臺灣博碩士論文加值系統

盧會之藥用歷史已有數千年，常用於美容醫療，目前常作”蘆
薈”兩字。許多研究指出庫拉索盧會有抗菌、抗癌、抗腫瘤、抑制
潰瘍及調節免疫等作用。庫拉索盧會為百合科植物，生長於高溫少
雨的區域。盧會中有多種有效成分，工業上需求量大，因此大量繁
殖庫拉索盧會作為商業用途有其必要性。
本研究主要目的有二： 1.建立庫拉索盧會大量繁殖之系統； 2.
利用酚－硫酸法、TLC、HPLC、MALDI-TOF 質譜儀測定六週齡之盧會
組織培養苗及野外生長兩年植株之甘露糖、盧會素、盧會大黃素及大黃素含量。
在庫拉索盧會的組織培養系統中，探討不同MS 鹽類濃度、蔗
糖濃度、各種cytokinins 及NAA 濃度、pH 值、活性碳、PVP
(polyvinylpyrrolidone)及各種有機添加物與不同凝膠物質，對庫
拉索盧會大量繁殖效率及發根之影響；栽培介質選擇以泥碳土、蛭
石及沙土混合而成，於溫室中觀察庫拉索盧會植株之生長情形。結
果顯示芽體誘導以全量MS 鹽類配合1 mg/L BA、3% 蔗糖、0.3%
gelrite、pH = 5.7 ± 0.1 之固體培養基，於25℃、光照38 μ
mol/m2s 的環境下培養為最佳條件；誘導芽體發根則以半量MS 鹽類
配合3% 蔗糖、0.9% agar、pH = 5.7 ± 0.1、10% 椰子汁的固體培養基，於25℃、光照38 μmol/m2s 的環境下培養為最佳條件；栽
培介質則以泥碳土：蛭石（1：1）效果最佳，移植兩個月後鮮重可
達21.5 g。
利用酚-硫酸法測定甘露糖結果顯示生長六週之庫拉索盧會組
織培養苗甘露糖含量為89.82 mg/g，而野外生長兩年庫拉索盧會之
甘露糖含量為30.09 mg/g。盧會素可在chloroform：methanol = 7：
3 混合展開液下觀察之，盧會大黃素及大黃素可在hexane：ethyl
acetate = 6：4 混合展開液下分離呈現。結果顯示生長六週之庫拉
索盧會組織培養苗及野外生長兩年植物體，在盧會素含量分別為
0.78 及0.21 mg/g，盧會大黃素含量為1.61 及3.02 mg/g，大黃
素的含量為2.01 及2.88 mg/g。

III
Summary
Aloe barbadensis Miller has been widely used in cosmetics, skin lotions or
medicine for thousand of years in different parts of the world. Many reports have
shown that Aloe species possesses anti-inflammatory, anti-tumor, anti-ulcer and
anti-cancer and anti-bacterial properties. A. barbadensis belongs to the family
Liliaceae. It is a xerophytic plant, but can be grown even in rain fed conditions. The
production of aloe leaves is insufficient to meet the industry demand currently;
hence, there is a need to develop alternative propagation methods which can boost
commercial production.
Present study was undertaken with the following objectives: (1) to standardize
micropropagation procedure in A. barbadensis, (2) to estimate the compounds like
mannose, aloin, aloe-emodin, and emodin by using Phenol-Sulfuric Acid method,
Thin Layer Chromatography (TLC), High Performance Liquid Chromatography
(HPLC) and MALDI-TOF Mass detection in 6 weeks tissue culture and 2 years wild
plants.
To meet the first objective, experiments were carried using different explants,
strengths of MS salts, sucrose, NAA, different cytokinins like Kinetin, BA and TDZ,
pH, organic supplements and two gelling agents (agar and gelrite) for induction of
optimum number of multiple shoots in A. barbadensis. Similarly, different factors
were tested for optimum rooting of shoots under in vitro conditions. Also, different
potting mixtures were tested for optimum survival of plants under green house
conditions.
The highest number of multiple shoots in A. barbadensis was obtained with
stem without terminal bud as an explant, half strength of MS salts supplemented
with 3% sucrose, 1 mg/L BA, 0.3% gelrite and pH = 5.7 ± 0.1. For in vitro rooting,
full strength MS salts, 3% sucrose, 0.9% agar, 10% coconut milk and pH = 5.7 ±
0.1 were the optimum conditions. Though 100% survival could be achieved on all
four potting mixtures tested, the highest fresh weight 21.5 g was obtained with peat
moss (Pm) and vermiculite used in 1:1 ratio after two month growth.
Results on mannose concentration in shoots measured by Phenol-Sulfuric Acid
method showed that the mannose content in leaves of six weeks tissue culture and
two years wild plants were 89.82 mg/g and 30.09 mg/g, respectively. Aloin,
aloe-emodin and emodin could be detected in leaf skin of A. barbadensis using TLC,
HPLC and MALDI-TOF Mass. Aloin in chloroform：methanol (7：3) , and
aloe-emodin and emodin in hexane：ethyl acetate (6：4) were suitable for detection
by TLC method. Quantities of compounds detected were as follows: Aloin 0.78mg/g and 0.21 mg/g, aloe-emodin 1.61 mg/g and 3.02 mg/g and emodin 2.01 mg/g
and 2.88 mg/g in tissue culture and wild plants, respectively.

摘要.............................................................................................................I
英文摘要................................................................................................. III
目錄........................................................................................................... V
表目錄......................................................................................................VI
圖目錄...................................................................................................VIII
縮寫表....................................................................................................... X
第壹章盧會之簡介.................................................................................. 1
第貳章藥用植物組織培養.................................................................... 10
第參章材料與方法................................................................................ 29
第肆章結果............................................................................................ 43
第伍章討論............................................................................................ 75
參考文獻.................................................................................................. 85
附錄.......................................................................................................... 96
Table 1. The yields of dried Aloe barbadensis Miller leaf (6.2g) from 2 years old wild type plant by silica gel column chromatography 34
Table 2. The yields of dried Aloe barbadensis Miller leaf (7.2g) from 6 weeks old in vitro plants by silica gel column chromatography 35
Table 3. In vitro shoot induction from different explant types of Aloe barbadensis Miller 53
Table 4. Effect of different MS salts strength on in vitro shoot multiplication in buds of Aloe barbadensis Miller 53
Table 5. Effect of different sucrose concentrations on in vitro shoot multiplication in buds of Aloe barbadensis Miller 54
Table 6. Effect of different BA concentrations on in vitro shoot multiplication in buds of Aloe barbadensis Miller 54
Table 7. Effect of different NAA concentrations with 1.0 mg/L BA on in vitro shoot multiplication in buds of Aloe barbadensis Miller 55
Table 8. Effect of different cytokinins on in vitro shoot multiplication in buds of Aloe barbadensis Miller 55
Table 9. Effect of different pH values on in vitro shoot multiplication in buds of Aloe barbadensis Miller 56
Table 10. Effect of different organic supplements on in vitro shoot multiplication in buds of Aloe barbadensis Miller 56
Table 11. Effect of different gelling agents on in vitro shoot multiplication in buds of Aloe barbadensis Miller 57
Table 12. Effect of different incubation periods on shoot /root growth parameters in Aloe barbadensis Miller 57
Table 13. Effect of different MS salts strength on in vitro rooting of shoot buds of Aloe barbadensis Miller 58
Table 14. Effect of different sucrose concentrations on in vitro rooting of shoot buds of Aloe barbadensis Miller 58
Table 15. Effect of different NAA concentrations on in vitro rooting of shoot buds and ex vitro survival rate in Aloe barbadensis Miller 59
Table 16. Effect of activated charcoal or PVP on in vitro rooting of shoot buds and ex vitro survival rate from Aloe barbadensis Miller buds 59
Table 17. Effect of organic supplements on in vitro rooting of shoot buds and ex vitro survival rate in Aloe barbadensis Miller 60
Table 18. Effect of gelling agents on in vitro rooting of shoot buds and ex vitro survival rate in Aloe barbadensis Miller 60
Table 19. Effect of the different potting mixtures on ex vitro survival rate of tissue culture plants of Aloe barbadensis Miller 61
Table 20. Mannose content in gels of wild and tissue culture plants of Aloe barbadensis Miller 61
Table 21. Aloin, aloe-emodin and emodin contents in wild and tissue culture plants of Aloe barbadensis Miller 61


圖目錄
Fig. 1. Illustration of Aloe barbadensis Miller. 4
Fig. 2. The relative amounts of auxins and cytokinins often required to bring about morphogenesis. 23
Fig. 3. The calibration curve of D-mannose. 32
Fig. 4. The calibration curve of aloin standard. 37
Fig. 5. The calibration curve of aloe-emodin standard. 38
Fig. 6. The calibration curve of emodin standard. 38
Fig. 7. The HPLC spectrum of aloin in the mobile phage. 39
Fig. 8. The HPLC spectrum of aloe-emodin and emodin in the mobile phage. 39
Fig. 9. The chemical structure of aloin. 41
Fig. 10. The chemical structure of aloe-emodin. 41
Fig. 11. The chemical structure of emodin. 42
Fig. 12. In vitro shoot induction from different explant types of Aloe barbadensis Miller. 62
Fig. 13. Effect of different MS salts strength on in vitro shoot multiplication in buds of Aloe barbadensis Miller. 62
Fig. 14. Effect of different sucrose concentrations on in vitro shoot multiplication in buds of Aloe barbadensis Miller. 62
Fig. 15. Effect of different BA concentrations on in vitro shoot multiplication in buds of Aloe barbadensis Miller. 63
Fig. 16. Effect of different NAA concentrations with 1.0 mg/L BA on in vitro shoot multiplication in buds of Aloe barbadensis Miller. 64
Fig. 17. Effect of different cytokinins on in vitro shoot multiplication in buds of Aloe barbadensis Miller. 64
Fig. 18. Effect of different pH values on in vitro shoot multiplication in buds of Aloe barbadensis Miller. 64
Fig. 19. Effect of different organic supplements on in vitro shoot multiplication in buds of Aloe barbadensis Miller. 65
Fig. 20. Effect of different gelling agents on in vitro shoot multiplication in buds of Aloe barbadensis Miller. 65
Fig. 21.1. Effect of different incubation periods on in vitro shoot growth parameters in Aloe barbadensis Miller. 66
Fig. 21.2. Effect of different incubation periods on root growth parameters in Aloe barbadensis Miller. 67
Fig. 22. Effect of different MS salts strength on in vitro rooting of Aloe barbadensis Miller shoot buds. 68
Fig. 23. Effect of different sucrose concentrations on in vitro rooting of Aloe barbadensis Miller shoot buds. 68
Fig. 24.1. Effect of different NAA concentrations on in vitro shoot growth parameters in Aloe barbadensis Miller. 69
Fig. 25.1. Effect of activated charcoal or polyvinilpyrrolidone (PVP) on in vitro shoot growth parameters in Aloe barbadensis Miller. 70
Fig. 25.2. Effect of activated charcoal or polyvinilpyrrolidone (PVP) on in vitro rooting of Aloe barbadensis Miller shoot buds. 70
Fig. 26. Effect of organic supplements on in vitro rooting of Aloe barbadensis Miller shoot buds. 70
Fig. 27. Effect of different gelling agents on in vitro rooting of Aloe barbadensis Miller shoot buds. 71
Fig. 28. Effect of the different potting mixtures on ex vitro growth of Aloe barbadensis Miller plants. 71
Fig. 29. Separated chromatogram of aloin in leaf skin of Aloe barbadensis Miller from different sources using TLC. 72
Fig. 30. Separated chromatogram of aloe-emodin and emodin in leaf skin of Aloe barbadensis Miller from different sources using TLC. 72
Fig. 31. The spectrum of aloin (�� arrow) by MALDI-TOF Mass detection. 73
Fig. 32. The spectrum of aloe-emodin (�� arrow) by MALDI-TOF Mass detection. 73
Fig. 33. The spectrum of emodin (�� arrow) by MALDI-TOF Mass detection. 74

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