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اثر بازدارندگی و مکانیسم نانو ذرات نقره در کنترل رشد اگروباکتریوم رایزوژنز پس از همکشتی و انتقال ژن به توتون | ||
زیست شناسی کاربردی | ||
مقاله 1، دوره 33، شماره 4 - شماره پیاپی 66، اسفند 1399، صفحه 9-25 اصل مقاله (795.3 K) | ||
نوع مقاله: مقاله پژوهشی | ||
شناسه دیجیتال (DOI): 10.22051/jab.2020.29972.1345 | ||
نویسندگان | ||
مصطفی خوشحال سرمست* 1؛ حسن صالحی2 | ||
1استادیار گروه علوم باغبانی، دانشکده تولیدات گیاهی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان، ایران | ||
2استاد گروه علوم باغبانی، دانشکده علوم کشاورزی، دانشگاه شیراز، شیراز، ایران | ||
چکیده | ||
هنوز تأثیر احتمالی نانو ذرات نقره در کمک به فرایند انتقال ژن در گیاه از طریق اگروباکتریوم مورد توجه قرار نگرفته است. در اینجا ما نشان میدهیم که رشد اگروباکتریوم در غلظت 10 میکروگرم بر میلی لیتر نانو ذرات نقره هم در شرایط رشد دینامیک و هم محیط کشت جامد LB سرکوب شد اما پس از همکشتی قطعات برگ توتون با A. rhizogenes، کنترل رشد مجدد این باکتری به طور موثری نیازمند غلظتهای بالاتری از نانو ذرات نقره بود. استفاده از غلظت 150 میکروگرم در میلی لیتر و بالاتر باعث آسیب به بافت برگ میگردد و غلظت 100 میکروگرم در میلی لیتر در کنترل کامل باکتری پس از همکشتی موثر عمل نمیکند. بدین منظور مصرف همزمان نانو ذرات نقره و سفوتاکسیم با غلظتهای مختلف مورد بررسی قرار گرفت و مشخص شد که استفاده از 100 میکروگرم در میلی لیتر نانو ذرات نقره به همراه 200 میلیگرم در لیتر سفوتاکسیم باعث کمترین آسیب به برگ و بیشترین درصد باززایی میشود و این روش کاربرد نه تنها باعث کاهش سمیت عناصر سنگین میشود بلکه باعث میشود از مصرف غلظتهای بالای آنتی بیوتیک طی انتقال ژن اجتناب نماییم. نتایج میکروسکوپ الکترونی TEM نشان داد که نانو ذرات نقره میتواند رشد اگروباکتریوم را با چسبیدن به دیواره باکتری و نفوذ به درون سلول باکتری و اختلال در کار اندامکهای مختلف متوقف نمایند. نتایج بدست آمده از این آزمایش میتواند راه را برای استفاده از نانو ذرات نقره با قطر کمتر جهت توقف رشد مجدد باکتری طی انتقال ژن در گیاهان مختلف هموار نماید. | ||
کلیدواژهها | ||
اگروباکتریوم رایزوژنز؛ میکروسکوپ الکترونی؛ توتون؛ مهندسی ژنتیک | ||
عنوان مقاله [English] | ||
Inhibitory effects and mechanism of silver nanoparticles in control of Agrobacterium rhizogenes growth after co-cultivation and genetic transformation in tobacco | ||
نویسندگان [English] | ||
Mostafa Khoshhal Sarmast1؛ hasan Salehi2 | ||
1Assistant Professor, Department of Horticulture, Faculty of Plant Production, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran | ||
2Professor, Department of Horticulture, Faculty of Agricultural Sciences, Shiraz University, Shiraz, Iran | ||
چکیده [English] | ||
Potential influence of AgNPs on plant genetic transformation through Agrobacterium has not yet been addressed. Here we showed that the growth of Agrobacterium was suppressed in 10 µg/ml AgNPs but controlling the overgrowth of these bacteria would effectively necessitate a higher concentration of AgNPs when tobacco explants have inoculated with A. rhizogenes. Research result indicated that applying more than 150 µg/ml AgNPs and more, resulted in leaf injury and application of 100 µg/ml of AgNPs was unable to suppress bacteria regrowth after co-cultivation with tobacco leaves. The concurrent application of the AgNPs and Cefotaxime with different concentration was investigated and results indicated that using 100 µg/ml of AgNPs along with 200 mg/l of cefotaxime lead to the lowest leaf injury and the highest regeneration potential. This application not only caused a reduction in heavy metal toxicity but also decreases excess concentrations of antibiotics during the course of transformation. TEM manifested that the AgNPs could suppress Agrobacterium growth by potentially anchoring to and penetrating the bacterial cell wall. Our results suggest that the simultaneous use of AgNPs along with Cefotaxime can suppress the overgrowth of Agrobacterium during plant transformation. The result of this experiment can open a new window for application of AgNPs with lower diameter in order to suppress bacteria overgrowth. | ||
کلیدواژهها [English] | ||
Agrobacterium rhizogenes. Electron Microscopy, Genetic engineering, Tobacco | ||
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