بهینه‌سازی شرایط رشد ریشه‌های مویین گیاه Stevia rebaudiana Bertoni حاصل تلقیح Agrobacterium rhizogenes

نوع مقاله : علمی پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد، گروه علوم باغبانی، دانشکده کشاورزی، دانشگاه مراغه، مراغه، ایران.

2 استادیار، گروه علوم باغبانی، دانشکده کشاورزی، دانشگاه مراغه، مراغه، ایران.

3 دانشیار، گروه علوم باغبانی، دانشکده کشاورزی، دانشگاه مراغه، مراغه، ایران و دانشیار پژوهشی مرکز پژوهش‌های مجلس شورای اسلامی، تهران، ایران.

چکیده

گیاه دارویی استویا Stevia rebaudiana Bertoni درختچه چند ساله کوچکی از خانواده Asteraceae است که بومی جنوب آمریکا، به‌ویژه برزیل و پاراگوئه، می‌باشد. این پژوهش به‌منظور بررسی تأثیر عوامل مختلف در القا و رشد ریشه‌های مویین توسط آگروباکتریوم رایزوژنز در گیاه استویا صورت گرفت. بذور استویا در شرایط درون‌شیشه‌ای کشت شدند. پس از جوانه‌زنی و رشد گیاهچه‌ها، ریزنمونه‌های قطعات ریشه، ساقه و برگ در سنین 30، 60 و 90 روزه از گیاهچه‌های درون شیشه‌ای به‌دست آمد. سویه‌های ATCC15834، R1000 وC58 آگروباکتریوم رایزوژنز (حاوی صفر و 100 میکرومولار استوسرینگون) در دو محیط کشت NYA و LB با روش غوطه‌وری و اسپری به‌مدت 72 ساعت در دمای 27 درجه سانتی‌گراد هم‌کشت گردیدند. ریزنمونه‌های تلقیح شده بعد از حذف باکتری اضافی به محیط 1/2MS و MS جامد حاوی 400 میلی‌گرم بر لیتر آنتی‌بیوتیک سفوتاکسیم منتقل گردیدند. به‌منظور بهینه‌سازی محیط‌کشت‌های مختلف بر رشد ریشه‌های مویین در محیط کشت مایع، از محیط1/2MS با غلظت‌های 5/1% و 3 % ساکارز و IBA 2/0 میلی‌گرم بر لیتر استفاده‌ شد. نتایج حاصل از این تحقیق نشان داد هر سه سویه آگروباکتریوم بر القای ریشه مویین در ریزنمونه برگ استویا مناسب بودند ولی، سویه ATCC15834 قابلیت بیشتری در القای ریشه‌های مویین داشت. بیشترین میزان فراوانی ریشه مویین در محیط سوسپانسیون NYA و محیط هم‌کشتی 1/2MS بود. همچنین در مرحله پرآوری ریشه‌های مویین محیط 1/2MS دارای 5/1درصد ساکارز مناسب-ترین محیط شناخته شد. لاین‌های حاصل از سویه ATCC15834 استویا، کلون برگ (C2 LA) پر‌رشدترین لاین بود.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Optimization of hairy root production for Stevia rebaudiana Bertoni through inoculation by Agrobacterium rhizogenes

نویسندگان [English]

  • Roghieh Sharifpour 1
  • Mohammadali Ali Aazami 2
  • Mohammad Bagher Hassanpouraghdam 3
1 M. Sc. Student , Department of Horticulture, Faculty of Agriculture, University of Maragheh, Maragheh, Iran.
2 M. Sc. Student, Department of Horticulture, Faculty of Agriculture, University of Maragheh, Maragheh, Iran.
3 Associate Professor, Department of Horticulture, Faculty of Agriculture, University of Maragheh, Maragheh, Iran. and Research Associate Professor, Islamic Parliament Research Center, Tehran, Iran
چکیده [English]

Stevia rebaudiana Bertoni is a perennial shrub from the Asteraceae; native of Brazil and Paraguai. This Plant is containing sweet steviosides; rebaudioside and isostevol. This experiment was conducted to study the effects of diverse factors on induction and growth of hairy roots induced by Agrobacterium rhizogenes. The seeds were surface sterilized and then cultured on solid MS medium under 250C and 16:8 h photoperiod. After germination and development of seedlings, the root, stem and leaf explant of three different ages (30, 60 and 90 days) were placed on NYA and LB suspension media. Agrobacterium rhizogenes strains (ATTCC15834, R1000 and C58) and acetosyringone were treated on the plant sample by floating and spray method for 72 hrs at 270C. The incubated explants were treated to remove any bacteria and were transferred to 1/2MS media containing 400 mgl-1 cefotaxime. To optimize the growing media for hairy root induction and growth, 1/2 MS medium was enriched with 3% sucrose and 0.2 mgl-1 Indole-3-butyric acid. The results revealed that all the Agrobacterium strains were effective on hairy root induction from root and leaf explant. The highest hairy root induction was belonged to NYA suspension medium co-cultured with 1/2 MS. Furthermore, for the roots proliferation, 1/2MS medium containing 1/5% sucrose was the selected one. Leaf clone (C2LA) stevia lines obtained from ATCC15834 strain had the highest growth.

کلیدواژه‌ها [English]

  • Stevia rebaudiana
  • Agrobacterium rhizogenes
  • Hairy root
  • medium

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