ﺷﻨﺎﺳﺎﯾﯽ ژن ﻫﺎي دﺧﯿﻞ در اﺳﺘﻘﺮار راﺑﻄﻪ اﻧﮕﻠﯽ در ﮔﻞ ﺟﺎﻟﯿﺰ ﻣﺼﺮي ﺑﺮ ﭘﺎﯾﻪ ﺗﺠﺰﯾﻪ و ﺗﺤﻠﯿﻞ دادهﻫﺎي ﺑﯿﺎن ژن

Identification of candidate genes involved in parasitic relationship in Orobanch aegyptiaca based on gene expression data

ﮔﻞ ﺟﺎﻟﯿﺰ ﮔﯿﺎه اﻧﮕﻠﯽ اﺳﺖ ﮐﻪ ﺑﺎﻋﺚ ﮐﺎﻫﺶ ﻗﺎﺑﻞ ﺗﻮﺟﻬﯽ در ﺗﻮﻟﯿﺪ ﻣﺤﺼﻮﻻت زراﻋﯽ ﻣﯽﺷﻮد و ﮐﻨﺘﺮل آن ﻧﯿﺰ در ﺷﺮاﯾﻂ ﻣﺰرﻋﻪ دﺷﻮار اﺳﺖ. در اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﺗﺠﺰﯾﻪ وﺗﺤﻠﯿﻞ دادهﻫﺎي RNA-seq ﻣﻮﺟﻮد در NCBI، ﻫﻤﺮاه ﺑﺎ ﺗﺎﯾﯿﺪ آزﻣﺎﯾﺸﮕﺎﻫﯽ ﺑﺨﺸﯽ از ﻧﺘﺎﯾﺞ ﺣﺎﺻﻞ ﮔﺰارش ﻣﯽﺷﻮد. ﭘﺲ از ارزﯾﺎﺑﯽ ﮐﯿﻔﯿﺖ ﺧﻮاﻧﺶﻫﺎي ﺧﺎم آﻟﻮﻣﯿﻨﺎ، ﺧﻮاﻧﺶﻫﺎي ﺑﺎ ﮐﯿﻔﯿﺖ ﻗﺎﺑﻞ ﻗﺒﻮل روي ﺗﺮﻧﺴﮑﺮﭘﺘﻮم ﮔﻞ ﺟﺎﻟﯿﺰ ﻧﻘﺸﻪﯾﺎﺑﯽ ﺷﺪﻧﺪ. ﭘﺲ از آﻧﺎﻟﯿﺰ ﺑﯿﺎن ژن ﺑﺎ اﺳﺘﻔﺎده از ﻧﺮم اﻓﺰار DESeq، 391 ژن داراي ﺑﯿﺎن ﻣﺘﻔﺎوت در ﻣﺮﺣﻠﻪ آﺑﻨﻮﺷﯽ ﺑﺬر ﮔﻞﺟﺎﻟﯿﺰ و ﻣﺮﺣﻞ اﺗﺼﺎل ﮔﻞ ﺟﺎﻟﯿﺰ ﺑﻪ رﯾﺸﻪ ﻣﯿﺰﺑﺎن ﺷﻨﺎﺳﺎﯾﯽ ﺷﺪ. ﺑﯿﺎن اورﺗﻮﻟﻮگ اﯾﻦ ژن ﻫﺎ در ﺧﻮﯾﺸﺎوﻧﺪان ﻧﺰدﯾﮏ ﮔﻞ ﺟﺎﻟﯿﺰ، ﮐﻪ ﻏﯿﺮ اﻧﮕﻞ ﯾﺎ ﻧﯿﻤﻪ اﻧﮕﻞ ﻫﺴﺘﻨﺪ، ﻧﯿﺰ ﺑﺮرﺳﯽ ﺷﺪ. از ﺑﯿﻦ 391 ژﻧﯽ ﮐﻪ ﺑﯿﻦ ﻣﺮﺣﻠﻪ آﺑﻨﻮﺷﯽ و اﺗﺼﺎل ﺑﻪ ﻣﯿﺰﺑﺎن در ﮔﻞ ﺟﺎﻟﯿﺰ ﺗﻔﺎوت ﺑﯿﺎن ﻣﻌﻨﯽ داري داﺷﺘﻨﺪ، 87 ژن در ﺧﻮﯾﺸﺎوﻧﺪ اﻧﮕﻞ ﮔﻞ ﺟﺎﻟﯿﺰ ﻧﯿﺰ ﺑﯿﺎن ﺑﺎﻻﯾﯽ در زﻣﺎن اﺗﺼﺎل ﺑﻪ ﻣﯿﺰﺑﺎن داﺷﺘﻨﺪ، در ﺣﺎﻟﯽ ﮐﻪ اﯾﻦ ژن ﻫﺎ در ﺧﻮﯾﺸﺎوﻧﺪ ﻏﯿﺮ اﻧﮕﻞ ﺑﯿﺎن ﻧﻤﯽ ﺷﺪﻧﺪ. ﺑﺮ اﯾﻦ اﺳﺎس اﯾﻦ 87 ژن ﺑﻪ ﻋﻨﻮان ژن ﻫﺎي ﮐﺎﻧﺪﯾﺪ دﺧﯿﻞ در اﺳﺘﻘﺮار راﺑﻄﻪ اﻧﮕﻠﯽ ﺑﺎ ﻣﯿﺰﺑﺎن در ﻧﻈﺮ ﮔﺮﻓﺘﻪﺷﺪﻧﺪ. در اداﻣﻪ اﻟﮕﻮي ﺑﯿﺎن ﻧُﻪ ژن از اﯾﻦ ژن ﻫﺎي ﮐﺎﻧﺪﯾﺪ اﻧﺘﺨﺎﺑﯽ در ﺑﺎﻓﺖ روﯾﺸﯽ، ﮔﻞ، و ﺑﺎﻓﺖ ﻣﺤﻞ اﺗﺼﺎل ﮔﻞﺟﺎﻟﯿﺰ ﺑﻪ ﮔﻮﺟﻪﻓﺮﻧﮕﯽ ﺑﺮرﺳﯽ ﺷﺪ. ﻧﺘﺎﯾﺞ ﻧﺸﺎن داد ﮐﻪ ژن ﮐﺪﮐﻨﻨﺪه ﺳﺮﯾﻦ ﮐﺮﺑﻮﮐﺴﯽﭘﭙﺘﯿﺪاز اﻟﮕﻮي ﺑﯿﺎن ﻣﻮرد اﻧﺘﻈﺎر ﺑﺮاي ژن ﻫﺎي دﺧﯿﻞ در اﯾﺠﺎد راﺑﻄﻪ اﻧﮕﻠﯽ را دارد ﺑﻪ ﻧﺤﻮي ﮐﻪ ﺗﻨﻬﺎ در ﺑﺎﻓﺖ ﻣﺤﻞ اﺗﺼﺎل ﺑﻪ ﻣﯿﺰﺑﺎن ﺑﯿﺎن ﻣﯽﺷﻮد. ﻧﺘﺎﯾﺞ اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﻣﯽﺗﻮاﻧﺪ ﺑﺮاي اﻫﺪاف ﻣﻬﻨﺪﺳﯽ ژﻧﺘﯿﮏ ﮔﻮﺟﻪﻓﺮﻧﮕﯽ ﺑﺮاي اﯾﺠﺎد ﻣﻘﺎوﻣﺖ ﺑﻪ ﮔﻞ جال ي از ﻃﺮﯾﻖ ﺧﺎﻣﻮﺷﯽ ژن در ﻣﯿﺰﺑﺎن ﻣﻮرد اﺳﺘﻔﺎده ﻗﺮار ﮔﯿﺮد .

Broomrape (Orobanche aegyptica) is a notorious parasitic plant that cause significant production loss. Here we report analysis of publicly available RNA-seq data for broomrape, coupled with experimental verification of part of the results. After quality control of raw illumine reads, qualified reads were mapped against Orobanch transcriptome. Differential gene expression analysis, performed by using DESeq package, identified 391 differentially expressed genes between seed imbibition and haustorium attachment stages. The expression of orthologs of these genes in close relatives of Orobanch, which are parasitic, hemi- or non-parasitic, was investigated. From 391 identified genes, 87 genes showed high levels of expression in parasitic relatives and not in non-parasitic ones. Based on these analyses the 87 genes were considered as candidate genes involved in establishment of parasitic interaction between Orobanch and its host. The expression of nine of these genes were checked experimentally in flower tissues of Orobanch and in tissues sampled from the attachment site on the host root. The expression of Or2094, which a putative serine-carboxy peptidase, was detected only in the attachment site, supporting the role of this gene in establishment of the parasitic interaction. The results of this work will pave the way for future genetic engineering projects to use host-induced gene silencing strategy to enhance resistance to Orobanch in host crop plants.