陳世銘臺灣大學:生物產業機電工程學研究所陳育菘Chen, Yu-SongYu-SongChen2010-05-052018-07-102010-05-052018-07-102008U0001-1302200815235200http://ntur.lib.ntu.edu.tw//handle/246246/180175近年來種苗生產技術之進步，組織培養在種苗產業中應用極廣。組織培養光環境控制上，發光二極體( Light-Emitting Diode, LED )為一新興的栽培光源，其低熱產生、波長固定與低消耗功率等優點，於研究與產業應用上，相較日光燈管，提供了相當大的便利。本研究以(陳，2005) LED光環境控制系統與自行開發的LED可調控系統，設計不同的光環境，進行星辰花與龍膽組培苗光環境栽培，並以田口法-信號雜訊比( Taguchi method - Signal of Noise Ratio )結合主成份分析( Principal component analysis, PCA )，將所得相關性狀指標結合為一反映品質與變異的綜合指標，並透過田口主效應分析( Main effect analysis )與反應曲面法( Response surface method, RSM )，探討上述兩種組培苗的最適光環境條件。 星辰花組培苗出瓶品質分析共進行了兩次試驗(光質、光度；光質、光度、光週期)，其中第二次試驗亦進行光質、光積值的分析。星辰花組培苗經由不同光環境栽培後，其葉片葉綠素螢光反應、葉片形態參數與植株生長參數皆有顯著反應。綜合出瓶指標( 株高要高、葉片葉綠素含量要高、葉片面積要大、植株乾重要重 )最適光環境條件為，光質：藍光比例20 % ~ 30 % ( R/B = 4 ~ R/B = 7/3 )、光強度：120 μmol m-2 s-1、光週期：16 hr/day、光積值：6.62 mol m-2。 龍膽組培苗出瓶品質分析亦同星辰花出瓶品質分析，且多進行了特殊光質試驗與補充試驗(光質、光度、光週期)。龍膽組培苗經由不同光環境栽培後，其葉片葉綠素螢光反應、葉片形態參數與植株生長參數皆有顯著反應。綜合出瓶指標(一)( 葉片葉綠素含量要高、節間長要短、葉片面積要大、植株乾重要重 )最適光環境條件為，光質：藍光比例70 % ~ 80 % (R/B = 3/7 ~ R/B = 1/4)、光強度：120 μmol m-2 s-1、光週期方面：16 hr/day、光積值：6.69 mol m-2。而於補充試驗中，綜合出瓶指標(二)( 葉片葉綠素含量要高、健壯商數要小、葉片面積要大、植株乾重要重 )最適光環境條件為，光質：藍光比例65 % (R/B = 0.55)、光強度：122.5 μmol m-2 s-1、光週期方面：16 hr/day，與綜合出瓶指標(一)分析結果相近，顯示以節間長或健壯商數於龍膽組培苗綜合出瓶指標最適光環境分析，皆是合適的。 而龍膽組培苗藥性成份分析，其試驗設計同龍膽組培苗出瓶品質分析。龍膽組培苗經由不同光環境栽培後，其Gentiopicroside與Swertiamarin藥性成份總量(mg/株)皆有顯著反應。綜合藥性指標(一)( Gentiopicroside與Swertiamarin藥性成份總量(mg/株)皆要高)最適光環境條件為，光質：藍光比例50 % ~ 80 % (R/B = 1 ~ R/B = 1/4)、光強度：120 μmol m-2 s-1、光週期：20 hr/day、光積值：8.64 mol m-2。而綜合藥性指標(二)( Gentiopicroside與Swertiamarin藥性成份含量(%)皆要高 )最適光環境條件為，光質：藍光比例20 % (R/B = 4)、光強度：40 μmol m-2 s-1、光週期：4 hr/day、光積值：1.23 mol m-2。Gentiopicroside藥性成份總量的最適光環境條件為，光質：藍光比例50 % ~ 80 % (R/B = 1 ~ R/B = 1/4)、光強度：120 μmol m-2 s-1、光週期方面：20 hr/day、光積值：8.64 mol m-2。而Swertiamarin藥性成份總量最適光環境條件則為，光質：藍光比例50 % (R/B = 1/4)、光強度：120 μmol m-2 s-1、光週期方面：20 hr/day、光積值：7.36 mol m-2。 綜上所論，本研究以LED人工光源，並透過試驗設計最適化分析，已成功分析星辰花與龍膽組培苗出瓶品質及龍膽組培苗藥性成份最適光環境條件(光質、光度、光週期、光積值)，未來可進一步地利用所得最適光環境條件與LED人工光源，實際應用於組培苗光環境栽培，以提升星辰花與龍膽組培苗出瓶品質及龍膽組培苗藥性成份。The development of seedlings production technology in recent years; tissue culture has been widely applied in seedlings industry. In the control of light environment of tissue culture, Light-Emitting Diodes (LEDs) are new culture light. In comparision between fluorescent lamps and LEDs, LEDs have lower heat generation, specified wavelength and lower power consumption in research and seedlings industry. In the research, LED light environment control system (LECS) (Chen, 2005) and LED Light Control System developed in this study were designed for different light environment, and they were used to in vitro growth of Limonium, and Gentiana. Taguchi Method - signal of noise ratio combine with Principal Component Analysis (PCA), a set of property indexs were transformed into a comprehensive index that can reflect quality and variability. To probe into the optimal light environment of Limonium, and Gentiana by using main effect analysis and Response Surface Method (RSM). he analysis of ex vitro quality of Limonium, there are two experiments (light quality, light intensity; light quality, light intensity, photoperiod). Among of them, the secondary experiment can do analysis of light quality and light integral. The leaf morphological parameters, leaf chlorophyll fluorescence and plant morphological parmeters of the Limonium were effected significantly after different light environment of culture. The optimal light environment condition of comprehensive ex vitro index (plant height is high, chlorophyll contents (SPAD-502 reading, CMR) is high, leaf area is big, dry weight is heavy), light quality: 80 % red +20 % blue ~ 70 % red +30 % blue (R/B ratio = 4~R/B ratio = 7/3), light intensity:120 μmol m-2 s-1, photoperiod:16 hr/day and light integral:6.62 mol m-2. he analysis of ex vitro quality of Gentiana and the analysis of ex vitro quality of Limonium are the same, and it also did specific light quality experiment and supplemental experiment (light quality, light intensity, photoperiod). The leaf morphological parameters, leaf chlorophyll fluorescence, and plant morphological parmeters of the Gentiana were effected significantly after different light environment of culture. The optimal light environment condition of comprehensive ex vitro index one (chlorophyll contents (SPAD-502 reading, CMR) is high, height of internode is short, leaf area is big, dry weight is heavy), light quality: 30 % red +70 % blue ~ 80 % red +20 % blue (R/B ratio = 3/7 ~ R/B ratio = 1/4), light intensity: 120 μmol m-2 s-1, photoperiod: 16 hr/day and light integral: 6.69 mol m-2. In supplementa1 experiment, the optimal light environment condition of comprehensive ex vitro index two (chlorophyll contents (SPAD-502 reading, CMR) is high, sturdiness quotient is small, leaf area is big, dry weight is heavy), light quality: 35 % red +65 % blue (R/B ratio = 0.55), light intensity: 122.5 μmol m-2 s-1, photoperiod: 16 hr/day.The optimal light environment condition of comprehensive ex vitro index two is close to comprehensive ex vitro index one. The result show that: height of internode or sturdiness quotient is appropriate to analysis of the optimal light environment condition of comprehensive ex vitro index of Gentiana.he analysis of medicinal properties of Gentiana and the analysis of ex vitro quality of Gentiana have the same experimental design. The overall medicinal contents (mg/plant) of Gentiopicroside and Swertiamarin were effected significantly after different light environment of culture. The optimal light environment condition of comprehensive medicinal index one (overall medicinal contents (mg/plant) of Gentiopicroside and Swertiamarin are high), light quality: 50 % red + 50 % blue ~ 80 % red + 20 % blue (R/B ratio = 1 ~ R/B ratio = 1/4), light intensity: 120 μmol m-2 s-1, photoperiod: 20 hr/day and light integral: 8.64 mol m-2. The optimal light environment condition of comprehensive medicinal index two (the medicine contents (mg/mg) of Gentiopicroside and Swertiamarin are high), light quality: 20 % red + 80 % blue (R/B ratio = 4), light intensity: 120 μmol m-2 s-1, photoperiod: 20 hr/day and light integral: 8.64 mol m-2. The optimal light environment condition of overall medicinal contents (mg/plant) of Gentiopicroside, light quality: 50 % red + 50 % blue ~ 80 % red + 20 % blue (R/B ratio = 1 ~ R/B ratio = 1/4), light intensity: 120 μmol m-2 s-1, photoperiod: 20 hr/day and light integral: 8.64 mol m-2. The optimal light environment condition of overall medicinal contents (mg/plant) of Swertiamarin, light quality: 50 % red + 50 % blue (R/B ratio = 1), light intensity: 120 μmol m-2 s-1, photoperiod: 20 hr/day and light integral: 7.36 mol m-2.s a conclusion, the study used Light-Emitting Diodes (LEDs), and it has successfully analyzed the optimal light environment condition (light quality, light intensity, photoperiod, light integral) of ex vitro quality of Limonium and Gentiana and medicinal properties of Gentiana by using experimental design and optimal analysis. With these research results, LEDs and the optimal light environment condition could be used to the light environment of tissue culture in the future. It is useful for ex vitro quality of Limonium and Gentiana and medicinal properties of Gentiana.誌 謝 i 要 iibstract iv錄 vii目錄 x目錄 xv一章 前言 1-1 前言 1-2 研究目的 4二章 文獻探討 5-1 星辰花和龍膽 5-1-1 星辰花 5-1-2 龍膽 7-2 組織培養技術 9-3 光與植物 12-3-1 光線與色素 13-3-2 光對於植物之影響 15-4 LED於植物栽培之應用 19-4-1 傳統光源與發光二極體 19-4-2 發光二極體於植物栽培之應用 20-5 試驗設計與最佳化 25三章 材料與方法 30-1 試驗儀器與設備 30-1-1 試驗設備 30-1-1-1 LED光環境控制系統 30-1-1-2 LED可調控系統 35-1-2 組培苗生理與形態檢測儀器 42-2 分析方法 47-2-1 複因子變異數分析( Factorial Analysis of Variance ) 48-2-2 田口法 ( Taguchi Method ) 50-2-3 主成份分析法 ( Principal Component Analysis ) 54-2-4 反應曲面法 ( Response Surface Method ) 57-2-5 分析流程 60-3 星辰花組培苗出瓶品質分析 63-3-1 培育方法及試驗設計 63-3-2 取樣及性狀量測 69-3-3 綜合出瓶指標分析 72-4 龍膽組培苗出瓶品質分析 74-4-1 培育方法及試驗設計 74-4-2 取樣及性狀量測 85-4-3 綜合出瓶指標(一)分析 88-4-4 綜合出瓶指標(二)分析 90-5 龍膽組培苗藥性成份分析 92-5-1 培育方法及試驗設計 92-5-2 取樣及性狀量測 93-5-3 綜合藥性指標(一)分析 95-5-4 綜合藥性指標(二)分析 96四章 結果與討論 97-1 星辰花組培苗出瓶品質分析 98-1-1 光環境對星辰花組培苗出瓶品質之影響 99-1-2 星辰花組培苗綜合出瓶指標最適光環境分析 112-1-3 星辰花組培苗出瓶品質綜論 129-2 龍膽組培苗出瓶品質分析 133-2-1 光環境對龍膽組培苗出瓶品質之影響 134-1-2 龍膽組培苗綜合出瓶指標最適光環境分析 152-2-2-1 綜合出瓶指標(一)分析 153-2-2-2 綜合出瓶指標(二)分析 178-2-3 龍膽組培苗出瓶品質綜論 184-3 龍膽組培苗藥性成份分析 192-3-1 光環境對龍膽組培苗藥性成份之影響 192-3-2 龍膽組培苗藥性成份最適光環境分析 197-3-2-1 綜合藥性指標(一)分析 198-3-2-2 綜合藥性指標(二)分析 215-3-2-3 藥性成份-Gentiopicroside分析 231-3-2-4 藥性成份-Swertiamarin 分析 241-3-3 龍膽組培苗藥性成份綜論 251五章 結論與建議 260-1 結論 260-2 建議 265六章 參考文獻 266application/pdf5761414 bytesapplication/pdfen-US發光二極體組織培養龍膽星辰花田口法主成分分析反應曲面法Light-Emitting DiodesTissue CultureGentianaLimoniumTaguchi MethodPrincipal Component AnalysisResponse Surface Methodthesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/180175/1/ntu-97-R94631003-1.pdf