Wolbachia Seminar Master [compatibility Mode]

‫داﻧﺸﮕﺎه ﺗﻬﺮان ‪ ،‬ﭘﺮدﻳﺲ ﻛﺸﺎورزي و ﻣﻨﺎﺑﻊ ﻃﺒﻴﻌﻲ‬ ‫ﮔﺮوه ﮔﻴﺎﻫﭙﺰﺷﻜﻲ‬

‫ﺳ ﻨﺎر ﮐﺎر ﻨﺎ ﯽ ار ﺪ‬

‫ا ﺘﺎﺘﺎد را ﺎﻤﺎ ‪ :‬د ﺮ ﻋ ﺿﺎ‬ ‫ﺮﺿﺎ ﻨﻨﺪا ﯽ‬ ‫ا ﺘﺎد ﺸﺎور ‪ :‬د ﺮ و ﯿﯿﺪ‬ ‫ارا ‪ :‬ﻤﺪ آﺑﺎدی‬

‫ﯽ ﻮه‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫ﻣﻘﺪﻣﻪ‪:‬‬

‫‪INTRUDECTION‬‬

‫ﺑﺎﻛﺘﺮي ﻫﺎي درون ﺳﻠﻮﻟﻲ اوﻟﻴﻦ ﺑﺎر درﺑﺎﻓﺖ ﻫﺎي ﺗﻮﻟﻴﺪ ﻣﺜﻠﻲ ﭘﺸﻪ ي‬ ‫ﺗﻮﺳﻂ ﻫﺮﺗﻴﮓ و وﻟﺒﺎخ در ﺳﺎل ‪ 1924‬ﮔﺰارش ﺷﺪﻧﺪ‪ ،‬و اﻳﻦ رﻳﻜﺘﺴﻴﺎﻫﺎ ﻧﻬﺎﻳﺘﺎ‬ ‫ﻧﺎﻣﻴﺪه ﺷﺪﻧﺪ‪.‬‬ ‫‪pipiense‬ﻧﺎ ﺪ‬

‫‪Culex pipiense‬‬ ‫‪Wolbachia‬‬

‫در دﻫﻪ ي ‪ 1950‬ﮔﻠﻠﻮوﻳﭻ و ﻻوان ﭘﻲ ﺑﺮدﻧﺪ ﻛﻪ ﺑﺮﺧﻲ از ﻛﺮاس ﻫﺎي ﺧﺎص درون ﮔﻮﻧﻪ‬ ‫داﺷﺘﻨﺪ و ﺎﻳﺎ‬ ‫ﻧﺘﺎج ﻛﻛﻤﺘﺮي ا‬ ‫ﺗﻮﻟﻴﺪ ﺎ‬ ‫آﻧﻬﺎ ﺎﻳﺎ ﻟ‬ ‫دﻳﮕﺮ آ ﺎ‬ ‫ﺑﻌﺒﺎرت ﮕ‬ ‫‪Culex‬ﻏﻴﺮ ﺎﺑﺎرور ااﺳﺖ‪ ،‬ﺎ‬ ‫‪C l‬‬ ‫ﻫﺎي‬ ‫ااي ﭘﺸﻪ ﺎ‬ ‫اﻳﻨﻜﻪ اﺻﻼ داراي ﻧﺘﺎج ﻧﺒﻮدﻧﺪ‪.‬‬ ‫ﻻوان اﻇﻬﺎر داﺷﺖ ﻛﻪ اﻳﻦ ﻓﺎﻛﺘﻮر ﻧﺎﺳﺎزﻛﺎري داراي اﻟﮕﻮي وراﺛﺖ ﺳﻴﺘﻮﭘﻼﺳﻤﻴﻚ اﺳﺖ‬ ‫)ﺑﻌﺒﺎرت دﻳﮕﺮ از ﻃﺮﻳﻖ ﺟﻨﺲ ﻣﺎده ﻧﻪ از ﻃﺮﻳﻖ ﺟﻨﺲ ﻧﺮ ﺑﻪ ﻧﺴﻞ ﺑﻌﺪ ﻣﻨﺘﻘﻞ ﻣﻲ ﺷﻮد( و‬ ‫ﻛﺮد‬ ‫ﺳﻴﺘﻮﭘﻼﺳﻤﻲ ﻧﺎﻣﮕﺬاري ﻛ د‬ ‫اﻳﻦ ﭘﺪﻳﺪه را ﻧﺎﺳﺎزﮔﺎري ﺳﻴﺘﻮﭘﻼﺳﻤ‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫آﻧﺘﻲ‬ ‫ﻛﺎرﺑﺮد آﻧﺘ‬ ‫را ﺎﺑﺎ ﻛﺎ‬ ‫ﻓﺮﺿﻴﻪ ا‬ ‫ﻣﺮﺗﺒﻂ ااﺳﺖ و ااﻳﻦ ﻓ ﺿ‬ ‫رﻳﻜﺘﺴﻴﺎﻳﻲ ﺗ ﻂ‬ ‫ﻋﻮاﻣﻞ ﻜﺘ ﺎ‬ ‫ﺣﻀﻮر ا ﻞ‬ ‫اﻇﻬﺎر ﻛﻛﺮد ﻛﻛﻪ ‪ CI‬ﺑﺎﺎ ﻀ‬ ‫وﺑﺎر)‪ ،(1971‬اﻇ ﺎ‬ ‫ﻳﻦ ﺎ‬ ‫ﺑﻴﻮﺗﻴﻚ ﺑﻤﻨﻈﻮر ﺣﺬف ﻋﻮاﻣﻞ ﻓﻮق اﺛﺒﺎت ﻛﺮدﻧﺪ‪.‬‬

‫‪INTRUDECTION‬‬

‫ﻣﺸﺎﻫﺪه ﮔﺮدﻳﺪ ﻛﻪ ﻧﺮﻫﺎﻳﻲ از اﺳﺘﺮﻳﻦ ﻫﺎي آﻟﻮده ﺑﺎ ﻣﺎده ﻫﺎﻳﻲ از ﻫﻤﺎن اﺳﺘﺮﻳﻦ ﻛﻪ ﺑﺎ ﻣﻮاد آﻧﺘﻲ‬ ‫ﺑﻴﻮﺗﻴﻜﻲ ﺗﻴﻤﺎر ﺷﺪه اﻧﺪ‪ ،‬ﻧﺎﺳﺎزﮔﺎرﻧﺪ‪ ،‬درﺣﺎﻟﻲ ﻛﻪ ﻛﺮاس ﻋﻜﺲ آن ﺳﺎزﮔﺎز ﺑﻮد)ﻧﺎﺳﺎزﮔﺎري ﻳﻜﻄﺮﻓﻪ(‪.‬‬ ‫ﭘﭘﺲ از آن ﻧﻤﻮﻧﻪ ﻫﺎﻳﻲ از ‪CI‬در ﻃﻴﻒ وﺳﻴﻌﻲ از ﺣﺸﺮات ﺷﺎﻣﻞ ‪ :‬ﺳﻮﺳﻚ آرد‪،‬ﺳﺮﺧﺮﻃﻮﻣﻲ‬ ‫ﻳﻮﻧﺠﻪ‪،‬زﻧﺒﻮرﻫﺎي ﭘﺎرازﻳﺘﻮﺋﻴﺪ‪،‬ﭘﺸﻪ ي ‪Aedes‬و ﻣﮕﺲ ﻣﻴﻮه ﻣﺸﺎﻫﺪه ﮔﺮدﻳﺪ‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫‪INTRUDECTION‬‬

‫‪CI‬اوﻟﻴﻦ ﻣﻮرد ﻛﺸﻒ ﺷﺪه ﺑﻌﻨﻮان ﻋﺎﻣﻞ ﻛﺎﻫﻨﺪه در ﺗﻌﺪاد ﻧﺘﺎج ﺣﺎﺻﻞ از ﻛﺮاس ﺑﻴﻦ ﮔﻮﻧﻪ ﻫﺎي ﻣﺸﺨﺺ ﺑﻮد‬ ‫ﻧﺸﺎن داده ﻣﻲﻲ ﺷﺪ‪.‬‬ ‫ﻛﺮاس‬ ‫س ﻫﺎي ﺑﺑﻌﺪي ن‬ ‫در ﺮ‬ ‫آن ر‬ ‫ﺳﻴﺘﻮﭘﻼﺳﻤﻲ ن‬ ‫ﺗﻮارث ﻴ ﻮﭘ ﻲ‬ ‫و ﻮر‬ ‫در ﺑﺮﺧﻲ ﻣﻮارد‪،‬ﺣﻀﻮر اﻳﻦ ﺑﺎﻛﺘﺮي در ﺗﺨﻤﺪان ﻳﺎ اﻧﺪام ﺗﻨﺎﺳﻠﻲ ﻧﺮ ﺑﺎ روش ﻫﺎي ﻣﻴﻜﺮوﺳﻜﭙﻲ ‪ ،‬ﺑﺎ ﻛﺎرﺑﺮد‬ ‫آﻧﺘﻲ ﺑﻴﻮﺗﻴﻚ ﻫﺎ ﻳﺎ ﺑﺎ ﺣﺮارت دﻫﻲ ﺗﺎﻳﻴﺪ ﺷﺪ‪ .‬ﺑﺎ اﻳﻦ ﺣﺎل ارﺗﺒﺎﻃﺎت ﻓﻴﻠﻮژﻧﺘﻴﻚ ﺑﻴﻦ ﺑﺎﻛﺘﺮي ﻫﺎي ‪CI‬ﻳﺎﻓﺖ‬ ‫ﻧﺎﺷﻨﺎﺧﺘﻪ ﺑﻮد‪.‬‬ ‫ﻣﺜﻠﻲ‬ ‫اواﻳﻞ دﻫﻪ ي ‪ 1990‬ﺎ ﺎ‬ ‫ﻣﺨﺘﻠﻒ ﺎﺗﺎ ا ا ﻞ‬ ‫ﻣﻴﺰﺑﺎن ﻠ‬ ‫ﺣﺸﺮات ﺎ‬ ‫ا‬ ‫ﺗﻮﻟﻴﺪ ﺜﻠ‬ ‫ﻫﺎي ﻟ‬ ‫ﺷﺪه در ﺎﺑﺎﻓﺖ ﺎ‬ ‫‪ 16‬ﻧﺸﺎن داد ﻛﻪ وﻟﺒﺎﺧﻴﺎ داراي ارﺗﺒﺎط ﻣﻮﻧﻮﻓﻴﻠﻴﺘﻴﻚ ﺑﺎ ﺳﺎﻳﺮ رﻳﻜﺘﺴﻴﺎ ﻫﺎ‬ ‫‪16SrDNA‬‬ ‫ﻓﻴﻠﻮژﻧﻲ ﺑﺮ اﺳﺎس ﺗﻮاﻟﻲ ‪SrDNA‬‬ ‫‪16‬ﻣﻲﻲ ﺑﺑﺎﺷﺪ‪.‬‬ ‫‪16SrDNA‬‬ ‫ﻲ ‪SrDNA‬‬ ‫ﺗﻮاﻟﻲ‬ ‫ﻳﻲ درر ﻮ‬ ‫واﮔﺮاﻳﻲ‬ ‫ﺣﺪود ‪ %2‬و ﺮ‬ ‫زﻳﺮﺷﺎﺧﻪ)‪ (A & B‬ﺑﺑﺎ و‬ ‫داراي دوو زﻳﺮ‬ ‫ﺲ ‪ Wolbachia‬ر‬ ‫ﻫﺴﺘﻨﺪ‪ .‬ﺟﺟﻨﺲ‬ ‫وﺟﻮد اﻳﻦ دو زﻳﺮ ﺷﺎﺧﻪ ﻫﻤﭽﻨﻴﻦ ﺑﺎ ژ ن ﻫﺎي ﻛﺪ ﻛﻨﻨﺪه ي ﭘﺮوﺗﺌﻴﻦ ﻧﻴﺰ ﺗﺎﻳﻴﺪ ﺷﺪﻧﺪ‪.‬‬ ‫‪ ،Wolbachia pipientis‬ﻳﻚ ﮔﻮﻧﻪ ي اﻟﻘﺎ ﻛﻨﻨﺪﻫﻲ ﻧﺎﺳﺎزﮔﺎري ﺳﻴﺘﻮﭘﻼﺳﻤﻲ اﺳﺖ ﻛﻪ ﮔﻮﻧﻪ ي ﺗﻴﭗ ﺟﻨﺲ‬ ‫‪ Wolbachia‬ﻣﻲ ﺑﺎﺷﺪ و ﻣﺘﻌﻠﻖ ﺑﻪ ﺷﺎﺧﻪ ي ‪ B‬در اﻳﻦ ﺟﻨﺲ اﺳﺖ‪.‬‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫‪WOLBACHIA‬‬ ‫ﺑﺎﻛﺘﺮي ﻫﺎي ﺟﻨﺲ ‪Wolbachia‬رﻳﻜﺘﺴﻴﺎﻫﺎﻳﻲ ﻫﺴﺘﻨﺪ ﻛﻪ از ﻃﺮﻳﻖ ﺳﻴﺘﻮﭘﻼﺳﻢ اﻧﺘﻘﺎل ﭘﻴﺪا ﻣﻲ ﻛﻨﻨﺪ و در‬ ‫ﺑﺎﻓﺖ ﻫﺎي ﺗﻮﻟﻴﺪ ﻣﺜﻠﻲ )ﺑﻴﻀﻪ ﻫﺎ و ﺗﺨﻤﺪان ﻫﺎ( ﻣﺤﺪوده ي وﺳﻴﻌﻲ از ﺑﻨﺪﭘﺎﻳﺎن وﺟﻮد دارﻧﺪ‪.‬‬ ‫در ﻣﻮارد اﺳﺘﺜﻨﺎﻳﻲ در ﺑﺎﻓﺖ ﻫﺎي ﺳﻮﻣﺎﺗﻴﻚ ﻧﻴﺰ ﻣﺸﺎﻫﺪه ﺷﺪه اﺳﺖ)ﺑﺎﻓﺖ ﻋﺼﺒﻲ ﻣﮕﺲ ﺳﺮﻛﻪ و ﺑﺎﻋﺚ‬ ‫ﻣﺮگ ﻗﺒﻞ از ﺑﻠﻮغ ﻣﻲ ﺷﻮﻧﺪ(‬ ‫وﻟﺒﺎﺧﻴﺎ ﻣﺎﻧﻨﺪ ﺳﺎﻳﺮ ﻣﻮﺟﻮدات زﻧﺪه ﺧﻮاﻫﺎن ﺑﻬﺮه ﮔﻴﺮي از ﻣﺤﻴﻂ زﻧﺪﮔﻲ ﺧﻮد ﺑﺮاي ﺣﺪاﻛﺜﺮ ﻛﺎراﻳﻲ ﻣﻲ‬ ‫ﺑﺎﺷﻨﺪ‪ .‬ﺑﻪ اﻳﻦ ﻣﻨﻈﻮر‬ ‫اﻳﻦ ﺑﺎﻛﺘﺮي در ﻣﻴﺰﺑﺎن ﺧﻮد ﺗﻐﻴﻴﺮاﺗﻲ را ﺑﻮﺟﻮد ﻣﻲ آورد ﺗﺎ ﺑﺘﻮاﻧﺪ ﺟﻤﻌﻴﺖ ﺧﻮد را اﻓﺰاﻳﺶ داده و ﺑﻪ‬ ‫ﭘﺮاﻛﻨﺶ ﺑﻴﺸﺘﺮي دﺳﺖ‬

MECHANISM OF WOLBACHIA

WOLBACHIA

MECHANISM OF WOLBACHIA

STRAIN

:‫اﺳﺘﺮﻳﻦ ﻫﺎي وﻟﺒﺎﺧﻴﺎ‬

‫ ﮔﮔﺎﻧﻪ اي ﺑﺮاي اﻳﻦ‬6 ‫ اﺳﺘﺮﻳﻦ ﻫﺎي‬16 16S S rDNA ‫ و‬fts Z ‫ﺑﺮاﺳﺎس ﺗﻌﻴﻴﻦ ﺗﻮاﻟﻲ ژن ﻫﺎي‬ .(A-F) ‫ﺑﺎﻛﺘﺮي ﺗﻌﻴﻴﻦ ﮔﺮدﻳﺪه اﺳﺖ‬

OF

A&B C&D E F

Arthropods

No specialist, Nonessential

Nematodes

Specialist, Essential

WOLBACHIA

Springtails

Termites

Unknown

‫‪MECHANISM OF WOLBACHIA‬‬

‫اﺳﺘﺮﻳﻦ ﻫﺎي وﻟﺒﺎﺧﻴﺎ‪:‬‬

‫ﺷﻜﻞ ‪ .1‬ﺑﺮاﺳﺎس ﺗﻌﻴﻴﻦ ﺗﻮاﻟﻲ ژن ﻫﺎي ‪ fts Z‬و ‪ 16S rDNA‬اﺳﺘﺮﻳﻦ ﻫﺎي ‪ 6‬ﮔﺎﻧﻪ اي ﺑﺮاي اﻳﻦ ﺑﺎﻛﺘﺮي ﺗﻌﻴﻴﻦ ﮔﺮدﻳﺪه‬ ‫اﺳﺖ )‪.(A-F‬‬

MECHANISM OF WOLBACHIA

STRAIN OF

: D & C ‫ﮔﺮوه‬

‫ﻫﻤﺰﻳﺴﺘﻲ‬ ( Mutualism) utua s ) ‫ﻲ‬ ‫راﺑﻄﻪ ﻤﺰﻳ‬ ‫داراي ر ﺑ‬ ‫ﺧﻮد ر ي‬ ‫ﻣﻴﺰﺑﺎنن ﻮ‬ ‫ﺑﺑﺎ ﻴﺰﺑ‬

•

‫ﻓﻘﻂ در ﻧﻤﺎﺗﺪ ﻫﺎ دﻳﺪه ﺷﺪه اﺳﺖ‬

•

Onchoceridea or filaria nematodes (pathogen in mamales mamales))

‫ﺑﺮاي ﻣﻴﺰﺑﺎن ﺧﻮد ﺿﺮوري اﺳﺖ‬

•

‫ﺗﺨﺼﺼﻲ ﻫﺴﺘﻨﺪ‬

•

ovaries Embryos and hypoderms ‫آﻟﻮدﮔﻲ‬ ovaries, ‫ﺑﺎﻋﺚ آﻟ دﮔ‬ ‫ﺎﻋﺚ‬

•

(vertical transmission ) ‫ﻓﻘﻂ اﻧﺘﻘﺎل ﻋﻤﻮدي‬

•

(embryogenesis . Larval development & adult survival )

WOLBACHIA

MECHANISM OF WOLBACHIA

D & C groups : Pre-treatment

Wolbachia in oocytes

Post-treatment empty uterus

degenerated oocytes

Figure2 Figure 2 . Vertical transverse of ovaries in a filarial nematode before and after tetracycline treatment and then staining

‫‪MECHANISM OF WOLBACHIA‬‬

‫‪STRAIN‬‬

‫ﮔﺮوه ﻫﺎي ‪ A‬و ‪: B‬‬

‫•‬ ‫•‬ ‫•‬ ‫•‬

‫ﺑﻨﺪ ﭘﺎﻳﺎن و ﺑﺨﺼﻮص ﺣﺸﺮات را آﻟﻮده ﻣﻲ ﻛﻨﻨﺪ‬ ‫ﺑﺮاي ﻣﻮﺟﻮد ﺿﺮوري ﻧﻴﺴﺘﻨﺪ ) اﺳﺘﺜﻨﺎ‪ .‬ﺗﺎﻛﻨﻮن دو ﻣﻮرد اﻳﻦ ﺑﺎﻛﺘﺮي در ﺣﺸﺮات ﺿﺮوري ﮔﺰارش‬ ‫‪.(( Coccotrypes‬‬ ‫‪C‬‬ ‫‪d‬‬ ‫‪dactyliperda‬‬ ‫)‪li d (col‬‬ ‫‪ Asorbara‬و )‪( l‬‬ ‫‪A b‬‬ ‫‪tabida,‬‬ ‫‪tabida‬‬ ‫‪bid , Hym‬‬ ‫‪H‬‬ ‫ﺷﺪه اﺳﺖ‪:‬‬ ‫در دوﮔﻮﻧﻪ ي ﻣﺬﻛﻮر ﻣﺎده ﻫﺎي ﺗﻴﻤﺎر ﺷﺪه ﺑﺎ آﻧﺘﻲ ﺑﻴﻮﺗﻴﻚ ﻗﺎدر ﺑﻪ ﺗﺨﻤﮕﺬاري ﻧﻴﺴﺘﻨﺪ‬ ‫ﺗﻐﻴﻴﺮ ﻣﻣﻲ دﻫﻨﺪ‬ ‫ﻣﻜﺎﻧﻴﺴﻢ ﻫﺎي ﻣﺘﻌﺪدي ﺗﻐ‬ ‫ﻣﻴﺰﺑﺎن را ﺑﺎ ﻣﻜﺎﻧ‬ ‫ﺗﻮﻟﻴﺪ ﻣﺜﻞ ﻣ ﺰﺑﺎن‬ ‫ﺗﻮﻟ ﺪ‬ ‫ﺑﻨﻈﺮ ﻧﻤﻲ رﺳﺪ ﻛﻪ ﻣﻴﺰﺑﺎن ﺗﺨﺼﺼﻲ داﺷﺘﻪ ﺑﺎﺷﻨﺪ‬ ‫ﺑﻌﻨﻮان ﻣﺜﺎل ‪:‬‬ ‫ﺑ ﻮ‬

‫‪OF‬‬

‫‪WOLBACHIA‬‬

‫)‪Leptopilina heterotoma (parasitic wasp‬‬ ‫)‪Drosophila simulanus (host‬‬ ‫‪host),‬‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫اﻧﺘﻘﺎل اﻓﻘﻲ ﻳﺎ ﺑﻴﻦ ﺗﺎﻛﺴﻮﻧﻲ )‪(Horizontal / Intertaxon Transmission‬‬

‫‪TRANSSMITION‬‬

‫ﻫﺴﺘﻨﺪ‪.‬‬ ‫وﻟﺒﺎﺧﻴﺎ ﻫ ﺘﻨﺪ‬ ‫ﺑﺎﻛﺘﺮي ﻟ ﺎﺧ ﺎ‬ ‫اﻓﻘﻲ ﺎﻛﺘ ي‬ ‫ﺑﺨﻮﺑﻲ ﻧﺸﺎن دﻫﻨﺪه ي اﻧﺘﻘﺎل اﻓﻘ‬ ‫ﻣﻮﻟﻜﻮﻟﻲ ﺨ‬ ‫ﺎت ﻟﻜ ﻟ‬ ‫ﻣﻄﺎﻟﻌﺎت‬ ‫ﻄﺎﻟ‬ ‫ﻳﻜﻲ از اﺳﺘﺮﻳﻦ ﻫﺎي ﺷﺎﺧﻪ ‪A‬اﻳﻦ ﺑﺎﻛﺘﺮي )‪ (Adm‬داراي اﻧﺘﻘﺎل اﻓﻘﻲ ﺑﺴﻴﺎروﺳﻴﻌﻲ اﺳﺖ‪.‬‬ ‫‪ Coleoptera‬و ‪.Hymenoptera‬‬ ‫‪Coleoptera,,‬‬

‫‪Lepidoptera‬‬

‫اﻧﺘﻘﺎل اﻓﻘﻲ در ‪ B-Wolbachia‬ﻧﻴﺰ ردﻳﺎﺑﻲ ﺷﺪه اﺳﺖ ﺑﻄﻮري ﻛﻪ در ﺑﺎﻛﺘﺮي ﻫﺎي ﺟﺪا ﺷﺪه از زﻧﺒﻮر‬ ‫ﻚ ﺷﺷﺠﺮه ااي‬ ‫ﻧﺰدﻳﻚ‬ ‫ﺑﺴﻴﺎر ﻧﺰ‬ ‫ﺖ ﺎ‬ ‫ﻗﺮاﺑﺖ‬ ‫داراي ﻗ ا‬ ‫ﻣﻴﺰﺑﺎﻧﺶ )‪ (Protocalliphora‬ا ا‬ ‫ﻣﮕﺲ ﺰ ﺎﻧﺶ‬ ‫ﭘﺎرازﻳﺘﻮﺋﻴﺪ ‪Nasonia giraulti‬و ﮕ‬ ‫ﺎ از ﺘ ﺋ ﺪ‬ ‫ﻫﺴﺘﻨﺪ‪.‬‬ ‫ﻣﻜﺎﻧﻴﺴﻢﻢ ﺒﺗﺒﺎدل اﻳﻦ‬ ‫ﻣﻴﺰﺑﺎﻧﺶ ﺑﺑﻪ ﻋﻨﻮان ﻳﻚ ﻴ‬ ‫ﺗﺎﻛﺴﻮﻧﻲ ﭘﭘﺎرازﻳﺖ و ﻴ ﺑ‬ ‫ﻲ‬ ‫ﻣﻴﺪارد ﻛﻪ اﻧﺘﻘﺎل ﺑﻴﺑﻴﻦ‬ ‫اﻇﻬﺎر ﻴ‬ ‫اﻳﻦ ﻳﺎﻓﺘﻪ ﻬ‬ ‫ﺑﺎﻛﺘﺮي اﻣﻜﺎن ﭘﺬﻳﺮ اﺳﺖ‪ .‬دﻳﮕﺮ روش ﻫﺎي داراي ﭘﺘﺎﻧﺴﻴﻞ اﻳﻦ ﺗﺒﺎدل ﺷﺎﻣﻞ ﺷﻜﺎرﮔﺮو ﺷﻜﺎر ﻣﻴﺒﺎﺷﺪ‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫ﭘﺮاﻛﻨﺶ ﺑﺎﻛﺘﺮي‬ ‫ﺳﺎل ‪ 1997‬ااز‬ ‫ﺎﺗﺎ ﺎل‬

‫وﻟﺒﺎﺧﻴﺎ )‪(Distribution‬‬

‫‪DISTRIBUTION‬‬

‫‪ 80‬ﮔﻮﻧﻪ ﺣﺸﺮه‬ ‫‪ 17‬ﮔﻮﻧﻪ ‪Isopode‬‬ ‫ﻳﻚ ﮔﻮﻧﻪ ﻛﻨﻪ‬ ‫ﻳﻚ ﮔﻮﻧﻪ ﻧﻤﺎﺗﺪ‬ ‫ﮔﺰارش ﺷﺪه اﺳﺖ‪...‬‬

‫ﻣﺤﺪودﻳﺘﻲ ﺑﺮاي ﭘﺮاﻛﻨﺶ اﻳﻦ ﺑﺎﻛﺘﺮي ﻫﺎ ﺷﻨﺎﺧﺘﻪ ﻧﺸﺪه اﺳﺖ !!!‬ ‫)آﻣﺎر ﻣﺮﺑﻮط ﺑﻪ ﺳﺎل‪1997‬ﻣﻲ ﺑﺎﺷﺪ و ﻣﺴﻠﻤﺎ ﺗﺎ ﻛﻨﻮن ﺑﺮ ﺗﻌﺪاد اﻳﻦ ﻣﻴﺰﺑﺎن ﻫﺎ اﻓﺰوده ﺷﺪه اﺳﺖ(‪.‬‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫ﺑﺮرﺳﻲ اﺧﻴﺮ ﺣﺸﺮات ‪ neotropical‬وﺟﻮد اﻳﻦ ﺑﺎﻛﺘﺮي را در ﺣﺪود ‪ 16‬درﺻﺪ ﮔﻮﻧﻪ ﻫﺎ‬ ‫را از راﺳﺘﻪ ﻫﺎي‪:‬‬

‫‪DISTRIBUTION‬‬

‫‪Coleoptera‬‬

‫‪Diptera‬‬ ‫‪Hemiptera/Homoptera‬‬ ‫‪Hymenoptera‬‬ ‫‪Lepidoptera‬‬ ‫‪O h‬‬ ‫‪Orthoptera‬‬

‫ﺣﺸﺮات‬ ‫)ﺑﺮرﺳﻲ ﺸ ا‬

‫‪neotemprate‬‬

‫ﺑﺎﻻرا ﺗﺎﺗﺎﻳﻴﺪ ﻛﻛﺮد((‪.‬‬ ‫ﻧﺘﺎﻳﺞ ﺎﻻ ا‬ ‫ﻧﻧﻴﺰ ﻧﺘﺎ‬

‫ﺳﺎﻳﺮ ﻧﻮاﺣﻲ ﺟﻐﺮاﻓﻴﺎﻳﻲ ؟؟؟‬ ‫‪ 10-30‬ﻣﻴﻠﻴﻮن ﮔﻮﻧﻪ ﺣﺸﺮات ﺑﻨﺎﺑﺮاﻳﻦ ﺗﻌﺪاد ﺣﺸﺮات آﻟﻮده ﺣﺪود ‪ 1.5-5‬ﻣﻴﻠﻴﻮن ﮔﻮﻧﻪ‬ ‫اﻳﻦ ﭘﺮاﻛﻨﺶ ﺑﺎﻛﺘﺮي وﻟﺒﺎﺧﻴﺎ را ﻓﺮاواﻧﺘﺮﻳﻦ ﺑﺎﻛﺘﺮي درﻣﻴﺎن ﺑﺎﻛﺘﺮي ﻫﺎي ﭘﺎرازﻳﺖ ﻗﺮار ﻣﻲ دﻫﺪ‪.‬‬

DIIST TRI IBU UTIION N

MECHANISM OF WOLBACHIA

‫ﻧﻤﺎﻳﻲ از اﻧﺘﻘﺎل ﺑﺎﻛﺘﺮي وﻟﺒﺎﺧﻴﺎ ﺗﻮﺳﻂ ﺟﻨﺲ ﻣﺎده در زﻧﺒﻮر‬

Nasonia vitripennis

‫‪MECHANISM OF WOLBACHIA‬‬

‫ﻧﺎﺳﺎﮔﺎري ﺳﻴﺘﻮﭘﻼﺳﻤﻲ‬

‫)‪(Cytoplasmic Incompatibility‬‬

‫‪CYTOPLASMIC‬‬ ‫ﻧﺎﺳﺎزﮔﺎري ﺣﺎﺻﻞ ازوﻟﺒﺎﺧﻴﺎ)اﺳﺘﺮﻳﻦ (‪ ،‬ﻳﻚ ﻧﺎﺳﺎزﮔﺎري ﺗﻮﻟﻴﺪﻣﺜﻠﻲ ﺑﻴﻦ اﺳﭙﺮم و ﺗﺨﻢ اﺳﺖ ﻛﻪ ﺑﻄﻮر ﻣﻌﻤﻮل در‬ ‫ﮔﻮﻧﻪ ﻫﺎي دﻳﭙﻠﻮﺋﻴﺪ ﻣﻨﺠﺮ ﺑﻪ ﻣﺮگ زﻳﮕﻮت و در ﮔﻮﻧﻪ ﻫﺎي ﻫﺎﭘﻠﻮ‪ -‬دﻳﭙﻠﻮﺋﻴﺪ ﻣﻨﺠﺮ ﺑﻪ ﻧﺮزاﻳﻲ ﻣﻲ ﮔﺮدد‪.‬‬ ‫‪INCOMPATIBILITUY‬‬ ‫‪CI‬‬

MECHANISM OF WOLBACHIA

• The simple rule of cytoplasmic incompatibility (CI) is...

if the male has an infection that is not p present in his mate, it’s an incompatible cross

X

W

CI

MECHANISM OF WOLBACHIA

W

CYTOPLASMIC W

X

W

Normal egg hatch rate

Normal egg hatch rate

X

INCOMPATIBILITUY X

W

Reduced egg hatch rate

C TOPLAS CYT SMI IC INCO I OMPAT TIB BILI ITUY Y

MECHANISM OF WOLBACHIA

(Nasonia vitripennis) ‫ﻧﺤﻮه ي اﻳﺠﺎد ﻧﺎﺳﺎزﮔﺎري ﻧﺎﺷﻲ از وﻟﺒﺎﺧﻴﺎ‬

C TOPLAS CYT SMI IC INCO I OMPAT TIB BILI ITUY Y

MECHANISM OF WOLBACHIA

Here is what a normal set of chromosomes h

But in an embryo resulting from an incompatible cross (ie., a CI mating) the p paternal chromosomes become abnormally heterochromatic, forming a ‘chromosomal clump.’

C TOPLAS CYT SMI IC INCO I OMPAT TIB BILI ITUY Y

MECHANISM OF WOLBACHIA

The ‘clump’ fails to form chromosomes, leading to Developmental arrest (ie., death) of diploid host organisms (e.g., mosquitoes).

In haplo-diploid insects, the loss of paternal chromosomes due to CI is not lethal, but leads to all male broods. all-male broods

C TOPLAS CYT SMI IC INCO I OMPAT TIB BILI ITUY Y

MECHANISM OF WOLBACHIA

‫ﺑﺎﻛﺘﺮي ﻫﺎ از ﻃﺮﻳﻖ ﺗﺨﻢﻢ ﻫﺎ )ﺟﻨﺲ ﻣﺎده( اﻧﺘﻘﺎل ﻣﻲ ﻳﺎﺑﻨﺪ و ﻧﻪ از ﻃﺮﻳﻖ اﺳﭙﺮمم )ﺟﻨﺲ ﻧﺮ(‪،‬اﻟﺒﺘﻪ‬ ‫اﺳﺘﺜﻨﺎﻫﺎﻳﻲ ﻧﻴﺰ وﺟﻮد دارد ﻛﻪ در ﻣﻮارد ﻧﺎدر اﻧﺘﻘﺎل از ﻃﺮﻳﻖ اﺳﭙﺮم ﻧﻴﺰ ﺻﻮرت ﻣﻲ ﭘﺬﻳﺮد‪.‬‬

‫ﻧﺤﻮه ي ﺗﺎﺛﻴﺮ وﻟﺒﺎﺧﻴﺎ در اﺳﭙﺮﻣﺎﺗﻮژﻧﺰ در ﻧﺮ آﻟﻮده و ﺗﻮﻟﻴﺪ اﺳﭙﺮم آﻟﻮده‬

‫‪C TOPLAS‬‬ ‫‪CYT‬‬ ‫‪SMI‬‬ ‫‪IC‬‬ ‫‪INCO‬‬ ‫‪I‬‬ ‫‪OMPAT‬‬ ‫‪TIB‬‬ ‫‪BILI‬‬ ‫‪ITUY‬‬ ‫‪Y‬‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫اﻧﻮاع ﻧﺎﺳﺎزﮔﺎري‪:‬‬

‫‪CYTOPLASMIC‬‬

‫‪ .1‬ﺗﻚ ﺳﻮﻳﻪ )‪(Monodirectional‬‬ ‫ﻧﺎﺳﺎزﮔﺎري ﺗﻚ ﺳﻮﻳﻪ ﺑﻄﻮر ﻣﻌﻤﻮل زﻣﺎﻧﻲ رخ ﻣﻲ دﻫﺪ ﻛﻪ اﺳﭙﺮم ﺣﺎﺻﻞ از ﻧﺮآﻟﻮده ﺑﻪ‬ ‫وﻟﺒﺎﺧﻴﺎ ﺑﺎ ﻳﻚ ﺳﻠﻮل ﺗﺨﻢ ﻏﻴﺮآﻟﻮده ﺗﻠﻘﻴﺢ اﻧﺠﺎم دﻫﺪ )ﺗﻠﻘﻴﺢ درﻋﻜﺲ اﻳﻦ ﺣﺎﻟﺖ‪ ،‬ﻳﻌﻨﻲ‬ ‫اﺳﭙﺮم ﻧﺮ ﻏﻴﺮ آﻟﻮده ﺑﺎ ﺳﻠﻮل ﺗﺨﻢ آﻟﻮده‪ ،‬ﻛﺎﻣﻼ ﺳﺎزﮔﺎر اﺳﺖ(‪.‬‬

‫‪ .2‬دوﺳﻮﻳﻪ )‪(Bidirectional‬‬ ‫ﻧﺎﺳﺎزﮔﺎري دوﺳﻮﻳﻪ زﻣﺎﻧﻲ اﺗﻔﺎق ﻣﻲ اﻓﺘﺪ ﻛﻪ ﻫﺮدو ﻓﺮد ﻧﺮو ﻣﺎده ﺑﻪ اﻳﻦ ﺑﺎﻛﺘﺮي )اﻟﺒﺘﻪ‬ ‫ﺳﻮﻳﻪ ﻫﺎي ﻣﺨﺘﻠﻒ( آﻟﻮده ﺑﺎﺷﻨﺪ و اﺳﭙﺮم و ﺗﺨﻤﻚ ﺣﺎﺻﻞ از اﻳﻦ اﻓﺮاد ﺑﺎ ﻳﻜﺪﻳﮕﺮ ﻛﺮاس‬ ‫اﻧﺠﺎم دﻫﻨﺪ) اﮔﺮﻫﺮدو ﻓﺮد ﺑﺎ ﺳﻮﻳﻪ ﻫﺎي ﻳﻜﺴﺎﻧﻲ از اﻳﻦ ﺑﺎﻛﺘﺮي آﻟﻮده ﺑﺎ ﺷﻨﺪ ‪ ،‬ﺗﻠﻘﻴﺢ ﺑﻴﻦ‬ ‫اﺳﺖ((‪.‬‬ ‫ﺳﺎزﮔﺎر ا ﺖ‬ ‫ﻛﺎﻣﻼ ﺎزﮔﺎ‬ ‫ﻫﺎ ﻛﺎ ﻼ‬ ‫آن ﺎ‬

‫‪INCOMPATIBILITUY‬‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫‪Monodirectional CI‬‬ ‫در ﻧﺎﺳﺎزﮔﺎري ﻳﻚ ﺳﻮﻳﻪ‬ ‫ﺣﺸﺮه ﻧﺮ ﻳﺎ ﻣﺎده و ﻳﺎ‬ ‫اﺳﺘﺮﻳﻦ‬ ‫ﺗﻨﻬﺎﺎ ﺑﺎ ﻳﻚ ا ﺘ ﻳ‬ ‫ﻫﺮدو ﺗﻨ‬ ‫ﻫ د‬ ‫وﻟﺒﺎﺧﻴﺎ آﻟﻮده ﻫﺴﺘﻨﺪ‪.‬‬ ‫ﻧﺎﺳﺎزﮔﺎري ﻳﻚ ﺳﻮﻳﻪ‬

‫‪CYTOPLASMIC‬‬ ‫‪INCOMPATIBILITUY‬‬

MECHANISM OF WOLBACHIA

Bidi ti Bidirectional l CI ‫در ﻧﺎﺳﺎزﮔﺎري دو ﺳﻮﻳﻪ‬ ‫ﺣﺸﺮه ﻧﺮﺮ و ﻣﺎده ﺑﺑﺎ‬ ‫ﺮ‬ ‫دواﺳﺘﺮﻳﻦ ﻣﺨﺘﻠﻒ از وﻟﺒﺎﺧﻴﺎ‬ .‫آﻟﻮده ﻫﺴﺘﻨﺪ‬

CYTOPLASMIC INCOMPATIBILITUY

MECHANISM OF WOLBACHIA

Additive Incompatibility p y

An insect can be infected with more than one infection type...

‫ﺣﺎﻟﺘﻲ ﻛﻪ در آن ﺣﺸﺮات ﻧﺮو ﻣﺎده‬ ‫ﺑﺎ ﺑﻴﺶ از ﻳﻚ اﺳﺘﺮﻳﻦ وﻟﺒﺎﺧﻴﺎ آﻟﻮده‬ .‫ﺘﻨﺪ‬ ‫ﻫﻫﺴﺘﻨﺪ‬ .... ‫ﺑﺎ اﺗﻜﺎ ﺑﻪ ﻗﺎﻧﻮن اﺻﻠﻲ ﻧﺎﺳﺎزﮔﺎري‬

CYTOPLASMIC INCOMPATIBILITUY CO U

‫‪MECHANISM OF WOLBACHIA‬‬

‫‪Additi Incompatibility‬‬ ‫‪Additive‬‬ ‫‪I‬‬ ‫‪tibilit‬‬ ‫ﺣﺎﻟﺘﻲ ﻛﻪ در آن ﺣﺸﺮات ﻧﺮو ﻣﺎده ﺑﺎ ﺑﻴﺶ از ﻳﻚ‬ ‫اﺳﺘﺮﻳﻦ وﻟﺒﺎﺧﻴﺎ آﻟﻮده ﻫﺴﺘﻨﺪ‪.‬‬

‫‪ Aedes albopictus‬ﻛﻪ ﺑﻄﻮر ﻣﻌﻤﻮل ﺑﺎ‬ ‫ﻫﺮدو اﺳﺘﺮﻳﻦ ‪wAlbA‬و ‪ wAlbB‬آﻟﻮده‬ ‫آﻟﻮدﮔﻲ‬ ‫ﺣﺎﻟﺖ آﻟ دﮔ‬ ‫رﻧﮓ ﺎﻟﺖ‬ ‫داﻳﺮه ددو ﻧﮓ‬ ‫ﻫﺴﺘﻨﺪ ) داﻳ‬ ‫ﻫ ﺘﻨﺪ‬ ‫ﻣﻀﺎﻋﻒ را ﻧﺸﺎن ﻣﻲ دﻫﺪ(‪.‬‬

‫‪PCR amplification‬‬

‫ﺑﻄﻮري ﻛﻪ ﻣﻼﺣﻈﻪ ﻣﻲ ﮔﺮدد‪ ،‬ﻧﺘﺎﻳﺞ ﺣﺎﺻﻠﻪ ﺑﺨﻮﺑﻲ‬ ‫ﻧﺸﺎن دﻫﻨﺪه ي اﻳﻦ آﻟﻮدﮔﻲ ﻣﻀﺎﻋﻒ در اﺳﺘﺮﻳﻦ‬ ‫آي‪.‬اچ اﻳﻦ ﭘﺸﻪ ﻣﻴﺒﺎﺷﺪ ‪.‬‬

‫‪CYTOPLASMIC‬‬ ‫‪INCOMPATIBILITUY‬‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫ﭼﮕﻮﻧﮕﻲ اﻧﺘﻘﺎل وﻟﺒﺎﺧﻴﺎ درون ﺗﺨﻢ ﻣﻴﺰﺑﺎن‬

‫ﻧﺤﻮه ي ﺣﺮﻛﺖ وﻟﺒﺎﺧﻴﺎ در ﺗﺨﻢ و اﻧﺘﻘﺎل آن ﺑﻪ ﻗﻄﺐ ﻋﻘﺒﻲ ﺗﺨﻢ‬ ‫ﻫﺎي درون ﺗﺨﻢ ااﺳﺖ‬ ‫ﺗﻮﺑﻮل ﺎ‬ ‫ل‬ ‫واﺑﺴﺘﻪ ﺑﻪ ﺷﺒﻜﻪ ﻣﻴﻜﺮو‬ ‫اﺣﺘﻤﺎﻻ ا‬ ‫ا ﺎﻻ‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫رﻧﮓ آﻣﻴﺰي ﺗﺨﻢ ﻫﺎي درون ﺗﺨﻤﺪان وﺟﻮد و اﻧﺘﻘﺎل‬ ‫ﻋﻘﺒﻲ ﺗﺨﻢ ﻧﺸﺎن ﻣﻣﻲ دﻫﺪ ‪000‬‬ ‫وﻟﺒﺎﺧﻴﺎ راا ددر ﻗﻄﺐ ﻋﻘﺒ‬

‫‪DAPI–F.I.S.H. staining‬‬

‫‪F.I.S.H. staining‬‬

‫‪STAINING TECHNIQUE‬‬

MECHANISM OF WOLBACHIA

‫ﭼﮕﻮﻧﮕﻲ ﺟﺎﻳﮕﻴﺮي وﻟﺒﺎﺧﻴﺎ در ﻃﻲ ﻣﺮاﺣﻞ ﻣﺨﺘﻠﻒ ﺟﻨﻴﻦ زاﻳﻲ‬ ... ‫و اﻧﺘﻘﺎل ﺑﻪ ﺑﻴﻀﻪ ﻫﺎ و ﺗﺨﻤﺪان در ﺟﻨﺲ ﻧﺮو ﻣﺎده‬ Presence of Wolbachia in transinfected C. embryos, ovaries, and testes.

capitata

(A) embryo ,Wolbachia localization at the mitotic spindles. (B) The posterior part of embryo. (C) Uniform distribution of Wolbachia bacteria in a postgastrulation embryo. (D) Distribution of Wolbachia during oogenesis (E) adult testes (F) A sperm cyst. [Scale bars: 40 µ m (A, B, and D– F) and 100 µ m (C).]

‫‪MECHANISM OF WOLBACHIA‬‬

‫‪(M‬‬ ‫‪d off A‬‬ ‫ﻞ ) ‪ti‬‬ ‫ﻣﻜﺎﻧﻴﺴﻢ ﻫﺎ‬ ‫ﻜﺎﻧ‬ ‫‪Mode‬‬ ‫‪Action‬‬ ‫ﻫﺎي ﻋﻋﻤﻞ‬

‫‪MODE‬‬

‫ﺳﻴﺘﻮﭘﻼﺳﻤﻲ‬ ‫ﻼ‬ ‫ﻧﺎﺳﺎزﮔﺎري‬ ‫اﻟﻒ( ﺎ ﺎ ﮔﺎ‬ ‫اﮔﺮﭼﻪ ﻫﻨﻮز ﻣﻜﺎﻧﻴﺴﻢ دﻗﻴﻘﻲ ﺑﺮاي اﻳﻦ ﻋﻤﻞ ﺑﺎﻛﺘﺮي ﻣﺸﺨﺺ ﻧﺸﺪه اﺳﺖ‪،‬اﻣﺎ ﻇﺎﻫﺮا اﻳﻦ ﺑﺎﻛﺘﺮي‬ ‫اﻧﺠﺎم ﻣﻲ دﻫﺪ‪:‬‬ ‫ﻛﻠﻲ اﻧ ﺎ‬ ‫ﺑﻄﻮر ﻛﻠ‬ ‫را ﻄ‬ ‫ﻋﻤﻞ ﻋﻤﺪه ا‬ ‫دو ﻞ‬

‫‪OF‬‬

‫‪ .1‬دﺳﺘﻜﺎري درﺳﺎﺧﺘﺎراﺳﭙﺮم در ﺗﺮ آﻟﻮده ﺑﻪ ﺑﺎﻛﺘﺮي‬

‫‪ACTION‬‬

‫)‪(modification of sperm‬‬

‫‪ .2‬ﺗﻮﻟﻴﺪ ﻋﻮاﻣﻞ ﺗﺮﻣﻴﻢ ﻛﻨﻨﺪﻫﻲ اﻳﻦ ﺗﻐﻴﻴﺮات در ﺳﻠﻮل ﻫﺎي ﺗﺨﻢ ﻣﺎده )‪،(rescue in the fertilized egg‬‬

MECHANISM OF WOLBACHIA

‫ﻓﺮﺿﻴﻪ ﻫﺎي اراﺋﻪ ﺷﺪه ﺑﺮاي ﻧﺎﺳﺎزﮔﺎري ﺳﻴﺘﻮﭘﻼﺳﻤﻲ‬

MODE Lock & key hypothesis

OF

Sink or Titration-Restitution h.

Slow-Motion h.

‫ﻛﻠﻴﺪ‬-‫ ﻓﺮﺿﻴﻪ ي ﻗﻔﻞ‬.1 ‫ﺑﺮﮔﺮداﻧﺪن‬-‫ ﻓﺮﺿﻴﻪ ﺑﺮدن‬.2

ACTION

‫ ﻓﺮﺿﻴﻪ ي ﺣﺮﻛﺖ آﻫﺴﺘﻪ‬.3

M DE O MOD OF ACT A TIO ON N

MECHANISM OF WOLBACHIA

(Lock & key hypothesis)

‫ﻛﻠﻴﺪ‬-‫ ﻓﺮﺿﻴﻪ ي ﻗﻔﻞ‬.1

M DE O MOD OF ACT A TIO ON N

MECHANISM OF WOLBACHIA

(Sink or Titration Titration-Restitution Restitution h. h)

‫برگرداندن‬-‫ﻓﺮﺿﻴﻪ بردن‬ ‫ ﻓ ﺿ‬.2

(Slow-Moution Slow Moution h. h)

AC CT TIO ON

MO OD DE OF

MECHANISM OF WOLBACHIA

‫ﺣﺮﻛﺖ آﻫ ﺘﻪ‬ ‫آﻫﺴﺘﻪ‬ ‫ﻓﺮﺿﻴﻪ ي ﺣ ﻛﺖ‬ ‫ ﻓ ﺿ ﻪ‬.3

MECHANISM OF WOLBACHIA

...‫جايگيري مي كند‬ ‫ولباخيا طي مراحل جنين زايي در سنتروزوم گ‬

MOLECULAR

IDENTIFICATION C O Fig. 6. Wolbachia localize to the centrosome during embryogenesis in N. vitripennis. Embryos from a female infected withWolbachia, fixed and stained for DNA (red) and tubulin (green). Inset, Wolbachia (arrow) can be seen clustered near the centrosomes (arrowhead) throughout the cell cycle.

‫‪MECHANISM OF WOLBACHIA‬‬

‫‪MODE‬‬ ‫‪OF‬‬ ‫‪ACTION‬‬ ‫ﺷﻜﻞ ‪ .5‬ﻓﻨﻮﺗﻴﭗ ﻧﺎﺳﺎزﮔﺎري اﻳﺠﺎد ﺷﺪه د رﺣﺸﺮات ﺗﻮﺳﻂ وﻟﺒﺎﺧﻴﺎ ﺑﻪ ﺳﺎﺧﺘﺎر‬ ‫ﻣﻨﺤﺼﺮ ﺑﻪ ﻓﺮد دوك ﻫﺎي ﮔﻮﻧﻮﻣﻴﻚ واﺑﺴﺘﻪ اﺳﺖ‪.‬‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫‪MODE‬‬ ‫‪OF‬‬ ‫‪ACTION‬‬ ‫ﺷﻜﻞ ‪ .4‬ﻣﺪﻟﻲ ﺑﺮاي ﭼﮕﻮﻧﮕﻲ ﺑﻬﺮه ﺑﺮي وﻟﺒﺎﺧﻴﺎ از دوك ﻫﺎي ﮔﻮﻧﻮﻣﺮﻳﻚ‬ ‫ﻣﺮگ در‬ ‫ﻧﺮزاﻳﻲ و گ‬ ‫ﺣﺎﻟﺖ ﻧ ا‬ ‫ﺗﻮﻟﻴﺪ ﻫﺮدو ﺎﻟ‬ ‫ﺑﻤﻨﻈﻮر ﺗ ﻟ‬ ‫ﻈ‬ ‫‪Nasonia sp.‬‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫‪MODE‬‬ ‫‪OF‬‬ ‫‪ACTION‬‬ ‫ﺷﻜﻞ ‪ .4‬ﻣﺪﻟﻲ ﺑﺮاي ﭼﮕﻮﻧﮕﻲ ﺑﻬﺮه ﺑﺮي وﻟﺒﺎﺧﻴﺎ از دوك ﻫﺎي ﮔﻮﻧﻮﻣﺮﻳﻚ ﺑﻤﻨﻈﻮر ﺗﻮﻟﻴﺪ ﻫﺮدو ﺣﺎﻟﺖ ﻧﺮزاﻳﻲ و ﻣﺮگ درﺟﻨﺲ‬ ‫‪Nasonia‬‬ ‫‪N‬‬ ‫‪i .‬‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫ب(‬

‫‪PI Wolbachia‬‬

‫‪STRAIN‬‬ ‫‪OF‬‬

‫اﺳﺘﻮﺗﺎﻣﺮ و ﻫﻤﻜﺎران )‪ (1990‬درﻳﺎﻓﺘﻨﺪ ﻛﻪ ﺗﻴﻤﺎر ﺑﺮﺧﻲ از ﻣﺎده ﻫﺎي اﺳﺘﺮﻳﻦ ﻫﺎي ﻣﺎده زا در ‪Trichogramma‬ﺑﺎ‬ ‫آﻧﺘﻲ ﺑﻴﻮﺗﻴﻚ ﻣﻨﺠﺮ ﺑﻪ ﺗﻐﻴﻴﺮ ﺣﺎﻟﺖ آن ﻫﺎ ﺑﻪ ﻧﺮ زاﻳﻲ ﻣﻲ ﮔﺮدد‪.‬‬

‫وﻟﺒﺎﺧﻴﺎ اﻛﻨﻮن در ﻣﺤﺪوده ي وﺳﻴﻌﻲ از زﻧﺒﻮر ﻫﺎي ﭘﺎرازﻳﺘﻮﺋﻴﺪ‪ ،‬ازﺟﻤﻠﻪ درﺟﻨﺲ ﻫﺎي‬ ‫‪ ،Aohytis, Encarcia, Leptopilina and Muscidifurax‬ﻳﺎﻓﺖ ﺷﺪه ﻛﺎرﻫﺎي ﺑﻌﺪي ﺻﻮرت ﮔﻔﺘﻪ در اﻳﻦ ﺧﺼﻮص‬ ‫‪Trichogramma,‬‬

‫‪WOLBACHIA‬‬

‫ﺎط ااﺳﺖ‪.‬‬ ‫وﻟﺒﺎﺧﻴﺎﺎ در اارﺗﺒﺎط‬ ‫ﻫﺎي ﻟ ﺎ‬ ‫ﺑﺎﻛﺘﺮي ﺎ‬ ‫ﺣﺎﻟﺖ ﺑﺎﺎ ﺎﻛ‬ ‫داد ﻛﻛﻪ ااﻳﻦ ﺗﻐﻴﻴﺮ ﺎﻟ‬ ‫ﻧﺸﺎن ا‬ ‫ﺎ‬

‫زاﻳﻲ در ااﻳﻦ‬ ‫زاﻳﻲ ﺑﻪ ﻧﻧﺮ زا‬ ‫ﺣﺎﻟﺖ از ﺎﻣﺎده زا‬ ‫ﺗﻐﻴﻴﺮ ﺎﻟ‬ ‫ﺑﺎﻋﺚ ﺗﻐ‬ ‫ﻫﺎ ﺎ ﺚ‬ ‫ﺑﺎﻛﺘﺮي ﺎ‬ ‫ﺣﺬف ااﻳﻦ ﺎﻛﺘ‬ ‫ﺑﻴﻮﺗﻴﻜﻲ ﺎﺑﺎ ﺬف‬ ‫آﻧﺘﻲ ﺗ ﻜ‬ ‫ﺣﺮارﺗﻲ و آﻧﺘ‬ ‫ﺗﻴﻤﺎرﻫﺎي ا ﺗ‬ ‫ﺗ ﺎ ﺎ‬ ‫زﻧﺒﻮرﻫﺎ ﻣﻲ ﮔﺮدد‪.‬اﻳﻨﻜﻪ آﻳﺎ ﺑﺎﻛﺘﺮي ﻫﺎي ‪ PI‬در دﻳﮕﺮ راﺳﺘﻪ ﻫﺎي ﺣﺸﺮات ﻏﻴﺮ از ﺑﺎل ﻏﺸﺎﺋﻴﺎن وﺟﻮد داردﻳﺎ‬ ‫ﮔﺰارﺷﻲ از وﺟﻮد اﻳﻦ ﺑﺎﻛﺘﺮي در ﺳﺮﺧﺮﻃﻮﻣﻲﻲ ‪Naupactus tesselatus‬اراﺋﻪ‬ ‫ﺧﻴﺮ‪ ،‬ﻫﻨﻮز ﻧﺎﺷﻨﺎﺧﺘﻪ اﺳﺖ‪،‬اﻟﺒﺘﻪ‬ ‫ﻲ‬ ‫ﺷﺪه اﺳﺖ‪.‬‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫ﻣﻜﺎﻧﻴﺴﻢ ﻋﻤﻞ ﺑﺎﻛﺘﺮي ﻫﺎي‬

‫‪PI‬‬

‫‪:‬‬

‫‪STRAIN‬‬ ‫‪OF‬‬

‫ﻣﻜﺎﻧﻴﺴﻢﻢ‬ ‫دﻗﻴﻘﻲ ﻣﻮرد ﻴﻧﻴﺎز اﺳﺖ‪،‬اﻣﺎ ﻴ‬ ‫ﺗﻜﺜﻴﺮ ﺷﺪن ﮔﺎﻣﺖ ﻣﻄﺎﻟﻌﺎت ﻴ ﻲ‬ ‫ﻣﻜﺎﻧﻴﺴﻢﻢ ﻴ‬ ‫ﺗﻌﻴﻴﻦ ﻴ‬ ‫درﺧﺼﻮص ﻴﻴ‬ ‫اﺣﺘﻤﺎﻟﻲ‪:‬‬ ‫رﺷﺘﻬﺎي دوﻛﻲﻲ ﺷﻜﻞﻞ‬ ‫اﺧﺘﻼل در ﺳﻨﺘﺮوزومم ﻫﺎ ﻳﺎ ﺗﺸﻜﻴﻞﻞ ﻬ‬ ‫اﺗﺼﺎل رﺷﺘﻪ اي دوﻛﻲ ﺑﻪ ﻛﺮوﻣﻮزم ﻫﺎ‪ ،‬ﻳﺎ ﻛﻴﻨﺘﻴﻚ دوك ﻫﺎ‬

‫‪WOLBACHIA‬‬

‫ﮔﻮﻧﻪ ﻫﺎي ﻫﺎﭘﻠﻮﺋﻴﺪ ﺑﺮاي ﻣﻮر ﺣﻤﻠﻪ ﻗﺮار ﮔﺮﻓﺘﻦ ﺗﻮﺳﻂ اﻳﻦ ﺑﺎﻛﺘﺮي ﻫﺎ ﻛﻪ ﺗﻮﻟﻴﺪ ﺳﺎزﮔﺎر ﺷﺪه اﻧﺪ‪ .‬در ﮔﻮﻧﻪ‬ ‫دﻳﭙﻠﻮﺋﻴﺪ) ﺑﺎ ﺗﻚ ﻟﻮﻛﻮس ﺗﻌﻴﻴﻦ ﻛﻨﻨﺪه ي ﺟﻨﺴﻴﺖ( ﺗﻜﺜﻴﺮ ﺷﺪن ﮔﺎﻣﺖ ﺑﻄﻮر ﻣﻌﻨﻲﻲ داري ﻛﻤﺘﺮ‬ ‫ﻫﺎﭘﻠﻮ ﭙ‬ ‫ﻫﺎي ﭘ‬ ‫رخ ﻣﻲ دﻫﺪ و اﻳﻦ ﺑﺨﺎﻃﺮ ﭘﺎﻛﺴﺎزي ﻧﺮﻫﺎ از ﺟﻤﻌﻴﺖ اﺳﺖ‪ .‬ﺑﻌﺒﺎرت دﻳﮕﺮ ﺗﻜﺜﻴﺮ ﺷﺪن ﮔﺎﻣﺖ در اﻳﻦ ﺣﺸﺮات‬ ‫رخ ﻧﻤﻲ دﻫﺪ ﭼﺮاﻛﻪ اﮔﺮ اﻳﻦ ﻋﻤﻞ ﺗﻔﺎق اﻓﺘﺪ‪ ،‬ﺑﺠﺎي ﺗﻮﻟﻴﺪ ﻣﺎده ﻫﺎي دﻳﭙﻠﻮﺋﻴﺪ‪ ،‬ﻧﺮ ﻫﺎي دﻳﭙﻠﻮﺋﻴﺪ ﺗﻮﻟﻴﺪ ﻣﻲ‬ ‫ﺷ ﻧﺪ‬ ‫ﺷﻮﻧﺪ‪.‬‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫ﻛﻨﺘﺮل ﻟ‬ ‫ل‬ ‫ﻛﺎرﺑﺮد ﺎﺑﺎﻛﺘﺮي ﺎ‬ ‫ﻓﻮاﻳﺪ ﺎ‬ ‫ا‬ ‫ﺑﻴﻮﻟﻮژﻳﻚ‪:‬‬ ‫ﻫﺎي ‪ PI‬در‬

‫‪STRAIN‬‬

‫)‪ .(i‬ﮔﻮﻧﻪ ﻫﺎي ﺑﻜﺮزا ﻧﺘﺎج ﺑﺎﻻﺗﺮي و رﺷﺪ ﺟﻤﻌﻴﺖ ﺑﻴﺸﺘﺮي دارﻧﺪ‪.‬‬

‫‪OF‬‬

‫)‪ .(ii‬در ﺗﻮﻟﻴﺪ ﻛﻠﻮﻧﻲ و ﭘﺎﻳﺪار ﺷﺪن آن ﻣﻮﻓﻖ ﺗﺮﻧﺪ‪،‬ﭼﺮاﻛﻪ ﻧﻴﺎزي ﺑﻪ ﺟﻨﺲ ﻧﺮو ﻣﺎده ﺑﺮاي‬

‫ﺟﻔﺖ ﮔﻴﺮي ﻧﻴﺴﺖ‪.‬‬ ‫)‪ .(iii‬در رﻫﺎﺳﺎي اﻧﺒﻮﻫﻲ ﺑﻬﺎي ﻛﻤﺘﺮي ﺻﺮف ﻣﻲ ﺷﻮد ﭼﺮاﻛﻪ از ﺗﻮﻟﻴﺪ ﻧﺮ ﻫﺎ ﻣﻤﺎﻧﻌﺖ ﺑﻌﻤﻞ ﻣﻲ آﻳﺪ‪.‬‬

‫‪WOLBACHIA‬‬

‫* ﻧﻜﺘﻪ ي ﻣﻬﻤﻲ ﻛﻪ در ﺧﺼﻮص ﺑﺎﻛﺘﺮي ﻫﺎي ‪PI‬وﺟﻮد دارد اﻳﻦ اﺳﺖ ﻛﻪ آﻳﺎ اﺣﺘﻤﺎل اﻧﺘﻘﺎل آن ﻫﺎ ﺑﻪ ﻣﻴﺰﺑﺎن‬ ‫ﭼﺮاﻛﻪ‬ ‫اﻧﺠﺎمم ﺷﺪه اﺳﺖ(‪.‬ﭼﺮ‬ ‫ي ‪ CI‬ﺠ‬ ‫ﺑﺎﻛﺘﺮي ﻫﺎي‬ ‫ﻣﻮرد ﺑ ﺮي‬ ‫در ﻮر‬ ‫آﻧﭽﻪ ﻛﻪ ر‬ ‫دارد)ﻣﺎﻧﻨﺪ ﭽ‬ ‫وﺟﻮد ر‬ ‫رﻳﺰﺗﺰرﻳﻖ وﺟﻮ‬ ‫ﻃﺮﻳﻖ رﻳﺰ ﺰرﻳﻖ‬ ‫از ﺮﻳﻖ‬ ‫ﺟﺪﻳﺪ ز‬ ‫ﺟ ﻳ‬ ‫اﻳﻨﻜﺎر ﻛﺎراﻳﻲ اﻳﻦ ﺑﺎﻛﺘﺮي ﻫﺎ د را در ﻛﻨﺘﺮل ﺑﻴﻮﻟﻮﻳﻚ دوﭼﻨﺪان ﻣﻲ ﻛﻨﺪ‪.‬‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫ج( ‪Feminizing Wolbachia‬‬

‫‪STRAIN‬‬

‫اﻳﻦ اﺳﺘﺮﻳﻦ ازﺑﺎﻛﺘﺮي وﻟﺒﺎﺧﻴﺎ از ﺳﺨﺖ ﭘﻮﺳﺘﺎن ﮔﺰارش ﺷﺪه اﺳﺖ و اﺧﻴﺮا در‬ ‫‪ Zyginidia pullula ( Hemiptera‬ﻳﺎﻓﺖ ﺷﺪه اﺳﺖ‪.‬‬ ‫‪ furnacalis (Lepidoptera: Crambidae‬و ))‪Hemiptera‬‬ ‫))‪Crambidae‬‬ ‫ﺑﻬﺘﺮﻳﻦ ﻧﻤﻮﻧﻪ در اﻳﻦ زﻣﻴﻨﻪ ‪ Armadillium vulgar‬اﺳﺖ ﻛﻪ ﻣﻄﺎﻟﻌﺎت زﻳﺎدي روي آن ﺻﻮرت‬ ‫ﮔﺮﻓﺘﻪ اﺳﺖ‪ .‬اﻳﻦ ﺑﺎﻛﺘﺮي دراﻳﻦ ﮔﻮﻧﻪ ﺑﺎ ﺳﺮﻛﻮب و ﻣﻤﺎﻧﻌﺖ از ﻳﻚ ﻏﺪه ي آﻧﺪروژﻧﻴﻚ ﻧﺮ‬ ‫ﻫﺎ راﺑﻪ ﻣﺎده ﻫﺎ ﺗﺒﺪﻳﻞ ﻣﻲ ﻛﻨﺪ)اﻟﺒﺘﻪ ﮔﻮﻧﻪ ﻫﺎي ﺣﺪواﺳﻂ ﻧﻴﺰ ﺣﺎﺻﻞ ﻣﻲ ﺷﻮد(‪.‬‬ ‫‪Ostrinia‬‬

‫‪OF‬‬

‫‪WOLBACHIA‬‬

‫‪MECHANISM OF WOLBACHIA‬‬

‫ﻣﻜﺎﻧﻴﺴﻢ ﻋﻤﻞ ﺑﺎﻛﺘﺮي ﻫﺎي ‪PI‬‬

‫ﻣﻜﺎﻧﻴﺴﻢ ﻋﻤﻞ اﻳﻦ ﺑﺎ ﻛﺘﺮي ﻫﺎ در‬

‫‪A. vulgar‬‬

‫‪STRAIN‬‬

‫ﻣﻮرد ﻣﻄﺎﻟﻌﻪ ﻗﺮار ﮔﺮﻓﺘﻪ اﺳﺖ‪:‬‬

‫‪OF‬‬

‫ﻫﺘﺮوﮔﺎﻣﺘﻲﻲ )‬ ‫ﭘﻮﺳﺖ ﻧﺸﺎنن داد ﻛﻪ ﻣﺎده ﻫﺎ ﺮو‬ ‫ﻳﻦ ﺳﺨﺖ ﭘﻮ‬ ‫ﺟﻨﺴﻴﺖ درر اﻳﻦ‬ ‫ﻴﻴﻦ ﺟ ﻴ‬ ‫ﻮص ﺗﻌﻴﻴﻦ‬ ‫اﺳﺘﺎﻧﺪارد درر ﺧﺼﻮص‬ ‫ر‬ ‫ي‬ ‫ﺗﺴﺖ ﻫﺎي‬ ‫ﻣﺎده ﻫﺎ ‪ ZW‬و ﻧﺮ ﻫﺎ ‪ (ZZ‬ﻫﺴﺘﻨﺪ‬ ‫ﻣﻨﺠﺮ ﺑﺑﻪ ﺶ‬ ‫ﻛﺎﻫﺶ‬ ‫ﻗﺎﺑﻞ ﻣﻼﺣﻈﻪ ﺠﺮ‬ ‫ﻓﺮاواﻧﻲ ﺑﻞ‬ ‫ﺳﺎزي ﺑﺑﺎ ﺮ و ﻲ‬ ‫ﻣﻮﻧﺚ زي‬ ‫ﻫﺎي ﻮ‬ ‫ﻓﺎﻛﺘﻮر ي‬ ‫دﻳﮕﺮ ﻮر‬ ‫ﺑﺎﻛﺘﺮي ﻫﺎ و ﻳﻳﺎ ﻳ ﺮ‬ ‫اﺑﻦ ﺑ ﺮي‬ ‫ﺣﻀﻮر ﺑﻦ‬ ‫ﻮر‬ ‫ﻛﺮوﻣﻮزوم ﺗﻌﻴﻦ ﻛﻨﻨﺪه ي ﺟﻨﺴﻴﺖ ﻧﺮ ﻣﻲ ﮔﺮدد)‪ (W‬و ﺑﻨﺎﺑﺮاﻳﻦ ﻣﻜﺎﻧﻴﺴﻢ ﺗﻌﻴﻴﻦ ﻛﻨﻨﺪه ي ﺟﻨﺴﻴﺖ را‬ ‫ﺗﻀﻌﻴﻒ ﻣﻲ ﻛﻨﺪ‬

‫‪WOLBACHIA‬‬

‫اﺛﺮ ﺛﺎﻧﻮﻳﻪ اﻳﻦ ﻋﻤﻞ ﺗﺠﻤﻊ ژن ﻫﺎي ﺳﺮﻛﻮﺑﮕﺮ ﺗﻌﻴﻴﻦ ﻛﻨﻨﺪه ي ﻧﺮﻫﺎ در ﺟﻤﻌﻴﺖ ﻫﺎ ﻣﻲ ﺑﺎﺷﺪ‪.‬‬

MECHANISM OF WOLBACHIA

Screening arthropod DNA samples for Wolbachia

MOLECULAR 1- arthropod collection and storage step 1 step 2- pre-extraction processing

step 3- DNA extraction and purification from arthropod and Wolbachia step 4- determination of infection status and template amplification control step 5- optional- typing of Wolbachia strains A and B using two diagnostic PCRs with strain specific primers and/or the restriction profile step 6- optional sequencing of Wolbachia positive arthropod 28s D2 rDNA PCR product for molecular identification of the arthropod, and 16s rRNA, wsp, groEL, or other Wolbachia genes for strain typing in Wolbachia.

IDENTIFICATION

MECHANISM OF WOLBACHIA

MOLECULAR

Wolbachia gene for PCR diagnostic:

16 s ribosomal RNA (16 16ss rDNA rDNA)) Wolbachia surface p protein (wsp wsp) p) Heat shock protein (groEL) groEL) or other Wolbachia genes for strain typing in Wolbachia.

Control gene:

IDENTIFICATION C O

28 s ribosomal ib l RNA (28 s rDNA) rDNA DNA)) DNA Mitochondrial cytochrom c oxidase subunit I, 5 segment (COI (COI--5) Forkhead:: Single copy Nuclear gene (fkh Forkhead fkh))

MECHANISM OF WOLBACHIA

Screening arthropod DNA samples for Wolbachia

MOLECULAR 28 s rDNA

16 16ss rRNA

If

+

+

If

-

+

If

+ -

-

Infection

√ √

*

IDENTIFICATION If

√ : infection

*: No infection ?: Error in PCR reaction

?

MECHANISM OF WOLBACHIA

Molecular evidence for wolbachia in rice insect

Wolbachia 16S rRNA gene amplification in rice

insects. Lane M, Lambda DNA marker; lane 1, rice gall midge; lane 2,yellow stem borer (adult); lane 3, yellow stem borer (larvae); lane 4, whitebacked planthopper; lane 5, 5 brown planthopper; lane 6, 6 rice leaffolder; lane 7, rice skipper; lane 8, rice mealy bug; lane 9, rice stink bug; lane 10, green leafhopper; lane 11, rice caseworm; lane 12, mosquito. ( Behera et al., 2001)

MECHANISM OF WOLBACHIA

MOLECULAR Microinjection technique

Figure 1. Microinjection material and device: ((a)) top p view and (b) ( ) schematic transverse section. Plastic box 5 cm in diameter,, with cover removed during the injection time.

TECHNIQUE