بهینه‌سازی متغیرهای انتقال ژن توسط تفنگ ژنی با انتقال ژن کدکننده‌ی موتان‌سوکراز (gtfI) به نیشکر

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجوی دکتری، گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم‌آباد، ایران

2 استاد، گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم‌آباد، ایران

3 دانشیار و مربی گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم‌آباد، ایران

4 مربی گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه لرستان، خرم‌آباد، ایران

چکیده

نیشکر (Saccharum officinarum) مهم‌ترین گیاه صنعتی تولیدکننده‌ی ساکارز در دنیا می‌باشد. کشت بافت و تراریختی این گیاه با چالش‌هایی مواجه است که نیازمند بهینه‌سازی است. در این تحقیق، ژن کدکننده‌ی موتان‌سوکراز (gtfI) از باکتری Streptococcus downei جداسازی، همسانه‌سازی و به نیشکر انتقال داده شد. در روش تفنگ ژنی فاصله 9 سانتی‌متر پرتاب ژن از ریزنمونه نسبت به فاصله 12 سانتی‌متر بهتر و اختلاف معنی‌داری (P≤0.01) را نشان داد. هم‌چنین استفاده از ترکیب سوربیتول و مانیتول برای حفظ فشار اسمزی کالوس‌ها درصد تراریختی را به‌طور معنی‌داری (P≤0.01) بالا برد. ترکیب هورمونی IBA و NAA بهترین کارآیی را در ریشه‌دار کردن گیاهان تراریخت نشان داد. تجزیه قند گیاهان تراریخت حاصل از دو مرحله زیرکشت، نشان داد که میانگین پارامتر پل (میزان ساکارز) در هر دو لاین تراریخت نسبت به شاهدهای مربوطه با کاهش قابل ملاحظه‌‌ای در حدود 30 درصد همراه بود. این موضوع نشان می‌دهد که آنزیم موتان‌سوکراز به خوبی در لاین‌های تراریخت نیشکر بیان شده و توانسته است این میزان قند را مصرف کند.

کلیدواژه‌ها


عنوان مقاله [English]

Optimization of Biolistic-mediated Gene Transfer Parameters by Transferring Mutansucrase (gtfI) Coding Gene in Sugarcane

نویسندگان [English]

  • Maryam Ahmadi 1
  • Farhad Nazarian Firoozabadi 2
  • Ahmad Ismaili 3
  • Bizhan Bajelan 4
1 PhD Student, Department of Agronomy and Plant Breeding, Faculty of Agriculture, University of Lorestan, Khoram Abad, Iran
2 Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, University of Lorestan, Khoram Abad, Iran
3 Associate Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, University of Lorestan, Khoram Abad, Iran
4 Lecturer, Department of Agronomy and Plant Breeding, Faculty of Agriculture, University of Lorestan, Khoram Abad, Iran
چکیده [English]

Sugarcane (Saccharum officinarum) is the most important industrial sugar producing plant. Both tissue culture practices and transformation are challenging and need to be optimized. A gene encoding a mutansucrase (gtfI) enzyme from Streptococcus downei bacterium was isolated and cloned in a binary expression vector and transferred to sugarcane cultivars, using gene gun and Agrobacterium-mediated transformation. Results of this study showed that Agrobacterium was not able to deliver gtfI gene to sugarcane plants cells, whereas Biolistic gene transfer was successful and resulted in almost 25.7% transformation efficiency. Distance between explant and rupture disc carrying gtfI construct had a significant effect on transformation efficiency with 9cm distance producing the higher number of transformants than 12 cm distance. A combination of sorbitol and mannitol osmoticums showed a profound effect on transformation efficiency. Furthermore, a combination of IBA and NAA auxins had a significant effect on root regeneration. Interestingly, mutansucrase was found active in transgenic sugarcane lines utilizing sucrose by almost 33%. Detailed sugar analysis of a few transgenic sugarcane plants revealed that transgenic plants had a significant lower sucrose (lower pol%) content than untransformed control plants.

کلیدواژه‌ها [English]

  • Sugar analysis
  • Transformation
  • Sugarcane
  • Gene transfer
  • Glycosyltransferase
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