اثر رقم و تیمار تنظیم کننده رشد گیاهی بر باززایی درون شیشه‌ای لیلیوم با استفاده از ریزنمونه‌های TCL

نوع مقاله : مقالات پژوهشی

نویسندگان

• احمد شریفی ¹
• فاطمه کیخا آخر ¹
• محبوبه یزدی ²
• عبدالرضا باقری ²

¹ جهاد دانشگاهی استان خراسان رضوی

² دانشگاه فردوسی مشهد

https://doi.org/10.22067/jhorts4.v31i3.55475

چکیده

لیلیوم به عنوان چهارمین گل شاخه بریده دنیا مطرح است و روش‌ کشت بافت، روش مناسبی را برای تکثیر این گیاه زینتی فراهم کرده است. در این پژوهش به منظور بررسی عوامل مؤثر بر باززایی لیلیوم از پیازک‌های تولید شده در شرایط درون شیشه‌ای دو آزمایش جداگانه انجام گرفت. در مرحله اول، اثر ریزنمونه‌های با لایه سلولی نازک (TCL) با ضخامت‌های 1، 3 و 5 میلی‌متر در محیط کشت پایه موراشیگ و اسکوگ (MS) حاوی ترکیب‌های تنظیم کننده رشد گیاهی BA، 2ip و Kin با غلظت‌ 1 میلی‌گرم در لیتر در ترکیب با 5/0 میلی‌گرم در لیتر NAA بررسی شد. نتایج این آزمایش نشان داد که در هر سه تیمارتنظیم کننده رشد گیاهی، بیشترین میزان اندام زایی در ریزنمونه‌هایی با ضخامت 3 میلی‌متر بدست آمد. در واقع نتایج برهمکنش اندازه ریزنمونه‌ با تنظیم کننده رشد گیاهی موجود در محیط کشت نشان داد درصد باززایی در ریزنمونه‌های با ضخامت 3 میلی‌متر بهتر افزایش داشته است. بیشترین تعداد برگ و وزن خشک گیاهچه‌ها در محیط کشت حاوی BA مشاهده شد که نسبت به سایر تیمارها تفاوت معنی‌داری داشت. در آزمایش دوم به منظور بررسی اثر رقم و نوع تنظیم کننده رشد گیاهی سیتوکینین، ریزنمونه‌های TCL ارقام Robina، Donato، Nymph، Lessoto و Roxana با ضخامت 3 میلی‌متر در محیط ‌کشت MS حاوی ترکیبات تنظیم کننده رشد گیاهی مختلف (BA، Kin، 2ip و TDZ) با غلظت 1 میلی‌گرم در لیتر در ترکیب با 5/0 میلی‌گرم در لیتر NAA کشت شدند. نتایج نشان داد که رقم به عنوان یکی از عوامل تأثیرگذار بر اندام زایی بود. بیشترین تعداد ریشه، وزن خشک، تعداد پیازک و طول گیاهچه در رقم Roxanaمشاهده شد. بررسی درصد باززایی و شاخه‌زایی در ریزنمونه‌ها نشان داد که ارقام و هورمون‌های مختلف بر میزان باززایی و ریشه‌زایی مؤثر بوده و رقم Nymph در حضور TDZ نسبت به سایر ارقام و تنظیم کننده های رشد گیاهی کمترین باززایی و ریشه‌زایی را نشان داد. همچنین مقایسه وزن خشک در تیمارهای مختلف نشان داد که وزن خشک گیاهچه‌های رقم Roxana بطور معنی‌داری نسبت به سایر تیمارها افزایش یافته بود.

کلیدواژه‌ها

• اکسین
• ریزازدیادی
• ریزنمونه فلس
• سیتوکینین
• کشت درون شیشه‌ای

20.1001.1.20084730.1396.31.3.4.0

عنوان مقاله [English]

Effect of Cultivar and Plant Growth Regulators on Invitro Regeneration of Lilium Spp. Utilizing Thin Cell Layer Explants

نویسندگان [English]

• Ahmad Sharifi ¹
• Fatemeh Keykha Akhar ¹
• Mahboobeh Yazdi ²
• Abdolreza Bagheri ²

¹ Academic Center for Education, Culture and Research, Branch of Khorasan Razavi

² Ferdowsi University of Mashhad

چکیده [English]

Introduction: Lily (Lilium spp.) is a genus of herbaceous flowering plants, which consisted of many beautiful ornamental species with large prominent flowers. Most species are native to the Northern hemisphere temperate, though their range extends into the Northern subtropics. Some specific hybrids of Lilium spp. have been developed as cut flower in controlled conditions and in some cases can be grown as pot plant. Propagation rate of lily in natural clonal propagation methods is very low and one year produces of 1-2 bulblets per bulb scale. There is also possibility of disease transmission; so that, tissue culture techniques has provided an efficient method for its micropropagation.
Materials and Methods: In this study, two separate experiments under In vitro conditions the bulblet regeneration from thin cell layer (TCL) explants of Lilium spp. was investigated. In the first experiment, after two months the effect of TCL explants with 1, 3 and 5 mm thickness on MS medium containing 1 mg/l BA, 2ip and kin in combination with 0.5 mg/l NAA on regeneration parameters were assayed. In the second experiment to determine the effect of cultivar and cytokinin types, 3mm thickness TCL explants of five cultivars (Robina, Donato, Nymph, Lessoto and Roxana) were tested on MS medium containing different plant growth regulator (PGR) compounds including BA, kin, 2ip and TDZ at concentration of 1 mg/l in combination with 0.5 mg/l NAA. The regeneration parameters were assayed after four months. In all experiments, the medium was adjusted to pH 5.8 and autoclaved at 121°C for 15 min. All cultures were incubated at 25 ± 2°C with a 16 h photoperiod under cool white flourescent lights (30 µmol/m2).
Results and Discussion: According to the first experiment results, plant growth regulator of BA in all of surveyed parameters except root number was better than other PGRs and explants with 3 mm thickness was the best in all of parameters. The interaction of PGR and explants was significant, however maximum bulblet regeneration was observed in TCL explants with 3 mm thickness in all of PGR treatments (100%). While 1 mm thickness TCL in 2ip and 1 and 5 mm thickness TCL in Kin had the least regeneration percentage. Results revealed that the interaction of explants and medium is a key factor for suitable establishment, regeneration and growth of TCLs. Bulb dormancy is one of the limiting factors in regeneration of bulbous crop species. It seems under In vitro condition explants size and PGR combination of media especially cytokinin affected on breaking of dormancy. Maximum number of leaves and dry weight of bulblets in medium containing BA was significantly higher compared with other treatments. Most of studies confirmed the positive effect of BA on regeneration of lily. The function of cytokinin in plant promoted cell division and differentiation, which lead to growth and maintaining cells in meristematic status.
Result of second experiment showed that cultivar was one of the effective factors on regeneration trait. Oriental lily cultivar "Roxana" had the highest number of roots, bulblets, dry weight and length of plantlets and "Nymph" cultivar showed the lowest percentage of regeneration, dry weight, length of plantlets and rooting obtained. In all of cultivars BA induced more organogenesis percentage and plantlet dry weight, while TDZ induced more rooting percentage.The interaction of cultivar and PGR treatments on percentage of regenerated bulblets and rooting were significant. "Nymph" cultivar had minimum percentage of regeneration and rooting in medium containing TDZ and Kin. Furthermore, "Roxana" cultivar in medium containing BA showed the best dry weight comparison to other treatments.
Conclusion Lily has widely used in the floral industry as a cut flower or potted plant. In recent years, tissue culture was developed as reliable and highly effective method to overcome its limitations of vegetative propagation. The most advantage of this method is high multiplication rate and disease free propagation. In this study, bulblet regeneration of lilium Spp. from TCL explants under in vitro condition was considered as a highly efficient procedure for its micropropagation. With optimization of TCL system some parameters such as exogenously applied plant growth regulators, cultivar, explants types were investigated. Favorable conditions for bulblet regeneration were achieved with 3 mm thickness TCLs in MS medium containing 1 mg/l BA with 0.5 mg/l NAA. This protocol can be used for rapid micropropagation of many cultivars.

کلیدواژه‌ها [English]

• Auxin
• Bulb explant
• Cytokinin
• In vitro culture
• Micropropagation

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