تولید گیاهان عاری از ویروس سیب توسرخ از طریق کشت مریستم و بررسی اثرات پکتین و فلوروگلوسینول بر ازدیاد آن

نوع مقاله : مقاله پژوهشی

نویسندگان

1 استاد، گروه علوم گیاهی، دانشکده علوم زیستی، دانشگاه خوارزمی، تهران، ایران.

2 استادیار، گروه علوم گیاهی، دانشکده علوم زیستی، دانشگاه خوارزمی، تهران، ایران.

3 دانش آموخته کارشناسی ارشد، گروه علوم گیاهی، دانشکده علوم زیستی، دانشگاه خوارزمی، تهران، ایران.

10.22103/jab.2026.25499.1725

چکیده

هدف: کشت مریستم یکی از روش­هایی است که برای تولید گیاهان عاری از ویروس به­کار می رود. مطالعه حاضر با هدف حذف ویروس­های شایع گیاه سیب توسرخ انجام شد. همچنین به منظور بهبود شاخه­زایی و ریشه­زایی، تأثیر دو ترکیب پکتین و فلوروگلوسینول مورد بررسی قرار گرفت.
مواد و روش‌ها: ریزنمونه­های جوانه جانبی و انتهایی در سه بازه زمانی در تیر، مرداد و شهریور برداشت شدند. استقرار مریستم در غلظت­های مختلف بنزیل آدنین (BA) مورد ارزیابی قرار گرفت. ریزنمونه­های حاصل از رشد مریستم به محیط پرآوری MS دارای هورمون BA و GA3 منتقل شدند. سپس ریزنمونه­ها به محیط بهینه حاوی پکتین و فلوروگلوسینول به منظور بررسی اثر این دو تیمار بر طول شاخساره­ها، تعداد شاخساره و توسعه برگی انتقال یافتند. ریشه­زایی از ساقه­های طویل شده در مرحله پرآوری در محیط MS½ دارای NAA و IBA انجام شد. برای تأیید وضعیت عاری بودن از ویروس، گیاهان باززایی شده از مریستم‌ها با استفاده از RT-PCR آزمایش شدند.
نتایج: نتایج نشان داد ریزنمونه ‌های برداشت شده در نیمه دوم تیرماه در محیط­کشت MS با غلظت 1 میلی­گرم بر لیتر BA بیشترین درصد استقرار ( 66/26 درصد) را نشان دادند. محیط MS با غلظت 1 میلی­گرم بر لیتر BA و 1 میلی­گرم بر لیتر GA3 بهترین محیط برای افزایش طول ساقه بود. بیشترین توسعه برگی در محیط MS با غلظت 1 میلی­گرم بر لیتر BA و بدون GA3 مشاهده شد. اثر متقابل BA و GA3 بر تعداد شاخه­های فرعی معنی­دار بود. اضافه کردن پکتین در مرحله پرآوری باعث افزایش طول ساقه (45 درصد) و افزایش تعداد شاخه (6/82 درصد) شد. با اضافه کردن فلوروگلوسینول به محیط، طول ساقه (24 درصد) افزایش یافت و تعداد شاخه (13/39 درصد) کاهش یافت. بیشترین درصد ریشه‌زایی (34 درصد) در محیط کشتMS ½ با تیمار 1 میلی‌گرم بر لیتر IBA بعلاوه‌ی 5/0 میلی‌گرم بر لیتر NAA مشاهده شد. حضور تیمار فلوروگلوسینول باعث کاهش درصد ریشه­زایی در کل گیاهان و بالا رفتن میانگین تعداد ریشه در هر گیاه شد. در بررسی‌های مولکولی با RT-PCR، هیچ باندی دال بر آلوده بودن گیاهان حاصل از کشت مریستم وجود نداشت.
نتیجه‌گیری: این مطالعه نشان داد که کشت مریستم ابزاری کارآمد برای تولید گیاهان عاری از ویروس در گیاه سیب توسرخ است. روش RT-PCR از دقت کافی جهت ردیابی ویروس برخوردار بود. با در نظر گرفتن تمام جوانب، استفاده از پکتین و فلوروگلوسینول را می­توان به­عنوان دو ترکیب قابل اعتماد در بهبود شاخه­زایی در کشت مریستم مورد استفاده قرار داد.

کلیدواژه‌ها

عنوان مقاله [English]

Production of Malus domestica L. virus free via meristem culture and the effects of pectin and phloroglucinol on its proliferation

نویسندگان [English]

  • Parissa Jonoubi 1
  • Zohreh Shirkhani 2
  • Maedeh Nosoohian 3
1 Professor, Department of Plant Sciences, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.
2 Assistant Professor, Department of Plant Sciences, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.
3 MSc graduate, Department of Plant Sciences, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.
چکیده [English]

Objective
Meristem culture is a method used to produce virus-free plants. This study aimed to eliminate common viruses affecting Malus domestica L. Additionally, the investigation focused on the effects of two compounds, pectin and phloroglucinol, on improving branching and rooting.  
Materials and methods
Lateral and apical bud explants were collected at three different time points in July, August, and September. The establishment of meristems was assessed using various concentrations of BA hormones. Explants derived from meristem growth were then transferred to an MS propagation medium supplemented with BA and GA3 hormones. After identifying the most suitable culture medium for propagation, the explants were moved to an optimal medium containing pectin and phloroglucinol to examine their effects on shoot length, shoot number, and leaf development. Rooting of elongated stems from the propagation stage was performed in MS½ medium with NAA and IBA hormones. Additionally, an experiment was conducted to evaluate the influence of phloroglucinol on rooting percentage. To confirm their virus-free status, regenerated plants from meristems were tested using RT-PCR.
Results
The results indicated that explants collected in the latter half of July in MS medium with a concentration of 1 mg L-1 BA showed the highest establishment percentage (26.66%). The optimal medium for promoting stem elongation contained 1 BA and 1 mg L-1 GA3. The highest leaf development was observed in MS medium with 1 mg L-1 BA and without GA3. The interaction between BA and GA₃ significantly affected the number of secondary branches, whereas no significant effect was observed on the number of primary branches. Adding pectin in the proliferation stage increased the stem length (45%) and the branch number (82.6%). Incorporation of phloroglucinol into the medium led to a 24% increase in stem length but a 39.13% decrease in branch quantity. The highest rooting percentage (34%) was observed in½ MS medium treated with 1 IBA plus 0.5 mg L-1 NAA. Phloroglucinol treatment diminished the rooting percentage while increasing the average number of roots per explant. RT-PCR analysis of the plant samples derived from meristem culture did not reveal any bands indicative of viral infection.
Conclusions
This research demonstrated that meristem culture is an effective technique for generating virus-free red-fleshed apple plants. Additionally, the RT-PCR method proved to be sufficiently precise for virus detection. Taking all factors into account, the application of pectin and phloroglucinol can serve as two dependable compounds to enhance branching during meristem culture.

کلیدواژه‌ها [English]

  • hormone
  • lateral bud
  • regeneration
  • root induction

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مجله بیوتکنولوژی کشاورزی
دوره 18، شماره 3
شهریور 1405
صفحه 27-54

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