مطالعه اثر جهت قرار دادن بساک، نوع و ترکیب محیط‌کشت و کشت توأم بساک با تخمدان روی القاء کالوس در کشت بساک کدوی تخم پوست‌کاغذی (Cucurbita pepo var. Styriaca)

نوع مقاله: علمی - پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه بوعلی‌سینا، همدان

2 دانشیار گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه بوعلی‌سینا، همدان

3 استادیار گروه بیوتکنولوژی کشاورزی، دانشکده کشاورزی، دانشگاه بوعلی‌سینا، همدان

4 استادیار گروه زراعت و اصلاح نباتات، دانشکده کشاورزی، دانشگاه بوعلی سینا، همدان

چکیده

در این تحقیق، کشت بساک گیاه کدوی تخم پوست‌کاغذی (Cucurbita pepo var. Styriaca) به‌منظور تولید کالوس و جنین هاپلوئید بررسی شد. آزمایشات سیتوژنتیکی نشان داد که غنچه‌های 5/1 سانتی‌متری حاوی بساک‌های10-9 میلی‌متری و میکروسپورهایی در مرحله تک هسته‌ای میانی تا انتهایی برای کشت بساک مناسب بودند. سه آزمایش فاکتوریل در قالب طرح کاملاً تصادفی با سه تکرار اجرا گردید. در آزمایش اول، اثر جهت قرار دادن بساک بر روی محیط‌کشت و نوع و غلظت تنظیم‌کننده‌هاِی رشد گیاهی NAA (صفر، 5/0، 1 میلی‌گرم در لیتر) و 2,4-D (صفر، 1، 2 میلی‌گرم در لیتر) روی صفت درصد کالوس‌زایی بررسی شد. در این آزمایش کشت بساک‌ها از جهت محدب در محیط حاوی 1 میلی‌گرم در لیتر NAA و 2 میلی‌گرم در لیتر 2,4-D، بیش‌ترین درصد کالوس‌زایی را ایجاد کرد. در آزمایش دوم، اثرات متقابل نوع محیط‌کشت (E20 و B5) و ترکیب تیمارهای هورمونی2,4-D  (صفر، 5/2، 5 میلی‌گرم در لیتر) و BAP (صفر، 5/0، 1 میلی‌گرم در لیتر) بر صفت درصد کالوس‌زایی کدو مورد مطالعه قرار گرفت که محیط‌کشت E20 حاوی 1 میلی‌گرم در لیتر BAP و 5/2 میلی‌گرم در لیتر 2,4-D، بیش‌ترین درصد کالوس‌زایی را در مقایسه با تیمارهای دیگر ایجاد کرد. در آزمایش سوم اثر کشت توأم بساک کدوی تخم پوست‌کاغذی با تخمدان گندم و تخمدان کدو بر درصد کالوس‌زایی بساک کدو در دو نوع محیط‌کشت جامد و مایع E20 بررسی شد. در این آزمایش کشت توأم بساک کدو و تخمدان گندم در محیط‌کشت جامد، نسبت به بقیه تیمارها بیش‌ترین درصد کالوس‌زایی را نشان داد.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

The Study on the Effect of Anther Orientation, Type and Composition of Culture Medium and Ovary Co-culture on Callus Induction from Cultured Anthers of Pumpkin (Cucurbita pepo var. Styriaca)

نویسندگان [English]

  • Zhale Mohseni Araghi 1
  • Mohammad Reza Abdollahi 2
  • Asghar Mirzaie Asl 3
  • Javad Hamzei 2
  • Seyed Saed Mousavi 4
1 MSc Student, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Bu Ali Sina University, Hamedan
2 Associate Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Bu Ali Sina University, Hamedan
3 Assistant Professor, Department of Agriculture Biotechnology, Faculty of Agriculture, Bu Ali Sina University, Hamedan
4 Assistant Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Bu Ali Sina University, Hamedan
چکیده [English]

The production of doubled haploid cucurbit plants can accelerate the breeding programs of these species. In this research, anther culture of Cucurbita pepo var. styriaca was studied to produce callus and haploid embryos. Cytological studies showed that 15 mm male buds containing 9-10 mm anthers in size and microspores at mid to late-uninucleate were desirable for anther culture experiments. Three experiments were conducted as factorial based on completely Randomized Design (CRD) with three replications. In first experiment, the effect of anther orientation on culture medium as well as type and concentration of the plant growth regulators of NAA (0, 0.5 and 1 mgl-1) and 2,4-D (0, 1 and 2 mgl-1) on callogenesis percentage of pumpkin anther cultures were studied. Results showed that the culturing of anthers from convex side in a culture medium supplemented with 2 mgl-1 2,4-D and 1 mgl-1 NAA resulted in the highest percentage of callogenesis. In second experiment, the interaction effects of medium type (B5 and E20) and combination treatments of 2,4-D (0, 2.5 and 5 mgl-1) and BAP (0, 0.5 and 1 mgl-1) on callogenesis percentage were studied. E20 culture medium supplemented with 2.5 mgl-1 2,4-D and 1 mgl-1 BAP produced the highest percentage of callogenesis compared to other treatments. In third experiment, the effect of pumpkin and wheat ovary conditioned medium on callogenesis percentage from pumpkin anthers were studied in two E20 culture media (Solid and liquid). In this experiment, co-culturing of pumpkin anthers with wheat ovaries in solid culture medium resulted in the highest percentage of callogenesis compared to other treatments. 

کلیدواژه‌ها [English]

  • Anther culture
  • Callogenesis
  • Plant growth regulator
  • Ovary co-culture
  • Haploid

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