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Effects of Phosphinothricin on Shoot Induction of Leaf Explants of Echinacea purpurea
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In vitro regeneration of Echinacea purpurea L.: enhancement of somatic embryogenesis by indolebutyric acid and dark pre-incubation. In Vitro Cell. Dev. Biol. -Plant 39: 605-612.||摘要:||
本試驗以中興大學 陳宗禮教授提供的cichoric acid 超過2.2 % 乾重的三個紫錐菊篩選株 (T5.9、T2.15、D7.4) 進行phosphinothricin (PPT) 葉片培養試驗 (所有培養基均含0.44 μM BA及0.54 μM NAA)。以各濃度PPT (0、0.5、1.0、1.5 μM) 處理1、2及4週之研究紫錐菊葉片培植體誘導芽體的效果。
持續四週各濃度PPT處理中，T5.9篩選株獲得的芽體較健康，T2.15篩選株獲得的芽體易有玻璃質化現象，D7.4篩選株於0.5 μM PPT處理則有發根現象。T5.9篩選株以未處理PPT獲得最高芽體誘導率 (70.9%)，而芽體數則以0.5 μM PPT處理較高 (2芽體數/培植體) 。T2.15以0.5 μM PPT處理下有較高誘導率 (69.8 %) 及芽體數 (2.4芽體數/培植體)。D7.4篩選株於PPT處理下未能形成大量芽體。1.0 μM PPT處理下僅T2.15篩選株有芽體誘導形成，1.5 μM PPT處理下則三個篩選株培植體皆無芽體形成。
葉片培植體以各濃度PPT處理第一週或前二週，觀察PPT的繼續作用效果。僅處理一週下，三個篩選株於高濃度 (1.0與1.5 μM) PPT處理下有提早形成芽體之趨勢。T5.9與T2.15篩選株於0.5 μM PPT處理可誘導出最多的芽體數 (2.4 與2.2個/培植體)，D7.4篩選株則仍以0 μM PPT處理有較佳的芽體誘導率。僅前二週處理下，發現三個篩選株於1.0 μM PPT處理下芽體誘導率上升，T5.9、T2.15與D7.4培植體形成芽體數達4.6、2.8與4.5個。
Leaves of three elite plants of Echinacea purpurea L. with cichoric acid over 2.2 % DW provided kindly by professor Chung-Li Chen were used to study the effect of phosphinothricin (PPT) on shoot formation. All the media in this study, here, contained 0.44 μM BA and 0.54 μM NAA. The leaves were cultured in media containing different concentrations of PPT (0, 0.5, 1.0, 1.5 μM) for 1 or 2 weeks and then transferred to medium without PPT to compare with 4 or 5-week-long PPT treatment.
For the 4-week-long PPT treatment, explants had shoots healthly, hyperhydric and rooting apart from shoot (at 0.5 μM PPT) for line T5.9, T2.15 and D7.4, respectively. For line T5.9, the best shoot induction rate (70.9 %) occurred at 0 μM PPT but the maxium shoots (2.0 shoots per explant) formed at 0.5 μM PPT. For line T2.15, both of shoot induction rate (69.8 %) and maxium shoots (2.4 shoots per explant) turned up at 0.5 μM PPT. However, it seemed no good response to PPT treatment for line D7.4 plant. Shoot formation can be found only on line T2.15 explant as PPT concentration high up to 1.0 μM and no shoot emerged on explants of all the three line at 1.5 μM PPT.
The carry-over effect of PPT on shoot formation were detected by disposing leaves of the three lines with different concentrations of PPT treatment only for the first one or two weeks. For the first-one-week PPT treatment, shoots formed earlier on leaf explants of three lines at high concentrations of PPT (1.0 and 1.5 μM) than those of 4-week PPT treatment. There were maxium shoots per explant (2.4 and 2.2) for T5.9 and T2.15 at 0.5 μM PPT, respectively, but the better response for line D7.4 was still at 0 μM PPT. For the first-two-week PPT treatment, shoot induction rates increased with media containing 1.0 μM PPT. There were 4.6, 2.8, and 4.5 shoots per explant for T2.15, T5.9 and D7.4, respectively.
The soluble protein contents of explants of different concentration PPT treatments were analysed in the first 9 days. Explants featured shoot formation later increased soluble protein contents in the first 9 days, but explants with no shoot formation responsed on the contrary. The results of SDS-PAGE indicated that Rubisco contents decreased in PPT treatments and the new band, nearby 36 kDa, appeared in line T5.9.
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