40
ﺪﻧﺩﻮﺑ ﻦﻴﺗﻻﮊ ﺏﻭﺫ ﺕﺭﺪﻗ ﺪﻗﺎﻓ ﻪﻛ (
ﺗﻻﮊ ﻴ ﺪﻨﻛ ﻪﺑ ﺍﺭ ﻦ ﻱ
ﻡﺎﻌﻃ ﻚﻤﻧ ﻞﻤﺤﺗ ﺕﺭﺪﻗ ﺪﻗﺎﻓ ﻭ ﺪﻧﺩﻮﻤﻧ ﺏﻭﺫ ٦
% ﻟﻭ ﺪﻧﺩﻮﺑ ﻲ
ﻞﻤﺤﺗ ﺭﺩ ٥
% ﺪﻧﺩﺍﺩ ﻥﺎﺸﻧ ﺕﻭﺎﻔﺗ ،ﻚﻤﻧ .
ﺶﻨﻛﺍﻭ ﺎﻫ ﻱ ﻟﻮﺗ ﻴ ﺪ
ﺯﺎﮔ ﺯﺍ ﺣﺍ ،ﺰﻛﻮﻠﮔ ﻴ
ﻧ ﺀﺎ ﻴ ﻩﺭﻭﺍ ،ﺕﺍﺮﺘ
،ﺯﺁ ﻟﻮﺗ ﻴ ﻭﺭ ﻪﻧﺍﺪﮕﻧﺭ ﺪ ﻱ
ﺤﻣ ﻴ ﻂ ﺎﻣﺩ ﺭﺩ ﺪﺷﺭ ﻭ YDC
ﻱ ٣٦ ﺘﻧﺎﺳ ﺔﺟﺭﺩ ﻲ
ﺭﺩ ،ﺩﺍﺮﮔ
ﻠﻛ ﻴ ﺔ ﺍﺪﺟ ﻳ ﻪ ﻔﻨﻣ ﺎﻫ ﻲ ﺩﻮﺑ . ﻥﺁ ﺎﻫ ﺭ ﻭ ﻱ ﺤﻣ ﻴ ﻂ ،EMB
ﻨﻠﻛ ﻲ ﺎﻫ ﻱ ﻟﺎﺘﻣ ﺰﺒﺳ ﻴ
ﻜﺸﺗ ﻚ ﻴ ﺎﻧﺍﻮﺗ ﻭ ﺪﻧﺩﺍﺩ ﻞ ﻲﻳ
ﻑﺮﺼﻣ
ﺳ ﻴ ﺤﻣ ﺭﺩ ﺍﺭ ﺕﺍﺮﺘ ﻴ
ﺳ ﻂ ﻴ ﻥﻮﻤ ﺳ –
ﻴ ﺪﻨﺘﺷﺍﺩ ﺭﺎﮔﺁ ﺕﺍﺮﺘ .
ﺍﺪﺟ ﺶﻨﻛﺍﻭ ﻳ
ﻪ ﻭﺭ ﺎﻫ ﻱ ﺷ ﻴ ﺮ ﻞﺴﻧﺭﻮﺗ ﺳﺍ ﺕﺭﻮﺼﺑ ﺭﺍﺩ ﻴ
ﺪ ﻱ
ﺯﺭﺍ ﻳ ﺑﺎ ﻲ ﺪﺷ .
ﻥﻮﻣﺯﺁ ﺎﻫ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ ﻱ
ﺭﺎﮔﺁ ﺭﺩ ﻪﻳﻮﺳﻭﺩ ﺖﺸﻧ ﻥﻮﻣﺯﺁ :
ﺍ ﻳ ﻠﻛ ﺖﺴﻧﺍﻮﺗ ﺵﻭﺭ ﻦ ﻴ
ﺔ
ﺍﺪﺟ ﻳ ﻪ ﺎﻫ ﻱ
E. amylovora
ﻪﺑ ﻪﻛ ﺍﺭ ﻥﺎﺑﺰﻴﻣ
ﺎﻫ ﻖﻃﺎﻨﻣ ﻭ
ﺎﺳﺎﻨﺷ ،ﺪﻨﺘﺷﺍﺩ ﻖﻠﻌﺗ ﻒﻠﺘﺨﻣ ﻲﻳ
ﺎﻤﻧ ﻳ ﺪ . ﺘﻧﺁ ﻲ ﻡﺮﺳ ﺎﻫ ﻱ ،ﺭﻮﺑﺰﻣ
ﻏ ﺶﻨﻛﺍﻭ ﻴ ﺻﺎﺼﺘﺧﺍﺮ ﻲ ﺎﺳ ﺎﺑ ﻳ ﺮ ﺲﻨﺟ ﺎﻫ ﻱ ﺮﺘﻛﺎﺑ ﻳﺎ ﻲﻳ ﻼﺒﻗ ﻪﻛ
ﻣﺎﺳﺍ ﻲ ﻥﺁ ﺎﻫ ﺪﺷ ﺮﻛﺫ ﺪﻧﺩﺍﺪﻧ ﻥﺎﺸﻧ .
ﺎﺒﻳﺩ ﻥﻮﻣﺯﺁ :
ﺍ ﺭﺩ ﻳ
،ﻥﻮﻣﺯﺁ ﻦ ﺘﻧﺁ
ﻲ ﺎﻬﻣﺮﺳ ﻱ ﻬﺗ ﻴ ﻩﺪﺷ ﻪ
ﺎﺳﺎﻨﺷ ﻪﺑ ﺭﺩﺎﻗ ﻲﻳ
ﻠﻛ ﻴ ﺔ ﺟ ﺍﺪ ﻳ ﻪ ﺎﻫ ﻱ
E. amylovora
ﻁﻮﺑﺮﻣ ﻪﺑ
ﻥﺎﺑﺰﻴﻣ ﺎﻫ ﻒﻠﺘﺨﻣ ﻖﻃﺎﻨﻣ ﻭ ﺪﻧﺩﻮﺑ
) ﻞﮑﺷ ١ .(
ﺳﺭﺮﺑ ﺭﺩ ﻲ
ﺻﺎﺼﺘﺧﺍ ﻲ ﺍ ﻥﺩﻮﺑ ﻳ ﺘﻧﺁ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﺵﻭﺭ ﻦ ﻲ
ﻕﺎﺤﻟﺍ ﻡﺮﺳ
ﻪﺑ ﻩﺪﺷ ﻫ HRP
ﻴ ﻌﻃﺎﻘﺗ ﺶﻨﻛﺍﻭ ﻪﻧﻮﮕﭽ ﻲ
ﺎﺳ ﺎﺑ ﻳ ﺮ ﺲﻨﺟ ﺎﻫ
ﺩﺮﮕﻧ ﻩﺪﻫﺎﺸﻣ ﻳ
ﺪ . ﺍﻳ ﻨﭽﻤﻫ ﻥﻮﻣﺯﺁ ﻦ ﻴ
ﺩﻮﺟﻭ ﺖﺴﻧﺍﻮﺗ ﻦ
ﺮﺘﻛﺎﺑ ﻱ ﻩﺭﺎﺼﻋ ﺭﺩ ﺍﺭ ﺎﻫ
ﻱ ﺓﺩﻮﻟﺁ ﻥﺎﺑﺰﻴﻣ ﺎﻫ ﻱ ﺩﺭ ﻒﻠﺘﺨﻣ ﻳ
ﺑﺎ ﻲ
ﺎﻤﻧ ﻳ ﺪ . ﻧﺎﻣﺯ ﻲ ﺮﺳ ﺯﺍ ﻪﻛ ﻱ ﺖﻗﺭ ﻱﺎﻫ ﺮﺘﻛﺎﺑ ﻒﻠﺘﺨﻣ ﻳﺎ
ﻲﻳ ﺖﻬﺟ
ﺳﺎﺴﺣ ﺶﺠﻨﺳ ﻴ
ﺍ ﺖ ﻳ ﺩﺮﮔ ﻩﺩﺎﻔﺘﺳﺍ ﺵﻭﺭ ﻦ ﻳ
ﺪ
، ﺩﺮﺑﺭﺎﻛ ﻥﻭﺪﺑ
ﻨﻏ ﺔﻠﺣﺮﻣ ﻲ ﺯﺎﺳ ﻱ ١٠٧
× ١ ﻣ ﺭﺩ ﻝﻮﻠﺳ ﻴ
ﻠ ﻲ ﻟ ﻴ ﺩﺭ ﻞﺑﺎﻗ ﺮﺘ ﻳ
ﺑﺎ ﻲ
ﻟﻭ ﺩﻮﺑ ﻲ ﻨﻏ ﺎﺑ ﻲ ﺯﺎﺳ ﻱ ٢٤ ، ٤٨ ﻭ ٧٢ ﺮﺘﻛﺎﺑ ﺔﺘﻋﺎﺳ ﻱ
ﺭﺩ ﺎﻫ
ﺤﻣ ﻴ ﺎﻣ ﻂ ﻳ ﻛ ﻊ ﻴ ﺍ ،ﺏ ﮓﻨ ﻳ ﺎﺗ ﺖﺴﻧﺍﻮﺗ ﺵﻭﺭ ﻦ ١٠
ﺭﺩ ﻝﻮﻠﺳ
ﻣ ﻴ ﻠ ﻲ ﻟ ﻴ ﺩﺭ ﺍﺭ ﺮﺘ ﻳ ﺑﺎ ﻲ ﺎﻤﻧ ﻳ ﺪ ) ﻞﮑﺷ ٢ .(
ﻁﻮﺑﺮﻣ E. amylovora ﻩﺪﻨﻳﺎﻤﻧ ﻱﺎﻫﻪﻳﺍﺪﺟ ﺺﻴﺨﺸﺗ ‐١ ﻞﻜﺷ ﻪﺑ ﻥﺎﺑﺰﻴﻣ ﺎﻫ ﺎﺒﻳﺩ ﻥﻮﻣﺯﺁ ﻂﺳﻮﺗ ﺭﻮﺸﻛ ﻒﻠﺘﺨﻣ ﻖﻃﺎﻨﻣ ﻭ
ﻒﻠﺘﺨﻣ ﻱﺎﻫﺖﻗﺭ ﻲﺑﺎﻳﺩﺭ ﺖﻬﺟ ﺎﺒﻳﺩ ﻥﻮﻣﺯﺁ ﺖﻴﺳﺎﺴﺣ ‐۲ ﻞﻜﺷ ﻪﻳﺍﺪﺟ ﻲﻨﻏ ﺎﺑ ﺏﺁ ﺭﺩ Eal
ﻲﻨﻏ ﻥﻭﺪﺑ ﻭ ﻱﺯﺎﺳ ﻱﺯﺎﺳ
ﻞﻴﻓﻭﺮﭘ ﭘ DNA
ﻱﺪﻴﻣﺯﻼ : ﺍﺪﺟ ﻳ ﺍﺪﺟ ﻭ ﻊﺟﺮﻣ ﻪ ﻳ
ﻪ ﺎﻫ ﻱ
ﻪﺑ ﻖﻠﻌﺘﻣ ﻥﺎﺑﺰﻴﻣ
ﺎﻫ ﻭ ﺣﺍﻮﻧ ﻲ ﻒﻠﺘﺨﻣ )
ﺎﻨﺜﺘﺳﺍ ﻪﺑ ﻱ
ﺍﺪﺟ ﻳ ﻪ ﺎﻫ ﻱ ،Ea38
،Ea39
ﻣﺯﻼﭘ ﺪﻧﺎﺑ ﻪﮐ Ea40
ﻴ ﺪ ﻱ ﺭﺩ ﻥﺁ ﺎﻫ
ﻑﺮﻌﻣ ﻥﺍﺭﺎﮑﻤﻫ ﻭ ﻩﺩﺍﺯ :
ﻲﺧﺮﺑ ﻲﻳﺁﺭﺎﻛ ﻲﺳﺭﺮﺑ ﺵﻭﺭ
ﺎﻫ ﺮﺑ ﻲﻨﺘﺒﻣ ﻭ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ ﻱ ﺭﺩ PCR
...
۶۱
ﺩﺮﮕﻧ ﻩﺪﻫﺎﺸﻣ ﻳ
ﺪ ( ﺍﺭﺍﺩ ﻱ ﻳ ﻣﺯﻼﭘ ﺭﺍﻮﻧ ﮏ ﻴ
ﺪ ﻱ ﻟﻮﮑﻟﻮﻣ ﻥﺯﻭ ﺎﺑ ﻲ
ﺪﻧﺩﻮﺑ ﻪﺑﺎﺸﻣ .
ﻩﺮﻴﺠﻧﺯ ﺶﻨﻛﺍﻭ ﺯﺍﺮﻤﻴﻠﭘ ﻱﺍ
(PCR) : ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ
DNA
ﻣﺯﻼﭘ ﻴ ﺪ ﻱ ﺳﺍ ﺍﺪﺟ ﺯﺍ ﻩﺪﺷ ﺝﺍﺮﺨﺘ ﻳ
ﻪ ﺎﻫ ﻱ ﺩﺭﻮﻣ ﺁ ﺭﻭﺩ ﺭﺩ ﻥﻮﻣﺯ
ﻝﻭﺍ ،PCR
ﺪﻧﺎﺑ ﺤﺿﺍﻭ ﹰﻼﻣﺎﻛ ﻲ
ﺓﺩﻭﺪﺤﻣ ﺭﺩ ﻳ
ﻛ ﻚ ﻴ ﺯﺎﺑﻮﻠ ﻱ
ﺩﺮﮔ ﻩﺪﻫﺎﺸﻣ ﻳ
ﺪ . ﻨﭽﻤﻫ ﻴ ﻦ ﺎﺑ ﻥﺩﻭﺰﻓﺍ ﻘﺘﺴﻣ ﻴ ﻢ ﻝﻮﻠﺳ ﻱﺎﻫ
E.
amylovora
ﺨﻣ ﻪﺑ ،ﺶﻨﻛﺍﻭ ﻁﻮﻠ ﻳ
ﻚ ﺪﻧﺎﺑ ﺻﺎﺼﺘﺧﺍ ﻲ ﺭﺩ
ﺓﺩﻭﺪﺤﻣ ﻳ ﻛ ﻚ ﻴ ﺯﺎﺑﻮﻠ ﻱ ﺪﺷ ﻩﺪﻫﺎﺸﻣ .
ﻠﻛ ﺭﺩ ﺭﻮﻛﺬﻣ ﺪﻧﺎﺑ ﻴ
ﺔ
ﺍﺪﺟ ﻳ ﻪ ﺎﻫ ﻱ ﺎﻣﺯﺁ ﻳ ﺶ ﻣ ﺓﺪﺷ ﻪﺑ ﻖﻠﻌﺘ ﻥﺎﺑﺰﻴﻣ
ﺎﻫ ﻖﻃﺎﻨﻣ ﻭ
ﺎﻨﺜﺘﺳﺍ ﻪﺑ ﺭﻮﺸﻛ ﻒﻠﺘﺨﻣ ﻱ
ﺍﺪﺟ ﻪﻳ ﺎﻫ ﻳ ﻲ ﻣﺯﻼﭘ ﺪﻗﺎﻓ ﻪﮐ ﻴ
ﺪ
،ﺪﻧﺩﻮﺑ ﺅﺭ ﻳ ﺩﺮﮔ ﺖ ﻳ ﺪ ) ﻞﮑﺷ ٣ .(
ﺎﺑ ﺮﺳ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﻱ
ﺖﻗﺭ ﻱﺎﻫ
ﺮﺘﻛﺎﺑ ﻒﻠﺘﺨﻣ ﻳﺎ
ﻲﻳ ﻬﺗ ﻴ ﺭﻭﺩ ،ﺏﺁ ﺭﺩ ﻩﺪﺷ ﻪ ﻝﻭﺍ
ﺖﺴﻧﺍﻮﺗ PCR
١٠٧
× ٢ ﻣ ﺭﺩ ﻝﻮﻠﺳ ﻴ
ﻠ ﻲ ﻟ ﻴ ﺩﺭ ﺍﺭ ﺮﺘ ﻳ ﺑﺎ ﻲ ﺎﻤﻧ ﻳ ﺑ ﻪﻛ ﺪ ﻪ ﺭﻮﻃ
ﻝﺩﺎﻌﻣ ﻂﺳﻮﺘﻣ ١٠٤
× ٢ ﻫ ﺭﺩ ﻝﻮﻠﺳ ﺩﻮﺑ ﺶﻨﻛﺍﻭ ﺮ
) ﻞﮑﺷ ٤ (.
ﻲﻳﺎﻳﺮﺘﻛﺎﺑ ﻱﺎﻫﻪﻳﺍﺪﺟ ﻢﻴﻘﺘﺴﻣ ﻲﺑﺎﻳﺩﺭ ﻭ ﺺﻴﺨﺸﺗ ‐٣ ﻞﻜﺷ
E.amylovora
ﻩﺪﻨﻳﺎﻤﻧ ﻥﺎﺑﺰﻴﻣ ﺎﻫ ﺭﻮﺸﻛ ﻒﻠﺘﺨﻣ ﻖﻃﺎﻨﻣ ﻭ
ﻲﺑﺎﻳﺩﺭ ﺖﻬﺟ PCR ﺖﻴﺳﺎﺴﺣ ﻥﻮﻣﺯﺁ ‐٤ ﻞﻜﺷ
ﺖﻗﺭ ﻪﻳﺍﺪﺟ ﻒﻠﺘﺨﻣ ﻱﺎﻫ ﺏﺁ ﺭﺩ Eal
ﺩﺭﻭﺁﺮﻓ ﻩ ﺎﻫ ﻱ ﻪﺑ ﻁﻮﺑﺮﻣ ﺳﻮﺳ
ﭙ ﺴﻧﺎ ﻴ ﻥﻮ ﺎﻫ ﻱ ﻗﺭ ﻴ ﻖ ﺗ ﺮ
ﺮﺘﻛﺎﺑ ﻱ ﻪﻛ ﻝﻭﺍ ﺭﻭﺩ ﺭﺩ ﺩﺭ ﻞﺑﺎﻗ PCR
ﻳ ﺑﺎ ﻲ ﻥﻮﻣﺯﺁ ﺭﺩ ،ﺪﻧﺩﻮﺒﻧ
Nested–PCR
ﻥﺍﻮﻨﻌﺑ ﺐﻟﺎﻗ DNA
ﺑ ﻪ ﺺﺨﺸﻣ ﻭ ﺪﻨﺘﻓﺭ ﺭﺎﻛ
ﺩﺮﮔ ﻳ ﺍ ﺎﺑ ﻪﻛ ﺪ ﻳ
ﻣ ﺵﻭﺭ ﻦ ﻲ
ﻥﺍﻮﺗ ﺎﺗ ﻳ ﮏ ﺮﻫ ﺭﺩ ﺍﺭ ﻝﻮﻠﺳ
ﺩﺭ ﺶﻨﻛﺍﻭ ﻳ
ﺑﺎ ﻲ ﺩﻮﻤﻧ . ﺪﻧﺎﺑ ﻱ ﺍ ﺭﺩ ﻪﻛ ﻳ ﻭﺭ ﻥﻮﻣﺯﺁ ﻦ ﻱ
ﻝﮊ
ﺯﻭﺭﺎﮔﺁ ﺩﺮﮔ ﻩﺪﻫﺎﺸﻣ ﻳ
ﺓﺩﻭﺪﺤﻣ ﺭﺩ ،ﺪ ٨
/ ٠ ﻛ ﻴ ﺯﺎﺑﻮﻠ ﻱ ﺭﺍﺮﻗ
ﺖﺷﺍﺩ ) ﻞﮑﺷ ٥ .(
ﺭﺩ ﻮﻣﺯﺁ ﺻﺎﺼﺘﺧﺍ ﻥ ﻲ
ﻥﺩﻮﺑ ﺍﺪﺟ PCR
ﻳ ﻪ ﺎﻫ ﻱ
ﺮﺘﻛﺎﺑ ﻳﺎ ﻲﻳ ﻪﺑ ﻖﻠﻌﺘﻣ ﺲﻨﺟ
ﺎﻫ ﻱ ﺩ ﻳ ﺭﻭﺩ ﺭﺩ ﺮﮕ ﻝﻭﺍ
PCR
ﻫ ﻴ ﻪﻧﻮﮕﭽ ﺪﻧﺎﺑ ﻱ ﺔﻘﻄﻨﻣ ﺭﺩ ﻳ
ﻛ ﻚ ﻴ ﺯﺎﺑ ﻮﻠ ﻱ ﻳ ﺣﺍﻮﻧ ﺎ ﻲ ﺩ ﻳ ﺮﮕ
ﺍﻳ ﺍﺪﺟ ﻂﻘﻓ ﻭ ﺪﻧﺩﻮﻤﻨﻧ ﺩﺎﺠ ﻳ
ﺔ ﺍ ﺭﺩ ﻪﺘﻓﺭ ﺭﺎﻜﺑ Ea1
ﻳ ﻥﻮﻣﺯﺁ ﻦ
ﺪﻧﺎﺑ ﺖﺴﻧﺍﻮﺗ ﻳ
ﻛ ﻚ ﻴ ﺯﺎﺑ ﻮﻠ ﻱ ﻭﺭ ﻱ ﮊ ﻝ ﺯﻭﺭﺎﮔﺁ ﺍ ﻳ ﺎﻤﻧ ﺩﺎﺠ ﻳ ﺪ
) ﻞﮑﺷ ٦ .(
ﻲﺑﺎﻳﺩﺭ ﺖﻬﺟ Nested-PCR ﺖﻴﺳﺎﺴﺣ ﻥﻮﻣﺯﺁ ‐٥ ﻞﻜﺷ ﺖﻗﺭ
ﻪﻳﺍﺪﺟ ﻒﻠﺘﺨﻣ ﻱﺎﻫ ﺏﺁ ﺭﺩ Eal
ﻞﻜﺷ ۶
‐ ﻥﺩﻮﺑ ﻲﺻﺎﺼﺘﺧﺍ ﻥﻮﻣﺯﺁ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ PCR
ﻥﻮﻴﺴﻧﺎﭙﺳﻮﺳ ﻪﻳﺍﺪﺟ
ﻲﻳﺎﻳﺮﺘﻛﺎﺑ ﻒﻠﺘﺨﻣ ﻱﺎﻫ :
1) Eal, 2) A. vitis, 3) B. Subtilis, 4) E. carotovora sub sp.
Carotovora, 5) E. carotovora sub sp. Wasabiae, 6) P.
fluorescens. 7) P. syringae p. v syringae, 8) X. campestris p.v. begoniae 9, 10) Water
ﻧﺎﻣﺯ ﻲ ﺮﺳ ﻪﻛ ﻱ ﺎﻫ ﻱ ﺖﻗﺭ
E. amylovora
ﺓﺭﺎﺼﻋ ﺭﺩ
ﻳ ﺖﻓﺎﺑ ﺓﺪﺷ ﺖﺧﺍﻮﻨﻜ ﮒﺮﺑ
ﺳ ﻴ ﻬﺗ ﺐ ﻴ ﺩﺮﮔ ﻪ ﻳ ﺪ
، ﻝﻭﺍ ﺭﻭﺩ ﺭﺩ
ﺎﺗ PCR
١٠٦
× ١ ﻝﻮﻠﺳ ) ﻳ ﻡﺪﺻ ﮏ ﺳﺎﺴﺣ ﻴ ﺩﺭ ﺖ ﻳ ﺑﺎ ﻲ ﺏﺁ ﺭﺩ (
ﻭ ﺭﺩ
Nested-PCR
١٠٠ ﻮﻠﺳ ﻝ )ﺑ ﻴ ﺯﺍ ﺶ ﻳ ﮏ ﺠﻨﭘ ﻢﻫﺎ
ﺳﺎﺴﺣ ﻴ ﺩﺭ ﺖ ﻳ ﺑﺎ ﻲ ﺏﺁﺭﺩ ( ﺩﺭ ﻞﺑﺎﻗ ﻳ ﺑﺎ ﻲ ﺩﻮﺑ .
