ﻥﺍﺭﺎﮑﻤﻫ ﻭ ﻲﻨﻴﺴﺣ :
ﺮﺑ ﻲﻨﺘﺒﻣ ﻲﻟﻮﮑﻟﻮﻣ ﻱﺎﻫﺮﮕﻧﺎﺸﻧ ﺩﺎﺠﻳﺍ ﺎﺑ ﻪﺘﺳﻮﻴﭘ EST
...
۱۸۷
ﻪﻠﺠﻣ ﻡﻮﻠﻋ ﮔ ﻴ ﻋﺍﺭﺯ ﻥﺎﻫﺎ ﻲ ﻥﺍﺮﻳﺍ
ﺩ ﻩﺭﻭ ۴۰ ﻩﺭﺎﻤﺷ ، ۲ ، ۱۳۸۸ ) ۱۹۶
‐ ۱۸۷ (
ﺮﮕﻧﺎﺸﻧ ﺩﺎﺠﻳﺍ ﺎﻫ
ﻱ ﻟﻮﮑﻟﻮﻣ ﻲ ﺮﺑ ﻲﻨﺘﺒﻣ ﺖﻣﻭﺎﻘﻣ ﺎﺑ ﻪﺘﺳﻮﻴﭘ EST
ﻪﺑ ﻣﻮﻳﺭﺍﺯﻮﻓ ﺖﻳﻼﺑ ﻲ
ﻪﻠﺒﻨﺳ ﻡﺪﻨﮔ ﺭﺩ
ﻨﻴﺴﺣ ﻪﻧﺎﻤﺛ
۱ﻲ , ﻱﺩﺮﻣ ﻦﺴﺤﻣ
*۲
, ﺭﺎﻔﻏ ﺎﺿﺭﺪﻤﺤﻣ
۳ﻱ , ﻳ ﻣﺎﻧ ﻒﺳﻮ
۴ﻲ , ﺮﮐ ﻪﻟﻻ ﻳ ﻤ ﻲ ﺩﺎﺳﺮﻓ ,۵
ﺳ ﻴ ﻔﻄﺼﻣ ﺪ ﻲ ﭘ ﻴ ﺳﺮ ﻴ ﺪ
۶ﻱ , ﺳ ﻴ ﺒﻋ ﺱﻭﺮ ﻧﺎﺸﻴﻣﺪ
۷ﻲ ﻭ ﻠﻋ ﻲ ﺮﻬﺷﻮﺑ ﺕﺎﺠﻧ ﻩﺎﺷ ﺮﺒﮐﺍ
۸ﻱ
۱ ، ۷ , ۸ , ،ﺪﺷﺭﺍ ﻲﺳﺎﻨﺷﺭﺎﮐ ﻖﺑﺎﺳ ﻱﻮﺠﺸﻧﺍﺩ ﻭ ﺩﺎﺘﺳﺍ
،ﺭﺎﻴﺸﻧﺍﺩ ﻱﺯﺭﻭﺎﺸﮐ ﺲﻳﺩﺮﭘ ﻲﻌﻴﺒﻃ ﻊﺑﺎﻨﻣ ﻭ
ﻩﺎﮕﺸﻧﺍﺩ ﻥﺍﺮﻬﺗ
۲ , ۳ , ۴ , ۵ , ۶ , ﺭﺎﻳﺩﺎﺘﺳﺍ , ﺪﺷﺭﺍ ﺱﺎﻨﺷﺭﺎﮐ ,
ﻮﺠﺸﻧﺍﺩ ﻱ ﻖﺑﺎﺳ ﺳﺎﻨﺷﺭﺎﮐ ﻲ ﺪﺷﺭﺍ , ﺱﺎﻨﺷﺭﺎﮐ ,
ﺪﺷﺭﺍ ﺱﺎﻨﺷﺭﺎﮐ ﺲﮑﻴﻣﻮﻧﮊ ﺕﺎﻘﻴﻘﺤﺗ ﺶﺨﺑ
, ﻴﺑ ﻩﺪﻜﺸﻫﻭﮋﭘ ﻱﺯﺭﻭﺎﺸﻛ ﻱﮊﻮﻟﻮﻨﻜﺗﻮ
) ﺖﻓﺎﻳﺭﺩ ﺦﻳﺭﺎﺗ :
۸ / ۱۲ / ۸۶
‐ ﺐﻳﻮﺼﺗ ﺦﻳﺭﺎﺗ :
۱ / ۸ / ۸۷ (
ﻩﺪﻴﮑﭼ
ﻱﺎﻫﺮﮕﻧﺎﺸﻧ ﺩﺎﺠﻳﺍ ﺖﻬﺟ
ﻟﻮﮑﻟﻮﻣ ﻲ ﺮﺑ ﻲﻨﺘﺒﻣ ﻥﮊ ﺎﺑ ﻪﺘﺳﻮﻴﭘ EST
ﻪﺑ ﺖﻣﻭﺎﻘﻣ ﻩﺪﻨﻨﻛ ﻝﺮﺘﻨﻛ ﻱﺎﻫ ﺭﺩ FHB
ﻚﻳﮊﻮﻟﻮﻴﺑ ﺩﺮﻜﻠﻤﻋ ،ﻡﺪﻨﮔ ٨٦٦٠
EST ، ﺎﻫ ﻚﻧﺎﺑ ﺭﺩ ﺩﻮﺟﻮﻣ ﻱ
ﺕﺎﻋﻼﻃﺍ ﻪﻌﻟﺎﻄﻣ
ﺪﺷ ﻭ ٢٦٠
EST ، ﺎﺑ
ﺘﻧﺍ ﺺﺨﺸﻣ ﺩﺮﻜﻠﻤﻋ ﺪﻧﺪﺷ ﺏﺎﺨ
. ﻉﻮﻤﺠﻣ ﺭﺩ ٦٢
ﻆﻔﺣ ﺮﻴﻏ ﻭ ﻩﺪﺷ ﻆﻔﺣ ﻁﺎﻘﻧ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﺮﮔﺯﺎﻏﺁ ﺖﻔﺟ
ﺪﺷ ﻲﺣﺍﺮﻃ ﻩﺪﺷ .
ﺞﻳﺎﺘﻧ ﻪﻳﺰﺠﺗ ﭘﻴ ﮕﺘﺳﻮ ﻲ
QTL ﻭ ﻩﺩﺍﺩ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﻞﺴﻧ ﺭﺩ ﻲﭙﻴﺗﻮﻨﻓ ﻭ ﻲﭙﻴﺗﻮﻧﮊ ﻱﺎﻫ
ﺎﻫ ﻱ
F٣
ﻭ
F٥ ﻲﻗﻼﺗ ﺯﺍ ﻞﺻﺎﺣ ﺎﻫ
ﻱ ﺎﻧﺎﺘﻧﻭﺮﻓ )ﻧ ﻴ ﻡﻭﺎﻘﻣ ﻪﻤ / ( ﺕﻼﻓ ) ﺱﺎﺴﺣ ( ﻪﻛ ﺩﺍﺩ ﻥﺎﺸﻧ ﻳ
ﮏ ﺣﺍﺮﻃ ﺮﮕﻧﺎﺸﻧ ﻲ
ﻩﺪﺷ
ﻥﮊ ﺱﺎﺳﺍ ﺮﺑ ﺮﺜﮐﺍﺪﺣ ﺖﺳﺍ ﺭﺩﺎﻗ NPR1
٦ / ٩ ﺖﻓﺮﺸﻴﭘ ﻲﻨﺤﻨﻣ ﺮﻳﺯ ﺢﻄﺳ ﺲﻧﺎﻳﺭﺍﻭ ﺕﺍﺮﻴﻴﻐﺗ ﺯﺍ ﺪﺻﺭﺩ
ﺮﺛﺍ ﺎﺑ ﺍﺭ ﻲﮔﺩﻮﻟﺁ ﺍﺰﻓﺍ
ﻳ ﺸ ﻲ ﺪﻨﻛ ﻪﻴﺟﻮﺗ ﻲﻔﻨﻣ .
ﺍﻳ ﻦ ﻭﺯﺎﺑ ﻱﻭﺭ ﺮﮕﻧﺎﺸﻧ ﻱ
ﻡﻭﺯﻮﻣﻭﺮﻛ ﻩﺎﺗﻮﮐ ٣A
ﻥﺎﮑﻣ ﻳ ﺑﺎ ﻲ ﺪﺷ .
ﻥﮊ ﺮﻫﺎﻈﺗ ﺵﻭﺭ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ NPR1
QRT-PCR
ﺑ ﺩﺍﺩ ﻥﺎﺸﻧ ﻴ ﺍ ﻥﺎ ﻳ ﺭﺩ ﻥﮊ ﻦ ٩٦ ﻮﻟﺁ ﺯﺍ ﺲﭘ ﺖﻋﺎﺳ ﮔﺩ
ﻲ ﺭﺩ
ﺎﺑﻮﺷ ﮓﻧﻭ ﻱ
ﺍﺰﻓﺍ ﺎﻧﺎﺘﻧﻭﺮﻓ ﻭ ﻳ
ﻣ ﺶ ﻲ ﻳ ﺪﺑﺎ . ﺍﺮﺑﺎﻨﺑ ﻳ ﻦ ﻥﮊ ﻟﻮﮑﻟﻮﻣ ﺮﮕﻧﺎﺸﻧ ﻭNPR1
ﻲ ﻨﺘﺒﻣ ﻲ ﻥﺁ ﺮﺑ ﻧﺍﻮﺗ ﻲﻣ ﻨ ﺪ ﺭﺩ
ﺎﻫ ﻪﻣﺎﻧﺮﺑ ﻱ ﻘﺤﺗ ﻴ ﺗﺎﻘ ﻲ ﺍﺰﻓﺍ ﻳ ﻪﺑ ﺖﻣﻭﺎﻘﻣ ﺶ ﺪﻧﻮﺷ ﻪﺘﻓﺮﮔ ﺮﻈﻧ ﺭﺩ ﻡﺪﻨﮔ ﺭﺩ FHB
.
ﻩﮊﺍﻭ
ﺎﻫ ﻱ ﻱﺪﻴﻠﮐ ،ﻡﺪﻨﮔ ،ﻪﻠﺒﻨﺳ ﻲﻣﻮﻳﺭﺍﺯﻮﻓ ﺖﻳﻼﺑ :
،NPR1
،EST QRT-PCR
.
ﻪﻣﺪﻘﻣ
ﻩﮊﻭﺮﭘ ﺩﻮﺟﻭ ﺎﻫ
ﻱ ﻟﺍﻮﺗ ﻦﻴﻴﻌﺗ ﻲ
ﺩﺍﺪﻌﺗ ﻡﻮﻧﮊ ﻱ
ﺕﺍﺩﻮﺟﻮﻣ ﺯﺍ
ﻩﺪﻧﺯ
، ﺩﺎﻳﺯ ﺭﺎﻴﺴﺑ ﺕﺎﻋﻼﻃﺍ ﻱ
ﺚﻋﺎﺑ ﻪﮐ ﺪﻧﺍ ﻩﺩﻮﻤﻧ ﻢﻫﺍﺮﻓ ﺍﺭ
ﺪﻳﺪﺟ ﺶﺨﺑ ﺩﺎﺠﻳﺍ ﻱ
ﮊﻮﻟﻮﻴﺑ ﺕﺎﻌﻟﺎﻄﻣ ﺯﺍ ﻱ
ﻥﺍﻮﻨﻋ ﺖﺤﺗ
ﻣﻮﻧﮊ ﻴ ﺭﺎﺘﺧﺎﺳ ﺲﮑ
١ﻱ ﺖﺳﺍ ﻩﺪﺷ .
ﺎﻬﻜﻴﻨﻜﺗ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﻩﺯﻭﺮﻣﺍ
ﻱﺭﻭﺁ ﻦﻓ ﻭ ﺎﻫ
ﻱ ﺪﻨﻧﺎﻣ ﺪﻳﺪﺟ
، cDNA Real Time PCR
ﻭ
ﺭ ﻳ ﺍﺭﺁ ﺰ ﻳ ﻟﺍ ﻪ ﻴ ﺌﻠﮐﻮﻧﻮﮕ ﺗﻮ ﻴ ﺪ ﻱ ﺳﺮﺘﺳﺩ ﺚﻋﺎﺑ ﻲ
ﺗﺎﻋﻼﻃﺍ ﻪﺑ ﻲ
ﺭﺩ
ﮕﻧﻮﮕﭼ ﺩﺭﻮﻣ ﻲ
ﻥﮊ ﺩﺮﮑﻠﻤﻋ ﻥﺍﻮﻨﻋ ﺖﺤﺗ ﺎﻫ
ﻣﻮﻧﮊ ﻴ ﺲﮑ
ﺩﺮﮑﻠﻤﻋ
٢ﻱ ﻧ ﻴ ﺰ ﺖﺳﺍ ﻩﺪﺷ .
ﻝﺎﺳ ﺭﺩ ﻱﮊﻮﻟﻮﻨﻜﺗ ﺮﻴﺧﺍ ﻱﺎﻫ
ﻱﺎﻫﺮﮕﻧﺎﺸﻧ ﻱﺎﻫﺮﮕﻧﺎﺸﻧ ﺖﻤﺳ ﻪﺑ ﻲﻟﺎﻋ ﻥﺎﻫﺎﻴﮔ ﺭﺩ DNA
1 . Structural Genomics
2 .Functional Genomics
ﻣ ﻮ ﻲﻟﺍﻮﺗ ﺮﮕﻳﺩ ﻭ ﻡﻮﺘﭙﻳﺮﻜﺴﻧﺍﺮﺗ ﺮﺑ ﻲﻨﺘﺒﻣ ﻲﻟﻮﻜﻟ ﻱﺎﻫ
ﺰﻣﺭ
ﻭﺭ ﻩﺪﻨﻨﻛ ﺖﺳﺍ ﻩﺩﺮﻛ
. ﮕﻧﺎﺸﻧ ﻥﺎﻴﻣ ﻦﻳﺍ ﺭﺩ ﻱﺩﺮﻜﻠﻤﻋ ﻱﺎﻫﺮ
ﻡﻮﻧﮊ ﻲﻣ ﺏﺎﺨﺘﻧﺍ ﻩﺎﻴﮔ ﺭﺩ ﺖﻔﺻ ﻚﻳ ﺭﻮﻬﻇ ﻱﺎﻨﺒﻣ ﺮﺑ ﻪﻛ
ﻚﻴﺘﻧﮊ ﻉﻮﻨﺗ ﻦﻴﻤﺨﺗ ﻪﻌﻟﺎﻄﻣ ﺭﺩ ﺪﻧﻮﺷ ﻚﻤﻛ ﻪﺑ ﺏﺎﺨﺘﻧﺍ ،
ﺮﮕﻧﺎﺸﻧ
، ﻱﺎﻨﺒﻣ ﺮﺑ ﻱﺯﺎﺳ ﻪﻧﺎﺴﻤﻫ ﻭ ﻱﺍﺪﻳﺪﻧﺎﻛ ﻱﺎﻬﻧﮊ ﻲﺳﺭﺮﺑ
ﻥﺎﮑﻣ ﻲﺑﺎﻳ
، ﻦﻴﻴﻌﺗ ﻲﻜﻴﺘﻧﮊﻮﻠﻴﻓ ﻪﻌﻟﺎﻄﻣ ﻭ ﺎﻫQTL
ﻭ
ﺭﺍﺮﻗ ﻩﺩﺎﻔﺘﺳﺍ ﺩﺭﻮﻣ ﻲﻜﻴﻣﻮﻧﻮﺴﻛﺎﺗ ﻲﻣ
ﺪﻧﺮﻴﮔ
(Mardi et al.,
2004)
. ﺭﺎﺘﺧﺎﺳ ﻪﺑ ﻁﻮﺑﺮﻣ ﺩﺎﻳﺯ ﺭﺎﻴﺴﺑ ﺕﺎﻋﻼﻃﺍ ﺩﻮﺟﻭ ﻪﺑ ﻪﺟﻮﺗ ﺎﺑ
ﺭﺩ ﺎﻫ ﻥﮊ ﺩﺮﮑﻠﻤﻋ ﻭ ﺎﻫ ﮏﻧﺎﺑ
ﻱ ﺗﺎﻋﻼﻃﺍ ﻲ
، ﺮﺿﺎﺣ ﻝﺎﺣ ﺭﺩ
ﻣ ﺮﺴﻴﻣ ﻥﺎﮑﻣﺍ ﻦﻳﺍ ﻲ
ﻪﮐ ﺪﺷﺎﺑ ﺎﻫﺮﮕﻧﺎﺸﻧ
ﻱ ﺭﺎﺘﺧﺎﺳ ﻱ ﻭ
ﺩﺮﮑﻠﻤﻋ ﻱ ﺎﻨﺒﻣ ﺮﺑ ﻩﺪﻧﺯ ﺩﻮﺟﻮﻣ ﻚﻳ ﻡﻮﻧﮊ ﻱ
ﺎﻫﺮﮕﻧﺎﺸﻧ ﻱ
ﻝﻮﺌﺴﻣ ﻩﺪﻨﺴﻳﻮﻧ *
: ﻱﺩﺮﻣ ﻦﺴﺤﻣ
ﻦﻔﻠﺗ : ۲۷۰۰۸۴۵
‐ ۰۲۶۱
E-mail: [email protected]
۱۸۸ ﻪﻠﺠﻣ
ﻲﻋﺍﺭﺯ ﻥﺎﻫﺎﻴﮔ ﻡﻮﻠﻋ ﺩ ،ﻥﺍﺮﻳﺍ
ﻩﺭﻭ ۴۰ ﻩﺭﺎﻤﺷ ، ۲ ، ۱۳۸۸
ﺭﺎﺘﺧﺎﺳ ﻱ ﺩﺮﮑﻠﻤﻋ ﻭ ﻱ
ﻳﺎﺳﺎﻨﺷ ﻩﺪﻧﺯ ﺕﺍﺩﻮﺟﻮﻣ ﺮﻳﺎﺳ ﻡﻮﻧﮊ ﻲ
ﺪﻧﻮﺷ . ﻩﮊﻭﺮﭘ ﻞﻴﻤﻜﺗ ﺎﺑ ﻩﺯﻭﺮﻣﺍ ﺎﻫ
ﻱ ﻒﻳﺩﺭ ﺭﺩ ﻡﻮﻧﮊ ﻲﺑﺎﻳ
ﺩﺍﺪﻌﺗ ﻱ ﻥﺎﻫﺎﻴﮔ ﺯﺍ
ESTs، ﺎﻫ
١ﻱ ﺮﻳﺎﺳ ﻪﻌﻟﺎﻄﻣ ﻱﺍﺮﺑ ﻲﻧﺍﻭﺍﺮﻓ
ﺖﺳﺍ ﻩﺪﺷ ﻢﻫﺍﺮﻓ ﻥﺎﻫﺎﻴﮔ
EST . ﻲﻟﺍﻮﺗ ﺎﻫ ﻩﺎﺗﻮﮐ ﻱﺎﻫ
٢٠٠
‐ ٣٠٠ ﻲﻟﺍﻮﺗ ﺎﺑ ﻪﮐ ﺪﻨﺘﺴﻫ ﻱﺯﺎﺑ ﺖﻔﺟ
Single passﻲﺑﺎﻳ
ﻪﻧﺎﺴﻤﻫ ﻱﺎﻫ ﻲﻣ ﺪﻴﻟﻮﺗ cDNA
ﺯﺍ ﻲﺒﺳﺎﻨﻣ ﻱﺎﻤﻧ ﻭ ﺪﻧﻮﺷ
ﻲﻟﺍﻮﺗ ﺮﻫﺎﻈﺗ ﻱﺎﻫ ﻪﺘﻓﺎﻳ
ﻥﮊ ﺎﻫ ﺖﻓﺎﺑ ﺭﺩ ﺖﺤﺗ ﻒﻠﺘﺨﻣ ﻱﺎﻫ
ﺶﻨﺗ ﻝﺎﻤﻋﺍ ﺯﺍ ﺪﻌﺑ ﺎﻳ ﻭ ﻲﻌﻴﺒﻃ ﻂﻳﺍﺮﺷ ﺘﺴﻳﺯ ﻱﺎﻫ
ﻲ ﻭ
ﺘﺴﻳﺯﺮﻴﻏ ﻲ ﻲﻣ ﺩﻮﺟﻮﺑ ﺪﻧﺭﻭﺁ
. ﺐﻠﻏﺍ ﻲﻣﻮﻧﮊ ﻥﺎﮑﻣ ﻭ ﺩﺮﮑﻠﻤﻋ
ﺖﺳﺍ ﻪﺘﺧﺎﻨﺷﺎﻧ ﺎﻫEST
. ﻁﻮﺑﺮﻣ ﺕﺎﻋﻼﻃﺍ ﺩﻮﺟﻭ ﻪﺑ ﻪﺟﻮﺗ ﺎﺑ
ﻥﮊ ﺯﺍ ﻱﺩﺎﻳﺯ ﺩﺍﺪﻌﺗ ﻪﺑ ﺩﺮﮑﻠﻤﻋ ﻦﻴﻴﻌﺗ ﻥﺎﮑﻣﺍ ،ﺎﻫ
ﺎﻫEST
ﺩﺭﺍﺩ ﺩﻮﺟﻭ
(Peng et al., 2003)
.
ﻨﺘﺒﻣ ﻱﺎﻫﺮﮔﺯﺎﻏﺁ ﻲﺣﺍﺮﻃ ﺎﺑ ﻲ
ﺮﺑ ﺑﺎﺨﺘﻧﺍ ﻱﺎﻫ EST
ﻲ ﻭ
ﺳﺭﺮﺑ ﻲ ﻠﮑﺷ ﺪﻨﭼ ﻲ ﺭﺩ ﺎﻬﻧﺁ ﻦﻴﺑ ﻡﻭﺎﻘﻣ ﻦﻳﺪﻟﺍﻭ /
ﻞﻤﺤﺘﻣ ﻭ
ﺖﻴﻌﻤﺟ ﻭ ﺱﺎﺴﺣ ﺎﻬﻧﺁ ﺯﺍ ﻞﺻﺎﺣ ﻕﺮﻔﺗ ﻝﺎﺣ ﺭﺩ ﻱﺎﻫ
، ﻁﺎﺒﺗﺭﺍ
ﺖﻣﻭﺎﻘﻣ ﺎﺑ ﺎﻬﻧﺁ /
ﻞﻤﺤﺗ ﻪﺸﻘﻧ ﻱﻭﺭ ﺮﺑ ﺎﻬﻧﺁ ﻲﻣﻮﻧﮊ ﻥﺎﮑﻣ ﻭ ﻱﺎﻫ
ﻣ ﺺﺨﺸﻣ ﺩﻮﺟﻮﻣ ﮏﻴﺘﻧﮊ ﻲ
ﺩﻮﺷ . ﺎﻫﺮﮕﻧﺎﺸﻧ ﻲﮕﺘﺳﻮﻴﭘ ﻱ
ﻨﺘﺒﻣ ﻲ
EST ﺮﺑ ﺖﻣﻭﺎﻘﻣ ﺎﺑ ﺎﻫ /
ﻞﻤﺤﺗ ﻪﺑ ﺶﻨﺗ ﺎﻫ ﻱ ﺘﺴﻳﺯ ﻲ ﻭ
ﺘﺴﻳﺯﺮﻴﻏ ﻲ
، ﻫﺎﮔﺁ ﺶﻳﺍﺰﻓﺍ ﺐﺟﻮﻣ ﻲ
ﻲﻳﺎﻴﻤﻴﺷﻮﻴﺑ ﻱﺎﻫﺮﻴﺴﻣ
ﻥﮊ ﺎﻫ ﻱ ﺖﻣﻭﺎﻘﻣ ﻩﺪﻨﻨﮐ ﻝﺮﺘﻨﮐ /
ﻞﻤﺤﺗ ﻥﺩﻮﻤﻧ ﻢﮐﺍﺮﺘﻣ ﻭ
ﻪﻃﺎﺣﺍ ﻱﺎﻫﺮﮕﻧﺎﺸﻧ ﻪﻠﺻﺎﻓ ﻩﺪﻨﻨﮐ
ﺎﻫQTL
ﻱ ﻩﺪﺷ ﻪﺘﺧﺎﻨﺷ
ﻣ ﻲ ﺩﻮﺷ . ﺕﺎﻋﻼﻃﺍ ﻥﮊ ﺩﺮﮑﻠﻤﻋ ﺮﺑ ﻲﻨﺘﺒﻣ ﻱﺎﻫﺮﮕﻧﺎﺸﻧ ﺪﻴﻟﻮﺗ
ﺎﻫﺮﮕﻧﺎﺸﻧ ﺮﮕﻳﺩ
DNA ﻱ ﺪﻨﻧﺎﻣ ﻭ AFLP
ﻞﻴﻤﮑﺗ ﺍﺭ SSR
ﻪﺸﻘﻧ ﺭﺩ ﺰﻴﻧ ﺍﺭ ﻩﺪﻨﻨﮐ ﺪﮐ ﻲﺣﺍﻮﻧ ﻭ ﻩﺩﻮﻤﻧ ﺭﺎﮑﺷﺁ ﮏﻴﺘﻧﮊ ﻱﺎﻫ
ﻲﻣ
(Mardi et al., 2004)ﺪﻨﮐ .
ﺎﻳ ﻪﻠﺒﻨﺳ ﻲﻣﻮﻳﺭﺍﺯﻮﻓ ﺖﻳﻼﺑ
FHB2
ﻡﺪﻨﮔ
L.)
Triticum aestivum
( ﻪﻛ ﻒﻠﺘﺨﻣ ﻱﺎﻫ ﻪﻧﻮﮔ ﻂﺳﻮﺗ
ﻡﻮﻳﺭﺍﺯﻮﻓ ﺲﻨﺟ
Fusarium spp.) (
ﺩﻮﺷ ﻲﻣ ﺩﺎﺠﻳﺍ
، ﻜﻳ ﻲ ﺯﺍ
ﻱﺭﺎﻤﻴﺑ ﺎﻫ ﻱ ﻡﺪﻨﮔ ﻢﻬﻣ ﺖﺳﺍ ﻥﺍﺮﻳﺍ ﺭﺩ
. ﺭﺎﻤﻴﺑ ﻦﻳﺍ ﻱ ﺚﻋﺎﺑ
ﺭﺩ ﺖﻴﻤﺳ ﺩﺎﺠﻳﺍ ﻭ ﻪﻧﺍﺩ ﺖﻴﻔﻴﻛ ،ﺩﺮﻜﻠﻤﻋ ﺪﻳﺪﺷ ﺶﻫﺎﻛ ﺩﻮﺷ ﻲﻣ ﻡﺍﺩ ﻙﺍﺭﻮﺧ ﻭ ﻥﺎﺴﻧﺍ ﻱﺍﺬﻏ
(Mardi et al., 2004)
.
ﻥﺎﮑﻣ ﻭ ﻲﻳﺎﺳﺎﻨﺷ ﻪﻨﻴﻣﺯ ﺭﺩ ﻲﻔﻠﺘﺨﻣ ﺕﺎﻌﻟﺎﻄﻣ ﻥﻮﻨﮐﺎﺗ ﻲﺑﺎﻳ
ﻱﺎﻫQTL
ﻩﺪﻨﻨﮐ ﻝﺮﺘﻨﮐ ٣
FHB ﻪﺑ ﻡﺪﻨﮔ ﺭﺩ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ
ﻱﺎﻫﺮﮕﻧﺎﺸﻧ ﺭﺎﺘﺧﺎﺳ
ﻱ ﻡﻮﻧﮊ ﻡﺎﺠﻧﺍ ﺖﺳﺍ ﻩﺪﺷ
(Buerstmayr
et al., 2002; Buerstmayr et al., 2003; Lin et al., 2004; Somers et al., 2003; Steiner et al., 2004;
Waldron et al., 1999) Mardi et al. .
(2005)
ﻦﻳﺮﺘﻤﻬﻣ
1. Expressed sequence tag sites
2. Fusarium head blight 3. Quantitative trait loci
ﻪﺑ ﺖﻣﻭﺎﻘﻣ ﻩﺪﻨﻨﻛ ﻝﺮﺘﻨﻛ ﻱﺎﻫ QTL
ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﺍﺭ FHB
ﺖﻴﻌﻤﺟ ﻲﻗﻼﺗ ﺯﺍ ﻞﺻﺎﺣ ﻕﺮﻔﺗ ﻝﺎﺣ ﺭﺩ ﻱﺎﻫ ﺕﻼﻓ
× ﮓﻧﻭ
ﺎﺑﻮﺷ ﻱ ﻡﻭﺯﻮﻣﻭﺮﻛ ﻱﻭﺭ ﺍﺭ ﺎﻫ
BS ﻱ ۳
DL ﻭ ۲ ﻳﺎﺳﺎﻨﺷ ﻲ
ﺪﻧﺩﻮﻤﻧ . ﻪﻳﺰﺠﺗ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ QTL
٢٧٣ ﺮﮕﻧﺎﺸﻧ ﻭ AFLP
ﺖﻴﻌﻤﺟ ﺭﺩ SSR
ﺎﻫ ﻱ ﺯﺍ ﻞﺻﺎﺣ ﻕﺮﻔﺗ ﻝﺎﺣ ﺭﺩ ﺕﻼﻓ
×
ﺎﻧﺎﺘﻧﻭﺮﻓ ﺎﻬﻣﻭﺯﻮﻣﻭﺮﮐ ﻪﻛ ﺩﺍﺩ ﻥﺎﺸﻧ
BL ﻱ
،۱ ۳AL AS ﻭ ۷
ﻔﻨﻣ ﺮﺛﺍ ﻦﻳﺮﺘﺸﻴﺑ ﻲ
ﻨﻌﻣ ﻭ ﻲ ﮔﺩﻮﻟﺁ ﺲﻧﺎﻳﺭﺍﻭ ﺮﺑ ﺍﺭ ﺭﺍﺩ ﻲ
ﺪﻨﺘﺷﺍﺩ
(Mardi et al., 2006)
. ﻣ ﻪﻳﺰﺠﺗ ﺞﻳﺎﺘﻧ ﻮ
ﻟﻮﮑﻟ ﻲ ١١٠
ﺮﮕﻧﺎﺸﻧ ﻭﺭ ﺮﺑ SSR
ﻱ ﺎﻫ ﺖﻴﻌﻤﺟ ﻱ
ﺯﺍ ﻞﺻﺎﺣ ﻕﺮﻔﺗ ﻝﺎﺣ ﺭﺩ
ﺕﻼﻓ
× ﺎﻣﻮﺳ ﻱ ٣ ﮏﻳ ﻪﮐ ﺩﺍﺩ ﻥﺎﺸﻧ ﻭﺭ QTL
ﻱ ﻡﻭﺯﻮﻣﻭﺮﮐ
۳BS
ﭙﻴﺗﻮﻨﻓ ﺲﻧﺎﻳﺭﺍﻭ ﻢﻬﺳ ﻦﻳﺮﺘﺸﻴﺑ ﻲ
ﮔﺩﻮﻟﺁ ﻲ ﺮﺛﺍ ﺎﺑ ﺍﺭ
ﻔﻨﻣ ﻲ ﻣ ﻪﻴﺟﻮﺗ ﻲ ﺪﻳﺎﻤﻧ
(Mardi et al., 2002)
.
ﺩﻭﺪﺤﻣ ﻖﻓﻮﻣ ﺕﺎﺷﺭﺍﺰﮔ ﻡﺪﻨﮔ ﺭﺩ ﻱ
ﺩﺮﺑﺭﺎﮐ ﺯﺍ
ﺎﻫﺮﮕﻧﺎﺸﻧ ﻱ ﺩﺮﮑﻠﻤﻋ ﻱ ﻪﻳﺰﺠﺗ ﺭﺩ ﻡﺪﻨﮔ ﻡﻮﻧﮊ ﺩﻮﺟﻭ QTL
ﺩﺭﺍﺩ
Lu et al. .
(2006)
ﻲﻜﻳﺰﻴﻓ ﻲﺑﺎﻳ ﻪﺸﻘﻧ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ
EST٥٤ ﻡﻭﺯﻮﻣﻭﺮﻛ ﻱﻭﺭ ﺮﺑ ﻡﺪﻨﮔ ﻥﮊ ﻩﺩﻭﺪﺤﻣ 5BL
Tsn1
ﮔ ﻪﺑ ﻡﺪﻨﮔ ﺖﻴﺳﺎﺴﺣ ﺐﺒﺳ ﻪﻛ ﺍﺭ ﻡﺪﻨ
Pto ToxA
ﻲﻣ ﺩﻮﺷ
ﺩﻭﺪﺣ ﻪﺑ ٠٨
/ ٠ ﻲﺘﻧﺎﺳ ﺶﻫﺎﻛ ﻥﺎﮔﺭﻮﻣ ﺪﻨﻫﺩ
.
Raedschelders et al.
(2004)
ﺮﺑ ﻲﻨﺘﺒﻣ ﺮﮕﻧﺎﺸﻧ ﺩﺎﺠﻳﺍ ﺎﺑ
ﻥﮊ ﺯﺍ PCR XAXI-like
ﻱﻭﺭﺮﺑ ﻥﮊ ﻦﻳﺍ ﻲﻳﺎﺳﺎﻨﺷ ﻪﺑ ﻖﻓﻮﻣ
ﻡﻭﺯﻮﻣﻭﺮﻛ ۳B
ﺪﻧﺪﺷ ﻡﺪﻨﮔ ﺭﺩ .
ﻦﻳﺍ ﻑﺪﻫ ﺎﺠﻳﺍ ﻖﻴﻘﺤﺗ
ﺩ
ﺮﺑ ﻲﻨﺘﺒﻣ ﺮﮕﻧﺎﺸﻧ ﻪﺑ ﺖﻣﻭﺎﻘﻣ ﺎﺑ ﻪﺘﺳﻮﻴﭘ EST
ﺖﻳﻼﺑ
ﻣﻮﻳﺭﺍﺯﻮﻓ ﻲ ﻪﻠﺒﻨﺳ ﻡﺪﻨﮔ ﺭﺩ ﺩﻮﺑ .
ﻣ ﺵ ﻭﺭ ﻭ ﺩﺍﻮ ﺎﻫ
ﻫﺎﻴﮔﺩﺍﻮﻣ ﻲ ﺑﺎﻳﺯﺭﺍ ﻭ ﻲ ﺎﻫ ﻱ ﻪﻋﺭﺰﻣ ﺍ ﻱ : ﺖﻴﻌﻤﺟ ﻱﺎﻫ
F٣:٤
ﻭ
F٣:٥
ﻲﻗﻼﺗ ﺯﺍ ﻞﺻﺎﺣ ﺕﻼﻓ
× ﺎﻧﺎﺘﻧﻭﺮﻓ ﺩﺭﻮﻣ ﻖﻴﻘﺤﺗ ﻦﻳﺍ ﺭﺩ
ﺪﻨﺘﻓﺮﮔ ﺭﺍﺮﻗ ﻩﺩﺎﻔﺘﺳﺍ .
ﺕﻼﻓ ﺏﻮﻠﻄﻣ ﺕﺎﻴﺻﻮﺼﺧ ﺩﻮﺟﻭ ﺎﺑ
ﺢﻄﺳ ﻭ ﻲﻋﺍﺭﺯ ﻪﺑ ﻥﺍﺮﻳﺍ ﺭﺩ ﺩﺎﻳﺯ ﺖﺸﻛ ﺮﻳﺯ
ﺭﺎﻴﺴﺑ FHB
ﻭ ﺱﺎﺴﺣ ﺎﻧﺎﺘﻧﻭﺮﻓ
) ﻞﻳﺯﺮﺑ ﺯﺍ ( ﻪﺑ ﻡﻭﺎﻘﻣ ﻪﻤﻴﻧ ﻲﻣ FHB
ﺪﻨﺷﺎﺑ
(Mardi at el., 2004)
. ﺖﻴﻌﻤﺟ ﻱﺎﻫ
F٣:٤
ﻭ
F٣:٥
ﻪﺑ ﻩﺍﺮﻤﻫ
ﻦﻳﻻ ﻭﺩ ﻭ ﻦﻳﺪﻟﺍﻭ ﺱﻮﻣﺭ
ﻭ ﺱﺎﺴﺣ ﺪﻫﺎﺷ ﻥﺍﻮﻨﻋ ﻪﺑ ﻥﺎﻤﻟﺁ ﺯﺍ
CM_82036
ﻪﻋﺭﺰﻣ ﺭﺩ ﻡﻭﺎﻘﻣ ﺪﻫﺎﺷ ﻥﺍﻮﻨﻋ ﻪﺑ ﻦﻴﭼ ﺯﺍ ﻭ
ﺼﻣ ﻲﮔﺩﻮﻟﺁ ﺖﺤﺗ ﺑﺎﻳﺯﺭﺍ ﺩﺭﻮﻣ ﻲﻋﻮﻨ
ﻲ ﭙﻴﺗﻮﻨﻓ ﻲ ﺪﻨﺘﻓﺮﮔ ﺭﺍﺮﻗ .
ﺎﻳ ﻲﮔﺩﻮﻟﺁ ﺖﻓﺮﺸﻴﭘ ﻲﻨﺤﻨﻣ ﺮﻳﺯ ﺢﻄﺳ
AUDPC
ﺎﻨﺒﻣ ﺮﺑ ﻱ
ﻪﻠﺒﻨﺳ ﻱﺭﺎﻤﻴﺑ ﺖﻓﺮﺸﻴﭘ ﺪﺻﺭﺩ ١٠
، ١٤ ، ١٨ ، ٢٢ ﻭ ٢٦ ﺯﻭﺭ
ﻞﻜﺷ ﺮﻴﻴﻐﺗ ﻝﻮﻣﺮﻓ ﻪﻠﻴﺳﻮﺑ ﻲﮔﺩﻮﻟﺁ ﺯﺍ ﺲﭘ ﻪﺘﻓﺎﻳ
Shaner &
Finney (1977)
ﻩﺯﺍﺪﻧﺍ ﺮﻴﮔ ﻱ ﺪﺷ . ﻩﺩﺍﺩ ﺎﻫ ﻱ ﭙﻴﺗﻮﻨﻓ ﻲ ﻦﻳﺍ
ﻂﺳﻮﺗ ﻖﻴﻘﺤﺗ
Mardi et al.
(2006)
ﺪﺷ ﻞﺻﺎﺣ .
ﻥﺍﺭﺎﮑﻤﻫ ﻭ ﻲﻨﻴﺴﺣ :
ﺮﺑ ﻲﻨﺘﺒﻣ ﻲﻟﻮﮑﻟﻮﻣ ﻱﺎﻫﺮﮕﻧﺎﺸﻧ ﺩﺎﺠﻳﺍ ﺎﺑ ﻪﺘﺳﻮﻴﭘ EST
...
۱۸۹
ﺎﻫﺮﮕﻧﺎﺸﻧ ﺩﺎﺠﻳﺍ ﻱ
ﻨﺘﺒﻣ ﻲ ﺮﺑ :EST
ﻪﻳﺰﺠﺗ ﺎﻫ ﻱ ﻟﻮﮑﻟﻮﻣ ﻲ
ﻲﺳﺭﺮﺑ ﺎﺑ ٥٨٠١٥٥
ﻡﺪﻨﮔ EST
) ﺦﻳﺭﺎﺗ ﺎﺗ ١٤
ﻪﻳﻮﻧﺍﮊ
٢٠٠٥ (
، ٨٦٦٠
EST، ﻪﻠﺒﻨﺳ ﺯﺍ ﻞﺻﺎﺣ ﻪﺑ ﻩﺩﻮﻟﺁ ﻡﺪﻨﮔ ﻱﺎﻫ
F. graminearum
ﺭﺩ ﺩﻮﺟﻮﻣ ﺏﺎﺨﺘﻧﺍ NCBI
ﻭ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ
ﻪﻣﺎﻧﺮﺑ ﺯﺍ
BLASTn
ﺎﻬﻧﺁ ﮏﻳﮊﻮﻟﻮﻴﺑ ﺩﺮﮑﻠﻤﻋ ﺳﺭﺮﺑ
ﻲ ﺷ ﺪ . ﺯﺍ
ﻉﻮﻤﺠﻣ
EST ٨٦٦٠
، ٢٦٠ ﻟﺍﻮﺗ ﻲ ﻞﻳﺪﺒﺗ ﺯﺍ ﺲﭘ ﻭ ﺏﺎﺨﺘﻧﺍ
ﺖﻣﺮﻓ ﻪﺑ
FASTA
ﻡﺮﻧ ﺎﺑ ﺭﺍﺰﻓﺍ
Genedoc
ﺪﻧﺪﺷ ﻒﻳﺩﺮﻤﻫ .
ﻦﻳﺍ ﺮﺘﺸﻴﺑ ﻥﮊ ﺀﺰﺟ ﺎﻫEST
ﻥﺎﻫﺎﻴﮔ ﺮﮕﻳﺩ ﻭ ﻡﺪﻨﮔ ﻱﺎﻫ
ﺪﻧﺩﻮﺑ ﻚﺗﻮﮕﻴﻟﺍ ﺭﺍﺰﻓﺍ ﻡﺮﻧ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ .
، ١
٦٢ ﺯﺍ ﺮﮔﺯﺎﻏﺁ ﺖﻔﺟ
ﻦﻳﺍ ﺭﺩ ﻩﺪﺷ ﻆﻔﺣ ﺮﻴﻏ ﻭ ﻩﺪﺷ ﻆﻔﺣ ﻁﺎﻘﻧ ﻲﺣﺍﺮﻃ ﺎﻫ EST
ﻛ ﺪﺷ ،ﺩﺍﺪﻌﺗ ﻦﻳﺍ ﺯﺍ ﻪ ٤٢
ﻲﻟﺍﻮﺗ ﺱﺎﺳﺍ ﺮﺑ ﺮﮔﺯﺎﻏﺁ ﺖﻔﺟ
ﻥﮊ ﻭ ﻡﺪﻨﮔ ﻱﺎﻫ ٢٠
ﻥﮊ ﻲﻟﺍﻮﺗ ﺱﺎﺳﺍ ﺮﺑ ﺮﮔﺯﺎﻏﺁ ﺖﻔﺟ ﺯﺍ ﻱﺎﻫ
ﺩﻮﺑ ﺲﻴﺴﭘﻭﺪﻴﺑﺍﺭﺁ ﻭ ﺕﺭﺫ ،ﻮﺟ ،ﺞﻧﺮﺑ .
ﺶﻨﻛﺍﻭ ﻩﺮﻴﺠﻧﺯ ﻱﺎﻫ ﻱﺍ
ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﺯﺍﺮﻤﻴﻠﭘ ﺖﻔﺟ ﻭ ﻮﮕﻟﺍ ﻥﺍﻮﻨﻋ ﻪﺑ ﻦﻳﺪﻟﺍﻭ DNA
ﺪﺷ ﻡﺎﺠﻧﺍ ﻩﺪﺷ ﻲﺣﺍﺮﻃ ﻱﺎﻫﺮﻤﻳﺍﺮﭘ .
ﺯﺍﺮﻤﻴﻠﭘ ﺶﻨﻛﺍﻭ ﻝﻮﺼﺤﻣ
ﻱﻭﺭ ﺪﻴﻣﻼﻳﺮﮐﺍ ﻝﮊ ٦
ﺪﺷ ﮏﻴﮑﻔﺗ ﺪﺻﺭﺩ .
ﻪﻳﺰﺠﺗ Real Time PCR :
ﺝﺍﺮﺨﺘﺳﺍ ﺭﻮﻈﻨﻣ ﻪﺑ
RNA
ﻥﺎﻣﺯ ﺭﺩ ﻪﻠﺒﻨﺳ ﺯﺍ ﻲﻫﺩ ﻢﭼﺮﭘ ﻪﻠﺣﺮﻣ ﺭﺩ ﻱﺮﻴﮔ ﻪﻧﻮﻤﻧ ﻱﺎﻫ
۰ ، ۲۴ ، ۴۸ ﻭ ۹۶ ﺪﻫﺎﺷ ﺭﺎﻤﻴﺗ ﻭﺩ ﺭﺩ ﻲﮔﺩﻮﻟﺁ ﺯﺍ ﺪﻌﺑ ﺖﻋﺎﺳ
ﭗﻴﺗﻮﻧﮊ ﻭ ﺎﻧﺎﺘﻧﻭﺮﻓ ﻭ ﺕﻼﻓ ﻢﻗﺭ ﻭﺩ ﺯﺍ ﺭﺍﺮﮑﺗ ﻪﺳ ﺭﺩ ﻩﺩﻮﻟﺁ ﻭ ﺎﺑﻮﺷ ﮓﻧﻭ ﻱ
ﻡﻭﺎﻘﻣ ﺪﻫﺎﺷ ﻥﺍﻮﻨﻌﺑ ﺖﻓﺮﮔ ﺕﺭﻮﺻ
. ﻦﻴﻨﭽﻤﻫ
ﻭ ﺎﻤﻟ ،ﻡﻮﻠﮔ ﺯﺍ ﻩﺎﻴﮔ ﺪﺷﺭ ﺯﺍ ﻪﻠﺣﺮﻣ ﻦﻴﻤﻫ ﺭﺩ ﻱﺮﻴﮔ ﻪﻧﻮﻤﻧ ﻱﺎﻫ ﻥﺎﻣﺯ ﺭﺩ ﻻﺎﭘ ۰
، ۱۲ ﻭ ۲۴ ﺮﻫ ﺯﺍ ﻲﮔﺩﻮﻟﺁ ﺯﺍ ﺪﻌﺑ ﺖﻋﺎﺳ
ﺪﺷ ﻡﺎﺠﻧﺍ ﺭﺍﺮﮑﺗ ﻪﺳ ﻭ ﻢﻗﺭ ﻭﺩ ﺮﻫ ﺭﺩ ،ﺭﺎﻤﻴﺗ ﻭﺩ .
ﻱﺎﻫ ﻪﻧﻮﻤﻧ
ﺝﺍﺮﺨﺘﺳﺍ ﻡﺎﮕﻨﻫ ﺎﺗ ﻊﻳﺎﻣ ﺕﺯﺍ ﺭﺩ ﺩﺎﻤﺠﻧﺍ ﺯﺍ ﺪﻌﺑ ﺭﺩ RNA
۸۰
‐ ﻱﺭﺍﺪﻬﮕﻧ ﺩﺍﺮﮔ ﻲﺘﻧﺎﺳ ﻪﺟﺭﺩ ﺪﻧﺪﺷ
. ﺝﺍﺮﺨﺘﺳﺍ ﺎﺑ RNA
ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ
TrizolTM(from Gibco BRL Life Technologies)
ﺪﺷ ﻡﺎﺠﻧﺍ .
ﺖﻴﻤﮐ ﻦﻴﻴﻌﺗ ﻱﺍﺮﺑ ﺵﻭﺭ ﺯﺍ RNA
ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ ﻱﺮﺘﻣﻮﺘﻓﻭﺮﺘﮑﭙﺳﺍ .
ﺖﻴﻔﻴﮐ ﻦﻴﻴﻌﺗ ﺖﻬﺟ
RNA
ﺯﺭﺎﮔﺁ ﻝﮊ ﺯﺭﻮﻓﻭﺮﺘﮑﻟﺍ ﺵﻭﺭ ﺯﺍ ۳
/ ۱ % ﺮﻓﺎﺑ ﺎﺑ
1X MOPS
ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ .
ﻲﻟﺍﻮﺗ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﻧﺎﺑ ﺭﺩ ﺩﻮﺟﻮﻣ ﻱﺎﻫ
ﻲﺗﺎﻋﻼﻃﺍ ﮏ
ﻥﮊ ﻪﺑ ﻁﻮﺑﺮﻣ NCBI
ﻱﺎﻫ
No.BT009617)NPR1
(
18s ﻭ
) rRNA No.AY049040
( ﻥﮊ ﺮﻫ ﻱﺍﺮﺑ ﺮﻤﻳﺍﺮﭘ ﺖﻔﺟ ﮏﻳ
ﻡﺮﻧ ﻪﻠﻴﺳﻮﺑ ﺭﺍﺰﻓﺍ
Beacon designer (Primer Biotech)
ﺶﻨﮐﺍﻭ ﻱﺍﺮﺑ
Real Time PCR
ﺪﻧﺪﺷ ﻲﺣﺍﺮﻃ .
ﺶﻨﮐﺍﻭ
Real Time PCR
ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ
MyiQTM Single Color
Real-Time PCR Detection System
ﺶﻨﮐﺍﻭ ﻢﺠﺣ ﺭﺩ
1. Oligotech
۲۵ ﺪﺷ ﻡﺎﺠﻧﺍ ﺮﺘﻴﻟﻭﺮﮑﻴﻣ .
ﺯﺍ ﺶﻨﮐﺍﻭ ﻦﻳﺍ ﺭﺩ
iQTM SYBR®
Green Supermix Bio Rad Cat.# 170-8882)
( ﻩﺩﺎﻔﺘﺳﺍ
ﺪﺷ . ﻩﺩﺍﺩ ﺰﻴﻟﺎﻧﺁ ﺭﻮﻈﻨﻣ ﻪﺑ ﻱﺎﻫ
Real-Time
ﺭﺍﺰﻓﺍ ﻡﺮﻧ ﺯﺍ
ﺖﮐﺮﺷ ﻂﺳﻮﺗ ﻩﺪﺷ ﻲﺣﺍﺮﻃ
Bio Rad
ﺭﺍﺪﻘﻣ ﺱﺎﺳﺍ ﺮﺑ
threshold cycle (Ct)
ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ .
ﺯﺍ ﻥﮊ
18s rRNA
ﻱﺍﺮﺑ
ﻩﺩﺍﺩ ﻥﺩﺮﮐ ﻝﺎﻣﺮﻧ ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ ﺎﻫ
. ﻨﭽﻤﻫ ﻴ ﻡﺮﻧ ﺯﺍ ﻦ ﺭﺍﺰﻓﺍ
ﻲﻨﻌﻣ ﻲﺳﺭﺮﺑ ﻱﺍﺮﺑ ﺰﻴﻧ SPSS
ﺭﺩ ﻲﮔﺩﻮﻟﺁ ﻥﺎﻣﺯ ﺮﺛﺍ ﻱﺭﺍﺩ
ﻥﮊ ﻥﺎﻴﺑ ﻥﺍﺰﻴﻣ
NPR1
ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ .
ﺎﻫ ﻪﻳﺰﺠﺗ ﻱ ﺭﺎﻣﺁ ﻱ : ﻩﺪﻫﺎﺸﻣ ﺕﺭﻮﺻ ﺭﺩ ﺭﺍﻮﻧ
،ﻞﮑﺷ ﺪﻨﭼ
ﺰﺠﺗ ﻳ ﻪ ﻟﻮﮑﻟﻮﻣ ﻲ ١٧٠ ﻩﺎﻴﮔ
F3:4
ﺖﻔﺟ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ )
ﺎﻫ ﻱ (
ﺮﮔﺯﺎﻏﺁ ﻠﮑﺷ ﺪﻨﭼ ﻩﺪﻨﻨﮐ ﺩﺎﺠﻳﺍ ﻲ
ﺪﺷ ﻡﺎﺠﻧﺍ .
ﻱﺎﻫﻮﮕﻟﺍ ﺭﺍﻮﻧ ﻱ
ﺵﻭﺭ ﻖﺒﻃ
Lander et al.
(1987)
ﺪﻧﺪﺷ ﻲﻫﺩﺯﺎﻴﺘﻣﺍ .
ﻪﺸﻘﻧ
ﺮﮕﻧﺎﺸﻧ ﻲﮕﺘﺳﻮﻴﭘ ﺎﻫ
ﺪﻨﭼ ﻱ ﺭﺩ ﺎﻬﻧﺁ ﻩﺩﺎﻔﺘﺳﺍ ﺭﻮﻈﻨﻣ ﻪﺑ ﻞﻜﺷ
ﻥﺎﻜﻣ ﻪﻳﺰﺠﺗ ﻡﺮﻧ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ،ﻲﺑﺎﻳ
ﺭﺍﺰﻓﺍ
MAPMAKER
ver 3.00
ﺎﺑ ﺯﺍ ﺮﺘﻤﻛ ﻪﻠﺻﺎﻓ ﻦﺘﻓﺮﮔ ﺮﻈﻧ ﺭﺩ ٥٠
ﻲﺘﻧﺎﺳ ﻭ ﻥﺎﮔﺭﻮﻣ
LOD ≥٣ ﺪﺷ ﻡﺎﺠﻧﺍ
(Utz & Melchinger,
1996)
. ﻥﺎﻜﻣ ﻪﻠﺻﺎﻓ ﻲﺑﺎﻳ ﻡﺮﻧ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﺐﻛﺮﻣ ﻱﺍ
ﺭﺍﺰﻓﺍ
PLABQTL
ﻦﺘﻓﺮﮔﺮﻈﻧ ﺭﺩ ﺎﺑ ٥
/
LOD ≥ ٢ ﻩﺩﺍﺩ ﻭ
ﺎﻫ ﻱ
ﻪﻋﺭﺰﻣ ﺍ ﻱ ﺖﻴﻌﻤﺟ ﺯﺍ ﻞﺻﺎﺣ ﻱﺎﻫ
F٣:٤
ﻭ
F٣:٥
ﺪﺷ ﻡﺎﺠﻧﺍ .
ﺚﺤﺑ ﻭ ﺞﻳﺎﺘﻧ
ﻥﺎﮑﻣ ﻭ ﺩﺎﺠﻳﺍ ﺑﺎﻳ
ﻲ ﻨﺘﺒﻣ ﺮﮕﻧﺎﺸﻧ ﻲ
ﻥﮊ ﺮﺑ ﺎﺑ ﻪﺘﺳﻮﻴﭘ NPR1
ﺖﻣﻭﺎﻘﻣ ﻪﺑ ﻡﺪﻨﮔ ﺭﺩ FHB
ﻉﻮﻤﺠﻣ ﺯﺍ ٦٢
ﺣﺍﺮﻃ ﺮﮔﺯﺎﻏﺁ ﺖﻔﺟ ﻲ
ﻩﺪﺷ ﺖﻔﺟ ،
ﺣﺍﺮﻃ ﺮﮔﺯﺎﻏﺁ ﻲ
ﻩﺪﺷ ﻥﮊ ﺱﺎﺳﺍ ﺮﺑ NPR1
(Non-expressor pathogenesis related protein)
ﺍﺭﺍﺩ ﻱ
ﻠﮑﺷ ﺪﻨﭼ ﻲ ﺱﺎﺴﺣ ﻦﻳﺪﻟﺍﻭ ﻦﻴﺑ ﺕﻼﻓ
ﻡﻭﺎﻘﻣ ﻪﻤﻴﻧ ﻭ ﺎﻧﺎﺘﻧﻭﺮﻓ
.ﺩﻮﺑ ﺰﺠﺗ ﻳ ﻪ ﻟﻮﮑﻟﻮﻣ ﻲ ﻥﺎﻫﺎﻴﮔ ﻭ ﻦﻳﺪﻟﺍﻭ
F٣:٤
ﻞﮑﺷ ﺭﺩ ۱
ﺖﺳﺍ ﻩﺪﺷ ﻩﺩﺍﺩ ﻥﺎﺸﻧ .
ﻞﮑﺷ ١
‐ ﻲﺻﺎﺼﺘﺧﺍ ﺮﮔﺯﺎﻏﺁ ﺯﺍ ﻞﺻﺎﺣ ﻩﺪﺷ ﺮﻴﺜﮑﺗ ﺕﺎﻌﻄﻗ ﮏﻴﮑﻔﺗ
ﻥﮊ ﻦﻳﺪﻟﺍﻭ ﺭﺩ NPR1
ﺎﻧﺎﺘﻧﻭﺮﻓ FR) ( ﻭ ﺕﻼﻓ F) ( ﻭ
١١ ﻩﺎﻴﮔ F٣:٤
ﻗﻼﺗ ﺯﺍ ﻞﺻﺎﺣ ﻲ
ﺎﻬﻧﺁ
۱۹۰ ﻪﻠﺠﻣ
ﻲﻋﺍﺭﺯ ﻥﺎﻫﺎﻴﮔ ﻡﻮﻠﻋ ﺩ ،ﻥﺍﺮﻳﺍ
ﻩﺭﻭ ۴۰ ﻩﺭﺎﻤﺷ ، ۲ ، ۱۳۸۸
ﻪﻳﺰﺠﺗ ﺞﻳﺎﺘﻧ ﭘ
ﻴ ﮕﺘﺳﻮ ﻲ ﻩﺩﺍﺩ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﺎﻫ
ﻱ ﺩﻮﺟﻮﻣ
٢٧٣ ﺮﮕﻧﺎﺸﻧ ﻭ AFLP
ﺖﻴﻌﻤﺟ ﺭﺩ SSR
ﺎﻫ ﻱ ﺯﺍ ﻞﺻﺎﺣ
ﺕﻼﻓ
× ﺎﻧﺎﺘﻧﻭﺮﻓ
(Mardi et al., 2006)
، ﺩﺍﺩ ﻥﺎﺸﻧ ﻪﮐ
ﺖﻔﺟ
ﺎﻫﺮﮔﺯﺎﻏﺁ ﻱ ﻥﮊ ﺱﺎﺳﺍ ﺮﺑ ﻩﺪﺷ ﻲﺣﺍﺮﻃ ﻱﻭﺭ ﺮﺑ NPR1
ﻭﺯﺎﺑ ﻱ ﺪﻨﻠﺑ ﻡﻭﺯﻮﻣﻭﺮﻛ ٣A
ﺩﺭﺍﺩ ﺭﺍﺮﻗ )
ﻞﻜﺷ ٢ .(
ﻪﻳﺰﺠﺗ ﺞﻳﺎﺘﻧ
ﺎﻫﺮﮔﺯﺎﻏﺁ ﺖﻔﺟ ﺩﺍﺩ ﻥﺎﺸﻧ QTL
ﻱ ﺱﺎﺳﺍ ﺮﺑ ﻩﺪﺷ ﻲﺣﺍﺮﻃ
ﻪﻴﺣﺎﻧ ﺭﺩ ﻥﮊ ﻦﻳﺍ ﻂﺳﻮﺗ ﻩﺪﺷ ﻲﻳﺎﺳﺎﻨﺷ QTL
Mardi et al.
(2006) Steiner et al. ﻭ
(2004)
ﺎﻓ ﺪﺣ ﺭﺩ ﻭ ﻥﺎﮑﻣ ﻞﺻ
ﺎﻫ ﻱ
ﻧﮊ ﻲ
Xgwm720
ﻭ
Xe87m39_5
ﻧﺎﮑﻣ ﻪﻠﺻﺎﻓ ﺎﺑ ﻲ
ﺗﺮﺗ ﻪﺑ ﻴ ﺐ
۸ / ۶ ﻭ ۳ ﺘﻧﺎﺳ ﻲ ﻥﺎﮔﺭﻮﻣ ﺩﺭﺍﺩ ﺭﺍﺮﻗ ) ﻝﻭﺪﺟ ۱ .(
ﺑﺎﻳ ﻥﺎﮑﻣ ﺎﺑ ﻁﺎﺒﺗﺭﺍ ﺭﺩ ﻲ
ﻥﮊ ﺎﻫ ﻱ ﺖﻣﻭﺎﻘﻣ ﻩﺪﻨﻨﮐ ﻝﺮﺘﻨﮐ
FHB ﻪﺑ ﺭﺩ ﺎﻧﺎﺘﻧﻭﺮﻓ ﺖﺳﺍ ﻩﺪﺷ ﻪﺋﺍﺭﺍ ﺵﺭﺍﺰﮔ ﻭﺩ ﻥﻮﻨﻛﺎﺗ .
Steiner et al.
(2004)
ﻭ
Mardi et al.
(2006)
ﻦﻳﺮﺘﻤﻬﻣ ﻭﺭ ﺍﺭ ﺖﻣﻭﺎﻘﻣ ﻩﺪﻨﻨﮐ ﻝﺮﺘﻨﮐ QTL
ﻱ ﻭﺯﺎﺑ ﻱ ﺪﻨﻠﺑ
ﻡﻭﺯﻮﻣﻭﺮﻛ ۳A
ﻳﺎﺳﺎﻨﺷ ﻲ ﺪﻧﺩﻮﻤﻧ . ﻥﺎﺸﻧ ﻖﻴﻘﺤﺗ ﻦﻳﺍ ﺞﻳﺎﺘﻧ
ﺣﺍﺮﻃ ﺮﮕﻧﺎﺸﻧ ﺩﺍﺩ ﻲ
ﻥﮊ ﺱﺎﺳﺍ ﺮﺑ ﻩﺪﺷ
NPR1
ﻥﻭﺭﺩ ﺎﻘﻴﻗﺩ
ﻩﺪﺷ ﻦﻴﻴﻌﺗ QTL
ﻭﺭ ﻱ ﻡﻭﺯﻮﻣﻭﺮﮐ ۳A
ﺩﺭﺍﺩ ﺭﺍﺮﻗ . ﻪﭼﺮﮔﺍ
ﻢﻬﺳ
R2
ﺣﺍﺮﻃ ﺮﮕﻧﺎﺸﻧ ﻲ
ﻥﮊ ﺱﺎﺳﺍ ﺮﺑ ﻩﺪﺷ ﺭﺩ NPR1
ﺲﻧﺎﻳﺭﺍﻭ ﻪﻴﺟﻮﺗ
AUDPC
ﺍ ،ﺩﻮﺑ ﺰﻴﭼﺎﻧ ﻔﻨﻣ ﺮﻴﺛﺄﺗ ﻦﻳﺍ ﺎﻣ
ﻲ
ﺩﻮﺑ . ﮊﺎﮑﻨﻴﻟ ﻪﺸﻘﻧ ﻥﺩﻮﻤﻧ ﻢﻛﺍﺮﺘﻣ ﺮﺑ ﻩﻭﻼﻋ ﺮﮕﻧﺎﺸﻧ ﻦﻳﺍ ﻱ
ﻣﻮﻧﮊ ﻪﻴﺣﺎﻧ ﻦﻳﺍ ﺭﺩ ﺩﻮﺟﻮﻣ ﻲ
ﺶﻘﻧ ، ﻟﺎﻤﺘﺣﺍ ﻲ ﻥﮊ ﺍﺭ NPR1
ﻪﺑ ﺖﻣﻭﺎﻘﻣ ﺩﺎﺠﻳﺍ ﺭﺩ ﺩﻮﻤﻧ ﺺﺨﺸﻣ ﻡﺪﻨﮔ FHB
.
Makander et al.
) ﺼﺨﺷ ﺕﺎﺒﺗﺎﮑﻣ ﻲ
( ﺎﺑ ﺳﺭﺮﺑ ﻲ ﻥﮊ ﻥﺎﻴﺑ
ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﻡﺪﻨﮔ ﺭﺩ ﺲﻴﺴﭘﻭﺪﻴﺑﺍﺭﺁ ﺯﺍ ﻩﺪﺷ ﺍﺪﺟ NPR1
ﺭ ﺯﺍﺪﻧﺍ ﻩﺍ ﻥﮊ ﻥﺎﻴﺑ ﻪﻛ ﺪﻧﺩﺍﺩ ﻥﺎﺸﻧ ،ﺕﺭﺫ Ubi1
NPR1
ﺲﻴﺴﭘﻭﺪﻴﺑﺍﺭﺁ
(AtNPR1)
ﻪﺑ ﺖﻣﻭﺎﻘﻣ ﺶﻳﺍﺰﻓﺍ ﺚﻋﺎﺑ
FHB
ﻲﻣ ﻚﻴﻧﮋﺴﻧﺍﺮﺗ ﻡﺪﻨﮔ ﺭﺩ ﺩﻮﺷ
Cao et al. .
(1998)
ﻥﺎﻴﺑ
ﻥﮊ ﻥﺎﻴﺑ ﻱﻻﺎﺑ ﺢﻄﺳ ﻪﻛ ﺪﻧﺩﺮﻛ ﻪﻛ ﺲﻴﺴﭘﻭﺪﻴﺑﺍﺭﺁ NPR1
ﺖﻣﻭﺎﻘﻣ ﻝﺎﻨﮕﻴﺳ ﻝﺎﻘﺘﻧﺍ ﺮﻴﺴﻣ ﺖﺒﺜﻣ ﻩﺪﻨﻨﻛ ﻢﻴﻈﻨﺗ ﻚﻳ ﺎﺑ ﻲﻣ ﻚﻴﻤﺘﺴﻴﺳ ﻲﺑﺎﺴﺘﻛﺍ ﻪﺑ ﺖﻣﻭﺎﻘﻣ ﺶﻳﺍﺰﻓﺍ ﺚﻋﺎﺑ ،ﺪﺷ
ﺩﻮﺷ ﻲﻣ ﺲﻴﺴﭘﻭﺪﻴﺑﺍﺭﺁ ﻲﭼﺭﺎﻗ ﻭ ﻲﻳﺎﻳﺮﺘﻛﺎﺑ ﻱﺎﻫ ﻥﮊﻮﺗﺎﭘ .
ﻝﻭﺪﺟ ۱
‐ ﻪﻳﺰﺠﺗ ﻲﮔﺩﻮﻟﺁ ﺖﻓﺮﺸﻴﭘ ﻲﻨﺤﻨﻣ ﺮﻳﺯ ﺢﻄﺳ ﻱﺍﺮﺑ QTL
AUDPC) ( ﻪﻠﺻﺎﻓ ﻲﺑﺎﻳ ﻥﺎﻜﻣ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﺎﻳ ﺐﻛﺮﻣ ﻱﺍ
CIM) (
ﺎﻫ ﻞﺴﻧ ﺭﺩ
۳ﻱ ﻭ F F۵
ﺯﺍ ﻲﻗﻼﺗ ﺯﺍ ﻞﺻﺎﺣ ﺕﻼﻓ
× ﺎﻧﺎﺘﻧﻭﺮﻓ
ﺮﺛﺍ ﻡﻭﺯﻮﻣﻭﺮﻛ LODR2 ﻞﺴﻧ
۲۴۱ / ۱
AL‐ ٣ ۵/۶ ۲/۵۶ F۳
۳۷۱ / ۱
AL‐ ٣ ۹/۶ ۳/۳۳ F۵
ﻥﮊ ﻥﺎﻴﺑ ﻞﻴﻓﻭﺮﭘ NPR1
ﻱﺎﻫﺪﻧﺎﺑ ﺕﻮﻳﺭﺎﮐﻮﻳ ﺭﺩ rRNA
ﺯﺍ ﺪﻨﺗﺭﺎﺒﻋ ﺎﻫ ﻭ 28s
18s
ﻲﻫﺎﮔ ﻭ ﻝﻮﮑﻟﻮﻣ ﺎﻬﻧﺁ ﻦﻴﺑ ﻭ ﺖﺳﺍ 5s
mRNA ﻱﺎﻫ ﺕﺭﻮﺼﺑ
ﻲﻣ ﻩﺪﻳﺩ ﻲﮔﺪﻴﺸﮐ ﺪﻧﻮﺷ
. ﻱﺎﻫﺪﻧﺎﺑ ﺩﻮﺟﻭ ﻭ rRNA
ﻲﮔﺪﻴﺸﮐ ﻱﺎﻫﺪﻧﺎﺑ ﺩﻮﺟﻭ ﻥﻭﺪﺑ RNA
ﮏﻳ ﺮﮕﻧﺎﻳﺎﻤﻧ ،DNA
ﻲﻣ ﻖﻓﻮﻣ ﺝﺍﺮﺨﺘﺳﺍ ﺪﺷﺎﺑ
. ﺩﻮﺟﻭ ﻩﺩﻭﺪﺤﻣ ﺎﺒﻟﺎﻏ ﺭﺩ RNA
ﻝﻮﻃ ﺎﺗ ﻝﮊ ﻱﻭﺭ ۶Kb
ﺩﻮﺷ ﻲﻣ ﻞﻣﺎﺷ ﺍﺭ .
ﺭﺩ ﻪﮐ ﻪﻧﻮﮕﻧﺎﻤﻫ
ﻞﮑﺷ ۳ ﻱﺎﻫﺪﻧﺎﺑ ،ﺖﺳﺍ ﻩﺪﺷ ﺺﺨﺸﻣ ﻪﺑ ﻲﻣﻭﺯﻮﺒﻳﺭ RNA
ﻨﭽﻤﻫ ﺪﻨﺘﺴﻫ ﻩﺪﻫﺎﺸﻣ ﻞﺑﺎﻗ ﺡﻮﺿﻭ ﻴ
ﻱﺎﻫﺪﻧﺎﺑ ﺩﺍﺪﻌﺗ ﻦ
RNA
ﺩﻮﺟﻭ ﻞﻴﻟﺩ ﻪﺑ ﻪﮐ ﺖﺳﺍ ﺪﻧﺎﺑ ﻭﺩ ﺯﺍ ﺶﻴﺑ ﻲﻣﻭﺯﻮﺒﻳﺭ ﻩﻭﻼﻋ ﻲﺘﺳﻼﭘﻭﺮﻠﮐ ﻱﺎﻫrRNA rRNA ﺮﺑ
ﺭﺩ ﻱﺍ ﻪﺘﺴﻫ ﻱﺎﻫ
ﺪﺷ ﺎﺑ ﻲﻣ ﺰﺒﺳ ﻱﺎﻫ ﺖﻓﺎﺑ .
ﻞﻜﺷ ٢
‐ ﺕﻼﭘ ﮊﺎﮑﻨﻴﻟ ﻩﻭﺮﮔ ﻭ LOD
ﻱ ﺎﺑ ﻂﺒﺗﺮﻣ ﻧﮊ ﻥﺎﮑﻣ ﻲ ﻂﺒﺗﺮﻣ
ﻥﮊ ﺎﺑ ﻳﺎﺳﺎﻨﺷ NPR1
ﻲ ﻭﺭ ﻩﺪﺷ ﻱ ﻡﻭﺯﻮﻣﻭﺮﮐ ۳A
ﻞﮑﺷ ۳ RNA‐ ﻡﺪﻨﮔ ﻩﺎﻴﮔ ﺯﺍ ﻩﺪﺷ ﺝﺍﺮﺨﺘﺳﺍ
ﺯﺭﺎﮔﺁ ﻝﮊ ﻱﻭﺭ ﺮﺑ ۳
/ ۱
%
ﻥﺍﺭﺎﮑﻤﻫ ﻭ ﻲﻨﻴﺴﺣ :
ﺮﺑ ﻲﻨﺘﺒﻣ ﻲﻟﻮﮑﻟﻮﻣ ﻱﺎﻫﺮﮕﻧﺎﺸﻧ ﺩﺎﺠﻳﺍ ﺎﺑ ﻪﺘﺳﻮﻴﭘ EST
...
۱۹۱
ﻥﮊ ﻥﺎﻴﺑ ﺕﺍﺮﻴﻴﻐﺗ ﻲﺳﺭﺮﺑ ﺭﻮﻈﻨﻣ ﻪﺑ ﺯﺍ NPR1
ﺖﻴﻤﮐ ﻧ ﻲﺠﻨﺳ
١ﻲﺒﺴ ﺯﺍ ﻥﻮﭼ ﻭ ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ ﻪﺑ RNA
ﻩﺮﻴﺠﻧﺯ ﺶﻨﮐﺍﻭ ﺭﺩ ﻪﻴﻟﻭﺍ ﻱﻮﮕﻟﺍ ﻥﺍﻮﻨﻋ ﻲﻤﻧ ﺯﺍﺮﻤﻴﻠﭘ ﻱﺍ
ﻥﺍﻮﺗ
ﺱﻮﮑﻌﻣ ﻲﺴﻳﻮﻧﻭﺭ ﻲﻃ ﻪﻠﺣﺮﻣ ﻦﻴﻟﻭﺍ ﺭﺩ ﺩﺮﮐ ﻩﺩﺎﻔﺘﺳﺍ
RNA
cDNA ﻪﺑ ﺲﭙﺳ ،ﺪﺷ ﻞﻳﺪﺒﺗ ﺭﺩ ﻱﺍ ﻪﺘﺷﺭ ﮏﺗ cDNA
ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ ﺯﺍﺮﻤﻴﻠﭘ ﻱﺍ ﻩﺮﻴﺠﻧﺯ ﺶﻨﮐﺍﻭ .
ﺵﻭﺭ ﺭﺩ ﻪﮐ ﺎﺠﻧﺁ ﺯﺍ
ﺖﻴﻤﮐ ﻲﺒﺴﻧ ﻲﺠﻨﺳ ﺭﺩ ﺕﺎﻫﺎﺒﺘﺷﺍ
ﻲﻤﮐ ﺖﺷﻮﻧﻭﺭ ﻱﺯﺎﺳ ﻱﺎﻫ
ﻕﺎﻔﺗﺍ ﺶﻨﮐﺍﻭ ﺩﺍﻮﻣ ﺭﺩ ﺮﻴﻴﻐﺗ ﻪﻧﻮﮔ ﺮﻫ ﺎﺑ ﻲﮔﺩﺎﺳ ﻪﺑ mRNA
ﻥﮊ ﻥﺎﻴﺑ ﻱﻮﮕﻟﺍ ﺯﺍ ﺖﺳﺭﺩﺎﻧ ﺮﻴﺒﻌﺗ ﺚﻋﺎﺑ ﻭ ﺪﺘﻓﺍ ﻲﻣ ﻑﺪﻫ ﻱﺎﻫ
ﻭ ﺕﺎﻫﺎﺒﺘﺷﺍ ﻦﻳﺍ ﻥﺩﺮﮐ ﻢﮐ ﻱﺍﺮﺑ ﻝﻮﺒﻗ ﺩﺭﻮﻣ ﺵﻭﺭ ،ﺩﻮﺷ ﻲﻣ ﻥﺎﻣﺰﻤﻫ ﺮﻴﺜﮑﺗ ،ﺕﺍﺮﻴﻴﻐﺗ ﺢﻴﺤﺼﺗ ﺍ RNA
ﻪﺑ ﻪﮐ ﺖﺳﺍ ﻱ
ﺎﮑﺑ ﻥﺩﺮﮐ ﻝﺎﻣﺮﻧ ﻱﺍﺮﺑ ﻲﻠﺧﺍﺩ ﻊﺟﺮﻣ ﻥﺍﻮﻨﻋ ﺪﺑ ،ﺩﻭﺭ ﺭ
ﻳ ﻦ
ﻥﮊ ﺯﺍ ﺭﻮﻈﻨﻣ
18s rRNA
ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ ﻊﺟﺮﻣ ﻥﮊ ﻥﺍﻮﻨﻋ ﻪﺑ
) ﻝﻭﺪﺟ ۲
(Kim et al., 2003) ( .
ﻝﻭﺪﺟ ۲
‐ ﺎﻫﺮﮔﺯﺎﻏﺁ ﻱ ﺣﺍﺮﻃ ﻲ ﻩﺪﺷ
ﻟﺍﻮﺗ ﻲ ﻡﺎﻧ
5'-AAAGGCTGCGTTCAAGGG-3' NPF
5'-CTTCTTCACCTGTTGCTCATC-3' NPR
5'-GTGACGGGTGACGGAGAATT-3' 18s F
5'-GACACTAATGCGCCCGGTAT-3' 18s R
ﺶﻨﮐﺍﻭ ﻥﺎﻣﺪﻧﺍﺭ ﺯﺍ ﻲﺒﺴﻧ ﻱﺎﻫ ﻲﺑﺎﻳﺯﺭﺍ ﺭﺩ ﻪﮐ ﻲﻳﺎﺠﻧﺁ ﺯﺍ ﺶﻫﺎﮐ ﺎﻳ ﺶﻳﺍﺰﻓﺍ ﺖﺒﺴﻧ ﻪﺒﺳﺎﺤﻣ ﺖﻬﺟ ﺯﺍﺮﻤﻴﻠﭘ ﻱﺍ ﻩﺮﻴﺠﻧﺯ ﻲﻓﺮﻃ ﺯﺍ ﻭ ﺩﻮﺷ ﻲﻣ ﻩﺩﺎﻔﺘﺳﺍ ﻒﻠﺘﺨﻣ ﻱﺎﻫﺭﺎﻤﻴﺗ ﺭﺩ ﻥﮊ ﻥﺎﻴﺑ ﺭﺩ ﺕﻭﺎﻔﺗ ﺖﻠﻋ ﻪﺑ ﻊﺟﺮﻣ ﻭ ﻑﺪﻫ ﻱﺎﻫ ﻲﻟﺍﻮﺗ ﺮﻴﺜﮑﺗ ﻥﺎﻣﺪﻧﺍﺭ ﻝﻮﺼﺤﻣ ﻩﺯﺍﺪﻧﺍ ﻭ ﻝﻮﺼﺤﻣ ﻲﻟﺍﻮﺗ ،ﺮﮔﺯﺎﻏﺁ ﻝﺎﺼﺗﺍ ﻲﺣﺍﻮﻧ ﻭ ﻖﻴﻘﺤﺗ ﻦﻳﺍ ﺭﺩ ﺪﻨﺘﺴﻫ ﺕﻭﺎﻔﺘﻣ ﺯﺍﺮﻤﻴﻠﭘ ﻱﺍ ﻩﺮﻴﺠﻧﺯ ﺶﻨﮐﺍ
ﻑﺪﻫ ﻲﻟﺍﻮﺗ ﻱﺍﺮﺑ ﺕﻭﺎﻔﺘﻣ ﺩﺭﺍﺪﻧﺎﺘﺳﺍ ﻲﻨﺤﻨﻣ ﻭﺩ ﺯﺍ
NPR1)
(
ﻊﺟﺮﻣ ﻲﻟﺍﻮﺗ ﻭ )
18s rRNA
( ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ .
ﺜﮑﺗ ﻥﺎﻣﺪﻧﺍﺭ ﻴ
ﺎﺑ ﺮ
ﺯ ﻝﻮﻣﺮﻓ ﻳ ﺪﺷ ﻪﺒﺳﺎﺤﻣ ﺮ
(QIAGEN, 2005)
:
2 10 10
10(1 )= ( 13.32)= 0.301≈
= −solop −−
efficiency
ﻩﺩﺍﺩ ﺮﻧ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﺵﻭﺭ ﻦﻳﺍ ﺯﺍ ﻞﺻﺎﺣ ﻱﺎﻫ ﺭﺍﺰﻓﺍ ﻡ
iCycler
ﺪﻧﺪﺷ ﻞﻴﻠﺤﺗ ﻭ ﻪﻳﺰﺠﺗ .
ﻱﺎﻫ ﻑﺍﺮﮔ ﺭﺍﺰﻓﺍ ﻡﺮﻧ ﻦﻳﺍ ﺭﺩ
ﻪﻧﺎﮔﺍﺪﺟ ﺭﻮﻃ ﻪﺑ ﻪﻃﻮﺑﺮﻣ ﺏﻭﺫ ﻲﻨﺤﻨﻣ ﻭ ﻥﮊ ﺮﻫ ﻪﺑ ﻁﻮﺑﺮﻣ ﺩﻮﺷ ﻲﻣ ﻢﻴﺳﺮﺗ .
ﻥﮊ ﻱﺍﺮﺑ ﻪﻠﺻﺎﺣ ﻱﺎﻫﺭﺍﺩﻮﻤﻧ ﺯﺍ ﻱﺍ ﻪﻧﻮﻤﻧ
ﻞﮑﺷ ﺭﺩ ﻑﺪﻫ ﻥﮊ ﻭ ﻊﺟﺮﻣ ۴
ﺩﻮﻤﻧ ﻩﺪﻫﺎﺸﻣ ﻥﺍﻮﺗ ﻲﻣ ﺍﺭ .
1. Relative Quantification
ﻞﮑﺷ ﺭﺩ ﻪﮐ ﺭﻮﻄﻧﺎﻤﻫ ۴
ﺎﻫﺭﺎﻤﻴﺗ ﻪﻴﻠﮐ ﺖﺳﺍ ﺺﺨﺸﻣ
ﻱﺍﺭﺍﺩ ﺭﺍﺪﻘﻣ
CT
ﺪﻫﺩ ﻲﻣ ﻥﺎﺸﻧ ﻦﻳﺍ ﺪﻨﺘﺴﻫ ﻲﻧﺎﺴﮑﻳ ﺎﺒﻳﺮﻘﺗ
ﭺﺭﺎﻗ ﺎﺑ ﻲﮔﺩﻮﻟﺁ ﻂﻳﺍﺮﺷ ﺭﺩ ﻊﺟﺮﻣ ﻥﮊ ﻥﺎﻴﺑ ﻪﮐ
F. graminearum
ﺖﺳﺍ ﻪﺘﺷﺍﺪﻧ ﻱﺮﻴﻴﻐﺗ .
ﻦﻴﻨﭽﻤﻫ
ﻒﻠﺘﺨﻣ ﻱﺎﻫﺭﺎﻤﻴﺗ ﺍﺭﺍﺩ ﺍﺭ ﺭﻮﮐﺬﻣ ﻥﮊ ﺯﺍ ﻲﺗﻭﺎﻔﺘﻣ ﻱﺎﻫCt
ﺭﺩ ﻥﮊ ﻦﻳﺍ ﻥﺎﻴﺑ ﺭﺩ ﺕﻭﺎﻔﺗ ﺩﻮﺟﻭ ﺮﮕﻧﺎﻴﺑ ﺮﻣﺍ ﻦﻳﺍ ﻪﮐ ﺪﻨﺘﺴﻫ ﺷﺎﺑ ﻲﻣ ﻲﮔﺩﻮﻟﺁ ﻒﻠﺘﺨﻣ ﻱﺎﻫ ﻥﺎﻣﺯ ﺪ
.
ﻥﮊ ﻱﺍﺮﺑ ﺯﺍﺮﻤﻴﻠﭘ ﻱﺍ ﻩﺮﻴﺠﻧﺯ ﺶﻨﮐﺍﻭ ﺮﻴﺜﮑﺗ ﻲﻨﺤﻨﻣ ‐۴ ﻞﮑﺷ ﻊﺟﺮﻣ )
18s rRNA
( ﻑﺪﻫ ﻥﮊ ﻭ
NPR1)
( ﺎﻤﻟ ﺭﺩ
ﻩﺪﺷ ﻲﺣﺍﺮﻃ ﻱﺎﻫﺮﮔﺯﺎﻏﺁ ﻥﺩﻮﺑ ﻲﺻﺎﺼﺘﺧﺍ ﻲﺑﺎﻳﺯﺭﺍ
ﻪﮐ ﻲﻳﺎﺠﻧﺁ ﺯﺍ
SYBR Green
ﻪﺘﺷﺭ ﻭﺩ ﻪﺑ ﻝﺎﺼﺗﺍ ﺎﺑ -ﻱﺍ
،ﻲﺻﺎﺼﺘﺧﺍ ﺮﻴﻏ ﻱﺎﻫﺪﻧﺎﺑ ﺮﮕﻳﺩ ﻭ ﺮﻤﻳﺍﺩ ﺮﻤﻳﺍﺮﭘ ﻞﺜﻣ ﻲﻳﺎﻫ ﺎﺘﻧ ﻪﮐ ﺩﻮﺷ ﻲﻣ ﺚﻋﺎﺑ ﺪﻧﻮﺷ ﺩﺭﻭﺁﺮﺑ ﻲﻠﺻﺍ ﺖﻈﻠﻏ ﺯﺍ ﺮﺘﺸﻴﺑ ﺞﻳ
ﺯﺍ ﺎﻫﺮﮔﺯﺎﻏﺁ ﻥﺩﻮﻤﻧ ﻞﻤﻋ ﻲﺻﺎﺼﺘﺧﺍ ﺪﻴﻳﺄﺗ ﻱﺍﺮﺑ ﻦﻳﺍﺮﺑﺎﻨﺑ ﺏﻭﺫ ﻱﺎﻫ ﻲﻨﺤﻨﻣ ﺰﻴﻟﺎﻧﺁ
(Melt Curve Analysis) ﻩﺩﺎﻔﺘﺳﺍ
ﺪﺷ . ﺪﻨﻳﺍﺮﻓ ﻡﺎﻤﺗﺍ ﺯﺍ ﺲﭘ ﺏﻭﺫ ﻲﻨﺤﻨﻣ ﻢﻴﺳﺮﺗ ﻡﺎﺠﻧﺍ PCR
ﺪﺷ . ﻲﻣﺍﺭﺁ ﻪﺑ ﺕﺭﺍﺮﺣ ﻪﺟﺭﺩ ﺏﻭﺫ ﻲﻨﺤﻨﻣ ﻢﺳﺭ ﺭﻮﻈﻨﻣ ﻪﺑ
ﺖﻓﺎﻳ ﺶﻳﺍﺰﻓﺍ .
ﺭﺍﺮﺣ ﻪﺟﺭﺩ ﺭﺩ ﺕ
ﻲﻣﺎﻤﺗ ﻦﻴﻳﺎﭘ ﻱﺎﻫ ﺕﻻﻮﺼﺤﻣ
ﻩﺮﻴﺠﻧﺯ ﺶﻨﮐﺍﻭ ﻭ ﻩﺩﻮﺑ ﻱﺍ ﻪﺘﺷﺭ ﻭﺩ ﺕﺭﻮﺻ ﻪﺑ ﺯﺍﺮﻤﻴﻠﭘ ﻱﺍ
SYBR Green
ﻥﺍﺰﻴﻣ ﻪﺠﻴﺘﻧ ﺭﺩ ،ﺖﺳﺍ ﻞﺼﺘﻣ ﺎﻬﻧﺁ ﻪﺑ
ﺕﺭﺍﺮﺣ ﻪﺟﺭﺩ ﺭﺩ ﻪﮐ ﻲﻟﺎﺣ ﺭﺩ ﺖﺳﺍ ﻻﺎﺑ ﺖﻨﺳﺭﻮﻠﻓ ﻻﺎﺑ ﻱﺎﻫ
ﻩﺮﻴﺠﻧﺯ ﺶﻨﮐﺍﻭ ﺕﻻﻮﺼﺤﻣ ﻪﺑ ﻭ ﻩﺩﺍﺩ ﻊﺒﻃ ﺮﻴﻴﻐﺗ ﺯﺍﺮﻤﻴﻠﭘ ﻱﺍ
ﺘﻧ ﺭﺩ ،ﺪﻨﻳﺁ ﻲﻣ ﺭﺩ ﻱﺍ ﻪﺘﺷﺭ ﮏﺗ ﺖﻟﺎﺣ ﻲﻌﻳﺮﺳ ﺶﻫﺎﮐ ﻪﺠﻴ
ﺩﻮﺷ ﻲﻣ ﺩﺎﺠﻳﺍ ﺖﻨﺳﺭﻮﻠﻓ ﻥﺍﺰﻴﻣ ﺭﺩ .
ﻥﺍﺰﻴﻣ ﺵﻭﺭ ﻦﻳﺍ ﺭﺩ
ﻩﺯﺍﺪﻧﺍ ﺕﺭﺍﺮﺣ ﻪﺟﺭﺩ ﺶﻳﺍﺰﻓﺍ ﺎﺑ ﻥﺎﻣﺰﻤﻫ ﺖﻨﺳﺭﻮﻠﻓ ﻱﺮﻴﮔ
ﻲﻣ ﺕﻼﭘ ﺕﺭﺍﺮﺣ ﻪﺟﺭﺩ ﻞﺑﺎﻘﻣ ﺭﺩ ﻭ ﻩﺪﺷ ﺩﻮﺷ
. ﺏﻭﺫ ﻪﻄﻘﻧ ﺭﺩ
Tm) ( ۵۰ % ﺭﺩ ﻲﻧﮊﻭﺭﺪﻴﻫ ﻱﺎﻫﺪﻧﻮﻴﭘ ﻱﺍ ﻪﺘﺷﺭ ﻭﺩ ﻱﺎﻫDNA
ﻴﻐﺗ ﻲﻧﺎﻬﮔﺎﻧ ﺭﻮﻄﺑ ﺖﻨﺳﺭﻮﻠﻓ ﻥﺍﺰﻴﻣ ﻭ ﻩﺪﺷ ﺍﺪﺟ ﻢﻫ ﺯﺍ ﺮﻴ
ﻲﻣ ﺪﺑﺎﻳ . ﮏﻴﭘ ﺯﺍ ﮏﻳ ﺮﻫ ﻲﻨﺤﻨﻣ ﻦﻳﺍ ﺭﺩ ﺮﮕﻧﺎﻳﺎﻤﻧ ﺎﻫ
Tm
ﮏﻳ
ﻝﻮﺼﺤﻣ ﺖﺳﺍ PCR
(QIAGEN, 2005)
. ﺎﺑ ﻦﻳﺍﺮﺑﺎﻨﺑ
NPR1 18s rRNA
ﻪﻧﺎﺘﺳﺁ ﺪﺣ
۱۹۲ ﻪﻠﺠﻣ
ﻲﻋﺍﺭﺯ ﻥﺎﻫﺎﻴﮔ ﻡﻮﻠﻋ ﺩ ،ﻥﺍﺮﻳﺍ
ﻩﺭﻭ ۴۰ ﻩﺭﺎﻤﺷ ، ۲ ، ۱۳۸۸
ﺏﻭﺫ ﻱﺎﻫ ﻲﻨﺤﻨﻣ ﺰﻴﻟﺎﻧﺁ ﺭﺩ ﮏﻴﭘ ﮏﻳ ﺎﻬﻨﺗ ﻩﺪﻫﺎﺸﻣ ﻊﺟﺮﻣ ﻥﮊ ﻱﺎﻫﺮﮔﺯﺎﻏﺁ ﻪﮐ ﺪﺷ ﺺﺨﺸﻣ )
18s rRNA
( ﻥﮊ ﻭ
ﻑﺪﻫ ) (NPR1
ﺪﻨﻫﺩ ﻲﻣ ﺮﻴﺜﮑﺗ ﺍﺭ ﻲﺻﺎﺼﺘﺧﺍ ﻝﻮﺼﺤﻣ
) ﻞﮑﺷ ۵ .(
ﺶﻨﮐﺍﻭ ﺕﻻﻮﺼﺤﻣ ﻦﻴﻨﭽﻤﻫ ﺯﺍﺮﻤﻴﻠﭘ ﻱﺍ ﻩﺮﻴﺠﻧﺯ
ﺯﺭﺎﮔﺁ ﻝﮊ ﻱﻭﺭ ﺮﺑ ۵
/ ۱ % ﺖﻓﺮﮔ ﺭﺍﺮﻗ ﻲﻨﻴﺑﺯﺎﺑ ﺩﺭﻮﻣ .
ﻦﻳﺍ
ﻩﺩﻭﺪﺤﻣ ﺭﺩ ﺪﻧﺎﺑ ﮏﻳ ﺎﻬﻨﺗ ﺰﻴﻧ ﻝﮊ ﻱﻭﺭ ﺮﺑ ﺎﻫﺮﮔﺯﺎﻏﺁ ۱۹۰
‐
۱۵۰ ﺪﻧﺩﻮﻤﻧ ﺩﺎﺠﻳﺍ ﺯﺎﺑ )
ﻞﮑﺷ ۶ .(
ﻞﮑﺷ ۵
‐ ﻊﺟﺮﻣ ﻥﮊ ﻱﺎﻫﺮﮔﺯﺎﻏﺁ ﺏﻭﺫ ﻲﻨﺤﻨﻣ )
18s rRNA (
ﻭ
ﻑﺪﻫ ﻥﮊ ) (NPR1 ﺖﻴﻤﮐ ﺵﻭﺭ ﺭﺩ ﺎﻬﻧﺁ ﻥﺩﻮﺑ ﻲﺻﺎﺼﺘﺧﺍ ﻭ
ﺒﺴﻧ ﻲﺠﻨﺳ ﻲ
ﺎﻤﻟ ﺭﺩ
ﻞﮑﺷ ۶
‐ ﻱﺍﺮﺑ ﻩﺪﺷ ﻲﺣﺍﺮﻃ ﻱﺎﻫﺮﮔﺯﺎﻏﺁ ﻥﺩﺮﮐ ﻞﻤﻋ ﻲﺻﺎﺼﺘﺧﺍ
ﻲﺒﺴﻧ ﻲﺠﻨﺳ ﺖﻴﻤﮐ ﺵﻭﺭ ﺯﺭﺎﮔﺁ ﻝﮊ ﻱﻭﺭ
۵ / ۱
%
ﻥﻮﻣﺯﺁ ﺞﻳﺎﺘﻧ Relative Quantification-PCR
ﻥﮊ ﻥﺎﻴﺑ ﺕﺍﺮﻴﻴﻐﺗ ﻲﺳﺭﺮﺑ ﻱﺍﺮﺑ ﻪﮑﻨﻳﺍ ﺯﺍ ﺪﻌﺑ NPR1
CT
ﻩﺩﺍﺩ ﺭﺍﺰﻓﺍ ﻡﺮﻧ ﺎﺑ ﺎﻫ
iCycler
ﺭﺍﺰﻓﺍ ﻡﺮﻧ ﻪﻠﻴﺳﻮﺑ ،ﺪﺷ ﺺﺨﺸﻣ
ﻩﺪﺷ ﻲﺣﺍﺮﻃ ﺖﮐﺮﺷ ﻂﺳﻮﺗ
Bio Rad
ﺭﺍﺪﻘﻣ ﺱﺎﺳﺍ ﺮﺑ
threshold cycle (Ct)
ﺯ ﻝﻮﻣﺮﻓ ﻖﺒﻃ ﺮﻴﺜﮑﺗ ﻥﺎﻣﺪﻧﺍﺭ ﻭ ﻳ
ﺮ
ﺪﻳﺩﺮﮔ ﻪﺒﺳﺎﺤﻣ ﻑﺪﻫ ﻥﮊ ﻥﺎﻴﺑ ﺶﻫﺎﮐ ﺎﻳ ﺶﻳﺍﺰﻓﺍ ﻥﺍﺰﻴﻣ
(Yuan et al., 2006)
.
) (
.
) (
arg
. arg
) (
) (
sample control ct
ref
sample control ct
et t
ref et t
E
Ratio E ∆ −
−
∆
=
ﺖﺒﺴﻧ ﻂﺳﻮﺗ ﻩﺪﺷ ﻲﺣﺍﺮﻃ ﺭﺍﺰﻓﺍ ﻡﺮﻧ ﺯﺍ ﻩﺪﻣﺁ ﺖﺳﺪﺑ ﻱﺎﻫ
ﺖﮐﺮﺷ
Bio Rad
ﻥﺎﻣﺯ ﺮﺛﺍ ﻱﺭﺍﺩ ﻲﻨﻌﻣ ﻲﺳﺭﺮﺑ ﺭﻮﻈﻨﻣ ﻪﺑ
ﻥﮊ ﻥﺎﻴﺑ ﻥﺍﺰﻴﻣ ﺭﺩ ﻲﮔﺩﻮﻟﺁ ﻼﻣﺎﮐ ﺡﺮﻃ ﺐﻟﺎﻏ ﺭﺩ NPR1
ﺭﺍﺰﻓﺍ ﻡﺮﻧ ﺎﺑ ﻲﻓﺩﺎﺼﺗ
SPSS
ﺰﺠﺗ ﻳ ﺪﻧﺪﺷ ﻪ . ﻥﺩﻮﺑ ﻝﺎﻣﺮﻧ ﻥﻮﻣﺯﺁ
ﺭﺍﺰﻓﺍ ﻡﺮﻧ ﻂﺳﻮﺗ ﻲﺸﻳﺎﻣﺯﺁ ﻱﺎﻄﺧ ﺲﻧﺎﻳﺭﺍﻭ
Minitab
ﺕﺭﻮﺻ
ﻞﻳﺪﺒﺗ ﺩﻮﺒﻧ ﻝﺎﻣﺮﻧ ﻲﺸﻳﺎﻣﺯﺁ ﻱﺎﻄﺧ ﻪﮐ ﻱﺩﺭﺍﻮﻣ ﺭﺩ ﻭ ﺖﻓﺮﮔ ﺖﻓﺮﮔ ﺕﺭﻮﺻ ﻩﺩﺍﺩ .
ﺴﻳﺎﻘﻣ ﻥﻮﻣﺯﺁ ﺎﺑ ﺎﻫ ﻩﺩﺍﺩ ﻦﻴﮕﻧﺎﻴﻣ ﻪ
ﻝﺎﻤﺘﺣﺍ ﺢﻄﺳ ﺭﺩ ﻦﮑﻧﺍﺩ ۵
% ﺖﻓﺮﮔ ﺕﺭﻮﺻ .
ﺭﺩ ﻪﻠﺻﺎﺣ ﺞﻳﺎﺘﻧ
ﺖﺳﺍ ﻩﺪﺷ ﻩﺩﺍﺩ ﻥﺎﺸﻧ ﺮﻳﺯ ﻱﺎﻫﺭﺍﺩﻮﻤﻧ )
ﻞﮑﺷ ۷ .(
ﺕﻼﻓ ﻢﻗﺭ ﺭﺩ )
ﺱﺎﺴﺣ ( ﻥﮊ ﺮﻫﺎﻈﺗ ﻥﺍﺰﻴﻣ ﻻﺎﭘ ﺭﺩ NPR1
ﻭ ﺖﺳﺍ ﻪﺘﺷﺍﺩ ﺶﻳﺍﺰﻓﺍ ﺪﻫﺎﺷ ﺭﺎﻤﻴﺗ ﻪﺑ ﺖﺒﺴﻧ ﻥﺎﻣﺯ ﻦﻴﻟﻭﺍ ﺭﺩ ﮔﺩﻮﻟﺁ ﻥﺎﻣﺯ ﺖﺷﺬﮔ ﺎﺑ ﺶﻳﺍﺰﻓﺍ ﻦﻳﺍ ﻲ
ﺍﺩ ﺶﻳﺍﺰﻓﺍ ﺭﺩ ﻭ ﻪﺘﺷ
ﻥﺎﻣﺯ ۲۴ ﺩﻮﺧ ﻥﺍﺰﻴﻣ ﻦﻳﺮﺗﻻﺎﺑ ﻪﺑ )
۵۲ / ۷ ﻪﺑ ﺖﺒﺴﻧ ﺮﺑﺍﺮﺑ
ﻝﺮﺘﻨﮐ ( ﺖﺳﺍ ﻩﺪﻴﺳﺭ .
ﻱﺎﻫ ﻥﺎﻣﺯ ﺭﺩ ﻥﮊ ﻦﻳﺍ ﺮﻫﺎﻈﺗ ۱۲
ﻭ
۲۴ ﻩﺩﺍﺩ ﻥﺎﺸﻧ ﻥﺎﻣﺯ ﻦﻴﻟﻭﺍ ﻪﺑ ﺖﺒﺴﻧ ﻱﺭﺍﺩ ﻲﻨﻌﻣ ﺶﻳﺍﺰﻓﺍ
ﺖﺳﺍ . ﺰﻴﻧ ﻡﻮﻠﮔ ﺭﺩ ﺕﻼﻓ ﻢﻗﺭ ﺭﺩ ﻱﺩﻮﻌﺻ ﺪﻧﻭﺭ ﻦﻴﻨﭼ ﻥﮊ ﻥﺎﻴﺑ ﻥﺎﻣﺯ ﻦﻴﻟﻭﺍ ﺭﺩ ﻪﮐ ﺕﻭﺎﻔﺗ ﻦﻳﺍ ﺎﺑ ﺖﺳﺍ ﻩﺪﺷ ﻩﺪﻫﺎﺸﻣ ﺭﺩ ﻲﻟﻭ ﺖﺳﺍ ﻪﺘﻓﺎﻳ ﺶﻫﺎﮐ ﺪﻫﺎﺷ ﺭﺎﻤﻴﺗ ﻪﺑ ﺖﺒﺴﻧ NPR1
ﻥﺎﻣﺯ ﻥﺎﻣﺯ ﻦﻴﻟﻭﺍ ﻭ ﺪﻫﺎﺷ ﺭﺎﻤﻴﺗ ﻪﺑ ﺖﺒﺴﻧ ﺶﻳﺍﺰﻓﺍ ﺪﻌﺑ ﻱﺎﻫ
ﺖﺳﺍ ﻩﺪﻳﺩﺮﮔ ﻩﺪﻫﺎﺸﻣ .
ﻪﺑ ﺖﺒﺴﻧ ﻥﮊ ﺮﻫﺎﻈﺗ ﺶﻳﺍﺰﻓﺍ ﺎﻤﻟ ﺭﺩ
ﻥﺎﻣﺯ ﻭ ﻥﺎﻣﺯ ﻦﻴﻟﻭﺍ ﺭﺩ ﺪﻫﺎﺷ ﺭﺎﻤﻴﺗ ۲۴
ﻩﺪﺷ ﻩﺩﺍﺩ ﻥﺎﺸﻧ
ﻥﺎﻣﺯ ﺭﺩ ﻲﻟﻭ ،ﺖﺳﺍ ۱۲
ﺖﺳﺍ ﻩﺪﺷ ﻩﺪﻳﺩ ﻥﮊ ﺮﻫﺎﻈﺗ ﺶﻫﺎﮐ .
ﻳﺍﺰﻓﺍ ﺩﻮﺟﻭ ﺎﺑ ﻥﺎﻣﺯ ﺭﺩ ﺶ
۲۴ ،ﺪﻫﺎﺷ ﺭﺎﻤﻴﺗ ﻪﺑ ﺖﺒﺴﻧ
ﺖﺳﺍ ﻩﺩﻮﺑ ﻥﺎﻣﺯ ﻦﻴﻟﻭﺍ ﺭﺩ ﻥﮊ ﺮﻫﺎﻈﺗ ﻥﺍﺰﻴﻣ ﻦﻳﺮﺘﺸﻴﺑ .
ﺭﺩ
ﺪﻧﻭﺭ ﻥﮊ ﺮﻫﺎﻈﺗ ﻥﺍﺰﻴﻣ ،ﺕﻼﻓ ﻢﻗﺭ ﻪﻠﺒﻨﺳ ﻱﺎﻫ ﻪﻧﻮﻤﻧ ﻥﺎﻣﺯ ﺎﺗ ﻲﺸﻳﺍﺰﻓﺍ ۴۸
ﺖﺳﺍ ﻩﺩﺍﺩ ﻥﺎﺸﻧ .
ﻥﺎﻣﺯ ﺭﺩ ﺎﻣﺍ ۹۶
ﮔﺩﻮﻟﺁ ﺯﺍ ﺲﭘ ﺖﻋﺎﺳ ﻲ
ﻥﺎﻣﺯ ﻪﺑ ﺖﺒﺴﻧ ۴۸
ﺮﻫﺎﻈﺗ ﻥﺍﺰﻴﻣ
ﺖﺳﺍ ﻪﺘﻓﺎﻳ ﺶﻫﺎﮐ ﻱﺭﺍﺩ ﻲﻨﻌﻣ ﺭﻮﻄﺑ )
۲۸۸ / ۰ ﺖﺒﺴﻧ ﺮﺑﺍﺮﺑ
ﺪﻫﺎﺷ ﺭﺎﻤﻴﺗ ﻪﺑ .(
ﺗﻮﻧﮊ ﺭﺩ ﻴ ﺭﺩ ﻥﮊ ﺮﻫﺎﻈﺗ ﻥﺍﺰﻴﻣ ﻱﺎﺑﻮﺷ ﮓﻧﻭ ﻡﻭﺎﻘﻣ ﭗ
ﺶﻳﺍﺰﻓﺍ ﺎﻤﻟ ﺭﺩ ﻥﺎﻣﺯ ﻦﻴﻟﻭﺍ ۰۷۸
/ ۲ ﺭﺎﻤﻴﺗ ﻪﺑ ﺖﺒﺴﻧ ﻱﺮﺑﺍﺮﺑ
ﻡﻮﻠﮔ ﻭ ﻻﺎﭘ ﺭﺩ ﻥﺎﻣﺯ ﻦﻴﻤﻫ ﺭﺩ ﻲﻟﻭ ﺖﺳﺍ ﻩﺩﺍﺩ ﻥﺎﺸﻧ ﻝﺮﺘﻨﮐ
18s rRNA
NPR1 18S
rRNA
NPR1
ﻥﺍﺭﺎﮑﻤﻫ ﻭ ﻲﻨﻴﺴﺣ :
ﺮﺑ ﻲﻨﺘﺒﻣ ﻲﻟﻮﮑﻟﻮﻣ ﻱﺎﻫﺮﮕﻧﺎﺸﻧ ﺩﺎﺠﻳﺍ ﺎﺑ ﻪﺘﺳﻮﻴﭘ EST
...
۱۹۳
ﺖﺳﺍ ﻩﺪﺷ ﻩﺪﻳﺩ ﺶﻫﺎﮐ .
ﺭﺩ ﻱﺮﻴﻴﻐﺗ ﻱﺪﻌﺑ ﻱﺎﻫ ﻥﺎﻣﺯ ﺭﺩ
ﺖﺳﺍ ﻩﺪﺸﻧ ﻩﺪﻫﺎﺸﻣ ﻥﮊ ﻥﺎﻴﺑ ﻥﺍﺰﻴﻣ .
ﻪﻠﺒﻨﺳ ﻱﺎﻫ ﻪﻧﻮﻤﻧ ﺭﺩ
ﺗﻮﻧﮊ ﻴ ﺖﺳﺍ ﻩﺪﺷ ﻩﺪﻳﺩ ﻥﮊ ﻥﺎﻴﺑ ﺶﻫﺎﮐ ﻱﺎﺑﻮﺷ ﮓﻧﻭ ﭗ .
ﺎﻣﺍ
ﻥﺎﻣﺯ ﺭﺩ ۹۶
ﻥﺎﻣﺯ ﻪﺑ ﺖﺒﺴﻧ ۴۸
ﺪﺷ ﻩﺪﻳﺩ ﺶﻳﺍﺰﻓﺍ .
ﺭﺩ
ﺖﺳﺍ ﻩﺩﻮﺒﻧ ﺭﺍﺩ ﻲﻨﻌﻣ ﺕﺍﺮﻴﻴﻐﺗ ﻦﻳﺍ ﻉﻮﻤﺠﻣ .
Pale a a
de
b d
e de
c
de de
0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0
0 12 24
Hours postinoculation
Fold increase over control
Falat Wangshuibai Frontana
Glume
a
b c
bc bc bc
0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0
0 12 24
Hours pos tinoculation
Fold increase over control
Falat Wangshuibai
Lemma
bc c
a
bc bc b
bc c
a
0.0 1.0 2.0 3.0 4.0 5.0
0 12 24
Hours postinoculation
Fold increase over control
Falat Wangshuibai Frontana
Spike
b a
ab ab
a
ab ab b
a
ab ab ab
0.0 0.5 1.0 1.5 2.0
0 24 48 96
Hours postinoculation
Fold increase over control
Falat Wangshuibai Frontana
ﻞﮑﺷ ٧
‐ ﻪﺴﻳﺎﻘﻣ ﻣ ﻴ ﮕﻧﺎ ﻴ ﻦ ﻥﮊ ﺮﻫﺎﻈﺗ ﻲﺒﺴﻧ ﺮﻫﺎﻈﺗ ﺭﺩ NPR1
ﺕﻼﻓ ﻡﺎﻗﺭﺍ ) ﺱﺎﺴﺣ ( ﺎﻧﺎﺘﻧﻭﺮﻓ ، ) ﻡﻭﺎﻘﻣ ﻪﻤﻴﻧ (
ﻮﺷ ﮓﻧﻭ ﻭ ﻱﺎﺑ ) ﻡﻭﺎﻘﻣ (
ﻥﮊ ﻥﺎﻴﺑ ﻥﺍﺰﻴﻣ ﻦﻳﺮﺗﻻﺎﺑ ﺎﻧﺎﺘﻧﻭﺮﻓ ﻢﻗﺭ ﺭﺩ NPR1
) ﻪﻤﻴﻧ
ﻡﻭﺎﻘﻣ ( ﻲﮔﺩﻮﻟﺁ ﺕﺎﻋﺎﺳ ﻦﻴﻟﻭﺍ ﺭﺩ )
۰ ، ۱۲ ﻭ ۲۴ ( ﻪﺑ
F. graminearum
) ۱۱ / ۴ ﻭ ۴۰۹ / ۲ ﺭﺎﻤﻴﺗ ﻪﺑ ﺖﺒﺴﻧ ﺮﺑﺍﺮﺑ
ﻻﺎﭘ ﻭ ﺎﻤﻟ ﺭﺩ ﺪﻫﺎﺷ (
ﺖﺳﺍ ﻩﺩﻮﺑ .
ﺮﻴﻴﻐﺗ ﻥﮊ ﺮﻫﺎﻈﺗ ﻪﻠﺒﻨﺳ ﺭﺩ ﻲﻨﻌﻣ
ﺖﺳﺍ ﻩﺩﺮﮑﻧ ﻱﺭﺍﺩ .
ﺗﻮﻧﮊ ﺪﻨﻧﺎﻣ ﺎﻣﺍ ﻴ
ﺑﻮﺷ ﮓﻧﻭ ﭗ ﻱﺎ
ﻥﺎﻣﺯ ﻪﺑ ﻁﻮﺑﺮﻣ ﻥﮊ ﻥﺎﻴﺑ ﻥﺍﺰﻴﻣ ﻦﻳﺮﺘﺸﻴﺑ ۹۶
ﺖﺳﺍ ﻩﺩﻮﺑ .
ﻥﺎﻣﺯ ﺭﺩ ﻉﻮﻤﺠﻣ ﺭﺩ ﻦﻴﻳﺎﭘ ﻱﺎﻫ
ﺕﻼﻓ ﺱﺎﺴﺣ ﻢﻗﺭ ﺭﺩ ﺮﺗ
ﻥﮊ ﻥﺎﻴﺑ ﺭﺩ ﻲﺸﻳﺍﺰﻓﺍ ﺪﻧﻭﺭ ﺖﺳﺍ ﻩﺪﺷ ﻩﺪﻫﺎﺸﻣ NPR1
.
ﻥﺎﻴﺑ ﻥﺍﺰﻴﻣ ﻦﻳﺮﺘﺸﻴﺑ ﻻﻮﻤﻌﻣ ﻦﻴﻳﺎﭘ ﻱﺎﻫ ﻥﺎﻣﺯ ﺭﺩ ﻪﮑﻳﺭﻮﻄﺑ ﺖﺳﺍ ﻩﺩﻮﺑ ﺕﻼﻓ ﺱﺎﺴﺣ ﻢﻗﺭ ﻪﺑ ﻁﻮﺑﺮﻣ ﻥﮊ .
ﻡﻭﺎﻘﻣ ﻡﺎﻗﺭﺍ ﺭﺩ
ﻘﻣ ﻪﻤﻴﻧ ﻭ ﻤﻫ ﺭﺩ ﻡﻭﺎ ﻴ
ﻩﺪﺷ ﻩﺪﻫﺎﺸﻣ ﻲﺸﻫﺎﮐ ﺪﻧﻭﺭ ﺎﻳ ﺎﻬﻧﺎﻣﺯ ﻦ
ﻥﮊ ﺮﻫﺎﻈﺗ ﻥﺍﺰﻴﻣ ﺭﺩ ﻱﺮﻴﻴﻐﺗ ﺎﻳ ﻭ ﺖﺳﺍ ﻩﺪﻫﺎﺸﻣ NPR1
ﺖﺳﺍ ﻩﺪﺸﻧ .
ﻥﺎﻣﺯ ﺭﺩ ﻥﮊ ﻥﺎﻴﺑ ﺭﺩ ﺕﻭﺎﻔﺗ ﻦﻳﺍ ۹۶
ﹰﻼﻣﺎﮐ
ﻥﺎﻣﺯ ﺭﺩ ﻪﮐ ﺕﻼﻓ ﻪﮑﻳﺭﻮﻄﺑ ﺖﺳﺍ ﻩﺪﺷ ﺲﮑﻋ ﻦﻴﻳﺎﭘ ﻱﺎﻫ
ﻥﮊ ﻥﺎﻴﺑ ﻦﻳﺮﺘﺸﻴﺑ
NPR1
ﻣ ﻦﻳﺮﺘﻤﮐ ،ﺖﺳﺍ ﻪﺘﺷﺍﺩ ﺍﺭ ﻴ
ﺍﺭ ﻥﺍﺰ
ﮓﻧﻭ ﺭﺩ ﻭ ﺖﺳﺍ ﻪﺘﺷﺍﺩ ﻥﺍﺰﻴﻣ ﻦﻳﺮﺘﺸﻴﺑ ﺎﻧﺎﺘﻧﻭﺮﻓ ﻭ ﻱﺎﺑﻮﺷ
ﺖﺳﺍ ﻩﺩﻮﺑ .
ﺍﺪﻳﺪﻧﺎﮐ ﻥﮊ ﻥﺎﻴﺑ ﻞﻴﻓﻭﺮﭘ ﺞﻳﺎﺘﻧ ﻱ
ﻪﺑ ﺖﻣﻭﺎﻘﻣ
) FHB
(NPR1
ﻡﻭﺎﻘﻣ ﻭ ﻡﻭﺎﻘﻣ ﻪﻤﻴﻧ ،ﺱﺎﺴﺣ ﻦﻳﺪﻟﺍﻭ ﺭﺩ
ﺕﺎﻌﻟﺎﻄﻣ ﺞﻳﺎﺘﻧ ﻪﺑﺎﺸﻣ
Golkari et al.
(2007)
ﻪﺑ ﻁﻮﺑﺮﻣ
ﺩﺍﺪﻌﺗ ﺎﺑ ﻪﻳﺍﺭﺁﺰﻳﺭ ﻪﻳﺰﺠﺗ ﻱ
ﻥﮊ ﺯﺍ ﻂﺳﻮﺗ ﻪﺘﻓﺎﻳ ﺀﺎﻘﻟﺍ ﻱﺎﻫ
F. graminearum
ﺩﻮﺑ ﻡﺪﻨﮔ ﺭﺩ .
ﻱﻮﮕﻟﺍ ﻖﻴﻘﺤﺗ ﻦﻳﺍ ﺭﺩ
ﻥﺎﮔﺭﺍ ﻱﺍﺮﺑ ﻥﮊ ﻥﺎﻴﺑ ﺮﮕﻳﺪﮑﻳ ﻪﺑ ﻲﮑﻳﮊﻮﻟﻮﻴﺑ ﻅﺎﺤﻟ ﺯﺍ ﻪﮐ ﻲﻳﺎﻫ
ﮏﻳﺩﺰﻧ ،ﺪﻧﺭﺍﺩ ﻱﺮﺘﺸﻴﺑ ﻪﺑﺎﺸﺗ ﺩﻮﺑ ﺮﺗ
. ﺩﺍﺩ ﻥﺎﺸﻧ ﺎﻣ ﺕﺎﻌﻟﺎﻄﻣ
ﻥﮊ ﺮﻫﺎﻈﺗ ﻱﻮﮕﻟﺍ ﻪﮐ ﻪﺑﺎﺸﺗ ﻦﻳﺮﺘﺸﻴﺑ ﻻﺎﭘ ﻭ ﻡﻮﻠﮔ ﺭﺩ NPR1
ﺪﻧﺭﺍﺩ ﺮﮕﻳﺪﮑﻳ ﻪﺑ ﺍﺭ
Golkari et al. .
(2007)
ﺪﻧﺩﻮﻤﻧ ﺵﺭﺍﺰﮔ
ﺭﺩ ﻲﮕﺘﺴﺒﻤﻫ ﻦﻳﺮﺘﺸﻴﺑ ﻦﻴﺑ ﻪﺘﻓﺎﻳ ﺀﺎﻘﻟﺍ ﻱﺎﻫ ﻥﮊ ﻥﺎﻴﺑ ﻱﻮﮕﻟﺍ
ﻣ ﻻﺎﭘ ﻭ ﻡﻮﻠﮔ ﻲ
ﺪﺷﺎﺑ .
ﺍﺪﻳﺪﻧﺎﮐ ﻥﮊ NPR1 ﻱ
ﻪﺑ ﺖﻣﻭﺎﻘﻣ ﻡﺪﻨﮔ ﺭﺩ FHB
ﺦﺳﺎﭘ ﺎﻫ ﻱ ﻋﺎﻓﺩ ﻲ ﻞﻣﺍﻮﻋ ﺮﻴﺛﺄﺗ ﺖﺤﺗ ﻪﮐ ﮏﻴﻤﺘﺴﻴﺳ
ﮑﻳﮊﻮﻟﻮﻴﺑ ﻲ ﻣ ﮏﻳﺮﺤﺗ ﻲ
ﺑﺎﺴﺘﮐﺍ ﺖﻣﻭﺎﻘﻣ ﻞﻣﺎﺷ ،ﺪﻧﻮﺷ ﻲ
ﮏﻴﻤﺘﺴﻴﺳ ﻭ
ﺋﺎﻘﻟﺍ ﮏﻴﻤﺘﺴﻴﺳ ﺖﻣﻭﺎﻘﻣ ﻲ
ﻲﻣ ﺪﻨﺷﺎﺑ .
ﺦﺳﺎﭘ ﺎﻫ ﻱ ﻋﺎﻓﺩ ﻲ ﺍ ﻪﮑﺒﺷ ﻂﺳﻮﺗ ﻱ
ﺯﺍ ﺎﻫﺮﻴﺴﻣ ﻱ ﻝﺎﻘﺘﻧﺍ
ﻣ ﻝﺮﺘﻨﮐ ﺪﻨﺘﺴﻫ ﻂﺒﺗﺮﻣ ﻢﻫ ﺎﺑ ﻪﮐ ﻝﺎﻨﮕﻴﺳ ﻲ
ﺪﻧﻮﺷ . ﺭﺩ
ﺦﺳﺎﭘ ﻦﻳﺍ ﺩﺎﺠﻳﺍ ﺎﻫ ﻥﻮﻣﺭﻮﻫ ﺎﻫ
ﻱ ﺪﻴﺳﺍ ﮏﻴﻠﻴﺴﻴﻟﺎﺳ
SA) ( ،
ﺪﻴﺳﺍ ﮏﻴﻧﻮﻤﺳﺎﺟ
JA) ( ﻠﺻﺍ ﺶﻘﻧ ﻦﻠﻴﺗﺍ ﻭ ﻲ
ﺀﺎﻔﻳﺍ ﺍﺭ
ﻣ ﻲ
(Manosalva et al., 2000) ﺪﻨﻨﮐ .
ﻝﺎﻨﮕﻴﺳ ﻝﺎﻘﺘﻧﺍ
SA
ﺍﺮﺑ ﻱ ﻥﮊ ﻥﺎﻴﺑ ﻥﺩﺮﮐ ﻝﺎﻌﻓ ﺎﻫ
١ ﻱ ﻞﻤﻋ ﻪﺑ ﺯﺎﻴﻧ PR NPR1
ﺩﺭﺍﺩ . ﻪﺑ ﻥﮊﻮﺗﺎﭘ ﻪﻠﻤﺣ ﻩﺎﻴﮔ
ﻥﺍﺰﻴﻣ ﺶﻳﺍﺰﻓﺍ ﺐﺟﻮﻣ ﻭ SA
ﺎﻫ ﻪﻄﺳﺍﻭ ﻊﻳﺮﺳ ﺪﻴﻟﻮﺗ ﻱ
ﻝﺎﻌﻓ ﻥﮋﻴﺴﮐﺍ ﺪﻨﻧﺎﻣ
ﻪﻧﮋﻴﺴﻛﺍ ﺏﺁ
ﻣ ﻲ ﺩﻮﺷ . ﺍﺮﺑ ﻱ ﺜﻨﺧ ﻲ ﺯﺎﺳ ﻱ ﻩﻮﻘﻟﺎﺑ ﺭﻮﻃ ﻪﺑ ﻪﮐ ﺕﺎﺒﻴﮐﺮﺗ ﻦﻳﺍ
ﻤﺳ ﻲ ﺎﻫ ﻩﺪﻨﻨﮐ ﻑﺬﺣ ،ﺪﻨﺘﺴﻫ
ROIﻱ ﻣ ﻝﺎﻌﻓ ﻲ ﺯﺍ ﻭ ﺪﻧﻮﺷ
1 . Pathogenesis related protein