بهینه سازی ریزازدیادی پایه GN15 (هیبرید هلو و بادام) در شرایط بیوراکتور غوطه‎وری موقت

نوع مقاله : مقاله پژوهشی

نویسندگان

گروه پژوهشی بیوتکنولوژی گیاهان باغبانی، پژوهشکده بیوتکنولوژی صنعتی، سازمان جهاد دانشگاهی خراسان رضوی، مشهد، ایران

چکیده

محیط کشت مایع به دلیل فراهم کردن محیط یکنواخت تر و عدم نیاز به جابجایی نمونه‎های کشت شده در هنگام تعویض محیط کشت، شرایط بهتری را برای ریزازدیادی گیاهان فراهم می‎کند. به‌منظور بررسی ریزازدیادی پایه GN15، ابتدا اثر ترکیب تنظیم کننده های مختلف بر استقرار ریزنمونه گره در محیط کشت نیمه جامد MS مورد ارزیابی قرار گرفت. سپس جهت مقایسه شرایط بیوراکتور غوطه‎وری موقت و کشت نیمه جامد بر ضریب تکثیر و سلامت گیاهان، در قالب سه آزمایش جداگانه، اثر دوره تناوب غوطه‎وری (غوطه‎وری 10 دقیقه در هر 6، 12 و 24 ساعت)، غلظت تنظیم کننده‎ رشد (غلظت‌های صفر، 5/0 و 1 میلی‌گرم در لیتر BA) و غلظت ساکارز (3، 4 و 5 درصد) در محیط کشت پایه MS مورد ارزیابی قرار گرفت. ریشه زایی گیاهچه‎های تکثیر شده نیز در محیط کشت MS حاوی 6/0 میلی‌گرم در لیترIBA در دو سیستم مورد بررسی قرار گرفت. استقرار ریزنمونه‎ها در محیط کشت به شدت تحت تاثیر انتخاب تنظیم کننده‎های رشد قرار گرفت، به‌طوری که محیط کشت MS حاوی 5/0 میلی‌گرم در لیتر BA بهترین نتیجه را نشان داد. نتایج آزمایش‎های تکثیر مشخص نمود که بیوراکتور غوطه‎وری موقت شرایط بهتری را برای رشد و تکثیر شاخساره‎های GN15 فراهم می‌کند. بهترین شرایط با دوره غوطه‎وری 10 دقیقه در هر شش ساعت، محیط کشت حاوی 5/0 میلی‌گرم در لیتر BA و سه درصد ساکارز به‌دست آمد. همچنین بیوراکتور غوطه‎وری موقت ارزیابی شده در این تحقیق برای ریشه زایی این پایه مناسب نبود و تمام گیاهچه‎ها در این شرایط شیشه‌ای شده و طی فرایند سازگاری از بین رفتند.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Micropropation of GN15 Rootstock (Hybrid of Almond and Peach) in Temporary Immersion Bioreactor

نویسندگان [English]

  • Ahmad Sharifi
  • Mahdiyeh Kharrazi
  • Azadeh khadem
Horticultural Plants Biotechnology Department, Research Institute for Industrial Biotechnology, Iranian Academic Centre for Education, Culture and Research (ACECR)- Khorasan Razavi Branch, Mashhad, Iran.
چکیده [English]

The liquid medium creates optimal conditions for plant micropropagation by providing a more consistent environment and eliminating the hassle of explant transferring during the subcultures. This study was focused on the improving of GN15 rootstock micropropagation. At the first, the impact of various plant growth regulator combinations on node explant establishment in semi-solid MS medium was investigated to select the best establishment medium. Then the comparision between two culture system including temporary immersion bioreactor system and conventional semi-solid was performed to evaluate propagation rate and plant health of GN15 rootstock. To investigate the effects of immersion times (10 minutes every 6, 12, and 24 h), plant growth regulator concentrations (0, 0.5, and 1 mg/l BA) and sucrose concentrations (3, 4, and 5 %) in MS medium three separate experiments were designed. Rooting of microcuttings was assessed in both systems with MS medium containing 0.6 mg/l IBA. The resulted revealed that successful establishment of explants was heavily dependent on plant growth regulators, MS medium containg 0.5 mg/L BA was the best. The results also demonstrated that the temporary immersion system with 10-minute immersions frequency every 6 hours, 0.5 mg/L BA, and 3% sucrose created optimal growth conditions for GN15 shoots. However, the rooting and acclimatization tests revealed that the temporary immersion system was not suitable for rooting the GN15 rootstock under the studied conditions. In overall, to maximize the benefits of the temporary immersion system in GN15 micropropagation, it is recommended to utilize a two-stage strategy. This includes carrying out the propagation in the temporary immersion system followed by rooting the microcuttings in semi-solid media.

کلیدواژه‌ها [English]

  • Humidity
  • Liquid medium
  • Temporary immersion
  • Ventilation
  • Vitrification

Extended Abstract

Introduction

The liquid medium creates a more consistent environment, eliminates explant transfer needs during medium replacement and enhances conditions for plant micropropagation. Moreover, in this culture method, larger vessels can be utilized compared to semi-solid medium, extending transfer times and fostering economical micropropagation. This study focused on the micropropagation of GN15 rootstock, a hybrid of almond and peach, recognized as the optimal rootstock for calcareous soils in Iran, conducted in temporary immersion bioreactor and semi-solid culture setups.

Materials and methods

At the first, the impact of various plant growth regulator combinations on node explant establishment in semi-solid MS medium was investigated to select the best establishment medium. Then the comparison between two culture systems including temporary immersion bioreactor system and conventional semi-solid was done to evaluate propagation indices and plant health of GN15 rootstock. To investigate the effects of immersion times (10 minutes every 6, 12, and 24 h), plant growth regulator concentrations (0, 0.5, and 1 mg/l BA) and sucrose concentrations (3, 4, and 5 %) in MS medium, three separate experiments were designed. Rooting of microcuttings was assessed in both systems with MS medium containing 0.6 mg/l IBA. The plantlets were then transferred to the pots filled with a mixture of cocopeat and vermiculite in a 1:1 ratio for acclimatization.

 

Results and Discussion

The successful establishment of explants in the medium was strongly influenced by the plant growth regulators, with the highest percentage of plantlet establishment observed in the medium containing 1 mg/L BA. This medium also exhibited a high vitrification percentage, although the other media showed lower establishment percentages, no vitrification was observed. Results from propagation experiments in both systems revealed that the temporary immersion system offered superior conditions for the growth and propagation of GN15 plantlets. In the survey of immersion duration, 10-minute immersions every 6 hours resulted in the highest number of shoots, although they showed symptoms of vitrification. As the duration between immersion cycles increased, the number of propagated plantlets decreased. Increasing the concentration of BA led to enhanced shoot proliferation and plantlet growth in both systems, especially in the temporary immersion system. However, higher BA concentrations resulted in vitrification in plantlets from the temporary immersion system. In case of sucrose, a 3% sucrose level was found to be optimal for plantlet propagation in both systems. On the other hand, at a 5% sucrose level, the quantity and quality of propagated plantlets significantly decreased. Rooting and acclimatization results for GN15 rootstock in the two systems indicated that the temporary immersion system was not suitable for rooting this rootstock, as the plantlets became vitrified and disappeared during acclimatization. The semi-solid culture system provided better conditions for rooting.

 

Conclusion

This study demonstrated that for the large-scale propagation of GN15 rootstock under in vitro conditions, shoot propagation using a temporary immersion system is achievable. The system utilized MS medium supplemented with 0.5 mg/L BA, 3% sucrose, and featured immersion cycles of 10 minutes every 6 hours. Additionally, the best rooting of regenerated shoots was achieved in a semi-solid medium.

 

مراجع

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علوم باغبانی ایران
دوره 55، شماره 4
دی 1403
صفحه 577-596

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  • تاریخ دریافت: 01 آذر 1402
  • تاریخ بازنگری: 24 تیر 1403
  • تاریخ پذیرش: 23 مرداد 1403

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