مطالعه نقش پیشبر مصنوعی SP-DDEE القاء‌پذیر توسط بیمارگرهای قارچی جهت بیان ژن مقاومت در گیاه Brassica napus

نوع مقاله: مقاله کامل پژوهشی

نویسندگان

1 تهران، پژوهشگاه ملی مهندسی ژنتیک و زیست فناوری، گروه بیوتکنولوژی کشاورزی

2 پژوهشگاه ملی مهندسی ژنتیک و زیست فناوری، گروه بیوتکنولوژی کشاورزی

چکیده

بیان ژن‌های مقاومت توسط پیشبرهایی که صرفا در پاسخ به بیمارگرها فعال می‌شوند مزیت بزرگی در تولید گیاهان مقاوم به بیماری محسوب می‌شود. در این بررسی ژن کیتیناز کایمری تحت پیشبر القاءپذیر با بیمارگر SP-DDEE به گیاه کلزا منتقل شد و پتانسیل این گیاهان در مهار رشد بیمارگرهای قارچی در محیط in vivo  مورد ارزیابی قرار گرفت. در این راستا، سه سازه ی pGDEC، pGMPC و pBISM2 به ترتیب حاوی پیشبر SP-DDEE ( شامل دوعنصر D و دو عنصر E17 بالادست پیشبر حداقل)، SP-MP (شامل فقط پیشبر حداقل) و CaMV35S ( شامل پیشبر دائمی) جهت تراریختی گیاهان مورد استفاده قرار گرفت. بررسی‌ها با استفاده از آزمون فعالیت آنزیمی مشخص نمود که پیشبر القائی مورد نظر نسبت به محرک متیل جاسمونات القاءپذیر بوده ولی پاسخ مشخصی به محرک سالیسیلیک اسید نشان نداد. بعلاوه، نتایج مشخص نمود عصاره دیسک‌های برگی گیاهان تراریخت حاوی پیشبر SP-DDEE تیمار شده با متیل جاسمونات به نسبت گیاهان شاهد قادر به مهار رشد بیمارگرهای قارچی Sclerotinia sclerotiorum و Rhizoctonia solani می‌باشند. در مجموع مشخص شد پیشبر SP-DDEE، نه تنها نسبت به محرک متیل جاسمونات به عنوان مولکول پیام رسان در مسیر مقاومت به بیمارگرهای نکروترف القاءپذیر می‌باشد، بلکه میزان بیان ژن کیتیناز کایمری، تحت کنترل این پیشبر نیز جهت افزایش مقاوت نسبت به بیمارگرهای قارچی مناسب به نظر می‌رسد.

کلیدواژه‌ها


عنوان مقاله [English]

Study of the role of the synthetic fungal pathogen inducible SP-DDEE promoter for expression of resistant gene in Brassica napus

نویسندگان [English]

  • M. Moradyar 1
  • M. Motallebi 2
  • M.R Zamani 1
  • R. Aghazadeh 1
چکیده [English]

The expression of defensive genes from a promoter that is specifically activated in response to pathogen invasion is highly desirable for engineering disease-resistant plants. In this study, chimeric chitinase gene under the control of SP-DDEE pathogen inducible promoter transformed to the canola plants and these plants were evaluated for their in vivo biocontrol potential against fungal pathogens. In this regard, three constructs, pGDEC, pGMPC, pBISM2 containing synthetic promoters, SP-DDEE (two D and two E elements + minimal promoter), SP-MP (minimal promoter), and the CaMV35S constitutive promoter, respectively were used. Enzyme activity assay demonstrated that the synthetic pathogen-inducible promoter was responsive to the Methyl jasmonate (MJ) elicitor, but not responsive to the salicylic acid. Moreover, results indicated that leaf total protein from transgenic lines harboring the SP-DDEE promoter treated with MJ, inhibited the growth of the fungal pathogens of Sclerotinia sclerotiorum and Rhizoctonia solani. Overall, data show that not only the SP-DDEE synthetic promoter is highly responsive to MJ, as an important chemical signal in necrotrophic pathogen defense, but the inducible expression of the chimeric chitinase gene, when controlled by the SP-DDEE promoter, is also seems to be appropriate to increased resistance to fungal pathogens.

کلیدواژه‌ها [English]

  • Canola
  • Chimeric chitinase
  • Rhizoctonia solani
  • Sclerotinia sclerotiorum
  • Synthetic promoter

 

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