گیاه پزشکی

گیاه پزشکی

GSh302: پروتئین نامزد افکتوری در قارچ Ascochyta rabiei، القاشونده در شرایط برهمکنش با گیاه میزبان، با ساختار پیشبری حامل جایگاه اتصال فاکتور رونویسی bZIP

نوع مقاله : علمی پژوهشی -انگلیسی

نویسندگان
1 دانش آموخته دکتری بیوتکنولوژی و به نژادی گیاهان زراعی، دانشکده کشاورزی، دانشگاه فردوسی مشهد، مشهد، ایران.
2 دانشیار گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه فردوسی مشهد، مشهد، ایران.
چکیده
شناسائی و درک کارکرد پروتئین‌های افکتوری ترشح‌شده توسط قارچ‌های نکروتروف برای توسعه روش­های دفاعی مؤثر در گیاهان و اصلاح ارقام مقاوم مورد توجه قرار می­گیرد. فراهم شدن داده­های ژنومی و ترانسکریپتومی مربوط به مراحل توسعه بیمارگر روی میزبان می­تواند با پیشرفت‌هایی که در ابزارهای واکاوی مبتنی بر روش­های بیوانفورماتیک انجام شده­است امکان شناسائی این ژن­های افکتوری را بطور چشمگیری افزایش دهد. در مطالعه حاضر با هدف شناسائی افکتورهای قارچ Ascochyta rabiei از ترکیبی از روش‌های این سیلیکو با تمرکز بر ویژگی­های ساختاری پروتئین­های افکتوری از جمله ترشحی بودن، طول کوتاه، توانائی تشکیل باندهای دی­سولفیدی استفاده شده است. همچنین به منظور تایید ارتباط کارکردی افکتور نامزد شده از روش واکاوی توالی پیشبر ژن کدکننده پروتئین افکتوری و بررسی حضور جایگاه­های تنظیمی مرتبط با فاکتورهای نسخه برداری دخیل در برهمکنش بیمارگر و میزبان استفاده شده است. همچین ردیابی ژن نامزد شده در سطح ژنوم و ترانسکریپتوم قارچ A. rabiei در شرایط رشد در محیط کشت و همچنین در شرایط برهمکنش با گیاهان مقاوم و حساس نخود مورد بررسی قرار گرفته­است. در این بررسی­ها یک پروتئین 159 اسیدآمینه­ای به عنوان نامزد افکتوری شناسائی شد و تحت نام GSh302 موقعیت توالی کدکننده و توالی پروتئین آن به ترتیب با شماره بازیابی OR875943 و WPV74376 در بانک ژن ثبت شد. واکاوی توالی بالادست ژن، حضور عناصر تنظیمی جایگاه اتصال فاکتورهای نسخه­برداری از جمله Sko1 (bZIP_1) را تایید نمود که در فعال­سازی بیان ژن­های ترشحی در مراحل برهمکنش قارچ با گیاه نقش کلیدی دارد. ردیابی توالی ژن در جدایه­های قارچ A. rabiei رشد یافته در محیط کشت تایید گردید ولی در سطح رونوشت آن­ها قابل ردیابی نبود. این ژن در گیاهان مقاوم و حساس نخود 96 ساعت پس از مایه زنی در سطح ژنوم و ترانسکریپتوم قارچ قابل ردیابی بود. روش ترکیبی بکارگرفته شده در این مطالعه، پروتئین GSh302 را به‌عنوان یک پروتئین نامزد افکتوری در قارچ A. rabiei که در زمان برهمکنش قارچ با گیاه نخود بیان می شود معرفی نمود.
کلیدواژه‌ها
موضوعات

عنوان مقاله English

GSh302: A candidate effector protein in Ascochyta rabiei, induced during host interaction, with a promoter structure harboring a bZIP transcription factor binding site

نویسندگان English

M. Hasani 1
F. Shokouhifar 2
M. Mamarabadi 2
1 Ph.D. of Plant Biotechnology, in Department of Biotechnology and Plant Breeding, Faculty of Agriculture, Ferdowsi University of Mashhad. Mashhad. Iran.
2 Associate Professor, Department of Plant Protection, Faculty of Agriculture, Ferdowsi University of Mashhad. Mashhad. Iran.
چکیده English

Effector proteins secreted by necrotrophic fungi have been considered essential to identify and understand their functions to develop effective defenses and breed resistant plant varieties. In recent years, bioinformatics-based analysis tools have increased the likelihood of identifying these effector genes by providing genomic and transcriptomic data on the stages of pathogen development within the host. In this study, a combination of in silico methods were used to identify the effectors of the fungus Ascochyta rabiei, focusing on the structural features of effector proteins, including secretion, short length, and disulfide-bond-forming ability. Also, the promoter sequence analysis of the gene encoding the effector protein and the presence of regulatory sites for transcription factors involved in the pathogen-host interaction were used to confirm the functional relationship of the candidate effector. Moreover, the candidate gene was tracked at the genome and transcriptome levels in A. rabiei under growth conditions in culture medium and during interactions with resistant and susceptible chickpea genotypes. A 159-amino-acid protein, GSh302, was identified as a candidate effector, and its coding sequence (accession number OR875943) and protein sequence (accession number WPV74376) were registered in GenBank. Analysis of the upstream sequence of the gene confirmed the presence of regulatory elements, including transcription factor binding sites and the Sko1 (bZIP_1) transcription factor, which plays a key role in activating the expression of secretory genes during the fungus-plant interaction. The gene sequence was tracked in A. rabiei isolates grown in culture medium, but it was not detectable at the transcript level. This gene was discernible at the genome and transcriptome levels of the fungus in resistant and susceptible chickpea cultivars 96 hours after inoculation. The combined method used in this study identified the GSh302 protein as a candidate effector in A. rabiei, expressed during the fungus-chickpea interaction.

کلیدواژه‌ها English

Computational Analyses
Gene Expression Tracking
In Silico Predictions
Molecular Fungal-Plant Interaction
Secretome
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