مطالعه برخی عوامل مؤثر بر جنین‌زایی میکروسپورها و باززایی گیاهچه‌ها در کلزا (Brassica napus L.)

نوع مقاله : علمی پژوهشی

نویسندگان

1 دانشیار دانشگاه آزاد اسلامی، واحد رشت، گروه زراعت و اصلاح نباتات، رشت، ایران

2 دانشیار گروه اصلاح نباتات و بیوتکنولوژی، دانشکدة کشاورزی، تهران، ایران.

چکیده

برخی عوامل موثر بر جنین‌زایی در کشت میکروسپورهای جداشده کلزا و باززایی گیاهچه‌ها از جنین‌های حاصله در آزمایش‌هایی جداگانه بررسی شدند. آزمایش‌ها به صورت فاکتوریل دو فاکتوره و در قالب طرح پایه کاملاً تصادفی انجام شدند. در تمام آزمایش‌ها، ارقام در سه سطح شامل گلوبال (Global)، اپشن (Option) و پی‌اف (PF7045/91) فاکتور اول را تشکیل می‌دادند. میکروسپورها از غنچه‌های به طول 4-3 میلی‌متر جدا و در محیط کشت NLN-13 کشت شدند. میکروسپورهای کشت‌شـده به مدت ۱۴ روز در معرض تنش دمایی ۳۰ درجه سانتی‌گراد و در شرایط تاریکی قرار گرفتند و پس از آن به اتاق رشدی با دمای ۲۵ درجه سانتی‌گراد و در داخل یک لرزاننده منتقل گردیدند. برای باززایی، جنین‌های 35-25 روزه به محیط کشت جامد B5 منتقل ‌شدند و بعد از حدود 20 تا 25 روز، گیاهچه‌های طبیعی، گیاهچه‌های غیر طبیعی، جنین‌های ریشه‌دار و جنین‌های تغییرنکرده شمارش شدند. در آزمایش اول جنین‌زایی اثر حجم محیط کشت، رقم و نیز اثرمتقابل آنها معنی‌دار بود. در رقم گلوبال، حجم 5/12 میلی‌لیتر محیط کشت با متوسط تولید 5/695 جنین در هر پتری‌دیش، بیشترین میزان جنین‌زایی را داشت. در آزمایش دوم جنین‌زایی، اثر زغال فعال و اثرمتقابل دو فاکتور معنی‌دار بود. در آزمایش سوم جنین‌زایی منابع مختلف هیدرات کربن با هم اختلاف معنی‌دار داشتند. در آزمایش اول باززایی، اسید جیبرلیک اثر معنی‌دار بر تعداد جنین‌های تغییرنکرده، گیاهچه‌های غیرطبیعی، گیاهچه‌های طبیعی و جنین‌های ریشه‌دار داشت و اثرمتقابل اسید جیبرلیک و رقم بر گیاهچه‌های طبیعی معنی‌دار بود. در آزمایش دوم باززایی، اثرمتقابل عامل ژلی و رقم بر گیاهچه‌های طبیعی معنی‌دار بود.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Effects of some factors on microspore embryogenesis and plantlet regeneration in Brassica napus L.

نویسندگان [English]

  • Peyman Sharifi 1
  • Ahmad Moieni 2
1 Associate Professor, Department of Agronomy and Plant Breeding, Rasht Branch, Islamic Azad University, Rasht, Iran
2 Associate Professor, Department of Plant Breeding and Biotechnology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran
چکیده [English]

The effect of some factors on embryogenesis from isolated microspore and plantlet regeneration from microspore-derived embryos was studied separately in Brassica napus. Experiments carried out in factorial based on completely randomized design. The first factor in all of the experiments was cultivars contained Global, Option and PF7045/91. The microspores were isolated from 3-4 mm buds and cultured on NLN-13 medium. Cultures incubated at 30˚C and darkness for 14 days, and then transferred to shaker in the growth chamber at 25˚C. In regeneration experiment, embryos with 20-25 days old were transferred to B5 medium and nearly 20-25 days after transferring embryos, normal regenerated plantlets, abnormal regenerated plantlets, rooted embryos and non differentiated embryos were counted. In the first embryogenesis experiments, the effects of medium volume, cultivar and interaction effects of two factors were significant. In Global cultivar, the highest values of embryos (695.5 per Petri) were observed in 12.5 ml medium volume. In the second embryogenesis experiment, the interaction between activated charcoal and cultivar on embryogenesis was significant. In the third embryogenesis experiment, the form of carbohydrate had a significant effect on embryo yield. In the first experiment of plantlet regeneration, GA3 had significant effect on normal regenerated plantlets, abnormal regenerated plantlets, rooted embryos and non differentiated embryos and the interaction between GA3 and cultivar was significant for normal regenerated plantlets. In the second experiment of plantlet regeneration, the interaction between gelling agent and cultivar was significant on normal regenerated plantlets.

کلیدواژه‌ها [English]

  • Embryogenesis
  • Haploid Plants
  • Microspore Culture
  • Rapeseed
  • Plant Regeneration

مراجع

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