حفاظت گیاهان, دوره (33), شماره (1), سال (2019-4) , صفحات (1-8)

عنوان : ( ردیابی همولوگ ژن توماتیناز در ژنوم قارچ Fusarium oxysporum f.sp. melonis )

نویسندگان: فاطمه زرندی , فرهاد شکوهی فر , بهرام شریف نبی , بدرالدین ابراهیم سید طباطبائی , ضیاء الدین بنی هاشم , ضیاء الدین بنی هاشم ,
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چکیده

Introduction : Phytopathogenic fungi exposed various proteins to overcome plant defense systems. Production of saponins likes α-Tomatine is one of the tomato preformed defences barriers which should be detoxificated by the pathogens. It has been revealed before which most of Fusarium species and forma specials could produce tomatinase, a glycosyl hydrolases protein, to de-glycosylate α-Tomatine. Fusarium oxysporium f. sp. melonis (FOM) wildly attached melon cultivars and at the time of designing this project there was only a report of which the gene sequence of tomatinase probably exists in the genome of FOM using southern blotting experiment. We have done this study to tracking the whole tomatinase gene sequence in the FOM genomic sequence and investigate the probability genetic variation of the gene in the nucleotide and protein sequences. Materials and methods: Fusarium oxysporium f. sp. melonis (Fom) race1 have been reported previously from Khourasan province of Iran was cultured in liquid medium and the mycelia were used for the genomic DNA isolation. Primers were designed based on conserved sequence in upstream and downstream of FoTom1 sequence (AJ012668). PCR carried out and amplified segments were bi-directional sequenced and results were analyzed by Vector NTi software. The sequencing result was aligned with FoTom1 sequence as Refseq and the single nucleotide variations were detected by CLC work bench software. The effects of the mutations on the protein structure were predicted by CLC work bench software. Result and discussion: Electrophoresis pattern of PCR products showed a single band of the expected size in the strain FomR1 that was on the same size with the band amplified from FoL genome. The designed primers based on the FoTom1 sequence amplified a specific segment in the Fom genome. Alignment the sequencing results with the Fo-Tom1 from Fusarium oxysporium f. sp. lycopercisi (Fol) in nucleotide level revealed 14 mutations which seven of them were appeared in the protein sequence. Three mutations have occurred in the functional domain of the protein. At the position 145 an acidic amino acid with a negative charge, substituted by a polar amino acid. Replacements at positions 218 and 236 have been the same in terms of polarity and at position 218; both amino acids have a positive charge. There were no introns in the coding sequence of FomTomR1 region as same as FoTom1. Pairwise alignment results showed some an-synonyms mutations between two sequences that made some changes in the secondary structure of the translated protein from FomTomR1. The first an synonym mutation, SNP15(E→G), inside the signal peptide, converts the alpha helix to a new beta sheet. SNP34(K→N) and SNP35(S→N) mutations shortened the alpha helix. The other mutations were happened out of the alpha helixes and beta sheets. To predict effects of the mutations in the function of FomTomR1, in-silico analyses have been done and the results revealed that three mutations happened in the functional domains of tomatinase in Fom. The mutations in the hydrolysis domain may affect the structure of FomTomR1and can be effective in the function of the protein. The presence of different active saponins components in Melon may be an evolutionary reason for some variation in sequence and structure of the FomTomR1 protein in Fom. To prove the differences in the function of the tomatinase, the interaction of proteins with various kinds of melon saponins components should be investigated using in the future studies. Conclusion: The results showed the tracked sequence could be homolog of the Tomatinse gene in the Fom genome. We named it Fom-TomR1 and the sequenced was submitted in the Genbank with accession number MF178403. In the future study the effect of the gene should to be investigated in the pathogenesis of FOM on melon cultivars and it could be considered as a general screening index using heterologous expression of the gene.

کلمات کلیدی

, Fusarium wilts disease, gene detection, single nucleotide polymorphism, secondary structure prediction.
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@article{paperid:1074754,
author = {فاطمه زرندی and شکوهی فر, فرهاد and بهرام شریف نبی and بدرالدین ابراهیم سید طباطبائی and ضیاء الدین بنی هاشم and ضیاء الدین بنی هاشم},
title = {ردیابی همولوگ ژن توماتیناز در ژنوم قارچ Fusarium oxysporum f.sp. melonis},
journal = {حفاظت گیاهان},
year = {2019},
volume = {33},
number = {1},
month = {April},
issn = {2008-4749},
pages = {1--8},
numpages = {7},
keywords = {Fusarium wilts disease; gene detection; single nucleotide polymorphism; secondary structure prediction.},
}

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%0 Journal Article
%T ردیابی همولوگ ژن توماتیناز در ژنوم قارچ Fusarium oxysporum f.sp. melonis
%A فاطمه زرندی
%A شکوهی فر, فرهاد
%A بهرام شریف نبی
%A بدرالدین ابراهیم سید طباطبائی
%A ضیاء الدین بنی هاشم
%A ضیاء الدین بنی هاشم
%J حفاظت گیاهان
%@ 2008-4749
%D 2019

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