بهينه سازي اندام زايي غيرمستقيم گياه آنتوريوم(Anthuriumscherzerianum)

Optimization of Indirect Regeneration in Anthuriumscherzerianum

آنتوریوم گلدانی(Anthuriumscherzerianum)یك گیاهزینتی چند سالهاست كه به دلیل مشكلات تكثیر سنتی در این گیاه، از كشت بافت به عنوان روشی مناسب برای تكثیر سریع و حذف بیماری¬های آن نام برده می شود. در این پژوهش اثر تنظیم كننده-های رشد گیاهی و نوع ریزنمونه بر روی باززایی غیرمستقیم A.scherzerianum مورد بررسی قرار گرفت. به منظور القای كالوس در ریزنمونه¬های برگ و دمبرگ آنتوریوم اثر سطوح مختلف تنظیم كننده¬های رشد BA و TDZ (5/0، 1 و 2 میلی گرم بر لیتر) در تركیب با NAA یا 2,4-D (5/0، 1 و 2 میلی گرم بر لیتر) در محیط كشت پایه MSدر شرایط تاریكی مورد بررسی قرار گرفت.برای القای باززایی در كالوس¬های تولید شده اثر سطوح پایین تر سایتوكنین¬ها و اكسین¬ها در محیط كشت پایه MS بررسی شد. برای ریشه¬زایی گیاهچه¬های، از محیط كشت1⁄2MS حاوی تركیبات هورمونی IBA و IAA استفاده شد. نتایج آزمایش¬ها نشان داد كه ریزنمونه دمبرگ بیشترین كالوس¬زایی، درصد زنده¬مانی و حجم كالوس را در محیط كشت حاوی 2 میلی گرم در لیتر BAو 5/0 میلی گرم در لیتر 2,4-D تولید كرد. همچنین نتایج آزمایش باززایی نشان داد كه در تركیب هورمونی 75/0 میلی گرم در لیتر BAو 05/0 میلی گرم در لیتر2,4-D بیشترین تعداد شاخساره (9/6)، طول شاخساره (5 سانتی متر) تولید شد. در آزمایش ریشه¬زایی نیز بیشترین درصد ریشه¬زایی (95 درصد)، تعداد ریشه (5/4) و طول ریشه (5/3 سانتی متر) در محیط كشت حاوی 2/0 میلی گرم در لیتر IBAبدست آمد. در نهایت گیاهچه¬های ریشه دار شده، برای سازگاری با شرایط برون شیشه¬ای در بستر كوكوپیت و پرلیت (2:1) 90 درصد سازگار شدند.

Introduction: Anthurium is a popular genus of the Araceae (order Spathiflorae).The flower consists of a protruding spadix containing numerous florets, subtended by a brightly colored modified leaf, the spathe. Anthuriums are bisexual and protogynous.Anthuriumscherzerianum as the most important species ofAnthurium genus is a potted perennial plant. Due to having beautiful, attractive and long-life flowers, A. scherzerianum can be used for the production of pot and cut flowers. Tissue culture is suggested as the most commonly method in order to rapid propagation and removing disease in a short period of time. This method also recommended for Anthuriumbecause of problems in classical propagation method of this flower..The three basic propagation methods for Anthuriumare propagation by seed, traditional vegetative and tissue culture.Micropropagation of Anthurium is using forcommercial production. Materials and Methods: In this study, the effect of plant growth regulators and explants on indirect regeneration of A. scherzerianumdetermined in separate experiments. In the first experiment, callogenesis was done by leaf explants on MS medium containing growth regulators, BA in three concentrations (0.5, 1.25 and 2 mg/l) in combination with 2, 4-D (0.5, 1.25 and 2 mg/l) or NAA (0.5, 1.25 and 2 mg/l) and the combinations of TDZ (0.5, 1.25 and 2 mg/l) with 2, 4-D (0, 0.5 mg/l). In the second experiment, regeneration was done on MS medium containing 0.75 mg/l BA with 0.05 mg/l 2, 4-D and 0.1 mg/l NAA and also in combination with TDZ (0.75mg/l). For rooting, MS medium containing different concentrations of IBA and IAA (0, 0.2 and 1 mg/l) were used. Callus induction, regeneration and rooting experiments were done based on completely randomized design, with 12, 6 and 6 replications, respectively.Data from all the schemes used in this study were analyzed with SAS statistical software. The comparison of means using Duncan 's multiple range test was evaluated at the 5% level. Results and Discussion: Analysis of variance showed that the effect of explant type and hormone combinations was significant on the percentage of callogenesis, callus volume and survival percentage. The interaction effect of explant type and combination of hormones was also significant on percentage of callogenesisand the volume of callus. Means comparisons showed that the highest callogenesis, viability and callus volume were achieved on MS medium containing 2 mg/l of BA and 0.5 mg/l of 2, 4-D. Petiole explants, also produced the highest percentage of callus (95%), survival rate (96%) and callus with dimensions of 6 mm2. Callus formation in leaf vein explants was higher than others. The effect of explant type and hormone combinations on regeneration, number of branches, number of leaves and leaf length was significant.The interaction of explant and hormone combinations on regeneration, number of branches, number of leaves and leaf length was also significant. Moreover, results of regeneration experiment indicated that the maximum number of shoots (6.9) and the maximum shoot length (5 cm), number of leaves (18) and the leaf length (2.8 mm) were achieved in 0.75 mg/l BA mg/l of and 0.05 mg/l 2, 4-D. In this study, petiole explants were also regenerated earlier than leaf explants.The effect of hormone combinations and concentrations was significant on rooting specially on the number of roots and root length.Furthermore, results of rooting experiment revealed that the highest rooting percentage (95%), the maximum number of roots (4.5 per plantlet) and the longest roots (3.5 cm) were produced in the medium containing 0.2 mg/l of IBA. Finally, the rooted plantlets were adapted (90%) in vivo condition by placing them on a mixture of cocopeat and perlite (2:1) substrate. Conclusion: In this study callugensis, regeneration and rooting of A.scherzerianum’s petiole and leaf explants were studied and different levels of plant growth regulators used for callugensis and regeneration. In this study petiole explants showed the highest callugenesis and regeneration. MS medium containing BA (2 mg/l) and 2, 4-D (0.5 mg/l), was the best for callugenesis. Also the highest percentage of regeneration was observed in medium containing BA (0.75 mg/l) and 2, 4-D (0.05 mg/l). Moreover low concentration (0.2 mg/l) of auxin has a better effect on rooting than high levels (1mg/l) so that the highest rooting percentage was produced in medium containing IBA (0.2 mg/l) and the lowest rooting percentage was produced in medium containing IAA (1 mg/l). Anthurium plantlets acclimized is cocopeat and perlite substrate (2: 1) with 90% acclimation.