[[alternative]]The Effects of Magnetic Fields, Magnetized Growth Medium, and Temporary Immersion System on the Organogenesis and Proliferation of Protea cynaroides L. Explants

系統編號: PD10207-0125計畫編號: NSC102-2313-B343-001執行機關: 南華大學自然生物科技學系研究期間: 10208~10307[[abstract]]帝王花（Protea cynaroides L.）在花卉栽培產業是熟知的切花。近年來，其產區已遍布全球，然而， 目前使用之增殖方法有所不足且已過時;不像其他高經濟之切花已有相當進步之增殖技術，傳統繁殖帝 王花之方法（如：插條及播種）很耗時，且多數成功率低。插條法之發根通常需要 6 個月，且成效不 一，此外，增殖者亦面臨難以克服生長緩慢、特定營養鹽需求以及生理失序之情形。不正確之施肥及 培植過程中莖部之酚類氧化常導致明顯之植株消耗。儘管近年來在建立帝王花試管內繁殖協定並減少 生理失序上已有長足進步，但在器官發生及增殖的速度上依然懸而未決。爰此，發展新方法以有效刺 激帝王花快速再生及增殖高品質的培植體是刻不容緩的。本研究擬探討磁場、磁化培養基及使用短暫 浸漬系統（TIS）對帝王花幼苗之器官發生及增殖效應，並由此開發一結合磁場及磁化培養基之短暫 浸漬系統以刺激帝王花培植體之器官發生及增殖效應。 在磁場影響之研究中，將培植體暴露於（1）恆定磁場、（2）變動磁場或（3）磁化培養基中。在 上述三項研究中，磁場大小為 0.1 mT、0.5 mT、1 mT、5 mT、10 mT、20 mT、30 mT 及 50 mT。在第 一項研究中，恆定磁場與帝王花幼芽之距離為 2 cm、5 cm 或 10 cm。在變動磁場之研究中，磁鐵將附 著於電力驅動之木製懸臂，使之以 5 rpm 之速度旋轉，且使植株接受暴露 2 s、4 s 或 6 s 並間隔 2 s。 此外，上述實驗中，南北極之影響亦將探討之。在磁化培養基之研究中，使用經磁化之 RO 水配製培 養基。在短暫浸漬系統研究中，培植體將每 1、2、3 或 4 小時浸漬於含不同濃度生長調節劑之 Plantima® 容器(A-Tech Bioscientific, Taiwan)中 5 min 或 10 min。上述研究中所得之最佳磁場及浸漬條件將用以開 發新的微繁殖系統。在所有實驗中，植株之根長、幼芽、葉片及其葉綠素含量、淨光合作用速率、培 植體之再生及增殖速率將記錄並比較之。[[abstract]]Protea cynaroides L. is a well-known cut flower in the floriculture industry. In recent years, its production areas have been expanding worldwide. However, the propagation methods currently used are inadequate and outdated. Unlike other economically-important cut flowers where propagation methods are highly advanced, traditional methods used to propagate P. cynaroides plants such as stem cuttings and seeds are time-consuming, and often result in low success rates. Root formation of stem cuttings usually take up to six months, with inconsistent results. Furthermore, propagators are faced with difficulties in overcoming the slow-growing characteristics, unique nutritional requirements, and physiological disorders of P. cynaroides plants. The application of incorrect fertilization and phenolic oxidation of stems during propagation often result in significant losses of plant material. Despite advances made in recent years in establishing propagation protocols and reducing physiological disorders of P. cynaroides explants in vitro, accelerating organogenesis and proliferation rates of plantlets remain unresolved. In light of these challenges, the development of new methods that will stimulate rapid regeneration and proliferation of high quality explants in an efficient manner is desperately needed to bring the production of P. cynaroides plants into the 21st century. The aim of this study is therefore to investigate the effects of magnetic fields, magnetized growth medium, and the use of temporary immersion systems (TIS), on the organogenesis and proliferation of P. cynaroides plantlets. In addition, a novel approach is proposed whereby a new temporary immersion system is developed in combination with magnetic fields and magnetized growth media to stimulate organogenesis and proliferation of P. cynaroides explants. In the magnetic field study, three separate investigations will be conducted. Explants will be exposed to: 1) a constant magnetic field; 2) an alternating magnetic field; 3) magnetized growth medium. In all three studies, different magnetic strengths will be used: 0.1 millitesla (mT); 0.5 mT; 1 mT, 5 mT, 10 mT, 20 mT, 30 mT, and 50 mT). In the first study, magnets will be placed at a distance of 2 cm, 5 cm or 10 cm from microshoots. In the second investigation, magnets will be placed on a wooden arm attached to an electric motor (5 rpm). Each explant will be exposed to a magnetic field for 2 s, 4 s or 6 s intervals, for a duration of 2 s. The North and South poles of each magnet will also be tested in both experiments. In the third study, RO water will first be magnetized, and then used to prepare the growth medium. In the TIS study, microshoots will be immersed in Plantima® containers (A-Tech Bioscientific, Taiwan), containing various concentrations of growth regulators for either 5 or 10 min, every 1 h, 2 h, 3 h, or 4 h. The optimum magnetic and immersion conditions will then be combined to develop a new micropropagation system. In all experiments, data for growth of roots, shoots, and leaves, their chlorophyll content and net photosynthetic rate, as well as the regeneration and proliferation rates of explants will be collected and analyzed.Protea cynaroides L. is a well-known cut flower in the floriculture industry. In recent years, its production areas have been expanding worldwide. However, the propagation methods currently used are inadequate and outdated. Unlike other economically-important cut flowers where propagation methods are highly advanced, traditional methods used to propagate P. cynaroides plants such as stem cuttings and seeds are time-consuming, and often result in low success rates. Root formation of stem cuttings usually take up to six months, with inconsistent results. Furthermore, propagators are faced with difficulties in overcoming the slow-growing characteristics, unique nutritional requirements, and physiological disorders of P. cynaroides plants. The application of incorrect fertilization and phenolic oxidation of stems during propagation often result in significant losses of plant material. Despite advances made in recent years in establishing propagation protocols and reducing physiological disorders of P. cynaroides explants in vitro, accelerating organogenesis and proliferation rates of plantlets remain unresolved. In light of these challenges, the development of new methods that will stimulate rapid regeneration and proliferation of high quality explants in an efficient manner is desperately needed to bring the production of P. cynaroides plants into the 21st century. The aim of this study is therefore to investigate the effects of magnetic fields, magnetized growth medium, and the use of temporary immersion systems (TIS), on the organogenesis and proliferation of P. cynaroides plantlets. In addition, a novel approach is proposed whereby a new temporary immersion system is developed in combination with magnetic fields and magnetized growth media to stimulate organogenesis and proliferation of P. cynaroides explants. In the magnetic field study, three separate investigations will be conducted. Explants will be exposed to: 1) a constant magnetic field; 2) an alternating magnetic field; 3) magnetized growth medium. In all three studies, different magnetic strengths will be used: 0.1 millitesla (mT); 0.5 mT; 1 mT, 5 mT, 10 mT, 20 mT, 30 mT, and 50 mT). In the first study, magnets will be placed at a distance of 2 cm, 5 cm or 10 cm from microshoots. In the second investigation, magnets will be placed on a wooden arm attached to an electric motor (5 rpm). Each explant will be exposed to a magnetic field for 2 s, 4 s or 6 s intervals, for a duration of 2 s. The North and South poles of each magnet will also be tested in both experiments. In the third study, RO water will first be magnetized, and then used to prepare the growth medium. In the TIS study, microshoots will be immersed in Plantima® containers (A-Tech Bioscientific, Taiwan), containing various concentrations of growth regulators for either 5 or 10 min, every 1 h, 2 h, 3 h, or 4 h. The optimum magnetic and immersion conditions will then be combined to develop a new micropropagation system. In all experiments, data for growth of roots, shoots, and leaves, their chlorophyll content and net photosynthetic rate, as well as the regeneration and proliferation rates of explants will be collected and analyzed