培養基添加物對蝴蝶蘭組培瓶苗生長發育之影響

Abstract

本研究主要使用蝴蝶蘭‘V3’ (Phalaenopsis Sogo Yukidian ‘V3’) 約帶3片葉之中母瓶苗為材料，而在奈米銀第二次試驗中則採用不易發根蝴蝶蘭Phalaenopsis Mei Dar Green ‘Shih-hua Green Apple’。在1/2 MS基礎培養基添加不同物質 (殼聚醣、氯化鈉、矽酸鹽、奈米銀及奈米金)，探討其對蝴蝶蘭組培苗之生長與發育的影響，期能獲得高品質瓶苗。 添加殼聚醣之試驗中使用的濃度為5-40 mg•L-1。組培苗培養至第12週後，以添加5 mg•L-1之處理可增加蝴蝶蘭‘V3’ (Phalaenopsis Sogo Yukidian ‘V3’) 組培苗根長度。另外於培養基中添加50-400 mg•L-1氯化鈉之試驗中，於培養第12週時，各處理之組培苗生長量並無顯著差異。 在0.33-12 μM矽酸鉀 (K2SiO3)、矽酸鈣 (CaSiO3) 與矽酸鈉 (Na2SiO3) 之試驗中，培養12週後，在培養基添加0.33 μM CaSiO3之處理可增加組培苗葉片硬度；而培養基中添加9.0 μM CaSiO3可顯著增加組培苗根徑。另外透過顯微觀察，組培苗葉片中具晶體累積，其晶體含有矽元素，且由數個小顆粒晶體聚集成球狀結構。在組培苗第一片葉中，1.0 μM CaSiO3之處理其葉片矽晶體形成率大於50%，且此處理可提高葉片中矽晶體分布程度、矽晶體直徑與矽晶體數量。 添加0.125-320 mg•L-1奈米銀之試驗中，蝴蝶蘭‘V3’組培苗在培養至第12週時，80 mg•L-1奈米銀處理可增加組培苗葉片數與根數；20和80 mg•L-1 奈米銀則可提高組培苗地下部鮮乾重。另外，添加奈米銀於不易發根蝴蝶蘭 Phalaenopsis Mei Dar Green ‘Shih-hua Green Apple’品種的培養基中，組培苗培養至第12週時，添加奈米銀顯著增加組培苗根數、葉片數與新生葉片數，且減少組培苗葉片黃化的徵狀。但各濃度處理並無顯著影響組培苗發根率，且奈米銀處理皆降低組培苗葉片長、根長與根尖活性。 於有或無活性碳之培養基中分別添加濃度0、3.75 × 10-4、1.13 × 10-3、3.39 × 10-3、1.01 × 10-2和3.03 × 10-2 nM奈米金。培養至第12週，無添加活性碳且奈米金濃度為3.39 × 10-3 nM之處理略促進根數、根徑與地下部鮮重。添加活性碳則可有效增加組培苗地下部之生長發育，其增加組培苗之根徑與地下部乾鮮重，但奈米金各濃度處理並無顯著差異。整體而言，添加活性碳處理有助於組培苗地下部生長，其影響效果較添加奈米金處理顯著。 於培養基中添加殼聚醣 (5-40 mg•L-1)、氯化鈉 (50-400 mg•L-1) 與奈米金 (3.75 × 10-4至 3.03 × 10-2 nM) 並未有效提升組培苗生長。矽酸鹽 (0.33-12 μM) 雖對組培苗整體生長量並無顯著影響，但透過顯微鏡可觀察1.0 μM CaSiO3之處理提高第一片葉片矽晶體形成率、矽晶體分布程度、矽晶體直徑與矽晶體數量。此外，添加低濃度奈米銀可增加組培苗根數，但奈米銀添加則降低根尖活性，使組培苗有較短根長和側根之發生。

In this study, the subculturing flask plantlets of Phalaenopsis Sogo Yukidian ‘V3’ with 3 leaves were mainly used as material, while Phalaenopsis Mei Dar Green ‘Shih-hua Green Apple’, which is a difficult rooting cultivar, was used in the second experiment of nanosilver addition. The plantlets were cultured in 1/2 MS with different additives (chitosan, sodium chloride, silicate, nanosilver, and nanogold). The effects of these different additives in medium on the growth and development of plantlets was explored in order to improve the quality of plantlets in vitro. In the experiment of adding 5-40 mg•L-1 chitosan, the root length of plantlets increased in the 5 mg•L-1 chitosan treatment after cultured for 12 weeks. In addition, in the experiment of adding 50-400 mg•L-1 NaCl, the growth increment of plantlet was not significantly different among all treatments after cultured 12 weeks. In the experiment of adding 0.33-12 μM K2SiO3, CaSiO3, or Na2SiO3, the leaf firmness of plantlets increased in 0.33 μM CaSiO3 treatment, and the root diameter of plantlets significantly increased in 9.0 μM Na2SiO3 treatment. Furthermore, the silicon-containing crystals which accumulated in leaves of plantlets were observed by microscope. These silica bodies were assembled by a lot of small particles and formed as spherical bodies. In the 1.0 μM CaSiO3 treatment, formation of silica bodies was above 50%, and index of silica bodies distributions, diameter of silica bodies, and number of silica bodies of the first leaf were increased. In the experiment of adding 0.125-320 mg•L-1 nanosilver, the number of leaf and number of root of plantlests increased in the 80 mg•L-1 nanosilver treatment after cultured for 12 weeks. The root fresh and dry weight increased in 20 and 80 mg•L-1 nanosilver treatments. Another experiment tested on a difficult-rooting cultivar, Phalaenopsis Mei Dar Green ‘Shih-hua Green Apple’, was designed. The number of root, leaf and new leaf significantly increased and the symptom of leaf yellowing reduced in adding nanosilver treatments. However, adding nanosilver treatments did not significantly affect rooting rate of plantlet and reduced the leaf length, root length and acitivity of root tip after cultured for 12 weeks. The experiment of adding nanogold was divided into two parts, with or without activated charcoal. Both parts were added with 0, 3.75 × 10-4, 1.13 × 10-3, 3.39 × 10-3, 1.01 × 10-2 and 3.03 × 10-2 nM of nanogold. Without activated charcoal, the number of roots, root diameter, and root fresh weight increased in the 3.39 × 10-3 nM nanogold treatment. In addition, with activated charcoal, the growth and development of root were promoted. Adding activated charcoal increased root diameter, root fresh weight, and dry weight, but there were not significant difference among various nanogold concentrations. As a whole, the root growth of plantles was improved in the medium with activated charcoal and the effects of adding activated charcoal were more significant than various concentrations of nanogold. The growth of plantlets was not significantly promoted in various concentrations of chitosan (5 to 40 mg•L-1), NaCl (50 to 400 mg•L-1), and nanogold (3.75 × 10-4 to 3.03 × 10-2 nM). Although there was no significant increase in the growth of plantlets in adding silicate (0.33 to 12 μM) medium, the formation and distribution of silica bodies, number of silica bodies, and diameter of silica bodies in the first leaf of plantlets were increased in 1.0 μM CaSiO3 treatment. In addition, number of roots increased in low concentration of nanosilver treatment, but high concentration of nanosilver treatments reduced the activity of the root tip, promoted lateral root formation, and decreased the root length.