كليدواژه :
باززايي غيرمستقيم , درون شيشه اي , سوخك , لاله واژگون , محيط كشت
چكيده فارسي :
ايران يكي از خاستگاه‌هاي لاله واژگون (Fritillaria imperialis L.) است. اين گياه زينتي در معـرض انقراض مي‌باشد. بنابراين افزايش آن از طريق كشت بافت ضروري است. اين پژوهش در چهار آزمايش: توليد پينه، سوخك‌زايي، ريشه‌زايي و سازگاري انجام گرديد. در آزمايش اول از دو ريزنمونه فلس و برگ اوليه در محيط كشت MS با غلظت هاي مختلف NAA و IBA اقدام به توليد پينه گرديد. نتايج نشان داد بيشترين وزن پينه از ريزنمونه فلس در محيط كشت MS حاوي 0/5 ميلي‌گرم در ليتر NAA و 0/5 ميلي‌گرم در ليتر IBA توليد گرديد. در آزمايش دوم از پينه هاي به‌دست آمده، در محيط كشت MS با غلظت‌هاي مختلف TDZ و KIN، پس از 6 و 12 هفته، تعداد سوخك مورد بررسي قرار گرفت. نتايج نشان داد، بيشترين تعداد سوخك بعد از 12 هفته از پينه حاصل از ريزنمونه فلس در محيط كشت MS حاوي 0/5 ميلي‌گرم در ليتر NAA و سپس در محيط كشت اندام زايي با 0/5 ميلي‌گرم در ليتر TDZ همراه با 0/5 ميلي‌گرم در ليتر KIN تشكيل شد. سپس سوخك‌ها به محيط كشت MS با غلظت‌هاي مختلف NAA (صفر، 0/5، 1 و 2 ميلي‌گرم در ليتر) جهت ريشه‌زايي منتقل شدند. نتايج نشان داد، بيشترين تعداد ريشه از سوخك‌هاي حاصل از ريزنمونه برگ اوليه، در محيط كشت حاوي 1 ميلي‌گرم در ليترNAA به‌دست آمد. در‌نهايت سوخك‌هاي ريشه‌دار شده پس از شستشو با آب مقطر جهت سازگاري به گلدان‌هاي حاوي پرليت،كوكوپيت، پرليت و كوكوپيت كه در اتوكلاو گندزدايي شده بودند، منتقل شدند. نتايج نشان داد سوخك‌هاي حاصل از ريزنمونه فلس در بستر كشت كوكوپيت، داراي بيشترين تعداد ريشه بود. اين تحقيق نشان داد كه استفاده از كشت بافت، نقش مهمي در جلوگيري از انقراض لاله واژگون دارد.
چكيده لاتين :
Background and Objectives
Iran is home of one of the most delightful and graceful flower of Fritillaria in the world. Its habitation is commonly expanded on the vast areas: growing in beds, borders, backgrounds and rock gardens. Besides, it is also suitable for naturalizing and excellent for cut flowers. Fritillaria imperialis as a medicinal plant has pharmaceutical purpose in which possesses a health beneficial essence that used in production of mucus and cough medicine. In addition, extractions of plant tissues contain certain of properties which affect to reduce blood pressure and blood sugar. Unfortunately, the habitation of this ornamental plant is under threat and majority of its habitation already being to ruin due to human activities and urbanization. The propagation of Fritillaria through offsets (bulblets) is prolonging approach which takes approximately 3-4 years. Whereas, using tissue culture technique for flower production is required less time probably within a few months and economically has a huge advantage due to low cost and a numerous plant production in a short time.
Materials and methods
This research was carried out in four experiments: callus formation, bulblet production, root formation and acclimatization. In the present study, a factorial experiment was assigned as completed randomized design (CRD) with three replications. In the first experiment, leaf primordia and scale explants were cultured on MS medium supplemented with different concentrations of NAA and IBA in order to produce callus formation. In the second experiment, the calli were cultured on organogenesis medium which were contained MS medium supplement, with kin. and TDZ, after 6 and 12 weeks. In the third experiment, the bulblets were transferred into rooting media, which were contained MS medium supplemented with different concentrations of NAA (0, 0.5, 1.0 and 2.0 mg/l). In the fourth experiment, after washing the bulblet whit warm distilled water (40ᵒC) that contain 2.0 mg/l benlate fungicide, they then were planted into the pots containing sterilized perlite, cocopeat and perlite along with cocopeat.
Results
In the first experiment, the results demonstrated that the highest weight of callus (3.29g) was sustained in scale explants on basal MS medium which was supplemented with 0.5mg/l NAA along with 0.5mg/l IBA. In second experiment, the result indicated that the numerous of bulblets were developed on MS medium supplemented with 0.5mg/l TDZ along with 0.5mg/l kin. Which of these, the callus scale were produced on MS medium supplemented with 0.5mg/l NAA. In the third experiment, in order to retain rooting the bulblets, they were cultured on MS medium supplemented with different concentration of NAA. The result elucidated that the large quantity of roots (7.7) were obtained from leaf primordia on MS medium supplemented with 1.0mg/l NAA. In the fourth experiment, rooted bulblets were washed with warm distilled water for 5-10min, and then they were transferred into pots with perlite, cocopeat and perlite + cocopeat.
Discussion
The results revealed information that the rooted bulblets from scale growing on cocopeat substrate had retained the most diameter (21.64mm) as compared to other treatments. The finding results under this investigation strongly suggested that using micropropagation it is definitely a possible trend to abolish the barriers which caused to deteriorate of Fritillaria imperiallis.
