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Genetic transformation of Persian melon (Cucumis melo L.) via Agrobacterium | ||
| Iranian Journal of Genetics and Plant Breeding | ||
| دوره 12، شماره 1 - شماره پیاپی 23، تیر 2023، صفحه 11-21 اصل مقاله (769.88 K) | ||
| نوع مقاله: Research paper | ||
| شناسه دیجیتال (DOI): 10.30479/ijgpb.2023.19087.1351 | ||
| نویسندگان | ||
| Davood Naderi1؛ Amir Mousavi* 2؛ Mahmoud Lotfi3 | ||
| 1Department of Horticulture, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran. | ||
| 2Department of Plant Molecular Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran. | ||
| 3Department of Horticulture, College of Aburaihan, University of Tehran, Pakdasht, Tehran, Iran. | ||
| تاریخ دریافت: 28 تیر 1402، تاریخ بازنگری: 18 آذر 1402، تاریخ پذیرش: 25 آذر 1402 | ||
| چکیده | ||
| A reliable Agrobacterium-mediated transformation and regeneration protocol was developed for commercially important endemic Persian melon cultigens (Cucumis melo L.) comprising ‘Eyvanaki’, ‘Samsoori’, and ‘Khatooni’. The effect of selective Murashige and Skoog (MS) medium containing various concentrations of 6-benzyl adenine (BA) (0, 0.5, 1, and 1.5 mg l-1) and 1 mg l-1 Gibberellic acid (GA3) on regeneration of cotyledon, hypocotyl, and cotyledonary petioles derived from 6-day-old in vitro grown seedlings of the three Persian melons were investigated. For transformation, the sensitivity to kanamycin (Km) concentrations (0, 50, 75, 100, 125 mg l-1 ), the effect of three A. tumefaciens strains (GV3103, LBA4404, and AGL0), inoculation time (0.5, 1, 5, and 30 min), and co-cultivation time (24, 48, and 72 h) on direct shoot regeneration of cotyledonary petiole of ‘Samsoori’ were investigated. Shoot regeneration from cotyledonary petiole explants received the highest attention. Cotyledonary petiole segments of ‘Samsoori’ and ‘Khatooni’ treated respectively with 1.0 mg l-1 and 1.5 mg l-1 BA exhibited the highest potential for shoot multiplication; while the regeneration rate of ‘Eyvanaki’ was drastically lower. Putative transgenic ‘Samsoori’ plantlets selected in 100 mg l-1 Km were subcultured on elongation MS medium composed of 100 mg l-1 Km, 0.1 mg l-1 BA, 1 mg l-1 GA3 plus 400 mg l-1 CTX, and then successfully rooted on growth regulator-free MS medium for two weeks. Using histochemical GUS assay along with genomic PCR screening for GusA and VirG genes, the efficiency of transformation was estimated to be 10% for AGL0 and 6% in LBA4404. | ||
| کلیدواژه‌ها | ||
| AGL0؛ Cucumis melo؛ Multiple buds؛ Organogenesis؛ Reporter genes؛ Transgenic plant | ||
| عنوان مقاله [English] | ||
| تراریختی ژنتیکی در خربزه ایرانی (.Cucumis melo L) به واسطه آگروباکتری | ||
| نویسندگان [English] | ||
| داود نادری1؛ امیر موسوی2؛ محمود لطفی3 | ||
| 1گروه باغبانی، واحد اصفهان (خوراسگان)، دانشگاه آزاد اسلامی، اصفهان، ایران. | ||
| 2گروه بیوتکنولوژی کشاورزی، پژوهشکده زیست فناوری کشاورزی، پژوهشگاه ملی مهندسی ژنتیک و زیست فناوری، تهران، ایران. | ||
| 3گروه باغبانی، دانشکده ابوریحان، دانشگاه تهران، پاکدشت، تهران، ایران. | ||
| چکیده [English] | ||
| یک پروتکل قابل اعتماد برای تراریختی و باززایی با واسطه آگروباکتری برای ارقام مهم تجاری خربزه بومی ایرانی (Cucumis melo L. )از جمله «ایوانکی»، «سمسوری» و «خاتونی» ایجاد شد. اثر محیط MS انتخابی حاوی غلظت‌های مختلف 6-بنزیلادنین (BA) 0، 0.5، 1 و 1.5 میلی‌گرم در لیتر) و 1 میلی‌گرم در لیتر اسید جیبرلیک (GA3) بر باززایی دمبرگ‌های لپه ای، هیپوکوتیل و لپه‌های حاصل از دانهال‌های 6 روزه رشد یافته در شرایط درون شیشه سه خربزه ایرانی مورد بررسی قرار گرفت. برای مراحل تراریختی، ابتدا حساسیت به غلظت های کانامایسین (Km) شامل 0، 50، 75، 100، 125 میلی گرم در لیتر، تأثیر سه سویه A. tumefaciens ، شامل GV3103، LBA4404، و AGL0، زمان تلقیح (5/0، 1، 5 و 30 دقیقه) و زمان همکشتی (24، 48 و 72 ساعت) روی زادآوری مستقیم اندام هوایی دمبرگ لپه‌ای رقم سمسوری بررسی شد. باززایی نوساقه ها از ریزنمونه های دمبرگ لپه ای بیشترین کارایی را از خود نشان داد. بخش‌های دمبرگ لپه‌ای «سمسوری» و «خاتونی» به‌ترتیب در 0/1 میلی‌گرم در لیتر و 5/1 میلی‌گرم در لیتر BA بیشترین پتانسیل را برای تکثیر شاخساره نشان دادند. در حالی که میزان باززایی «ایوانکی» به شدت کمتر بود. گیاهچه‌های تراریخته فرضی «سمسوری» گزینش شده در 100 میلی‌گرم در لیتر کانامایسین بر روی محیط کشت MS طویل شدن جوانه متشکل از 100 میلی‌گرم در لیتر در کانامایسین، 1/0 میلی‌گرم در لیتر BA، 1 میلی‌گرم در لیتر GA3 به اضافه 400 میلی‌گرم در لیتر CTX قرار داده شده و سپس به مدت دو هفته با موفقیت روی محیط کشت MS بدون تنظیم کننده رشد ریشه دار شدند. با استفاده از سنجش هیستوشیمیایی GUS همراه با غربالگری ژنومیک بوسیله PCR برای ژن‌های gusA و virG، کارایی تراریختی برای AGL0 ، ده درصد و در LBA4404 ، شش درصد برآورد شد. | ||
| کلیدواژه‌ها [English] | ||
| جوانه ها, اندام زایی, گزارش گر, انتقال, باززایی | ||
| مراجع | ||
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