ﻥﺍﺭﺎﮑﻤﻫ ﻭ ﻱﺪﻤﺤﻣ :
ﻳﺎﺳﺎﻨﺷ ﻭ ﻥﺍﺮﻬﺗ ﻥﺎﺘﺳﺍ ﺕﺭﺫ ﻉﺭﺍﺰﻣ ﺭﺩ ﺐﻟﺎﻏ ﻲﺗﻮﭘ ﻲ ...
۴۳
ﻪﻠﺠﻣ ﺶﻧﺍﺩ ﮑﺷﺰﭙﻫﺎﻴﮔ ﻲ ﻥﺍﺮﻳﺍ
ﺩ ﻩﺭﻭ ۴۰ ﻩﺭﺎﻤﺷ ، ۱ ، ۱۳۸۸ ) ۵۳
‐ ۴۳ (
ﻥﺍﺮﻬﺗ ﻥﺎﺘﺳﺍ ﺕﺭﺫ ﻉﺭﺍﺰﻣ ﺭﺩ ﺐﻟﺎﻏ ﺱﻭﺮﻳﻭ ﻲﺗﻮﭘ ﻲﻳﺎﺳﺎﻨﺷ
ﻲﺳﺭﺮﺑ ﻭ ﻩﺭﺎﭘ
ﻥﺁ ﺕﺎﻴﺻﻮﺼﺧ ﺯﺍ ﻱﺍ
ﻱﺪﻤﺤﻣ ﺎﺿﺭﺪﻤﺤﻣ
*۱
ﻲﺒﻴﺒﺣ ﻲﻫﻮﻛ ﺎﻨﻴﻣ ، ﻭ ۲
ﻱﺪﻤﺤﻣ ﻲﺒﺣﺎﺼﻣ ﻦﻴﺴﺤﻣﻼﻏ ۳
۱ ، ﻲﺑﺮﻣ ، ﻭ ﺪﺣﺍﻭ ﻲﻣﻼﺳﺍ ﺩﺍﺯﺁ ﻩﺎﮕﺸﻧﺍﺩ ،ﻱﺯﺭﻭﺎﺸﻛ ﻩﺪﻜﺸﻧﺍﺩ ﻦﻴﻣﺍﺭ
۲ ، ۳
، ﻭ ﺭﺎﻳﺩﺎﺘﺳﺍ ﺸﻧﺍﺩ
ﺭﺎﻴ
، ﻱﺯﺭﻭﺎﺸﻛ ﺲﻳﺩﺮﭘ ﻲﻌﻴﺒﻃ ﻊﺑﺎﻨﻣ ﻭ
ﻥﺍﺮﻬﺗ ﻩﺎﮕﺸﻧﺍﺩ
) ﺖﻓﺎﻳﺭﺩ ﺦﻳﺭﺎﺗ :
۲۹ / ۱۱ / ۸۴
‐ ﺐﻳﻮﺼﺗ ﺦﻳﺭﺎﺗ :
۸ / ۳ / ۸۷ (
ﻩﺪﻴﮑﭼ
ﻝﺎﺳ ﻲﻋﺍﺭﺯ ﻞﺼﻓ ﻲﻃ ١٣٨٢
ﺩﺍﺪﻌﺗ ٢٣١ ﻧﻮﻤﻧ ﻪ ﮔﺮﺑ ﻲ ﺕﺭﺫ ﺯﺍ ﻭ ﺱﻭﺮﻳﻭ ﻪﺑ ﻩﺩﻮﻟﺁ ﻱﺎﻫ ﻢﻳﻼﻋ ﻱﺍﺭﺍﺩ
ﻚﻴﺋﺍﺯﻮﻣ ﺑ ﻩﺍﺮﻤﻫ ﻚﻴﺋﺍﺯﻮﻣ ﻢﻳﻼﻋ ﻱﺩﺭﺍﻮﻣ ﺭﺩ ﻭ ﻲﮕﻟﻮﺗﻮﻛ ﺎ
ﻥﺍﺮﻬﺗ ﻥﺎﺘﺳﺍ ﻒﻠﺘﺨﻣ ﻖﻃﺎﻨﻣ ﺕﺭﺫ ﻉﺭﺍﺰﻣ ﺯﺍ ،
ﻊﻤﺟ ﺪﻳﺩﺮﮔ ﻱﺭﻭﺁ .
ﻲﺘﻧﺁ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﻡﺮﺳ
ﺎﻫ ﻱ ﻠﭘ ﻲ ﺭﺎﻬﭼ ﻝﺎﻧﻮﻠﮐ ﻲﺗﻮﭘ
ﺱﻭﺮﻳﻭ
،MDMV SCMV
،
SrMV
ﻭ ﻪﻧﻮﻤﻧ ﻱﻭﺭ ﺮﺑ JGMV
ﺎﻫ ﻥﻮﻣﺯﺁ ﺎﻫ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ ﻱ
DAS-ELISA
،
DIBA
ﻭ ﺪﻳﺩﺮﮔ ﻡﺎﺠﻧﺍ TPIA
. ﺭﺩ
،ﻥﺎﻳﺎﭘ ﻪﻧﻮﻤﻧ ﻲﻣﺎﻤﺗ ﺎﺑ ﺎﻫ
ﻲﺘﻧﺁ ﻡﺮﺳ ﺱﻭﺮﻳﻭ ﺮﻜﺸﻴﻧ ﻚﻴﺋﺍﺯﻮﻣ
(SCMV)
ﺪﻧﺩﺍﺩ ﺯﻭﺮﺑ ﻱﻮﻗ ﺶﻨﻛﺍﻭ ﻲﺸﻨﻛﺍﻭ ﻭ
ﺎﺑ ﻲﺘﻧﺁ ﻡﺮﺳ ﺪﻳﺩﺮﮕﻧ ﻩﺪﻫﺎﺸﻣ ﺮﮕﻳﺩ ﺱﻭﺮﻳﻭ ﻲﺗﻮﭘ ﻪﺳ .
ﻪﻧﻮﻤﻧ ﺎﻫ ﺭﺩ ﻢﻴﺳﺎﺘﭘ ﺕﺎﻔﺴﻓ ﺮﻓﺎﺑ ١
/ ٠ ﺭﻻﻮﻣ
pH) ٧ (
ﻱﻭﺎﺣ ٢ ﻲﻠﭘ ﺪﺻﺭﺩ ﻥﻭﺪﻴﻟﻭﺮﻳﺎﭘ ﻞﻴﻨﻳﻭ
PVP) ( ﻩﺭﺎﺼﻋ ﺮﻴﮔ ﻭ ﻱ ﻪﻳﺍﺪﺟ ﺎﻫ ﻦﻳﺮﻴﺷ ﺕﺭﺫ ﻱﻭﺭ ﺮﺑ ﻪﻧﺎﺨﻠﮔ ﺭﺩ
ﻢﻗﺭ ٤٠٣ ﺍﺩ ﻡﻮﮔﺭﻮﺳ ﻭ ﺱﺭﺎﭘ ﻪﻧ
ﺎﻴﻤﻴﻛ ﻢﻗﺭ ﻱﺍ ﻪﻳﺎﻣ
ﻲﻧﺯ ﻭ ﺮﻴﺜﻜﺗ ﺪﻧﺪﻳﺩﺮﮔ . ﺭﺩ ﻥﻮﻣﺯﺁ ﺎﻫ ﺩﺪﺠﻣ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ ﻱ
ﻥﺎﻫﺎﻴﮔ ﻱﻭﺭ ﺮﺑ ﻪﻳﺎﻣ
ﻧﺯ ﺭﻮﻀﺣ ﺎﻬﻨﺗ ﺰﻴﻧ ﻩﺪﺷ ﻲ ﺪﻳﺩﺮﮔ ﺯﺮﺤﻣ ﺎﻬﻧﺁ ﺭﺩ SCMV
. ﻌﻟﺎﻄﻣ ﺭﺩ ﻪ ﻨﻣﺍﺩ ﻪ ،ﻲﻧﺎﺑﺰﻴﻣ
ﺍﺪﺟ ﻪﻳ ﺎﻫ ﺐﺨﺘﻨﻣ ﻱ ﻱﻭﺭ ﻪﺑ SCMV
Avena sativa Panicum miliaceum ،
Pennisetum americanum ،
Triticum aestivum Hordeum vulgare ،
Setaria italicaﻭ ﺪﻧﺪﺸﻧ ﻞﻘﺘﻨﻣ
. ﺺﻟﺎﺧ ﻱﺯﺎﺳ ﺱﻭﺮﻳﻭ ﺵﻭﺭ ﻪﺑ
Minipurification
ﻱﻭﺭ ﺮﺑ ٢ ﻩﺩﻮﻟﺁ ﻲﻫﺎﻴﮔ ﺖﻓﺎﺑ ﺯﺍ ﻡﺮﮔ ﻡﺎﺠﻧﺍ
ﻭ ﺺﻟﺎﺧ ﺱﻭﺮﻳﻭ ﺖﻈﻠﻏ ﻭ ﺖﺒﺴﻧ
A260/280
ﺐﻴﺗﺮﺗ ﻪﺑ ﻥﺁ ﻱﺍﺮﺑ
mg/ml
٤٥ / ١١ ﻭ ٢ / ١ ﺪﺷ ﻪﺒﺳﺎﺤﻣ .
ﻲﺳﺭﺮﺑ ﺭﺩ ﻪﺑ ﻲﭘﻮﻜﺳﻭﺮﻜﻴﻣ ﻥﻭﺮﺘﻜﻟﺍ
ﺵﻭﺭ ﻭﺮﺳ ﻖﻴﻔﻠﺗ ﻲﭘﻮﻜﺳﻭﺮﻜﻴﻣ ﻥﻭﺮﺘﻜﻟﺍ ﻭ ﻱﮊﻮﻟ
ISEM) ( ﺎﺑ ﻥﻮﻴﺳﺍﺭﻮﻛﺩ ﻭ ﻲﺘﻧﺁ
SCMVﻡﺮﺳ ﻩﺮﻜﻴﭘ ، ﻱﺎﻫ
ﻪﺘﺷﺭ ﻞﺑﺎﻗ ﻱﺍ ﻑﺎﻄﻌﻧﺍ ﺭﺩ SCMV
ﻪﻧﻮﻤﻧ ﺎﻫ ﺲﻜﻋ ﻭ ﻩﺪﻫﺎﺸﻣ ﺪﺷ ﻡﺎﺠﻧﺍ ﻱﺭﺍﺩﺮﺑ
. ﻡﺎﺠﻧﺍ ﺎﺑ ﻥﻮﻣﺯﺁ ﺎﻫ
SDS-ﻱ
PAGE
ﻭ
Western blotting
ﻪﻧﻮﻤﻧ ﻱﻭﺭ ﺮﺑ ﻥﻮﻴﺴﻧﺎﭙﺳﻮﺳ ﻭ ﻩﺩﻮﻟﺁ ﻲﻫﺎﻴﮔ ﻱﺎﻫ
ﻥﺯﻭ ،ﺺﻟﺎﺧ ﺱﻭﺮﻳﻭ ﻱﺎﻫ
ﻲﻟﻮﻜﻟﻮﻣ ﻲﺸﺷﻮﭘ ﻦﻴﺌﺗﻭﺮﭘ ﺩﻭﺪﺣ ﺭﺩ SCMV
KDa
٣٨ ‐ ٣٧ ﺩﻮﺑ . ﻮﻣﺯﺁ
RT-PCR ﻥ ﺖﻔﺟ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ
ﺮﻤﻳﺍﺮﭘ
SCMVF3 SCMV R3 ﻭ
ﻡﺎﺠﻧﺍ ﺩﻭﺪﺣ ﺭﺩ ﻪﺘﻓﺎﻳ ﺮﻴﺜﻜﺗ ﺕﺎﻌﻄﻗ ﻭ ﺖﻓﺮﮔ ٩٠٠bp
ﺪﻧﺩﻮﺑ ﻱﺪﻴﻳﺎﺗ ﻪﻛ
ﺱﻭﺮﻳﻭ ﺭﻮﻀﺣ ﺮﺑ ﻒﻋﺎﻀﻣ
SCMV
ﺭﺩ ﻪﻧﻮﻤﻧ ﺎﻫ ﺪﻣﺁ ﺖﺳﺪﺑ .
ﻚﻴﺋﺍﺯﻮﻣ ﺱﻭﺮﻳﻭ ﺮﺿﺎﺣ ﻖﻴﻘﺤﺗ ﺱﺎﺳﺍ ﺮﺑ
ﻲﺗﻮﭘ ﺮﻜﺸﻴﻧ ﺮﻳﻭ
ﺱﻭ ﻊﻳﺎﺷ ﺐﻟﺎﻏ ﻭ ﻲﮕﻟﻮﺗﻮﻛ ﻚﻴﺋﺍﺯﻮﻣ ﻭ ﻚﻴﺋﺍﺯﻮﻣ ﻲﻠﺻﺍ ﻞﻣﺎﻋ ﻭ ﻥﺎﺘﺳﺍ ﺕﺭﺫ ﻉﺭﺍﺰﻣ ﺭﺩ
ﻥﺍﺮﻬﺗ ﺪﺷﺎﺑ ﻲﻣ .
ﻩﮊﺍﻭ ﻱﺪﻴﻠﻛ ﻱﺎﻫ ﻥﺍﺮﻬﺗ ﻥﺎﺘﺳﺍ ،ﺍﺰﻳﻻﺍ ،ﺮﻜﺸﻴﻧ ﻚﻴﺋﺍﺯﻮﻣ ﺱﻭﺮﻳﻭ ،ﺕﺭﺫ ،ﺱﻭﺮﻳﻭ ﻲﺗﻮﭘ :
.
ﻝﻮﺌﺴﻣ ﻩﺪﻨﺴﻳﻮﻧ *
: ﻱﺪﻤﺤﻣ ﺎﺿﺭﺪﻤﺤﻣ
ﻦﻔﻠﺗ 4220650 : - 0291
E-mail: m_rmohammadi@yahoo.com
۴۴ ﻪﻠﺠﻣ
ﮑﺷﺰﭙﻫﺎﻴﮔ ﺶﻧﺍﺩ ﻲ
ﻩﺭﻭﺩ ،ﻥﺍﺮﻳﺍ ۴۰
ﻩﺭﺎﻤﺷ ، ۱ ، ۱۳۸۸
ﻪﻣﺪﻘﻣ
ﻲﻤﻠﻋ ﻡﺎﻧ ﺎﺑ ﺕﺭﺫ ﻩﺎﻴﮔ
Zea mays L.
ﻚﺗ ﻲﻫﺎﻴﮔ
ﻪﭙﻟ ﻱﺍ ﻪﻳﺎﭘ ﻚﻳ ، ﺩﺍﻮﻧﺎﺧ ﺯﺍ ﻪﻟﺎﺴﻜﻳ ﻭ
ﻩ ﺕﻼﻏ ﻪﻨﻴﻣﺍﺮﮔ ﺎﻳ
1
ﻲﻣ ﺪﺷﺎﺑ ﻪﻛ ﺭﺎﻨﻛ ﺭﺩ ﻠﻏ ﻝﻮﺼﺤﻣ ﻪﺳ ﺞﻧﺮﺑ ﻭ ﻡﺪﻨﮔ ﻪ
ﺭﺎﻴﺴﺑ
ﻲﻣ ﻞﻴﻜﺸﺗ ﺍﺭ ﻥﺎﻬﺟ ﻢﻬﻣ ﺪﻫﺩ
ﻝﻮﺼﺤﻣ ﻚﻳ ﻥﺍﻮﻨﻌﺑ ﻭ
ﻥﺎﻬﺟ ﺭﺩ ﻥﺁ ﺪﻴﻟﻮﺗ ﻭ ﺖﺸﻛ ﺮﻳﺯ ﺢﻄﺳ ﻚﻳﮋﺗﺍﺮﺘﺳﺍ ﺭﺩ ﻭ
ﺍﻳ ﻥﺍﺮ ﺖﺳﺍ ﺵﺮﺘﺴﮔ ﻪﺑ ﻭﺭ
Mirhadi, 2001) (
. ﺪﻴﻟﻮﺗ ﺶﻳﺍﺰﻓﺍ
ﺎﺑ ﻡﺃﻮﺗ ﻥﺎﻬﺟ ﻁﺎﻘﻧ ﺯﺍ ﻱﺭﺎﻴﺴﺑ ﺭﺩ ﺕﺭﺫ ﺖﺸﻛﺮﻳﺯ ﺢﻄﺳ ﻭ ﻳﺍﺪﻴﭘ ﻱﺎﻬﻳﺭﺎﻤﻴﺑ ﻥﺎﻴﻐﻃ ﺎﻳ ﻭ ﺪﻳﺪﺟ ﻲﺳﻭﺮﻳﻭ ﻱﺎﻬﻳﺭﺎﻤﻴﺑ ﺶ ﻩﺯﻭﺮﻣﺍ ﺍﺬﻟ ﻭ ﺖﺳﺍ ﻩﺩﻮﺑ ﻝﻮﺼﺤﻣ ﻦﻳﺍ ﺭﺩ ﺖﻴﻤﻫﺍ ﻢﻛ ﻲﺳﻭﺮﻳﻭ
ﻪﻣﺎﻧﺮﺑ ﺭﺩ ﻌﺳﻮﺗ ﻱﺎﻫ ﻪ
ﺱﻭﺮﻳﻭ ﻥﺎﻬﺟ ﺭﺩ ﺕﺭﺫ ﺖﺸﻛ ﺶﻴﺑ ﺎﻫ
ﻩﺩﺮﻛ ﺍﺪﻴﭘ ﺖﻴﻤﻫﺍ ﺶﻴﭘ ﺯﺍ
(Izadpanah, 1985) ﺪﻧﺍ .
ﻣ ﺯﺍ ﻴ ﻥﺎ
ﻭ ﻳ ﺱﻭﺮ ﺎﻫ ﻱ ﺑ ﻴ ﺭﺎﻤ ﻳ ﺕﺭﺫ ﺭﺩ ﺍﺰ ﻪﻧﻮﮔ
ﻱﺎﻫ ﺲﻨﺟ
Potyvirus
ﺯﺍ
ﺖﻴﻤﻫﺍ ﻩﮋﻳﻭ ﺪﻧﺭﺍﺩﺭﻮﺧﺮﺑ ﻱﺍ .
ﺮﺑ ﻦﻳﺮﺧﺁ ﺞﻳﺎﺘﻧ ﺱﺎﺳﺍ
،ﺕﺎﻘﻴﻘﺤﺗ ﺭﺎﻬﭼ
ﺲﻨﺟ ﺯﺍ ﺱﻭﺮﻳﻭ ﻪﻧﻮﮔ
Potyvirus
ﻱﻭﺭ
ﺯﺍ ﺪﻨﺗﺭﺎﺒﻋ ﻪﻛ ﺪﻨﺘﺴﻫ ﺍﺰﻳﺭﺎﻤﻴﺑ ﺕﺭﺫ :
ﻚﻴﺋﺍﺯﻮﻣ ﺱﻭﺮﻳﻭ
ﺕﺭﺫ ﻲﮕﻟﻮﺗﻮﻛ
Maize Dwarf Mosaic Virus: MDMV) (
،
ﺮﻜﺸﻴﻧ ﻚﻴﺋﺍﺯﻮﻣ ﺱﻭﺮﻳﻭ
(Sugarcane Mosaic Virus:
SCMV)
ﻡﻮﮔﺭﻮﺳ ﻚﻴﺋﺍﺯﻮﻣ ﺱﻭﺮﻳﻭ ،
(Sorghum Mosaic
Virus: SrMV)
ﻕﺎﻴﻗ ﻚﻴﺋﺍﺯﻮﻣ ﺱﻭﺮﻳﻭ ﻭ
:JGMV)
Johnsongrass Mosaic Virus
.(.
ﺱﻭﺮﻳﻭ ﻲﺗﻮﭘ ﻦﻳﺍ ﻪﻛ ﺎﻫ
ﺱﻭﺮﻳﻭ ﻲﺗﻮﭘ ﻪﺑ ﺕﻼﻏ ﻱﺎﻫ
ﺪﻧﺭﻮﻬﺸﻣ2
ﻭ ﻭ ﻚﻴﺋﺍﺯﻮﻣ ﻱﺭﺎﻤﻴﺑ
ﺮﮕﻳﺩ ﻭ ﺮﻜﺸﻴﻧ ،ﻡﻮﮔﺭﻮﺳ ،ﺕﺭﺫ ﺭﺩ ﺍﺭ ﻝﻮﺼﺤﻣ ﺕﺭﺎﺴﺧ ﺩﺍﻮﻧﺎﺧ ﻥﺎﻫﺎﻴﮔ ﻩ
ﻲﻣ ﺐﺒﺳ ﻪﻨﻴﻣﺍﺮﮔ ﺎﻳ ﺕﻼﻏ ﺪﻧﻮﺷ
، ﺭﺩ ﻚﻳ
ﻩﻭﺮﮔ ﺮﻳﺯ ﻩﻭﺮﮔ ﺮﻳﺯ ﻡﺎﻨﺑ
3SCMV
ﺯﺍ ﺲﻨﺟ
Potyvirus
ﻱﺎﺟ
ﻩﺪﺷ ﻩﺩﺍﺩ ﺪﻧﺍ
Shukla et al., 1989) (
. ﻥﺎﻴﻣ ﺯﺍ ﺍ ﻳ ﺭﺎﻬﭼ ﻦ
ﺱﻭﺮﻳﻭ ﻲﺗﻮﭘ
SCMV ،
MDMV ﻭ ﺎﻴﻧﺩ ﺮﺳﺎﺗﺮﺳ ﺭﺩ ﻊﻳﺎﺷ
ﺪﻨﺷﺎﺑ ﻲﻣ ﻪﻜﻴﻟﺎﺣ ﺭﺩ
ﻭ ﻩﺪﺤﺘﻣ ﺕﻻﺎﻳﺍ ﻭ ﺎﻴﻟﺍﺮﺘﺳﺍ ﺯﺍ JGMV
ﻩﺪﺤﺘﻣ ﺕﻻﺎﻳﺍ ﺯﺍ SrMV
ﻦﻴﭼ ﺯﺍ ﺍﺮﻴﺧﺍ ﻭ ﻩﺪﺷ ﺵﺭﺍﺰﮔ
ﺖﺳﺍ
Shukla et al., 1989; Chen et al., 2002;) .(
ﺭﺩ ﻝﺎﺳ ﺎﻫ ﻡﺎﻧ ﺎﺑ ﺰﻴﻧ ﻲﺗﻭﺎﻔﺘﻣ ﻭ ﺪﻳﺪﺟ ﻪﻧﻮﮔ ﺮﻴﺧﺍ ﻱ
ZeMV (Zea Mosaic Virus)
ﻦﻳﺍ ﻪﺑ ﻭ ﻪﻌﻟﺎﻄﻣ ﻞﻴﻳﺍﺮﺳﺍ ﺭﺩ
ﺖﺳﺍ ﻩﺪﺷ ﻪﻓﺎﺿﺍ ﻩﻭﺮﮔ ﺮﻳﺯ
Seifers et al., 2000) .(
ﺱﻭﺮﻳﻭ ﻥﻮﻨﻛ ﺎﺗ ﺰﻴﻧ ﻥﺍﺮﻳﺍ ﺭﺩ ﻱﻭﺭ ﺯﺍ ﻱﺩﺪﻌﺘﻣ ﻱﺎﻫ
ﺵﺭﺍﺰﮔ ﺕﺭﺫ ﻩﺪﺷ
ﺪﻧﺍ ﻭﺪﺑﺍﺭ ﻪﻛ ﺱﻭﺮﻳﻭ
ﺕﺭﺫ ﻲﻧﺍﺮﻳﺍ ﻚﻴﺋﺍﺯﻮﻣ
،ﻩﺩﻮﺑ ﺎﻬﻧﺁ ﻦﻳﺮﺘﻤﻬﻣ ﺕﺭﺫ ﺮﺑﺯ ﻲﮕﻟﻮﺗﻮﻛ ﺱﻭﺮﻳﻭ ﻲﺠﻴﻓ ﻭ
1. Poaceae
2. Poaceous potyviruses 3. SCMV subgroup
ﺕﺭﺎﺴﺧ ﺎﻫ ﻬﻧﺁ ﻱ ﺖﺳﺍ ﻩﺪﺷ ﺵﺭﺍﺰﮔ ﻖﻃﺎﻨﻣ ﻲﺧﺮﺑ ﺯﺍ ﺎ
(Afsharifar, 1991; Moini & Izadpanah, 2001)
. ﺯﺎﻏﺁ ﺯﺍ
ﻪﻫﺩ ۱۳۶۰ ﺵﺭﺍﺰﮔ ﻲﻳﺎﻫ ﻧﻴ ﺰ ﺯﺍ ﻩﺪﻫﺎﺸﻣ ﻩﺮﮑﻴﭘ ﺎﻫ ﻱ ﻪﺘﺷﺭ ﻱﺍ
ﺮﻳﺍ ﻒﻠﺘﺨﻣ ﻖﻃﺎﻨﻣ ﺯﺍ ﺕﺭﺫ ﻱﻭﺭ ﺭﺩ ﻲﺳﻭﺮﻳﻭ ﻲﺗﻮﭘ ﺩﻮﺟﻭ ﻥﺍ
ﺍﺩ
(Izadpanah, 1982; Izadpanah, 1985; ﺖﺷ
Afsharifar, 1991; Izadpanah & Kamran, 1995;
Masumi & Izadpanah, 1995)
ﺍ ﻭ ﻳ ﻭ ﻦ ﻳ ﻪﺘﺷﺭ ﺱﻭﺮ ﺍ
ﻱ
ﹰﺎﺗﺪﻤﻋ ﺮﺘﺳﺍ ﻳ ﻨ ﻲ ﺯﺍ ﻡﺎﻧ ﺎﺑ SCMV
SCMV-MS (Maize
Shiraz)
ﺨﺸﺗ ﻴ ﺭﺩ ﻪﮐ ﺩﻮﺑ ﻩﺪﺷ ﻩﺩﺍﺩ ﺺ ﻝﺎﺳ
ﺎﻫ ﻱ ﺧﺍ ﻴ ﺎﺑ ﺮ
ﺧﺮﺑ ﻪﺑ ﻪﺟﻮﺗ ﻲ
ﺕﻭﺎﻔﺗ ﺩ ﺎﺑ ﺎﻫ ﻳ ﺮﮕ ﺍﺪﺟ ﻪﻳ ﺎﻫ
SCMVﻱ ﻁﺎﻘﻧ ﺯﺍ
ﺩ ﻳ ﻧﺩ ﺮﮕ ﻴ ﺎﻧﺍﻮﺗ ﺖﻠﻋ ﻪﺑ ﻪﻠﻤﺟ ﺯﺍ ﺎ ﻲﻳ
ﺯﺎﺳ ﻩﺩﻮﻟﺁ ﺭﺩ ﻥﺁ ﻱ
ﮔ ﻴ ﻩﺎ
ﻗ ﻴ ﻕﺎ ﻨﭽﻤﻫ ﻴ ﻟﺩ ﻪﺑ ﻦ ﻴ ﺧﺮﺑ ﻞ ﻲ ﺎﻬﺗﻭﺎﻔﺗ ﻱ ﮊﻮﻟﻭﺮﺳ ﻳ ﮑ ﻲ ﻭ
ﻟﺍﻮﺗ ﺭﺩ ﺕﻭﺎﻔﺗ ﻲ
،ﻡﻮﻧﮊ ﻭ ﻳ ﺎﺷ ﺱﻭﺮ ﻳ ﺍ ﺭﺩ ﻊ ﻳ ﻪﻧﻮﮔ ﻥﺍﻮﻨﻋ ﻪﺑ ﻥﺍﺮ
ﺪﺟ ﻳ ﻡﺎﻧ ﺎﺑ ﺪ
IJMV) (
Iranian Johnsongrass Mosaic
Virus
ﻓﺮﻌﻣ ﻲ ﺖﺳﺍ ﻩﺪﺷ
(Masumi & Izadpanah, 2001;
Masumi et al., 2001)
. ﺧﺮﺑ ﻲ ﻘﻘﺤﻣ ﻴ ﻧ ﻦ ﻴ ﻪﺘﺴﻧﺍﻮﺗ ﺰ ﺪﻧﺍ
ﺎﺳﺎﻨﺷ ﻪﺑ ﻡﺍﺪﻗﺍ ﻲﻳ
ﺯﺎﺳﺍﺪﺟ ﻭ
MDMV ﻱ ﺧﺮﺑ ﺭﺩ ﺕﺭﺫ ﺯﺍ
ﻲ
ﺍ ﻖﻃﺎﻨﻣ ﻳ ﻤﻧ ﻥﺎﺘﺳﺯﻮﺧ ﻭ ﻥﺍﺭﺪﻧﺯﺎﻣ ﺭﺩ ﻪﻠﻤﺟ ﺯﺍ ﻥﺍﺮ ﺎﻳ
ﺪﻨ
(Moini & Izadpanah, 2001; Zare et al., 2004a;
Zare et al., 2004b)
. ﺎﻤﻠﺴﻣ ﻦﻳﺍ ﺩﻮﺟﻭ ﺯﺍ ﻲﻫﺎﮔﺁ ﻲﺗﻮﭘ
ﺱﻭﺮﻳﻭ ﻭ ﺎﻫ ﺎﺳﺎﻨﺷ ﻲﻳ ﻗﺩ ﻴ ﻌﻄﻗ ﻭ ﻖ ﻲ
ﻨﮑﺗ ﺎﺑ ﺎﻬﻧﺁ ﻴ
ﮏ ﺎﻫ ﻱ
ﺪﺟ ﻳ ﻧ ﻭ ﺪ ﻴ ﻫﺎﮔﺁ ﺰ ﻲ ﺯﺍ ﻭ ﺎﻬﻧﺁ ﻲﮔﺩﺮﺘﺴﮔ ﻭ ﺶﻨﻛﺍﺮﭘ ﻥﺍﺰﻴﻣ
ﺕﺭﺎﺴﺧ ﺎﻫ ﺕﺍﺮﻄﺧ ﻭ ﺎﻬﻧﺁ ﻲﻟﺎﻤﺘﺣﺍ
ﻲﻓﺮﻌﻣ ﻦﻴﻨﭽﻤﻫ ﻭ
ﺵﻭﺭ ﺕﺭﻭﺮﺿ ﻚﻳ ﺮﺛﻮﻣ ﻭ ﺪﻣﺁﺭﺎﻛ ،ﻊﻳﺮﺳ ﻲﻳﺎﺳﺎﻨﺷ ﻱﺎﻫ
ﻭ ﻡﻭﺎﻘﻣ ﻡﺎﻗﺭﺍ ﺪﻴﻟﻮﺗ ﻭ ﺡﻼﺻﺍ ﻱﺎﻫ ﻪﻣﺎﻧﺮﺑ ﺭﺩ ﺎﺗ ﺖﺳﺍ ﻢﻬﻣ ﺎﻳ
ﭘ ﻲﺘﺣ ﺶﻴ ﻱﺮﮕﻳﺩ ﻱﺮﻴﮕﺸﻴﭘ ﻭ ﻝﺮﺘﻨﻛ ﺵﻭﺭ ﻪﻧﻮﮔ ﺮﻫ ﻲﻨﻴﺑ
ﺱﻭﺮﻳﻭ ﻦﻳﺍ ﻥﺎﻴﻐﻃ ﻲﻟﺎﻤﺘﺣﺍ ﺮﻄﺧ ﺎﺑ ﻪﻠﺑﺎﻘﻣ ﺖﻬﺟ ﺭﺩ ﺭﺩ ﺎﻫ
ﻥﺩﻮﺑ ﺎﭘﻮﻧ ﻪﺑ ﻪﺟﻮﺗ ﺎﺑ ﻪﻛ ،ﻩﺪﻨﻳﺁ ﺖﻋﺍﺭﺯ ﻭ ﺖﺸﻛ
ﺕﺭﺫ ) ﺯﺍ ﻪﮐ
ﻪﻫﺩ ١٣٦٠ ﻟﻮﺗ ﻭ ﺖﺸﮐ ﻴ
ﺖﺳﺍ ﻩﺩﺎﻬﻧ ﻪﻌﺳﻮﺗ ﻪﺑ ﻭﺭ ﻥﺁ ﺪ (
ﺭﺩ
ﻩﺩﺎﻔﺘﺳﺍ ﺩﺭﻮﻣ ،ﺖﺴﻴﻧ ﺭﺎﻈﺘﻧﺍ ﺯﺍ ﺭﻭﺩ ﻥﺍﺪﻨﭼ ﻥﺁ ﻉﻮﻗﻭ ﻥﺍﺮﻳﺍ ﮔ ﺭﺍﺮﻗ ﺩﺮﻴ . ﺖﻬﺟ ﺭﺩ ﻭ ﺵﺮﮕﻧ ﻉﻮﻧ ﻦﻳﺍ ﺎﺑ ﺰﻴﻧ ﺮﺿﺎﺣ ﻖﻴﻘﺤﺗ
ﻑﺍﺪﻫﺍ ﻦﻳﺍ ﻪﺑ ﻥﺪﻴﺳﺭ ﻉﺭﺍﺰﻣ ﻩﺩﻭﺪﺤﻣ ﺭﺩ ﻊﻣﺎﺟ ﻲﺳﺭﺮﺑ ﺎﺑ ﻭ
ﻥﺎﺘﺳﺍ ﺯﺍ ﻲﻜﻳ ﻥﺍﻮﻨﻌﺑ ﻥﺍﺮﻬﺗ ﻥﺎﺘﺳﺍ ﻭﺮﺸﻴﭘ ﻱﺎﻫ
ﺡﺮﻄﻣ ﻭ ﺭﺩ
ﺕﺭﺫ ﺪﻴﻟﻮﺗ ﻭ ﺖﺸﻛ ﻪﺻﺮﻋ ﺪﻳﺩﺮﮔ ﻡﺎﺠﻧﺍ
ﺖﺳﺍ ﻩ .
ﺵﻭﺭ ﻭ ﺩﺍﻮﻣ ﺎﻫ
ﻪﻧﻮﻤﻧ ﻱﺭﺍﺩﺮﺑ ﻝﺎﺳ ﻲﻋﺍﺭﺯ ﻞﺼﻓ ﻲﻃ ١٣٨٢
ﺩﺍﺪﻌﺗ ٢٣١ ﻪﻧﻮﻤﻧ ﺮﺑ ﻲﮔ ﺯﺍ
ﺕﺭﺫ ﻲﺳﻭﺮﻳﻭ ﻲﮔﺩﻮﻟﺁ ﻪﺑ ﻙﻮﻜﺸﻣ ﻱﺎﻫ ﻢﻳﻼﻋ ﻱﺍﺭﺍﺩ ﻭ
ﻥﺍﺭﺎﮑﻤﻫ ﻭ ﻱﺪﻤﺤﻣ :
ﻳﺎﺳﺎﻨﺷ ﻭ ﻥﺍﺮﻬﺗ ﻥﺎﺘﺳﺍ ﺕﺭﺫ ﻉﺭﺍﺰﻣ ﺭﺩ ﺐﻟﺎﻏ ﻲﺗﻮﭘ ﻲ ...
۴۵
ﻚﻴﺋﺍﺯﻮﻣ ﻥﺎﺘﺳﺍ ﺏﻮﻨﺟ ﻭ ﺏﺮﻏ ﻒﻠﺘﺨﻣ ﻖﻃﺎﻨﻣ ﻉﺭﺍﺰﻣ ﺯﺍ
ﻥﺍﺮﻬﺗ ) ﺕﺭﺫ ﺖﺸﻛ ﺮﻳﺯ ﻖﻃﺎﻨﻣ (
ﻩﺩﻭﺪﺤﻣ ﺭﺩ ﻥﺎﺘﺳﺮﻬﺷ
ﺎﻫ ﻱ
ﻦﻴﮕﺸﻣ ،ﻍﻼﺒﺟﻭﺎﺳ ،ﺝﺮﻛ ،ﻦﻴﻣﺍﺭﻭ ،ﻥﺍﺮﻬﺗ
،ﺖﺷﺩ
،ﺮﻬﺷﺪﻤﺤﻣ
ﺮﻬﺸﻣﻼﺳﺍ ﻭ ﺭﺎﻳﺮﻬﺷ ،ﺩﺮﮕﺘﺸﻫ ﻊﻤﺟ
ﺭﻭﺁ ﺪﺷ ﻱ . ﻪﻧﻮﻤﻧ ﺎﻫ ﺲﭘ
ﻝﺎﻘﺘﻧﺍ ﺯﺍ ﻱﺎﻣﺩ ﺭﺩ ﻩﺎﮕﺸﻳﺎﻣﺯﺁ ﻪﺑ ٤˚C
ﺭﺩ ﻱﺭﺍﺪﻬﮕﻧ ﻝﺎﭽﺨﻳ
ﺪﻧﺪﺷ ﻱﺍﺮﺑ ﻭ ﻪﻧﻮﻤﻧ ﺕﺪﻣ ﻲﻧﻻﻮﻃ ﻱﺭﺍﺪﻬﮕﻧ ﺯﺍ ﻱﺭﺍﺪﻘﻣ ،ﺎﻫ
ﻪﻧﻮﻤﻧ ﺮﻳﺍﺭﺩ ﺰﻳﺮﻓ ﻩﺎﮕﺘﺳﺩ ﺭﺩ ﺎﻫ ﺖﺑﻮﻃﺭ 1
ﻭ ﻩﺪﺷ ﻱﺮﻴﮔ ﺭﺩ
ﺭﺰﻳﺮﻓ ٢٠˚C
‐ ﺪﻧﺪﺷ ﻱﺭﺍﺪﻬﮕﻧ .
ﻪﻳﺎﻣ ﻲﻧﺯ ﻭ ﺮﻴﺜﻜﺗ ﻪﻧﺎﺨﻠﮔ ﺭﺩ ﺱﻭﺮﻳﻭ ﻊﺑﺎﻨﻣ
ﻪﻌﻟﺎﻄﻣ ﻦﻳﺍ ﺭﺩ ﻢﻗﺭ ﻦﻳﺮﻴﺷ ﺕﺭﺫ
٤٠٣ ﻡﻮﮔﺭﻮﺳ ﻭ ﺱﺭﺎﭘ
ﻪﻧﺍﺩ ﻱﺍ ﻢﻗﺭ ﺎﻴﻤﻴﻛ ﻥﺍﻮﻨﻌﺑ ﻥﺎﺑﺰﻴﻣ ﻱﺮﻴﺜﻜﺗ ﻱﺎﻫ ﻩﺪﻧﺭﺍﺪﻬﮕﻧ ﻭ
2
ﺐﺳﺎﻨﻣ ﺪﻧﺪﺷ ﺏﺎﺨﺘﻧﺍ .
ﺭﺩ ﻡﻮﮔﺭﻮﺳ ﻭ ﻦﻳﺮﻴﺷ ﺕﺭﺫ ﺭﻭﺬﺑ
ﻱﻭﺎﺣ ﻙﺎﺧ ﻁﻮﻠﺨﻣ ﻪﺳﺎﻣ ﻭ ﻲﻣﺍﺩ ﺩﻮﻛ ،ﻲﻟﻮﻤﻌﻣ ﺱﺭ ﻙﺎﺧ
ﻪﺑ ﺖﺒﺴﻧ ٣ : ٢ : ١ ﻭ ﺭﺩ ﺏﺁ ﺭﺎﺨﺑ ﺎﺑ ﻱﺯﺎﺳ ﻞﻳﺮﺘﺳﺍ ﺯﺍ ﺲﭘ
ﺪﺷﺭ ﺯﺍ ﺲﭘ ﻭ ﺪﻧﺪﺷ ﻪﺘﺷﺎﻛ ،ﻭﻼﻛﻮﺗﺍ ﻩﺎﮕﺘﺳﺩ ﻠﺣﺮﻣ ﺭﺩ
ﻪ ٢ ﺎﺗ
٣ ﺑ ﻲﮔﺮ ﻪﻳﺎﻣ ﻲﻧﺯ ﺪﻧﺪﺷ . ﻪﻧﻮﻤﻧ ﻪﺑ ﺎﻫ ﺖﺒﺴﻧ ١:١٠
W/V) ( ﺭﺩ
ﻢﻴﺳﺎﺘﭘ ﺕﺎﻔﺴﻓ ﺮﻓﺎﺑ ١
/ ٠ ﺭﻻﻮﻣ
pH) ٧ ( ﻱﻭﺎﺣ ٢ ﺪﺻﺭﺩ
W/V) ( ﺩﺎﻣ ﺯﺍ ﻩ ﻥﻭﺪﻴﻟﻭﺮﻳﺎﭘ ﻞﻴﻨﻳﻭ ﻲﻠﭘ )3
(PVP
ﻩﺭﺎﺼﻋ ﻱﺮﻴﮔ
ﻩﺪﺷ ﻭ ﺖﻓﺎﺑ ﻩﺭﺎﺼﻋ ﮒﺮﺑ ﻱﻭﺭ ﺮﺑ ﻩﺩﻮﻟﺁ ﻱﺎﻫ
ﻱﺎﻫ ﺑﺭﺎﻛ ﻮ ﻡﻭﺪﻧﺍﺭ
ﺪﺷ ﻩﺪﻴﻟﺎﻣ ﻡﻮﮔﺭﻮﺳ ﻭ ﺕﺭﺫ ﻩﺪﺷ ﻲﺷﺎﭘ .
ﻪﻳﺎﻣ ﻲﻧﺯ ﹰﺎﺒﻟﺎﻏ ﺎﻫ ﺭﺩ
ﺏﺎﺘﻓﺁ ﺏﻭﺮﻏ ﻭ ﺮﺼﻋ ﻡﺎﮕﻨﻫ ﻭ
ﻪﺑ ﻭ ﻚﻨﺧ ﻲﻳﺎﻣﺩ ﻂﻳﺍﺮﺷ ﺭﺩ
ﻡﺎﺠﻧﺍ ﺏﺎﺘﻓﺁ ﻢﻴﻘﺘﺴﻣ ﺭﻮﻧ ﺯﺍ ﺭﻭﺩ ﺪﻳﺩﺮﮔ
.
ﻥﻮﻣﺯﺁ ﺎﻫ ﻱ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ
ﻪﻧﻮﻤﻧ ﻱﺎﻫ ﻪﻋﺭﺰﻣ ﻱﺍ ﺰﻴﻧ ﻭ ﻩﺎﮕﺸﻳﺎﻣﺯﺁ ﻪﺑ ﻝﺎﻘﺘﻧﺍ ﺯﺍ ﺲﭘ
ﻪﻧﺎﺨﻠﮔ ﺭﺩ ﺩﻮﺟﻮﻣ ﻥﺎﻫﺎﻴﮔ ﻭﺩ
ﻪﺘﻔﻫ ﺯﺍ ﺲﭘ ﻪﻳﺎﻣ ﻧﺯ ﻲ ﺮﻈﻧ ﺯﺍ
ﻪﺑ ﻲﮔﺩﻮﻟﺁ ﺩﺭﻮﻣ ﺱﻭﺮﻳﻭ ﻲﺗﻮﭘ ﺭﺎﻬﭼ ﺯﺍ ﻚﻳ ﺮﻫ
ﻪﻌﻟﺎﻄﻣ
SCMV)
MDMV،
SrMV ،
JGMVﻭ ( ﻡﺎﺠﻧﺍ ﺎﺑ ﻥﻮﻣﺯﺁ ﺎﻫ ﻱ
ﻲﻜﻳﮊﻮﻟﻭﺮﺳ
DAS-ELISA DIBA ،
TPIA ﻭ ﻲﺳﺭﺮﺑ ﺩﺭﻮﻣ
ﺪﻨﺘﻓﺮﮔ ﺭﺍﺮﻗ .
ﻡﺎﺠﻧﺍ ﻱﺍﺮﺑ ﻥﻮﻣﺯﺁ
ﺎﻫ ﺯﺍ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ ﻱ
ﻲﺘﻧﺁ ﻡﺮﺳ ﻝﺎﻧﻮﻠﻛ ﻲﻠﭘ ﻱﺎﻫ ﻪﻌﻟﺎﻄﻣ ﺩﺭﻮﻣ ﺱﻭﺮﻳﻭ ﻲﺗﻮﭘ ﺭﺎﻬﭼ
،
ﺮﺘﻨﻳﻭ ﻥﺎﻔﺘﺳﺍ ﺮﺘﻛﺩ ﻂﺳﻮﺗ ﻩﺪﺷ ﺍﺪﻫﺍ ﺯﺍ
ﺶﺨﺑ
ﺱﻭﺮﻳﻭ ﻲﻫﺎﻴﮔ ﻲﺳﺎﻨﺷ ﺴﺳﺆﻣ
DSMZ ﻪ ﻥﺎﻤﻟﺁ ﻩﺩﺎﻔﺘﺳﺍ
ﺪﻳﺩﺮﮔ .
ﻥﻮﻣﺯﺁ ﺍﺰﻳﻻﺍ 4
ﻥﻮﻣﺯﺁ ﺍﺰﻳﻻﺍ ﻞﻤﻌﻟﺍﺭﻮﺘﺳﺩ ﺱﺎﺳﺍ ﺮﺑ ﻝﻮﻤﻌﻣ
Clark &
Adams (1976)
ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﻭ ﻲﺘﻧﺁ
ﻡﺮﺳ ﻝﺎﻧﻮﻠﻛ ﻲﻠﭘ ﻱﺎﻫ
1. Freeze-drier
2. Propagation & maintenance hosts 3. Polyvinyl pyrrolidon
4. Enzyme Linked Immuno Sorbent Assay (ELISA)
ﻪﻌﻟﺎﻄﻣ ﺩﺭﻮﻣ ﺱﻭﺮﻳﻭ ﻲﺗﻮﭘ ﺭﺎﻬﭼ ﻱﺍﺰﻳﻻﺍ ﺕﺭﻮﺻ ﻪﺑ ﻭ
ﻪﻓﺮﻃﻭﺩ ﭻﻳﻭﺪﻧﺎﺳ ﺎﻳ ﻢﻴﻘﺘﺴﻣ ﻲﺘﻧﺁ
٥ﻱﺩﺎﺑ
DAS-ELISA) (
ﻡﺎﺠﻧﺍ ﻭ ﻣ ﻦﻳﺮﺧﺁ ﺭﺩ ﻠﺣﺮ
ﻪ ﻠﺻﺎﻓ ﺭﺩ ﻭ ﺍﺰﻳﻻﺍ ﻪ
١٥ ﺎﺗ ٦٠ ﻪﻘﻴﻗﺩ
ﻚﻫﺎﭼ ﻪﺑ ﺍﺮﺘﺴﺑﻮﺳ ﻝﻮﻠﺤﻣ ﻥﺩﻭﺰﻓﺍ ﺯﺍ ﺪﻌﺑ ﺪﻫﺎﺸﻣ ،ﺎﻫ
ﻭ ﻩ
ﺮﻴﺴﻔﺗ ﺠﻴﺘﻧ ﻪ ﺪﻫﺎﺸﻣ ﺎﺑ ﻥﻮﻣﺯﺁ ﻩ
ﻚﻫﺎﭼ ﮓﻧﺭ ﺮﻴﻴﻐﺗ ﺯﺍ ﺎﻫ
ﺭﺩ ﮓﻧﺭ ﺮﻴﻴﻐﺗ ﻥﺍﺰﻴﻣ ﺎﺑ ﺎﻬﻧﺁ ﻪﺴﻳﺎﻘﻣ ﻭ ﺩﺭﺯ ﻪﺑ ﻦﺷﻭﺭ ﻚﻫﺎﭼ ﺎﻫﺪﻫﺎﺷ ﻪﺑ ﻁﻮﺑﺮﻣ ﻱﺎﻫ )
ﻚﻫﺎﭼ ﻱﺎﻫ ﻭ ﻲﻔﻨﻣ ،ﺖﺒﺜﻣ
ﺮﻓﺎﺑ ( ﻩﺯﺍﺪﻧﺍ ﺎﺑ ﺎﻳ ﻭ ﺮﻴﮔ
ﻱﺭﻮﻧ ﺏﺬﺟ ﻱﺩﺪﻋ ﻥﺍﺰﻴﻣ ﺖﺒﺛ ﻭ ﻱ
ﻚﻫﺎﭼ ﺝﻮﻣ ﻝﻮﻃ ﺭﺩ ﺎﻫ ٤٠٥
ﺮﺘﻣﻮﻧﺎﻧ ) ﻩﺎﮕﺘﺳﺩ ﺭﺩ ﺍﺰﻳﻻﺍ
٦ﺭﺪﻳﺭ ﻝﺪﻣ
Statefax–2100
ﺖﻛﺮﺷ ﺖﺧﺎﺳ
Awareness
Technology
ﺎﻜﻳﺮﻣﺁ ( ﺴﻳﺎﻘﻣ ﻭ ﻪ ﻪﺑ ﻁﻮﺑﺮﻣ ﺩﺍﺪﻋﺍ ﺎﺑ ﺎﻬﻧﺁ
ﻚﻫﺎﭼ ﺪﻫﺎﺷ ﻱﺎﻫ ﻭ
ﻪﻄﺑﺍﺭ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ
R ≥ X+3sd
X) : ﻪﻧﻮﻤﻧ ﺏﺬﺟ ﻦﻴﮕﻧﺎﻴﻣ ،ﻢﻟﺎﺳ ﻱﺎﻫ
:sd
ﺍ ،ﺭﺎﻴﻌﻣ ﻑﺍﺮﺤﻧ :R
ﺖﺒﺜﻣ ﻪﻧﻮﻤﻧ ﺏﺬﺟ (
≥2X ﺎﻳ ﺪﺷ ﻡﺎﺠﻧﺍ .
ﻥﻮﻣﺯﺁ
٧TPIA
ﻲﻔﻴﺻﻮﺗ ﺵﻭﺭ ﺯﺍ ﻥﻮﻣﺯﺁ ﻦﻳﺍ ﻡﺎﺠﻧﺍ ﻱﺍﺮﺑ
Sambrook et al.
(1989) Dijkstra & deJager ﻭ
(1998)
ﺪﻳﺩﺮﮔ ﻩﺩﺎﻔﺘﺳﺍ .
ﻪﻧﻮﻤﻧ ﺯﺍ ﻲﻌﻄﻘﻣ ﻥﻮﻣﺯﺁ ﻡﺎﺠﻧﺍ ﻱﺍﺮﺑ ﻱﺎﻫ
ﺰﻟﻮﻠﺳﻭﺮﺘﻴﻧ ﺀﺎﺸﻏ ﺎﺑ ﻥﺩﺍﺩ ﺱﺎﻤﺗ ﺯﺍ ﺲﭘ ﻭ ﻩﺪﺷ ﻪﻴﻬﺗ ﻩﺩﻮﻟﺁ ،٨
ﻏ ﺕﺪﻣ ﻪﺑ ﺀﺎﺸ ١
ﺎﺗ ٥ ﮓﻨﻴﻛﻼﺑ ﻝﻮﻠﺤﻣ ﻞﺧﺍﺩ ﺭﺩ ﻪﻘﻴﻗﺩ
) ﻱﻭﺎﺣ ۱ ﺪﺻﺭﺩ ﻞﻜﻟﺍ ﻞﻴﻨﻳﻭ ﻲﻠﭘ (
ﺕﺪﻣ ﻪﺑ ﺪﻌﺑ ﻭ ﻩﺩﺍﺩ ﺭﺍﺮﻗ
٥ ﺭﺩ ﻪﻘﻴﻗﺩ ١٠
ﻲﻠﻴﻣ ﺮﻓﺎﺑ ﺮﺘﻴﻟ ﺎﻳ ﻮﺸﺘﺴﺷ ﻮﺸﺘﺴﺷ PBST
ﺪﺷ . ﺕﺪﻣ ﻪﺑ ﺰﻟﻮﻠﺳﻭﺮﺘﻴﻧ ﺬﻏﺎﻛ ﺲﭙﺳ ٢
ﺭﺩ ﺖﻋﺎﺳ ٥
ﻲﻠﻴﻣ ﺮﺘﻴﻟ ﺮﻓﺎﺑ ﺝﻭﺩﺰﻣ ﻪﻛ ﻲﺘﻧﺁ ﻱﺩﺎﺑ ﺝﻭﺩﺰﻣ ﻦﻴﻟﺎﻜﻟﺁ ﺎﻳ
ﺭﺍﺩﺯﺎﺗﺎﻔﺴﻓ
IgG-AP) ( ﻥﺁ ﺭﺩ ﺪﺷ ﻖﻴﻗﺭ ﺭﺩ ﺩﻮﺑ ﻩ ﻕﺎﺗﺍ ﻱﺎﻣﺩ
ﺮﻜﻴﺷ ﻩﺎﮕﺘﺳﺩ ﻱﻭﺭ ﻭ ﺪﺷ ﻩﺩﺍﺩ ﺭﺍﺮﻗ ﻢﻳﻼﻣ ﺖﻛﺮﺣ ﺭﻭﺩ ﺎﺑ
.
ﺯﺍ ﺲﭘ ﻮﺸﺘﺴﺷ ﺭﺎﺑﻭﺩ ﻱ
ﺀﺎﺸﻏ ﺭﺩ ﺮﻓﺎﺑ ﺖﻳﺎﻬﻧ ﺭﺩ PBST
ﻱﺍﺮﺘﺴﺑﻮﺳ ﻝﻮﻠﺤﻣ
Fast red NBT/BCIP ﺎﻳ
ﻱﻭﺭ ﺮﺑ ﺀﺎﺸﻏ
ﻧﻴ ﺰﻟﻮﻠﺳﻭﺮﺘ ﺕﺪﻣ ﻪﺑ ﻭ ﻪﻓﺎﺿﺍ ١٥
ﺎﺗ ٣٠ ﻂﻴﺤﻣ ﺭﺩ ﻪﻘﻴﻗﺩ
ﻨﻛﺍﻭ ﺎﺗ ﺪﺷ ﻩﺩﺍﺩ ﺭﺍﺮﻗ ﻚﻳﺭﺎﺗ ﮓﻧﺭ ﺶ
ﺩﺩﺮﮔ ﻡﺎﺠﻧﺍ ﻱﺰﻴﻣﺁ .
ﻥﻮﻣﺯﺁ
٩DIBA
ﻥﻮﻣﺯﺁ ﻦﻳﺍ ﻡﺎﺠﻧﺍ ﻱﺍﺮﺑ ﺰﻴﻧ
ﻲﻔﻴﺻﻮﺗ ﺵﻭﺭ ﺯﺍ
Sambrook
et al.
(1998) Dijkstra & de Jagerﻭ
(1998)
ﻲﻛﺪﻧﺍ ﺎﺑ
ﺕﺍﺮﻴﻴﻐﺗ ﺪﻳﺩﺮﮔ ﻩﺩﺎﻔﺘﺳﺍ .
ﻩﺭﺎﺼﻋ ﺍﺪﺘﺑﺍ ﻪﻧﻮﻤﻧ ﻱﺎﻫ
ﻩﺩﻮﻟﺁ ﻱﺎﻫ
ﻩﺭﺎﺼﻋ ﺮﻓﺎﺑ ﺭﺩ ﻩﺭﺎﺼﻋ ﻥﺩﺍﺩ ﺱﺎﻤﺗ ﺯﺍ ﺲﭘ ﻭ ﻪﻴﻬﺗ ﻱﺮﻴﮔ
ﺎﺑ ﺎﻫ
5. Double Antibody Sandwich - ELISA
6. ELISA-reader
7. Tissue Print Immunobinding Assay 8. Nitrocellulose membrane
9. Dot Immunobinding Assay
۴۶ ﻪﻠﺠﻣ
ﮑﺷﺰﭙﻫﺎﻴﮔ ﺶﻧﺍﺩ ﻲ
ﻩﺭﻭﺩ ،ﻥﺍﺮﻳﺍ ۴۰
ﻩﺭﺎﻤﺷ ، ۱ ، ۱۳۸۸
ﺮﺘﻴﻧ ﺀﺎﺸﻏ ،ﺰﻟﻮﻠﺳﻭ ﺕﺪﻣ ﻪﺑ ﺀﺎﺸﻏ ١
ﺎﺗ ٥ ﻝﻮﻠﺤﻣ ﻥﻭﺭﺩ ﻪﻘﻴﻗﺩ
ﺪﺷ ﻩﺩﺍﺩ ﺭﺍﺮﻗ ﮓﻨﻴﻛﻼﺑ .
ﺀﺎﺸﻏ ﺎﺑ ﻮﺸﺘﺴﺷ ﺯﺍ ﺲﭘ ﻪﺑ PBST
ﻥﻭﺭﺩ ٥ ﺎﺗ ١٠ ﻲﻠﻴﻣ ﺮﺘﻴﻟ ﻪﻛ ﺖﻴﮔﻮﺠﻧﺎﻛ ﺮﻓﺎﺑ ﻲﺘﻧﺁ
ﻱﺩﺎﺑ
ﺭﺍﺩ ﺯﺎﺗﺎﻔﺴﻓ ﻦﻴﻟﺎﻜﻟﺁ
IgG-AP) ( ﺩﻮﺑ ﻩﺪﺷ ﻖﻴﻗﺭ ﻥﺁ ﺭﺩ
ﺕﺪﻣ ﻪﺑ ﻭ ﻞﻘﺘﻨﻣ ٢
ﺭﺍﺮﻗ ﺮﻜﻴﺷ ﻱﻭﺭ ﻭ ﻕﺎﺗﺍ ﻱﺎﻣﺩ ﺭﺩ ﺖﻋﺎﺳ
ﺪﺷ ﻩﺩﺍﺩ . ﺯﺍ ﺲﭘ ﺐﻴﺗﺮﺗ ﻪﺑ ﻥﺁ ٢
ﺕﺪﻣ ﻪﺑ ﺭﺎﺑ ﺮﻫ ﻭ ﺭﺎﺑ ٥
ﺎﺑ ﻪﻘﻴﻗﺩ ﺮﻓﺎﺑ ﻭ PBST
ﻱﺍﺮﺑ ﹰﺎﺘﻳﺎﻬﻧ ﻭ ﺪﺷ ﻩﺩﺍﺩ ﻮﺸﺘﺴﺷ AP
ﺪﻫﺎﺸﻣ ﻩ ﺠﻴﺘﻧ ﻪ ﻱﺍﺮﺘﺴﺑﻮﺳ ﻝﻮﻠﺤﻣ ،ﻥﻮﻣﺯﺁ
Fast red
ﺎﻳ
NBT/BCIP
ﻪﺑ ﺀﺎﺸﻏ ﻱﻭﺭ ﺪﺷ ﻪﻓﺎﺿﺍ
.
ﺪﻴﻣﻼﻳﺮﻛﺍ ﻲﻠﭘ ﻝﮊ ﻱﺩﻮﻤﻋﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍ )1
SDS-PAGE (
ﻞﻤﻌﻟﺍﺭﻮﺘﺳﺩ ﺱﺎﺳﺍ ﺮﺑ ﻥﻮﻣﺯﺁ ﻦﻳﺍ
Sambrook et al.
(1998)
ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﻭ ﺪﻨﻨﻛ ﺰﻳﺎﻤﺘﻣ ﻝﮊ ﺯﺍ
ﻩ ١٥ ﻝﮊ ﻭ ﺪﺻﺭﺩ
ﺪﻨﻨﻛ ﻢﻛﺍﺮﺘﻣ ﻩ
٥ ﺪﺻﺭﺩ ﻡﺎﺠﻧﺍ ﺪﺷ . ﻩﺭﺎﺼﻋ ﻪﻧﻮﻤﻧ ﺎﻫ ﺮﻓﺎﺑ ﺭﺩ
ﻩﺭﺎﺼﻋ ﻱﺮﻴﮔ ) ﺮﻓﺎﺑ ٥٠ ﻲﻠﻴﻣ ﺭﻻﻮﻣ
Tris-HCl
، ٥ ﻲﻠﻴﻣ ﺭﻻﻮﻣ
Boric acid
، ٥ ﻭ ﻝﻭﺮﺴﻴﻠﮔ ﺪﺻﺭﺩ ٥
/ ١ ﺪﺻﺭﺩ (PVP
ﻪﻴﻬﺗ
ﻩﺭﺎﺼﻋ ﺲﭙﺳ ﻭ ﻩﺪﺷ ﻩﺩﻮﻣﺁ ﻭ ﺎﻫ
ﺎﺑ ﺺﻟﺎﺧ ﺱﻭﺮﻳﻭ ﻱﺎﻫ ﻢﺠﺣ
ﺮﻓﺎﺑ ﺯﺍ ﻱﻭﺎﺴﻣ ﻲﻠﻤﻟﺭﺎﺑ ﻭﺩ
) ٠٢ / ٠ ﺭﻻﻮﻣ
Tris-HCl
ﻭ ٢
ﻲﻠﻴﻣ ﺭﻻﻮﻣ ﻱﻭﺎﺣ EDTA
٢ ﺪﺻﺭﺩ ،SDS
٢٠ ﺪﺻﺭﺩ
ﻦﻴﺴﻴﻠﻴﮔ ﻭ ٠٥ / ٠ ﻮﻠﺑ ﻞﻨﻓﻮﻣﺮﺑ ﺪﺻﺭﺩ (
ﻭ ﻁﻮﻠﺨﻣ ﻪﻧﻮﻤﻧ
ﻱﺎﻫ
ﻲﻨﻴﺌﺗﻭﺮﭘ ﺮﻛﺭﺎﻣ ﻩﺍﺮﻤﻫ ﻪﺑ ﻩﺪﺷ ﻩﺩﺎﻣﺁ )
ﻥﺯﻭ ﺩﺭﺍﺪﻧﺎﺘﺳﺍ ﺮﻛﺭﺎﻣ
ﻲﻟﻮﻜﻟﻮﻣ ( ﺕﺪﻣ ﻪﺑ ٥ ﺵﻮﺟ ﻝﺎﺣ ﺭﺩ ﺏﺁ ﺭﺩ ﻪﻘﻴﻗﺩ ﺕﺭﺍﺮﺣ
ﺑ ﻭ ﻩﺩﺍﺩ ﺦﻳ ﻱﻭﺭ ﻪﺑ ﻪﻠﺻﺎﻓﻼ ﺪﺷ ﻞﻘﺘﻨﻣ
ﺪﻧ . ﺭﺍﺪﻘﻣ ٢٠ ﺎﺗ ٣٠
ﻪﻧﻮﻤﻧ ﺯﺍ ﺮﺘﻴﻟﻭﺮﻜﻴﻣ ﻚﻫﺎﭼ ﺭﺩ ﺎﻫ
ﻝﮊ ﻱﺎﻫ ﻪﺘﺨﻳﺭ ﻭ ﻥﺎﻳﺮﺟ
ﺖﺑﺎﺛ ﮊﺎﺘﻟﻭ ﺎﺑ ﻪﺘﻴﺴﻳﺮﺘﻜﻟﺍ ١٠٠
ﺕﺪﻣ ﻪﺑ ۳ ﺎﺗ ۴ ﺖﻋﺎﺳ ﺭﺍﺮﻗﺮﺑ
ﺪﺷ . ﻝﻮﻠﺤﻣ ﺭﺩ ﺐﺷ ﻚﻳ ﺕﺪﻣ ﻪﺑ ﻝﮊ ﻥﺁ ﺯﺍ ﺲﭘ ﮓﻧﺭ
ﺰﻴﻣﺁ ﻱ
ﻮﻠﺑ ﻱﺯﺎﻣﻮﻛ )
٥٠ ،ﻝﻮﻧﺎﺘﻣ ﺪﺻﺭﺩ ١٠
،ﻚﻴﺘﺳﺍ ﺪﻴﺳﺍ ﺪﺻﺭﺩ
٣٥ / ٠ ﺪﺻﺭﺩ ﺖﻧﺎﻴﻠﻳﺮﺑ ﻱﺯﺎﻣﻮﻛ ﻭ ﻮﻠﺑ
٤٠ ﺮﻄﻘﻣ ﺏﺁ ﺪﺻﺭﺩ (
ﻱﺮﺒﮕﻧﺭ ﻝﻮﻠﺤﻣ ﺭﺩ ﺲﭙﺳ ﻩﺪﺷ ﻩﺩﺍﺩ ﺭﺍﺮﻗ )
٢٥ ﺪﺻﺭﺩ
،ﻝﻮﻧﺎﺘﻣ ١٠ ﻭ ﺪﻴﺳﺍ ﻚﻴﺘﺳﺍ ﺪﺻﺭﺩ ٦٥
ﺮﻄﻘﻣ ﺏﺁ ﺪﺻﺭﺩ (
ﮓﻧﺭ ﺪﺷ ﻲﻳﺍﺩﺯ .
ﺕﻼﺑ ﻥﺮﺘﺳﻭ ﻥﻮﻣﺯﺁ )2
(EBIA
ﻥﻮﻣﺯﺁ ﻦﻳﺍ ﺰﻴﻧ
ﻪﺑ ﻲﻔﻴﺻﻮﺗ ﺵﻭﺭ
Sambrook et al.
(1998) Dijkstra & de Jager ﻭ
(1998)
ﺮﮔ ﻡﺎﺠﻧﺍ ﺪﻳﺩ .
ﻩﺭﺎﺼﻋ ﻲﻫﺎﻴﮔ ﻱﺎﻫ ﺎﻳ ﻭ
ﻩﺩﻮﻣﺁ ﻱﺎﻫ ﺮﺑ ﺍﺪﺘﺑﺍ ﺺﻟﺎﺧ ﺱﻭﺮﻳﻭ
ﻝﮊ ﻱﻭﺭ
SDS-PAGE
ﻪﻓﺮﻃ ﻭﺩ ﻩﺪﺷ ﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍ
ﺲﭙﺳ
ﻝﮊ ﺯﺍ ﻲﻜﻳ ﮓﻧﺭ ﺎﻫ
ﻱﺮﺒﮕﻧﺭ ﻭ ﻱﺰﻴﻣﺁ ﺎﺗ ﺪﺷ
ﻚﻴﻜﻔﺗ ﺯﺍ
1. Sodium Dodecyl Sulphate – Polyacrylamid Gel
Electrophoresis
2. Electroblot Immunoassay or Western Blotting
ﻞﺻﺎﺣ ﻥﺎﻨﻴﻤﻃﺍ ﻝﮊ ﻭﺩ ﺮﻫ ﺭﺩ ﻲﻨﻴﺌﺗﻭﺮﭘ ﻱﺎﻫﺪﻧﺎﺑ ﺏﻮﻠﻄﻣ ﺩﻮﺷ . ﺰﻟﻮﻠﺳﻭﺮﺘﻴﻧ ﺀﺎﺸﻏ ﻥﺩﺍﺩ ﺭﺍﺮﻗ ﺯﺍ ﺪﻌﺑ ﻡﻭﺩ ﻝﮊ ﻱ
ﻱﻭﺭ ﺮﺑ
ﻪﺑ ﻥﺁ ﺎﺗ ﻚﻧ ﻥﺮﺘﺳﻭ ﻝﺎﻘﺘﻧﺍ ﺮﻓﺎﺑ ﻱﻭﺎﺣ ﺕﻼﺑ ﻥﺮﺘﺳﻭ ﺮﻓﺎﺑ ﺎﻳ
) ﻞﻣﺎﺷ ٤٨ ﻲﻠﻴﻣ ﺭﻻﻮﻣ
Tris-base
، ٣٩ ﻲﻠﻴﻣ ﺭﻻﻮﻣ
Glycine
،
٠٣٧٥ / ٠ ﺪﺻﺭﺩ ﻭ SDS
٢٠ ﺮﻄﻘﻣ ﺏﺁ ﺭﺩ ﻝﻮﻧﺎﺘﻣ ﺪﺻﺭﺩ (
ﻞﻘﺘﻨﻣ ﺎﻳﺮﺟ ﻭ ﻥ ﺖﺑﺎﺛ ﻪﺘﻴﺴﻳﺮﺘﻜﻟﺍ ٣٥٠
ﺮﭙﻣﺁ ﻲﻠﻴﻣ ﺕﺪﻣ ﻪﺑ
۵ / ۱ ﺎﺗ ۲ ﺖﻋﺎﺳ ﺪﻳﺩﺮﮔ ﺭﺍﺮﻗﺮﺑ .
ﺝﺭﺎﺧ ﻭ ﻥﺎﻳﺮﺟ ﻊﻄﻗ ﺯﺍ ﺲﭘ
ﻭ ﻝﮊ ﻥﺩﻮﻤﻧ ﺕﺎﻴﻠﻤﻋ ﻝﮊ ﻱﻭﺭ ﺮﺑ ﺀﺎﺸﻏ
ﮓﻧﺭ ﮓﻧﺭ ﻭ ﻱﺰﻴﻣﺁ
ﻦﻴﺌﺗﻭﺮﭘ ﻞﻣﺎﻛ ﻝﺎﻘﺘﻧﺍ ﺯﺍ ﺎﺗ ﺖﻓﺮﮔ ﺕﺭﻮﺻ ﻱﺮﺑ ﻱﻭﺭ ﻪﺑ ﺎﻫ
ﺩﻮﺷ ﻞﺻﺎﺣ ﻥﺎﻨﻴﻤﻃﺍ ﺀﺎﺸﻏ .
ﺀﺎﺸﻏ ﺰﻟﻮﻠﺳﻭﺮﺘﻴﻧ ﻪﻜﻧﺁ ﺯﺍ ﺲﭘ
ﮓﻨﻴﻛﻼﺑ ﻝﻮﻠﺤﻣ ﺭﺩ ﺐﺷ ﻚﻳ ﺕﺪﻣ ﻪﺑ )
٢ ﺮﻴﺷ ﺪﺻﺭﺩ
ﺭﺩ ﻲﺑﺮﭼ ﻥﻭﺪﺑ ﻚﺸﺧ (PBST
ﺭﺩ ﻭ ﻱﺎﻣﺩ ٤ °C
ﺭﺩ
ﺪﺷ ﻩﺩﺍﺩ ﺭﺍﺮﻗ ﻝﺎﭽﺨﻳ
، ﺭﺩ
PBSTﺮﻓﺎﺑ ﻭﺩ ﺕﺪﻣ ﻪﺑ ﻭ ﻮﺸﺘﺴﺷ
ﺭﺩ ﺖﻋﺎﺳ ١٠
ﻲﻠﻴﻣ ﺮﺘﻴﻟ ﺮﻓﺎﺑ ﻱﻭﺎﺣ PBST
ﻲﺘﻧﺁ ﻱﺩﺎﺑ
ﺱﻭﺮﻳﻭ ﻲﺻﺎﺼﺘﺧﺍ
IgG) ( ﻭ ﺪﺷ ﻩﺩﺍﺩ ﺭﺍﺮﻗ ﺮﻜﻴﺷ ﻱﻭﺭ .
ﺲﭘ
ﺯﺍ ﺭﺎﺑ ﻪﺳ ﻮﺸﺘﺴﺷ
PBST ﺭﺩ ﺖﻋﺎﺳ ﻚﻳ ﺕﺪﻣ ﻪﺑ ﺀﺎﺸﻏ
ﺭﺩ ١٠ ﻲﻠﻴﻣ ﺮﺘﻴﻟ ﻱﻭﺎﺣPBST
ﻲﺘﻧﺁ ﻱﺩﺎﺑ ﺖﻴﺑﺭ ﻲﺘﻧﺁ ﺕﻮﮔ
3
ﺯﺎﺗﺎﻔﺴﻓ ﻦﻴﻟﺎﻜﻟﺁ ﻢﻳﺰﻧﺁ ﻪﺑ ﻞﺼﺘﻣ
GAR-AP) (
ﺪﺷ ﻩﺩﺍﺩ ﺭﺍﺮﻗ .
ﺎﺑ ﹰﺍﺩﺪﺠﻣ ﺀﺎﺸﻏ ﻭ ﻮﺸﺘﺴﺷ PBST
١٠ ﻲﻠﻴﻣ ﺮﺘﻴﻟ ﺮﻓﺎﺑ
ﺯﺎﺗﺎﻔﺴﻓ ﻦﻴﻟﺎﻜﻟﺁ ﺮﻓﺎﺑ ﺎﻳ ﺍﺮﺘﺴﺑﻮﺳ
(AP- buffer) ﻪﻓﺎﺿﺍ ﻥﺁ ﻪﺑ
ﺩﺮﮔ ﻳ ﺪ ﻭ ﺖﻬﺟ ﮓﻧﺭ ﺰﻴﻣﺁ ﻱ ﺯﺍ ﺩﺎﻣ ﻭﺩ ﻩ ﻲﮕﻧﺭ ) NBT
٢٢
ﺮﺘﻴﻟﻭﺮﻜﻴﻣ (
BCIP ﻭ ) ١٦ ﺮﺘﻴﻟﻭﺮﻜﻴﻣ (
ﻩﺩﺎﻔﺘﺳﺍ ﺪﺷ . ﺯﺍ ﺲﭘ
١٥ ﺎﺗ ٣٠ ،ﺎﻫﺪﻧﺎﺑ ﺭﻮﻬﻇ ﺎﺑ ﻪﻘﻴﻗﺩ ﺀﺎﺸﻏ
ﺮﻄﻘﻣ ﺏﺁ ﻞﺧﺍﺩ ﻪﺑ
ﻣ ﻞﻘﺘﻨ ﺪﺷ .
ﻥﻮﻣﺯﺁ RT-PCR
٤
ﻥﻮﻣﺯﺁ
‐ PCR
) RT
ﺶﻨﻛﺍﻭ ﻩﺮﻴﺠﻧﺯ ﻱﺍ ﺵﻭﺭ ﻪﺑ ﺯﺍﺮﻤﻴﻠﭘ
ﺱﻮﻜﻌﻣ ﻱﺭﺍﺩﺮﺑ ﻪﺨﺴﻧ (
ﻲﺻﺎﺼﺘﺧﺍ ﻱﺎﻫﺮﻤﻳﺍﺮﭘ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ
ﻂﺳﻮﺗ ﻩﺪﺷ ﻲﺣﺍﺮﻃ
Yang & Mirkov (1997)
ﺎﺑ ﻲﻟﻭ
ﻲﺷﻭﺭ ﹰﺎﺒﻳﺮﻘﺗ ﺖﻓﺮﮔ ﺕﺭﻮﺻ ﺎﻬﻧﺁ ﺵﻭﺭ ﺯﺍ ﺕﻭﺎﻔﺘﻣ .
ﺮﻤﻳﺍﺮﭘ ﻭﺩ
ﻢﻴﻘﺘﺴﻣ
SCMV F3
ﻭ ﺱﻮﻜﻌﻣ
SCMV R3
ﺍ ﺮﺑ ﻪﻛ ﺱﺎﺳ
ﻲﻟﺍﻮﺗ ﺖﻇﺎﻔﺣ ﻱﺎﻫ ﻩﺪﺷ
ﻱﺍ ﻱﺎﻬﺘﻧﺍ ﺭﺩ ٣′
ﻡﻮﻧﮊ ﺭﺩ ﻭ SCMV
ﻘﻄﻨﻣ ﻪ ﺪﻨﻨﻛﺪﻛ ﻩ ﻲﺸﺷﻮﭘ ﻦﻴﺌﺗﻭﺮﭘ
CP) ( ﻂﺳﻮﺗ
Yang &
Mirkov (1997)
ﻲﺣﺍﺮﻃ ﻩﺪﺷ ﺪﻧﺍ ﹰﻼﻣﺎﻛ ﺭﻮﻄﺑ ، ﻲﺻﺎﺼﺘﺧﺍ
ﻂﻘﻓ ﺨﺸﺗ ﻪﺑ ﺭﺩﺎﻗ ﻴ
ﺺ ﻦﻳﺮﺘﺳﺍ
SCMV ﻱﺎﻫ ﻣ ﻲ ﺪﻨﺷﺎﺑ .
SCMVF3:5′ TTTCTCACCAAGCTGGAA - 3′
SCMVR3:5′AGCTGTGTGTCTCTCTGTATTCTC - 3′
3. Goat anti-rabbit
4. Reverse Transcription-Polymerase Chain Reaction
ﻥﺍﺭﺎﮑﻤﻫ ﻭ ﻱﺪﻤﺤﻣ :
ﻳﺎﺳﺎﻨﺷ ﻭ ﻥﺍﺮﻬﺗ ﻥﺎﺘﺳﺍ ﺕﺭﺫ ﻉﺭﺍﺰﻣ ﺭﺩ ﺐﻟﺎﻏ ﻲﺗﻮﭘ ﻲ ...
۴۷
ﺶﻨﻛﺍﻭ RT-PCR ﻱﺎﻫ
ﺮﻠﻜﻳﺎﺳﻮﻣﺮﺗ ﻩﺎﮕﺘﺳﺩ ﺭﺩ
Palmcycler GP001ﻝﺪﻣ
) ﻲﻧﺎﭙﻤﻛ ﺖﺧﺎﺳ
Corbett
Reaserch
ﺎﻴﻟﺍﺮﺘﺳﺍ ( ﺖﻓﺮﮔ ﻡﺎﺠﻧﺍ .
،ﻥﻮﻣﺯﺁ ﻡﺎﺠﻧﺍ ﺯﺍ ﻞﺒﻗ
RNA Total
ﻲﻫﺎﻴﮔ ﺍ ﺩﺭﺍﻭ ﺶﻨﻛﺍﻭ ﻁﻮﻠﺨﻣ ﺭﺩ ﻭ ﺝﺍﺮﺨﺘﺳ
ﺪﺷ . ﻱﺍﺮﺑ ﺝﺍﺮﺨﺘﺳﺍ
RNA Total
ﻲﻫﺎﻴﮔ ﺮﻓﺎﺑ ﻭﺩ ﺯﺍ
RNAWIZ
) ﻝﻮﺼﺤﻣ
Antibion
( ﻭ ﺮﻓﺎﺑ
Trireagent
) ﻝﻮﺼﺤﻣ
(Sigma
ﺖﻴﻛ ﺯﺍ ﻦﻴﻨﭽﻤﻫ ﻭ
RNeasy Plant
Mini Kit
) ﻝﻮﺼﺤﻣ
QIAGEN
( ﻞﻤﻌﻟﺍﺭﻮﺘﺳﺩ ﺱﺎﺳﺍﺮﺑ ﺎﻨﻴﻋ
ﻩﺪﻨﻨﻛ ﺪﻴﻟﻮﺗ ﻱﺎﻫ ﻲﻧﺎﭙﻤﻛ ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ
.
ﺭﺩ ﻠﺣﺮﻣ ﻪ ﺖﺧﺎﺳ ﺱﻮﻜﻌﻣ ﻱﺭﺍﺩﺮﺑ ﻪﺨﺴﻧ ﺎﻳ cDNA
RT) (
،ﺶﻨﻛﺍﻭ ﻁﻮﻠﺨﻣ ﻲﻳﺎﻬﻧ ﻢﺠﺣ ٢٠
ﺮﻈﻧ ﺭﺩ ﺮﺘﻴﻟﻭﺮﻜﻴﻣ
ﺐﻴﺗﺮﺗ ﻪﺑ ﻁﻮﻠﺨﻣ ﺀﺍﺰﺟﺍ ﻭ ﻪﺘﻓﺮﮔ ﻝﻭﺪﺟ
۱ ﺪﺷ ﻁﻮﻠﺨﻣ ﺪﻧ
.
ﻥﻭﺪﺑ ﺶﻨﻛﺍﻭ ﻁﻮﻠﺨﻣ ﻥﺩﻭﺰﻓﺍ
ﻢﻳﺰﻧﺁ ﺏﻮﻴﺗ ﻥﻭﺭﺩ ،RT
ﻱﺎﻫ
٢ / ٠ ﻲﻠﻴﻣ ﺮﺘﻴﻟ ﻭ ﻞﻳﺮﺘﺳﺍ ﻱ
RNase-free
ﺕﺪﻣ ﻪﺑ ﻭ ﻪﻴﻬﺗ ٢
ﻱﺎﻣﺩ ﺭﺩ ﻪﻘﻴﻗﺩ ٧٢˚C
ﺖﻓﺮﮔ ﺭﺍﺮﻗ ﺮﻠﻜﻳﺎﺳﻮﻣﺮﺗ ﻩﺎﮕﺘﺳﺩ ﺭﺩ .
ﺯﺍ ﺲﭘ ﻪﺑ ﺎﻣﺩ ﺶﻫﺎﻛ ٤٢˚C
ﻢﻳﺰﻧﺁ ، ﺶﻨﻛﺍﻭ ﻁﻮﻠﺨﻣ ﻪﺑ RT
ﻪﻓﺎﺿﺍ ﺖﺧﺎﺳ ﺖﻬﺟ ﻭ
cDNA
ﻱﺎﻣﺩ ، ٤٢˚C
ﻣ ﻪﺑ ﺕﺪ ٤٥ ﺎﺗ
٦٠ ﺪﺷ ﻪﺘﻓﺮﮔ ﺮﻈﻧ ﺭﺩ ﻪﻘﻴﻗﺩ .
ﻝﻭﺪﺟ ١
‐ ﺶﻨﻛﺍﻭ ﻁﻮﻠﺨﻣ ﺀﺍﺰﺟﺍ RT
ﻩﺩﺎﻣ ﺖﻈﻠﻏ
ﺭﺍﺪﻘﻣ
ﺁ ﺏ ﻞﻳﺮﺘﺳﺍ ﻭ ﻩﺰﻴﻧﻮﻳﺩ ٦µl ‐
5X First strand buffer ٤µl ‐
DTT ١٠٠mM (٠/١M)
٢µl
dNTP mix ١٠
mM (each) ١µl
total RNA
‐ ٥µl
Reverse primer (SCMV R3) ١٠٠pmole /µl
١µl
RNase inhibitor enzyme ٤٠ u/ µl
٥ / ٠ µl
MMLV reverse transcriptase(RT) ٢٠٠u /µl
٥ / ٠ µl
ﻪﻠﺣﺮﻣ ﺭﺩ ﻩﺮﻴﺠﻧﺯ ﺶﻨﻛﺍﻭ ﺎﻳ ﺮﻴﺜﻜﺗ
ﺯﺍﺮﻤﻴﻠﭘ ﻱﺍ ﻢﺠﺣ 1
ﺶﻨﻛﺍﻭ ﻁﻮﻠﺨﻣ ﻲﻳﺎﻬﻧ ٥٠
ﺀﺍﺰﺟﺍ ﻭ ﻪﺘﻓﺮﮔ ﺮﻈﻧ ﺭﺩ ﺮﺘﻴﻟﻭﺮﻜﻴﻣ
ﻝﻭﺪﺟ ﺡﺮﺷ ﻪﺑ ﻥﺁ ۲
ﺪﻳﺩﺮﮔ ﻁﻮﻠﺨﻣ ﺩﺭﺍﻭ .
ﺶﻨﻛﺍﻭ ﻁﻮﻠﺨﻣ
ﺁ ﻥﺩﺮﻛ ﻪﻓﺎﺿﺍ ﻥﻭﺪﺑ ﻢﻳﺰﻧ
ﺏﻮﻴﺗ ﻥﻭﺭﺩ ،Taq
ﻱﺎﻫ ٢ / ٠
ﻲﻠﻴﻣ ﻭ ﻞﻳﺮﺘﺳﺍ ﻱﺮﺘﻴﻟ
RNase Free
ﻝﺎﻘﺘﻧﺍ ﺯﺍ ﺲﭘ ﻭ ﻪﻴﻬﺗ
ﻱﺭﺍﺮﻗﺮﺑ ﻭ ﺮﻠﻜﻳﺎﺳﻮﻣﺮﺗ ﻩﺎﮕﺘﺳﺩ ﻪﺑ ﻱﺎﻣﺩ
٩٤˚C
ﺕﺪﻣ ﻪﺑ ٢
ﻢﻳﺰﻧﺁ ،ﻪﻘﻴﻗﺩ ﺪﻳﺩﺮﮔ ﻪﻓﺎﺿﺍ Taq
. ﺲﭙﺳ ٣٥ ﻞﻜﻴﺳ ۴
ﻪﻘﻴﻗﺩ ﻞﻣﺎﺷ ﻱﺍ ۱
ﻱﺎﻣﺩ ﻪﻘﻴﻗﺩ ٩٤˚C
، ۵ / ۱ ﻱﺎﻣﺩ ﻪﻘﻴﻗﺩ
1. Polymerase chain reaction
۵۵˚C
ﻭ ۵ / ۱ ﻱﺎﻣﺩ ﻪﻘﻴﻗﺩ ۷۲˚C
ﺭﺩ ﻩﺎﮕﺘﺳﺩ ﺩﺮﮔ ﺭﻮﻈﻨﻣ ﻳ
. ﺪ
ﺭﺩ ﻠﺣﺮﻣ ﻚﻳ ﺰﻴﻧ ﻥﺎﻳﺎﭘ ﻪ
ﻲﻳﺎﻬﻧ ﻂﺴﺑ ﻲﻧﻻﻮﻃ 2
) ﻱﺎﻣﺩ
۷۲˚C
( ﺕﺪﻣ ﻪﺑ ١٠
ﻭ ﻥﻮﻴﺳﺍﺰﻳﺮﻤﻴﻠﭘ ﻞﻴﻤﻜﺗ ﻱﺍﺮﺑ ﻪﻘﻴﻗﺩ
ﻪﺘﺷﺭ ﻞﻣﺎﻛ ﺰﺘﻨﺳ ﺪﺷ ﻝﺎﻤﻋﺍ ﺎﻫ
.
ﻝﻭﺪﺟ ٢
‐ ﺶﻨﻛﺍﻭ ﻁﻮﻠﺨﻣ ﺀﺍﺰﺟﺍ PCR
ﻩﺩﺎﻣ ﺖﻈﻠﻏ
ﻣ ﺭﺍﺪﻘ
ﺏﺁ ﻞﻳﺮﺘﺳﺍ ﻭ ﻩﺰﻴﻧﻮﻳﺩ ٥ ‐
/ ٣٤ µl
10X First Strand Buffer ٥µl ‐
MgCl2 ٥٠mM
٢µl
dNTP mix ١٠
mM (each) ١µl
Forward primer(SCMV F3) ١٠٠pmole /µl
١µl
Reverse primer(SCMV R3) ١٠٠pmole /µl
١µl
cDNA
‐ ٥µl
Taq‐polymerase ٥ u/µl
٥ / ٠ µl
ﻥﻮﻣﺯﺁ ﻲﻳﺎﻬﻧ ﻝﻮﺼﺤﻣ
RT-PCR
ﺎﺑ ﻝﮊ ﻲﻘﻓﺍ ﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍ
ﺯﺭﺎﮔﺁ ١ ﺮﻓﺎﺑ ﺭﺩ ﺪﺻﺭﺩ
0/5X TBE
) ٧٥ / ١٠ ﺲﻳﺮﺗ ﻡﺮﮔ
،ﻱﺯﺎﺑ ٥ / ٥ ﻭ ﻚﻳﺭﻮﺑﺪﻴﺳﺍ ﻡﺮﮔ ٩٣
/ ٠ ﻡﺮﮔ ﻚﻳ ﺭﺩ EDTA
ﺮﻄﻘﻣ ﺏﺁ ﺮﺘﻴﻟ
pH ﻭ ٨ ( ﺩﺭﻮﻣ ﺖﻓﺮﮔ ﺭﺍﺮﻗ ﻩﺪﻫﺎﺸﻣ .
ﺺﻟﺎﺧ ﺱﻭﺮﻳﻭ ﻱﺯﺎﺳ
ﺺﻟﺎﺧ ﻱﺍﺮﺑ ﺱﻭﺮﻳﻭ ﻱﺯﺎﺳ
ﺵﻭﺭ ﺯﺍ Minipurification
ﻂﺳﻮﺗ ﻩﺪﺷ ﻩﺩﺍﺩ ﺡﺮﺷ
Jensen et al.
(1986)
ﻲﻛﺪﻧﺍ ﺎﺑ
ﺪﺷ ﻩﺩﺎﻔﺘﺳﺍ ﺕﺍﺮﻴﻴﻐﺗ .
٢ ﺩ ﻩﺯﺎﺗ ﻲﮔﺮﺑ ﺖﻓﺎﺑ ﺯﺍ ﻡﺮﮔ ﺭ
١٥
ﻲﻠﻴﻣ ﺮﺘﻴﻟ ﻱﺯﺎﺳ ﺺﻟﺎﺧ ﺮﻓﺎﺑ ﺯﺍ )
ﻢﻳﺪﺳ ﺕﺍﺮﺘﻴﺳ ١
/ ٠ ﺭﻻﻮﻣ ﺎﺑ
٥ pH
/ ٦ ﻭ ﻱﻭﺎﺣ ٢٥ / ٠
،ﻝﻮﻧﺎﺗﺍﻮﺘﭘﺎﻛﺮﻣ ﺪﺻﺭﺩ ١
/ ٠ ﺪﺻﺭﺩ
Na-DIECA
ﻭ ١ ﺪﺻﺭﺩ (PVP
ﻩﺭﺎﺼﻋ ﺮﻴﮔ ﻱ ﻪﻜﻧﺁ ﺯﺍ ﺲﭘ ﻭ
ﺪﺷ ﻩﺩﺍﺩ ﺭﻮﺒﻋ ﻪﻳﻻ ﺪﻨﭼ ﻞﻤﻠﻣ ﻪﭼﺭﺎﭘ ﺯﺍ
، ﺕﺪﻣ ﻪﺑ ٢٠ ﻪﻘﻴﻗﺩ
ﮊﻮﻔﻳﺮﺘﻧﺎﺳ ﻩﺎﮕﺘﺳﺩ ﺭﺩ
Beckman
ﻝﺪﻣ
J-21B
) ﺖﺧﺎﺳ
ﻳﺮﻣﺁ ﺎﻜ ( ﺖﻋﺮﺳ ﺎﺑ ١٠٠٠٠ rpm
ﺪﺷ ﮊﻮﻔﻳﺮﺘﻧﺎﺳ .
ﻲﻳﻭﺭ ﻪﻳﺎﻣ
ﮓﻧﺭ ﻢﻛ ﺰﺒﺳ ﻩﺩﺍﺩ ﺭﻮﺒﻋ ﻪﻳﻻ ﺪﻨﭼ ﻞﻤﻠﻣ ﻪﭼﺭﺎﭘ ﺯﺍ ﻩﺭﺎﺑﻭﺩ 3
ﺪﺷ ﻭ ﻩ ﺎﺑ ٤ ﻲﻠﻴﻣ ﻝﻮﻠﺤﻣ ﺯﺍ ﺮﺘﻴﻟ ٣٣
ﺪﺻﺭﺩ
Triton X-100
ﺮﻄﻘﻣ ﺏﺁ ﺭﺩ
V/V) ( ﺪﺷ ﻁﻮﻠﺨﻣ ﻲﺑﻮﺧ ﻪﺑ .
ﺭﺎﺼﻋ ﻩ ﻩﺩﺎﻣﺁ
ﻩﺪﺷ ﺸﻟﺎﺑ ﻱﻭﺭ ﺮﺑ ﺘ
ﺯﺍ ﻲﻜ ﻲﻠﻴﻣ ﺞﻨﭘ ﺯﺍ ﺮﺘﻴﻟ
ﻝﻮﻠﺤﻣ ٢٠ ﺪﺻﺭﺩ
ﺯﻭﺮﻛﻮﺳ
W/V) ( ﻢﻳﺪﺳ ﺕﺍﺮﺘﻴﺳﺮﻓﺎﺑ ﺭﺩ ١
/ ٠ ﺭﻻﻮﻣ
pH) ٥ / ٦ (
ﺕﺪﻣ ﻪﺑ ﻭ ٢
ﺎﺗ ٣ ﺖﻋﺮﺳ ﺎﺑ ﺖﻋﺎﺳ ٢٤٠٠٠rpm
ﻩﺎﮕﺘﺳﺩ ﺭﺩ
2. Final extension
3. Supernatant
۴۸ ﻪﻠﺠﻣ
ﮑﺷﺰﭙﻫﺎﻴﮔ ﺶﻧﺍﺩ ﻲ
ﻩﺭﻭﺩ ،ﻥﺍﺮﻳﺍ ۴۰
ﻩﺭﺎﻤﺷ ، ۱ ، ۱۳۸۸
ﮊﻮﻔﻳﺮﺘﻧﺎﺳ
Beckman
ﻝﺪﻣ
Spinco L50
ﺪﺷ ﮊﻮﻔﻳﺮﺘﻧﺎﺳ .
ﺭﺩ ﮊﻮﻔﻳﺮﺘﻧﺎﺳ ﻥﺎﻳﺎﭘ ﺭﺩ ﻩﺪﻣﺁ ﺖﺳﺪﺑ ﮓﻧﺭ ﻱﺮﻴﺷ ﺏﻮﺳﺭ ٣٠٠ ﻢﻳﺪﺳ ﺕﺍﺮﺘﻴﺳ ﺮﻓﺎﺑ ﺯﺍ ﺮﺘﻴﻟﻭﺮﻜﻴﻣ ١
/ ٠ ﺭﻻﻮﻣ
pH) ٥ / ٦ (
ﺭﺰﻳﺮﻓ ﺭﺩ ﻭ ﻞﺣ ٢٠˚C
‐ ﺪﺷ ﻱﺭﺍﺪﻬﮕﻧ .
ﺶﻔﻨﺑ ﺀﺍﺭﻭﺎﻣ ﺏﺬﺟ ﻥﺍﺰﻴﻣ
UV) ( ﺱﻭﺮﻳﻭ ﻥﻮﻴﺴﻧﺎﭙﺳﻮﺳ
ﻱﺎﻫ ﺝﻮﻣ ﻝﻮﻃ ﺭﺩ ﺺﻟﺎﺧ ٢٦٠
ﻭ ٢٨٠ ﻩﺎﮕﺘﺳﺩ ﺭﺩ ﺮﺘﻣﻮﻧﺎﻧ
ﺮﺘﻣﻮﺘﻓﻭﺮﺘﻜﭙﺳﺍ ﻝﺪﻣ LKB
ﺖﻛﺮﺷ ﺖﺧﺎﺳ 4050 Biochrom
ﻱﺮﻴﮔ ﻩﺯﺍﺪﻧﺍ ﻥﺎﺘﺴﻠﮕﻧﺍ ﺖﺒﺴﻧ ﻭ
A260/280
ﻭ ﺖﻈﻠﻏ
ﻝﻮﻣﺮﻓ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﻥﻮﻴﺴﻧﺎﭙﺳﻮﺳ ﺭﺩ ﺱﻭﺮﻳﻭ )
ﺟﺭﺩ ﻪ ﺖﻗﺭ ×
A260/EC (C =
ﺪﺷ ﻪﺒﺳﺎﺤﻣ .
ﻲﭘﻮﻜﺳﻭﺮﻜﻴﻣ ﻥﻭﺮﺘﻜﻟﺍ
ﻪﻧﻮﻤﻧ ﺯﺍ ﻱﺩﺍﺪﻌﺗ ﻱﺎﻫ
ﻪﻋﺭﺰﻣ ﻭ ﻱﺍ ﻪﻧﺎﺨﻠﮔ ﻪﺑ ﻩﺩﻮﻟﺁ ﻱﺍ
ﺱﺮﻳﻭ ﻴﻬﺗ ﻱﺍﺮﺑ ﻪ
ﺲﻜﻋ ﻪﺑ ﻲﻧﻭﺮﺘﻜﻟﺍ ﭖﻮﻜﺳﻭﺮﻜﻴﻣ ﻱﺎﻫ
ﺴﺳﻮﻣ
DSMZ ﻪ ﻥﺎﻤﻟﺁ
ﻝﺎﺳﺭﺍ ﺲﻜﻋ ﻭ ﺪﺷ ﻱﺎﻫ
ﺱﻭﺮﻳﻭ ﺶﺨﺑ ﺭﺩ ﻲﻧﻭﺮﺘﻜﻟﺍ ﭖﻮﻜﺳﻭﺮﻜﻴﻣ ﻲﺳﺎﻨﺷ
ﻥﺁ ﻲﻫﺎﻴﮔ
،ﻪﺴﺳﻮﻣ ﻂﺳﻮﺗ ﺮﺘﻨﻳﻭ ﻥﺎﻔﺘﺳﺍ ﺮﺘﻛﺩ ﻥﺍﺭﺎﻜﻤﻫ ﻭ
ﺳﺍ ﺎﺑ ﻩﺩﺎﻔﺘ
ﻝﺪﻣ ﺲﻳﺍﺯ ﻲﻧﻭﺮﺘﻜﻟﺍ ﭖﻮﻜﺳﻭﺮﻜﻴﻣ ﺯﺍ ۹۰۶
ﺪﺷ ﻪﻴﻬﺗ .
ﻨﻣﺍﺩ ﻲﺳﺭﺮﺑ ﻪ
ﻲﻧﺎﺑﺰﻴﻣ
ﺭﺎﻬﭼ ﺪﺟ ﻳﺍ ﻪ ﺱﻭﺮﻳﻭ ﻥﺍﻮﻨﻌﺑ ﻥﺎﺘﺳﺍ ﻒﻠﺘﺨﻣ ﻖﻃﺎﻨﻣ ﺯﺍ
ﻪﻟﻭﺰﻳﺍ ﺎﻫ ﻨﻣﺍﺩ ﺩﻭﺪﺤﻣ ﻲﺳﺭﺮﺑ ﻱﺍﺮﺑ ﭗﻴﺗ ﻱ ﻪ
ﻲﻧﺎﺑﺰﻴﻣ ﺱﻭﺮﻳﻭ
ﺪﻳﺩﺮﮔ ﺏﺎﺨﺘﻧﺍ ﻥﺍﺮﻬﺗ ﻥﺎﺘﺳﺍ ﺭﺩ ﺐﻟﺎﻏ .
ﺭﺎﺼﻋ ﺭﻮﻈﻨﻣ ﻦﻳﺍ ﻪﺑ ﻩ
ﻪﺑ ﻩﺩﻮﻟﺁ ﻱﺎﻬﮔﺮﺑ ﻟﻭﺰﻳﺍ
ﻪ ﺎﻫ ﺭﺩ ﺭﻮﻛﺬﻣ ﻱ ﻢﻴﺳﺎﺘﭘ ﺕﺎﻔﺴﻓ ﺮﻓﺎﺑ
١ / ٠ ﺭﻻﻮﻣ
pH) ٧ ( ﻘﻳﺮﻃ ﻪﺑ ﻭ ﺝﺍﺮﺨﺘﺳﺍ ﻪ
ﻱﻭﺭ ﻪﺑ ﻲﻜﻴﻧﺎﻜﻣ
ﺍﻳ ﻦ ﻥﺎﻫﺎﻴﮔ ﻪﻳﺎﻣ
ﺪﺷ ﻲﻧﺯ :
Panicum miliaceum
،
Pennisetum americanum
،
Avena sativa
،
Hordeum
vulgare
،
Setaria italica
،
Triticum aestivum
. ﻝﻮﻃ ﺭﺩ
ﺕﺪﻣ ۲۰ ﺯﺍ ﺪﻌﺑ ﺯﻭﺭ ﻪﻳﺎﻣ
ﻧﺯ ﻥﺎﻫﺎﻴﮔ ﻲ ﻪﻳﺎﻣ
ﻧﺯ ﻩﺪﺷ ﻲ ﺮﻈﻧ ﺯﺍ
ﻡﺎﺠﻧﺍ ﺎﺑ ﺰﻴﻧ ﻭ ﻢﻳﻼﻋ ﺭﻮﻬﻇ ﻥﻮﻣﺯﺁ
ﺎﻫ ﺩﺭﻮﻣ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ ﻱ
ﺪﻨﺘﻓﺮﮔ ﺭﺍﺮﻗ ﻲﺳﺭﺮﺑ .
ﺞﻳﺎﺘﻧ
ﻪﻧﻮﻤﻧ ﻱﺭﺍﺩﺮﺑ
ﻊﻳﺎﺷ ﻥﺎﺘﺳﺍ ﺕﺭﺫ ﻉﺭﺍﺰﻣ ﺭﺩ ﻪﻛ ﻲﺳﻭﺮﻳﻭ ﻢﻳﻼﻋ ﻦﻳﺮﺗ
ﻲﻃ ﻥﺍﺮﻬﺗ ﻪﻧﻮﻤﻧ
ﻱﺭﺍﺩﺮﺑ ﻝﺎﺳ ﻱﺎﻫ ١٣٨٢
،ﺪﺷ ﻩﺪﻫﺎﺸﻣ
ﻢﻳﻼﻋ ﻚﻴﺋﺍﺯﻮﻣ ﺩﻮﺑ ﻚﻴﻤﺘﺴﻴﺳ
ﻱﺩﺭﺍﻮﻣ ﺭﺩ ﻪﻛ ﺕﺭﻮﺼﺑ
ﻲﻳﺎﻫﺭﺍﻮﻧ ﻭﺩ ﺎﻳ ﻚﻳ ﺭﺩ1
ﮒﺮﺑ ﻲﻧﺎﻴﻣ ﮒﺮﺒﮔﺭ ﻑﺮﻃ ﺭﺩ ﻭ ﺎﻫ
1. Stripe
ﺩﻮﺑ ﻚﻴﻤﺘﺴﻴﺳ ﻚﻴﺋﺍﺯﻮﻣ ﻪﺑ ﻥﺪﺷ ﻞﻳﺪﺒﺗ ﻝﺎﺣ .
ﻱﺩﺭﺍﻮﻣ ﺭﺩ
ﻪﺗﻮﺑ ﻲﮕﻟﻮﺗﻮﻛ ﻢﻳﻼﻋ ﺎﺑ ﻡﺃﻮﺗ ﻚﻴﺋﺍﺯﻮﻣ ﻢﻳﻼﻋ ﺰﻴﻧ ﺕﺭﺫ ﻱﺎﻫ
2
ﺪﻳﺩﺮﮔ ﻩﺪﻫﺎﺸﻣ .
ﺕﺭﺫ ﻉﺭﺍﺰﻣ ﻲﻣﺎﻤﺗ ﺭﺩ ﺖﺷﺎﻛ ﺩﺭﻮﻣ ﻢﻗﺭ
ﺱﺍﺮﻛ ﻞﮕﻨﻴﺳ ٧٠٤
ﺰﺟ ﻪﺑ ﻭ ﺩﻮﺑ ﻥﺍﺰﻴﻣ ﻪﻛ ﺩﺭﻮﻣ ﺪﻨﭼ
ﻮﻟﺁ ﻥﺍﺰﻴﻣ ﹰﺍﺮﺜﻛﺍ ،ﺩﻮﺑ ﻻﺎﺑ ﻲﮔﺩﻮﻟﺁ ﺩﻮﺑ ﻦﻴﻳﺎﭘ ﻲﮔﺩ
. ﻉﺭﺍﺰﻣ ﺭﺩ
ﻴﺷﺎﺣ ﹰﺎﺗﺪﻤﻋ ﺕﺭﺫ ﻪ
ﻪﺑ ﻭ ﺩﻮﺑ ﻲﮔﺩﻮﻟﺁ ﻲﻠﺻﺍ ﻥﻮﻧﺎﻛ ﻉﺭﺍﺰﻣ
ﻲﻧﺍﻭﺍﺮﻓ ﺯﺍ ﻪﻋﺭﺰﻣ ﺰﻛﺮﻣ ﺖﻤﺳ ﻪﺑ ﺞﻳﺭﺪﺗ ﻪﻧﻮﻤﻧ
ﺎﻫ ﻩﺩﻮﻟﺁ ﻱ
ﻲﻣ ﻪﺘﺳﺎﻛ ﺪﺷ
.
ﻪﻧﻮﻤﻧ ﻱﺭﺍﺪﻬﮕﻧ ﻩﺎﮕﺸﻳﺎﻣﺯﺁ ﺭﺩ ﺎﻫ
ﻪﻧﺎﺨﻠﮔ ﻭ
ﻪﻧﻮﻤﻧ ﻱﺎﻫ ﻪﻋﺭﺰﻣ ﻱﺍ ﺮﺜﻛﺍﺪﺣ ﻲﻧﺎﻣﺯ ﻪﻠﺻﺎﻓ ﺎﺗ ٣
ﺎﺗ ٤ ﻪﺘﻔﻫ
ﻱﺎﻣﺩ ﺭﺩ ٤˚C
ﻝﺎﭽﺨﻳ ﺭﺩ ﻞﺑﺎﻗ
ﻱﺭﺍﺪﻬﮕﻧ ﻭ ﺪﻧﺩﻮﺑ
ﻦﻳﺍ ﺭﺩ
ﺭﺩ ﻲﺒﺳﺎﻨﻣ ﻲﻳﺍﺭﺎﻛ ﺯﺍ ،ﺕﺪﻣ ﻪﻳﺎﻣ
ﻧﺯ ﻭ ﻲ ﻥﻮﻣﺯﺁ ﺎﻫ ﻱ
ﺪﻧﺩﻮﺑ ﺭﺍﺩﺭﻮﺧﺮﺑ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ .
ﻪﻧﻮﻤﻧ ﻥﺩﺍﺩ ﺭﺍﺮﻗ ﺎﻫ
ﺭﺰﻳﺮﻓ ﺭﺩ
ﺭﺰﻳﺮﻓ ﺎﻳ ﻭ ﻲﻟﻮﻤﻌﻣ ٢٠˚C
‐ ﺶﻫﺎﻛ ﺚﻋﺎﺑ ﺪﻳﺪﺷ
ﻑﺬﺣ ﻭ
ﻱﺍﺮﺑ ﺎﻬﻧﺁ ﻲﻳﺍﺭﺎﻛ ﻪﻳﺎﻣ
ﻲﻧﺯ ﻭ ﻥﻮﻣﺯﺁ ﺎﻫ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ ﻱ
ﻲﻣ ﺪﻳﺩﺮﮔ . ﻪﻳﺎﻣ ﻡﺎﺠﻧﺍ ﻲﻧﺯ
ﻚﻨﺧ ﻱﺍﻮﻫ ﺭﺩ ﺎﻫ ﺭﻮﻧ ﺯﺍ ﺭﻭﺩ ﻪﺑ ﻭ
ﻆﻔﺣ ﺰﻴﻧ ﻭ ﺏﺎﺘﻓﺁ ﻢﻴﻘﺘﺴﻣ ﻲﻋﻮﻨﺼﻣ
ﻂﻳﺍﺮﺷ ﻦﻳﺍ )
ﺎﺑ
ﺮﻴﺼﺣ ﻪﻠﻴﺳﻮﺑ ﻱﺍ ﻪﺸﻴﺷ ﻒﻘﺳ ﻥﺪﻧﺎﺷﻮﭘ (
ﺴﺗ ﺐﺟﻮﻣ ﺭﺩ ﻊﻳﺮ
ﻲﻣ ﻢﻳﻼﻋ ﺭﻮﻬﻇ ﺪﺷ
. ﻭ ﻚﻴﺗﻭﺮﻠﻛ ﻁﺎﻘﻧ ﺕﺭﻮﺼﺑ ﻢﻳﻼﻋ ﺭﻮﻬﻇ
ﺪﻋﺎﻗ ﺭﺩ ﻱﺮﺳﺎﺗﺮﺳ ﻭ ﻪﺘﺳﻮﻴﭘ ﺎﻳ ﻭ ﻩﺪﻨﻛﺍﺮﭘ ﻦﺷﻭﺭ ﻩ
ﮒﺮﺑ ﻭﺩ ﺎﺗ ﻚﻳ ﻲﻃ ﺞﻳﺭﺪﺗ ﻪﺑ ﻪﻛ ﺩﻮﺑ ﻩﺪﺷ ﻞﻴﻜﺸﺗ ﻩﺯﺎﺗ ﻱﺎﻫ
ﻭ ﻚﻴﺋﺍﺯﻮﻣ ﻪﺑ ﺪﻌﺑ ﺯﻭﺭ ﭘ
ﻴ
3ﮏﺴ ﻱﺮﺳﺍﺮﺳ )
ﻚﻴﻤﺘﺴﻴﺳ (
ﻲﻣ ﻞﻳﺪﺒﺗ ﺪﺷ
) ﻞﻜﺷ ۱ .(
ﻥﻮﻣﺯﺁ ﺎﻫ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ ﻱ
ﻪﻧﻮﻤﻧ ﻲﺳﺭﺮﺑ ﺎﺑ ﻱﺎﻫ
ﻊﻤﺟ ﺭﻭﺁ ﻝﺎﺳ ﺭﺩ ﻩﺪﺷ ﻱ ١٣٨۲
ﺯﺍ
ﻖﻳﺮﻃ ﻥﻮﻣﺯﺁ ﺎﻫ
ELISA ﻱ
TPIA ،
DIBA ﻭ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﻭ
ﻲﺘﻧﺁ ﻡﺮﺳ ﻱﺎﻫ ﻠﭘ ﻲ ﻝﺎﻧﻮﻠﮐ ٤ ﺱﻭﺮﻳﻭ ﻲﺗﻮﭘ ﺮﻜﺸﻴﻧ ﻚﻴﺋﺍﺯﻮﻣ
(SCMV)
، ﺕﺭﺫ ﻲﮕﻟﻮﺗﻮﻛ ﻚﻴﺋﺍﺯﻮﻣ
(MDMV)
، ﻚﻴﺋﺍﺯﻮﻣ
(JGMV) ﻕﺎﻴﻗ ﻭ
ﻡﻮﮔﺭﻮﺳ ﻚﻴﺋﺍﺯﻮﻣ
(SrMV)
، ﻡﺎﻤﺗ
ﻪﻧﻮﻤﻧ ﻱﺎﻫ ﻊﻤﺟ ﺭﻭﺁ ﺎﻬﻨﺗ ﻩﺪﺷ ﻱ ﺎﺑ
ﻲﺘﻧﺁ ﻡﺮﺳ ﺱﻭﺮﻳﻭ
ﺪﻧﺩﺍﺩ ﻥﺎﺸﻧ ﺶﻨﻛﺍﻭSCMV
ﭻﻴﻫ ﺭﺩ ﻭ ﻪﻧﻮﻤﻧ ﺯﺍ ﻚﻳ
،ﺎﻫ ٣
ﺪﻧﺪﺸﻧ ﻲﻳﺎﺳﺎﻨﺷ ﺮﮕﻳﺩ ﺱﻭﺮﻳﻭ ﻲﺗﻮﭘ .
ﺕﺎﺸﻳﺎﻣﺯﺁ ﺞﻳﺎﺘﻧ
ﺳ ﻪﻧﻮﻤﻧ ﻱﻭﺭ ﺮﺑ ﻲﻜﻳﮊﻮﻟﻭﺮ ﻱﺎﻫ
ﻪﻳﺎﻣ ﻧﺯ ﺪﺷ ﻲ ﻩ ﺭﺩ ﺩﻮﺟﻮﻣ
ﺩﻮﺟﻭ ﺎﻬﻨﺗ ﺰﻴﻧ ﻪﻧﺎﺨﻠﮔ ﺩﺍﺩ ﻥﺎﺸﻧ ﺍﺭ SCMV
) ﻞﻜﺷ ٢ ﻭ ٣ .(
2. Dwarf mosaic
3. Mottling
ﻥﺍﺭﺎﮑﻤﻫ ﻭ ﻱﺪﻤﺤﻣ :
ﻳﺎﺳﺎﻨﺷ ﻭ ﻥﺍﺮﻬﺗ ﻥﺎﺘﺳﺍ ﺕﺭﺫ ﻉﺭﺍﺰﻣ ﺭﺩ ﺐﻟﺎﻏ ﻲﺗﻮﭘ ﻲ ...
۴۹
ﻪﻌﺳﻮﺗ ‐۱ ﻞﻜﺷ
ﻲﻜﻴﺋﺍﺯﻮﻣ ﻱﺎﻫﺭﺍﻮﻧ ﻭ ﻚﻴﻤﺘﺴﻴﺳ ﻚﻴﺋﺍﺯﻮﻣ ﻢﻳﻼﻋ
ﻢﻗﺭ ﻦﻳﺮﻴﺷ ﺕﺭﺫ ﻱﻭﺭ ﺮﺑ ٤٠٣
ﻪﻳﺎﻣ ﻝﺎﺒﻧﺩ ﻪﺑ ﺱﺭﺎﭘ
ﺎﺑ ﻲﻧﺯ
ﻪﻟﻭﺰﻳﺍ ﺎﻫ ﻪﻋﺭﺰﻣ ﻱ
SCMVﻱﺍ
SCMVﻡﺮﺳﻲﺘﻧﺁ ﺎﺑ TPIA ﺎﻳ ﺕﻼﺑﻮﺸﻴﺗ ﻥﻮﻣﺯﺁ ﻪﺠﻴﺘﻧ ‐۲ﻞﻜﺷ ) ﮓﻧﺭ ﺰﻴﻣﺁ ﻱﺍﺮﺘﺴﺑﻮﺳ ﻝﻮﻠﺤﻣ ﺎﺑ ﻱ
NBT/BCIP E5) (
E9 ﺎﺗ :
ﻲﻔﻨﻣ يﺎﻫﺪﻫﺎﺷ و D1
: D2
ﺖﺒﺜﻣ يﺎﻫ ﺪﻫﺎﺷ (
SCMVﻡﺮﺳﻲﺘﻧﺁ ﺎﺑ DIBA ﺎﻳ ﺕﻼﺑ ﺕﺍﺩ ﻥﻮﻣﺯﺁ ﻪﺠﻴﺘﻧ ‐٣ ﻞﻜﺷ ) ﮓﻧﺭ ﺰﻴﻣﺁ ﻱﺍﺮﺘﺴﺑﻮﺳ ﻝﻮﻠﺤﻣ ﺎﺑ ﻱ
Fast Red I3(
I6 ﺎﺗ : ﻱﺎﻫﺪﻫﺎﺷ
ﻲﻔﻨﻣ ) ﻩﺭﺎﺼﻋ ﻩﺎﻴﮔ ﻱﺎﻫ ﻢﻟﺎﺳ
( ﻭ I7
:I8
ﺖﺒﺜﻣ ﻱﺎﻫﺪﻫﺎﺷ :I12
ﺺﻟﺎﺧ ﺱﻭﺮﻳﻭ
ﻲﻠﭘ ﻝﮊ ﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍ ﻞﻳﺮﻛﺍ
ﺁ ﺪﻴﻣ
ﻲﻠﭘ ﻝﮊ ﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍ ﻞﻳﺮﻛﺍ
ﺁ ﺪﻴﻣ
SDS-PAGE
ﺭﺎﺼﻋ ﻩ
ﻥﺎﻫﺎﻴﮔ ﺴﻳﺎﻘﻣ ﻭ ﻩﺩﻮﻟﺁ ﻪ
ﻱﺎﻫﺪﻧﺎﺑ ﺎﺑ ﺎﻬﻧﺁ ﻲﻨﻴﺌﺗﻭﺮﭘ ﻱﺎﻫﺪﻧﺎﺑ
ﻧﻮﻤﻧ ﻲﻨﻴﺌﺗﻭﺮﭘ ﻪ
ﺍﺭ ﻱﻮﻗ ﻲﻓﺎﺿﺍ ﺪﻧﺎﺑ ﻚﻳ ﺩﻮﺟﻭ ،ﻢﻟﺎﺳ ﻩﺎﻴﮔ
ﺖﻴﻌﻗﻮﻣ ﺭﺩ
KDa
٣٨ ‐ ٣٧ ﺩﺍﺩ ﻥﺎﺸﻧ .
ﻡﺎﺠﻧﺍ ﺎﺑ
SDS-PAGE
ﺮﺑ ﻱﻭﺭ ﻩﺩﻮﻣﺁ ﻱﺎﻫ ﻢﻬﻧﺁ ،ﺪﻧﺎﺑ ﻚﻳ ﺎﻬﻨﺗ ﺰﻴﻧ ﺺﻟﺎﺧ ﺱﻭﺮﻳﻭ
ﺖﻴﻌﻗﻮﻣ ﺭﺩ
KDa
٣٨ ‐ ٣٧ ﺪﺷ ﻩﺪﻫﺎﺸﻣ )
ﻞﻜﺷ ٤ (.
ﺕﻼﺑ ﻥﺮﺘﺳﻭ
ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﺕﻼﺑ ﻥﺮﺘﺳﻭ ﻥﻮﻣﺯﺁ ﻡﺎﺠﻧﺍ ﻲﺘﻧﺁ
ﻡﺮﺳ
ﺭﺩ ﺰﻟﻮﻠﺳﻭﺮﺘﻴﻧ ﺀﺎﺸﻏ ﻱﻭﺭ ﺭﺩ ﻱﻮﻗ ﻱﺎﻫﺪﻧﺎﺑ ﺭﻮﻬﻇ ﻭ SCMV
ﺖﻴﻌﻗﻮﻣ
KDa
٣٨
‐ ٣٧ ﺩﻮﺟﻭ ، ﻪﻧﻮﻤﻧ ﺭﺩ ﺍﺭ SCMV
ﻫ ﻱﺎ
ﻥﺯﻭ ﺐﻴﺗﺮﺗ ﻦﻳﺪﺑ ﻭ ﺩﺮﻛ ﺪﻴﻳﺄﺗ ﻩﺩﻮﻟﺁ ﻲﻟﻮﻜﻟﻮﻣ
ﺌﺗﻭﺮﭘ ﻴ ﻦ
ﺸﺷﻮﭘ ﻲ ﻩﺯﺍﺪﻧﺍ ﺰﻴﻧ ﺱﻭﺮﻳﻭ ﺪﺷ ﻱﺮﻴﮔ
. ﻡﺎﺠﻧﺍ
SDS-PAGE
ﻭ
ﻱﻭﺭ ﺮﺑ ﺕﻼﺑ ﻥﺮﺘﺳﻭ ﺪﺟ
ﻪﻳﺍ ﺎﻫ ﻒﻠﺘﺨﻣ ﻱ ﻖﻃﺎﻨﻣ ﺯﺍ SCMV
ﻥﺎﺘﺳﺍ ﻒﻠﺘﺨﻣ ﻲﻟﻮﻜﻟﻮﻣ ﻥﺯﻭ ﺭﺩ ﺍﺭ ﻲﻓﻼﺘﺧﺍ ،ﻥﺍﺮﻬﺗ
ﺌﺗﻭﺮﭘ ﻴ ﻦ
ﺸﺷﻮﭘ ﻲ ﺩﺍﺪﻧ ﻥﺎﺸﻧ ﺎﻬﻧﺁ )
ﻞﻜﺷ ۵ .(
ﻞﻜﺷ ۴
‐ ﺠﻴﺘﻧ ﻪ ﻥﻮﻣﺯﺁ
SDS-PAGE
) ١ ﻭ ٨ : ﻩﺭﺎﺼﻋ ﻩﺎﻴﮔ ﻱﺎﻫ
ﻢﻟﺎﺳ ٢
، ٣ ﻭ ٧ : ﻩﺭﺎﺼﻋ ﻪﺑ ﻩﺩﻮﻟﺁ ﻩﺎﻴﮔ ﻱﺎﻫ ٤SCMV
ﻭ ٥ : ﺱﻭﺮﻳﻭ
ﺺﻟﺎﺧ
(SCMV)
٦ : ﻲﻨﻴﺌﺗﻭﺮﭘ ﺩﺭﺍﺪﻧﺎﺘﺳﺍ ﺮﻛﺭﺎﻣ )
ﻝﻮﺼﺤﻣ
Fermentas
( ( ﻦﻴﻳﺎﭘ ﻪﺑ ﻻﺎﺑ ﺯﺍ :
۱۱۶
‐ ۲ / ۶۶
‐ ۴۵
‐ ۳۵
‐ ۲۵ ﻮﻠﻴﻛ
ﻥﻮﺘﻟﺍﺩ
ﻡﺮﺳﻲﺘﻧﺁ ﺎﺑ EBIA ﺎﻳ ﺕﻼﺑ ﻥﺮﺘﺳﻭ ﻥﻮﻣﺯﺁ ﻪﺠﻴﺘﻧ ‐۵ﻞﻜﺷ ) SCMV
١ : ﻲﻨﻴﺌﺗﻭﺮﭘ ﺩﺭﺍﺪﻧﺎﺘﺳﺍ ﺮﻛﺭﺎﻣ ﻥﺮﺘﺳﻭ ﻪﺑ ﻁﻮﺑﺮﻣ ﻝﮊ ﺯﺍ
ﺕﻼﺑ ، ٥ ﻭ ٦ : ﺺﻟﺎﺧ ﺱﻭﺮﻳﻭ ،
٢ ، ٣ ، ٤ ، ٨ ﻭ ٩ : ﻩﺭﺎﺼﻋ ﻲﻫﺎﻴﮔ ﻱﺎﻫ
ﺱﻭﺮﻳﻭ ﻪﺑ ﻩﺩﻮﻟﺁ
، ٧ : ﺭﺎﺼﻋ ﻩ ﻢﻟﺎﺳ ﻩﺎﻴﮔ (
ﻥﻮﻣﺯﺁ RT-PCR
ﻥﻮﻣﺯﺁ
RT-PCR
ﺮﻤﻳﺍﺮﭘ ﺖﻔﺟ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ
SCMV
ﻭ F3 SCMV R3
ﻲﺻﺎﺼﺘﺧﺍ ﻪﻛ ﻦﻳﺮﺘﺳﺍ ﻭ SCMV
ﻥﺁ ﻱﺎﻫ
ﺩﻮﺟﻭ ،ﺪﻨﺘﺴﻫ ﺍﺭ SCMV
ﺭﺩ ﻪﻧﻮﻤﻧ ﺎﻫ ﺩﺮﻛ ﺪﻴﻳﺎﺗ ﻩﺩﻮﻟﺁ ﻱ .
ﺭﺩ
۵۰ ﻪﻠﺠﻣ
ﮑﺷﺰﭙﻫﺎﻴﮔ ﺶﻧﺍﺩ ﻲ
ﻩﺭﻭﺩ ،ﻥﺍﺮﻳﺍ ۴۰
ﻩﺭﺎﻤﺷ ، ۱ ، ۱۳۸۸
ﻥﻮﻣﺯﺁ ﻝﻮﺼﺤﻣ ﻲﺑﺎﻳﺯﺭﺍ
RT-PCR
ﻲﻘﻓﺍ ﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍ ﻖﻳﺮﻃ ﺯﺍ
ﺯﺭﺎﮔﺁ ﻝﮊ ١
ﻌﻄﻗ ﻪﺑ ﻁﻮﺑﺮﻣ ﺪﻧﺎﺑ ﺪﺻﺭﺩ ﻪ
ﻚﻳﺩﺰﻧ ﻩﺪﺷ ﺮﻴﺜﻜﺗ
bp ﻪﺑ ٩٠٠ ﺪﺷ ﺮﻫﺎﻇ )
ﻞﻜﺷ ٦ ( . ﻲﻟﻭ ﺭﺩ ﻲﺧﺮﺑ ﺎﻫﺪﻧﺎﺑ ﺩﺭﺍﻮﻣ
ﺰﻴﻧ ﻲﺻﺎﺼﺘﺧﺍ ﺮﻴﻏ ﺕﺎﻌﻄﻗ ﻭ ﻩﺪﻫﺎﺸﻣ
ﺖﺸﮔ .
RT-PCRﻥﻮﻣﺯﺁ ﻪﺠﻴﺘﻧ‐٦ﻞﻜﺷ
١ : ﻲﻔﻨﻣ ﺪﻫﺎﺷ )
ﻢﻟﺎﺳ ﻩﺎﻴﮔ ( ، ٢
، ٣
، ٤
، ٥ : ﻪﻧﻮﻤﻧ ﺱﻭﺮﻳﻭ ﻪﺑ ﻩﺩﻮﻟﺁ ﻱﺎﻫ
٦ : ﺺﻟﺎﺧ ﺱﻭﺮﻳﻭ ﻥﻮﻴﺴﻧﺎﭙﺳﻮﺳ
:L1 Kb ﺭﺪﻟ ١ ﻞﺻﺍﻮﻓ ﺎﺑ ٢٥٠bp
) ﺪﻧﺎﺑ ﻦﻴﺑﺎﻣ ﺮﻈﻧ ﺩﺭﻮﻣ ﻱﺎﻫﺪﻧﺎﺑ ٧٥٠
ﻭ ١٠٠٠ ﻩﺪﺷ ﺮﻫﺎﻇ ﺪﻧﺍ (
:L2 Kb ﺭﺪﻟ ١ ﻞﺻﺍﻮﻓ ﺎﺑ ١٠٠bp
ﻱﺯﺎﺳ ﺺﻟﺎﺧ SCMV
ﻩﺪﻣﺁ ﺖﺳﺪﺑ ﺏﻮﺳﺭ ﺺﻟﺎﺧ ﻥﺎﻳﺎﭘ ﺭﺩ
ﻱﺯﺎﺳ ﺭ ﻱﺮﻴﺷ ﮓﻧ
ﮓﻧﺭ ﺯﺍ ﻱﺭﺎﻋ ﻭ ﻧﺍﺩ
ﻪ ﻲﺼﻟﺎﺧﺎﻧ ﻭ ﺰﺒﺳ ﻱﺎﻫ ﻭ ﺩﻮﺑ ﻲﻫﺎﻴﮔ ﻱﺎﻫ
ﻝﮊ ﺭﺩ ﻩﺪﻣﺁ ﺖﺳﺪﺑ ﺺﻟﺎﺧ ﺱﻭﺮﻳﻭ ﻥﻮﻴﺴﻧﺎﭙﺳﻮﺳ ﺯﺭﻮﻓﻭﺮﺘﻜﻟﺍ ﻲﻠﭘ ﻞﻳﺮﻛﺍ ﺁ ﺯﺍ ﻥﺩﻮﺑ ﻱﺭﺎﻋ ﻭ ﺱﻭﺮﻳﻭ ﻱﻻﺎﺑ ﺖﻈﻠﻏ ،ﺪﻴﻣ
ﻦﻴﺌﺗﻭﺮﭘ ﺩﺍﺩ ﻥﺎﺸﻧ ﺍﺭ ﻲﻫﺎﻴﮔ ﻱﺎﻫ )
ﻞﻜﺷ ٨ ( . ﺖﺒﺴﻧ
٢٨٠ / ٢٦٠ ﻩﺩﻮﻣﺁ ﻱﺍﺮﺑ ﺩﻭﺪﺣ ﺭﺩ ﺺﻟﺎﺧ ﺱﻭﺮﻳﻭ ﻱﺎﻫ
٢ / ١
ﺮﻳﻭ ﺖﻈﻠﻏ ﻭ ﺩﻮﺑ ﺯﺍ ﻩﺩﺎﻔﺘﺳﺍ ﺎﺑ ﺰﻴﻧ ﻥﻮﻴﺴﻧﺎﭙﺳﻮﺳ ﺭﺩ ﺱﻭ
ﻝﻮﻣﺮﻓ ٤٥ / ١١ ﺮﺑ ﻡﺮﮔ ﻲﻠﻴﻣ ﻲﻠﻴﻣ
ﺮﺘﻴﻟ ﻪﺒﺳﺎﺤﻣ ﺪﺷ .
ﻲﭘﻮﻜﺳﻭﺮﻜﻴﻣ ﻥﻭﺮﺘﻜﻟﺍ
ﺲﻜﻋ ﺵﻭﺭ ﻪﺑ ﻩﺪﺷ ﻪﻴﻬﺗ ﻲﻧﻭﺮﺘﻜﻟﺍ ﭖﻮﻜﺳﻭﺮﻜﻴﻣ ﻱﺎﻫ
ﺎﺑ ﻥﻮﻴﺳﺍﺭﻮﻛﺩ ﻭ ISEM
ﻲﺘﻧﺁ
SCMV ﻡﺮﺳ ، ﺭﻮﻀﺣ
ﻩﺮﻜﻴﭘ ﻪﺘﺷﺭ ﻱﺎﻫ ﻑﺎﻄﻌﻧﺍ ﻱﺍ
ﻪﻧﻮﻤﻧ ﺭﺩ ﺍﺭ ﺱﻭﺮﻳﻭ ﺮﻳﺬﭘ ﻱﺎﻫ
ﺩﺍﺩ ﻥﺎﺸﻧ ﻩﺩﻮﻟﺁ )
ﻞﻜﺷ ٧
(.
ﻞﻜﺷ ٧
‐ ﻲﻧﻭﺮﺘﻜﻟﺍ ﭖﻮﻜﺳﻭﺮﻜﻴﻣ ﻱﺎﻬﺴﻜﻋ ﺯﺍ ﻩﺪﺷ ﻪﻴﻬﺗ
ﻪﻧﻮﻤﻧ ﻱﺎﻫ
ﻪﺑ ﻩﺩﻮﻟﺁ
SCMV
ﻩﺪﺷ ﻝﺎﺳﺭﺍ ﺴﺳﻮﻣ ﻪﺑ
DSMZ ﻪ ﻥﺎﻤﻟﺁ
ﻨﻣﺍﺩ ﻲﺳﺭﺮﺑ ﻪ
ﻲﻧﺎﺑﺰﻴﻣ SCMV
ﻪﻳﺎﻣ ﺭﺩ ﻲﻧﺯ ۴ ﻪﻳﺍﺪﺟ ﺐﻟﺎﻏ SCMV
ﺭﺩ ﻥﺎﺘﺳﺍ ﻥﺍﺮﻬﺗ
ﻥﺍﻮﻨﻌﺑ ﺪﺟ ﻪﻳﺍ ﺎﻫ ﻱ ﭗﻴﺗ ﻱﻭﺭ ﻪﺑ
Panicum miliaceum
،
Setaria italica
،
Pennisetum americanum
،
Avena Sativa
،
Hordeum Vulgare
،
Triticum Sativum
ﻪﻳﺎﻣ ﻲﻟﻭ ﺪﻳﺩﺮﮔ ﻡﺎﺠﻧﺍ ﺭﺍﺮﻜﺗ ﻭﺩ ﺭﺩ ﻲﻧﺯ ﺮﺑ ﻲﻤﻳﻼﻋ ﻪﻧﻮﮕﭽﻴﻫ
ﻪﻳﺎﻣ ﻥﺎﻫﺎﻴﮔ ﻱﻭﺭ ﺎﺗ ﻩﺪﺷ ﻲﻧﺯ
٢٠ ﻭ ﺪﻳﺩﺮﮕﻧ ﻩﺪﻫﺎﺸﻣ ﺪﻌﺑ ﺯﻭﺭ
ﻪﻧﻮﻤﻧ ﻪﻳﺎﻣ ﻱﺎﻫ ﺭﺩ ،ﻩﺪﺷ ﻲﻧﺯ
ﻥﻮﻣﺯﺁ ﺎﻫ ﺎﺑ ﻲﻜﻳﮊﻮﻟﻭﺮﺳ ﻱ
ﻲﺘﻧﺁ
SCMVﻡﺮﺳ ﺪﻧﺩﻮﺑ ﺶﻨﻛﺍﻭ ﺪﻗﺎﻓ .
ﺚﺤﺑ
ﻲﻃ ﻝﺎﺳ ﻲﻋﺍﺭﺯ ﻞﺼﻓ ١٣٨٢
ﻥﺍﺮﻬﺗ ﻥﺎﺘﺳﺍ ﺕﺭﺫ ﻉﺭﺍﺰﻣ
ﺱﻭﺮﻳﻭ ﻲﺗﻮﭘ ﺭﺎﻬﭼ ﻪﺑ ﻲﮔﺩﻮﻟﺁ ﺮﻈﻧ ﺯﺍ
،SCMV
، DMV
ﻭ SrMV
) JGMV
ﺱﻭﺮﻳﻭ ﻲﺗﻮﭘ ﻪﺑ ﻡﻮﺳﻮﻣ ﺕﻼﻏ ﻱﺎﻫ
(
ﺪﻨﺘﻓﺮﮔ ﺭﺍﺮﻗ ﻲﺳﺭﺮﺑ ﺩﺭﻮﻣ .
ﻊﻳﺎﺷ ﺭﺩ ﻪﻛ ﻲﺳﻭﺮﻳﻭ ﻢﻳﻼﻋ ﻦﻳﺮﺗ
ﻚﺴﻴﭘ ﻭ ﻚﻴﺋﺍﺯﻮﻣ ﻢﻳﻼﻋ ﺪﻳﺩﺮﮔ ﻩﺪﻫﺎﺸﻣ ﺎﻫﺪﻳﺩﺯﺎﺑ ﺩﻮﺑ
ﻭ ﺭﺩ
