類別:http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/862024-03-28T16:38:48Z2024-03-28T16:38:48Z黃連木葉子精油抑制黑色素生成之活性Ju-Yu Cheng鄭如彧http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/215592021-06-08T03:37:55Z2019-01-01T00:00:00Z標題: 黃連木葉子精油抑制黑色素生成之活性; Antimelanogensis Activity of Leaf Essential Oil
of Pistacia chinensis
作者: Ju-Yu Cheng; 鄭如彧
摘要: 本研究使用水蒸餾法萃取黃連木(Pistacia chinensis )葉子精油,以氣相層析法分析其中成分,其中最主要的成分為Limonene,占精油總量之57.06%,而精油中其他含量較多的化合物,依序為3-Carene(12.36%)、Selinene(5.65%)、Caryophyllene(5.08%)、Caryophyllene oxide(2.68%)等化合物。Limonene為黃連木葉子精油中最主要成分,而在自然界中以兩種不同光學異構物存在,右旋構型的D-Limonene及左旋構型的L-Limonene,本研究分別以黃連木葉子精油、D-Limonene及L-Limonene評估其黑色素生成抑制效果。
蕈類酪胺酸活性抑制分析,以L-Tyrosine或L-DOPA為基質時,黃連木葉子精油、D-Limonene及L-Limonene皆能產生良好的抑制效果,以L-Tyrosine為基質時,黃連木葉子精油及D-Limonene為對於酪胺酸酶為混合型抑制劑,L-Limonene則為競爭型抑制物;以 L-DOPA 為基質時,黃連木葉子精油及D-Limonene 為競爭型抑制劑,L-Limonene 則為混合型抑制物。由斑馬魚胚胎黑色素生成的評估結果,黃連木葉子精油、D-Limonene及L-Limonene在不對斑馬魚胚胎生長發育造成影響的情況下,對其體內的黑色素生成有明顯的抑制效果,IC50值依序為38.16 μg/mL 、44.83 μg/mL及72.87 μg/mL。
綜合上述結果,黃連木葉子精油與其成分D-Limonene及L-Limonene,皆對於黑色素生成有顯著的抑制效果,相當具有發展潛能之天然成分,希望未來能夠進一步研究作用機制,將其應用在相關領域當中。; In this study, the leaf essential oil of Pistacia chinensis was extracted by water distillation. The components were analyzed by gas chromatography, and the main component was Limonene with the relative content of 57.06%. Other compounds with a high relative content was 3-Carene (12.36%), Selinene (5.65%), Caryophyllene (5.08%), and Caryophyllene oxide (2.68%). Limonene is the main component of P. chinensis leaf essential oil, and it exists in nature in two different optical isomers, D-Limonene in the right-handed configuration and L-Limonene in the left-handed configuration. Therefore, the inhibitory effect of melanin is evaluated by the leaf essential oil of P. chinensis, D-Limonene and L-Limonene.
Results showed that when L-Tyrosine or L-DOPA was used as the substrate in the antityrosinase activity test, the leaf essential oil of P. chinensis, D-limonene and L-limonene exhibited great inhibitory effects. In antimelanogenesis test of zebrafish embryos, the leaf essential oil of P. chinensis, D-limonene and L-limonene had excellent inhibitory effect on the melanin production of zebrafish embryo without affecting its body growth and development, and IC50 values were 38.16 μg/mL (essential oil of P. chinensis), 44.83 μg/mL (D-limonene) and 72.87 μg/mL (L-limonene). Accordingly, the leaf essential oil of P. chinensis is a melanin inhibitor with development potential, and we hope that it can be applied in related fields in the future.2019-01-01T00:00:00Z黃連木葉子精油抑制黃嘌呤氧化酶之活性研究Chi-Ya Huang黃琪雅http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/170242021-06-07T23:53:35Z2013-01-01T00:00:00Z標題: 黃連木葉子精油抑制黃嘌呤氧化酶之活性研究; Studies on Xanthine Oxidase Inhibitory Activity of Pistacia chinensis Bunge Leaf oil
作者: Chi-Ya Huang; 黃琪雅
摘要: 本研究之目的為探討黃連木(Pistacia chinensis)枝條、葉子精油與抽出物抑制黃嘌呤氧化酶之活性,期盼開發黃連木於醫療保健領域的新用途。研究結果顯示黃連木枝條與葉子的乙醇抽出物、丙酮抽出物、熱水抽出物與精油中,以葉子精油抑制黃嘌呤氧化酶的效果最為顯著。 葉子精油的6個分離部中,以E1分離部最能抑制黃嘌呤氧化酶的催化;經由成分分析顯示,葉子精油與分離部E1的主要成分均為Limonene,相對含量分別為23.44%和61.06%,其他次要成分尚有cis-β-Ocimene、Terpinen-4-ol、α-Thujene、α-Pinene、4-Carene與3-Carene等。精油主成分Limonene經定量分析顯示,(+)-Limonene之含量明顯高於(-)-Limonene。成分中具有減少尿酸形成的化合物為Myrcene、(-)-Limonene與(+)-Limonene,使用次黃嘌呤為受質時,化合物的半數抑制濃度分別為36.88、33.53和37.69 μg/mL,而抑制酵素催化黃嘌呤為尿酸之IC50值則介於41.05-48.04 μg/mL。 精油、E1分離部與有效成分進行酵素動力學分析,發現E1分離部、Myrcene與(-)-Limonene是以競爭型方式阻礙酵素催化次黃嘌呤生成尿酸,而黃連木葉子精油、Camphene、(+)-Limonene和γ-Terpinene則為混合型態;當受質為黃嘌呤時,全部試樣皆為混合型的方式影響黃嘌呤氧化酶與黃嘌呤的親和力。綜合上述結果,可知黃連木葉子精油具有潛力發展為治療痛風的天然藥物。; The aim of this study was to evaluate the xanthine oxidase inhibitiory activities of oils and extracts from Pistacia chinensis leaf and twig. According to xanthine oxidase inhibition results, leaf oil exhibited the most excellent performance whether hypoxanthine or xanthine was used as the substrate. Among six fractions of leaf oil, E1 fraction possessed the best xanthine oxidase inhibition activity. Results from the chemical composition analysis of leaf oil and E1 fraction, both specimens showed limonene was the main compound with the relative content of 23.44% and 61.06%, respectively. The other compounds from leaf oil had cis-β-ocimene, terpinen-4-ol, α-thujene, α-pinene, 4-carene and 3-carene. In quantitative analysis of limonene, both leaf oil and E1 fraction had a much higher content of (+)-limonene than that of (-)-limonene. When the substrate was hypoxanthine, myrcene, (-)-limonene and (+)-limonene were the best active compunds against xanthine oxidase with IC50 values of 36.88, 33.53 and 37.69 μg/mL, respectively. When the substrate was xanthine, IC50 values of myrcene, (-)-limonene and (+)-limonene ranged from 41.05 to 48.04 μg/mL. In the enzyme kinetic studies, E1 fraction, myrcene and (-)-limonene exhibited the competitive-type inhibition against xanthine oxidase; and leaf oil, camphene, (+)-limonene and γ-terpinene exhibited the mix-type inhibition when using hypoxanthine as a substrate. When the substrate was xanthine, all active specimens presented the mix-type inhibition. Accordingly, leaf oil from Pistacia chinensis is a natural remedy with a great potential for the treatment of gout.2013-01-01T00:00:00Z麻竹組織培養培植體消毒及癒傷組織誘導之探討Ying-Chun Chen陳盈君http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/189872021-06-08T01:41:36Z2016-01-01T00:00:00Z標題: 麻竹組織培養培植體消毒及癒傷組織誘導之探討; Study on explants decontamination and callus induction for Dendrocalamus latiflorus
作者: Ying-Chun Chen; 陳盈君
摘要: 麻竹(Dendrocalamus latiflorus)為臺灣分布最廣泛且適應力良好之竹種,也是臺灣重要的竹類經濟作物。但麻竹遭受竹嵌紋病毒(Bamboo mosaic virus, BaMV)危害嚴重,阻礙了臺灣的竹產業發展。再者,現今常用的育苗法具有諸多限制,使麻竹產業發展面臨瓶頸。
至今已有諸多學者成功建立麻竹組織培養系統。其中因組織培養需於無菌環境中進行,良好的消毒條件成為建立系統的首要環節。但因麻竹培植體消毒困難,以常規的消毒藥劑難以將附於培植體表面之微生物及內生菌完全去除,因故多數學者使用消毒能力佳的HgCl2作為消毒藥劑。然而HgCl2易造成環境汙染,因此本研究選用對環境傷害較小的消毒藥劑,重新尋找麻竹組織最佳的消毒方式。並將培植體用於誘導癒傷組織,尋得癒傷組織誘導與增生之最適條件。
本試驗以麻竹含芽枝條進行試驗,結果顯示使用高濃度藥劑搭配短時間消毒,並且培養於含抗生素培養基,能有效地降低感染率至20%以下,同時不影響芽生長。進一步以麻竹筍作為試材,誘導麻竹癒傷組織。結果顯示,單獨使用1% NaOCl消毒45 min後以無菌水潤洗,培養在1/2 MSp680 + 2.41 mg/L Picloram培養基避光培養可得麻竹癒傷組織。同時也發現添加100 μg/ml Cefotaxime + 100 μg/ml Timetin於培養基可降低感染率。另以麻竹已抽芽枝條經2% NaOCl消毒30 min,輔以超音波震盪消毒15 min後,經3次無菌水潤洗後,置於添加50 μg/ml Cefotaxime及50 μg/ml Timetin培養基中,可得良好的消毒效果。培植體在不照光條件下,培養於1/2 MSp680 + 2.21 mg/L 2,4-D培養基結果最佳,誘導率達82.8%。然而癒傷組織生長仍需依附於培植體,單獨培養皆未能持續增生。
本結果可作為未來進一步研究麻竹癒傷組織形成及再生之基礎,並於可見之未來應用於深入探討細胞生長變化及其機制,提供未來竹類生長相關研究及應用之基石。; Ma bamboo (Dendrocalamus latiflorus) is an important commercial crop of Taiwan and it is the most distributed and well adapted bamboo specie. However, it is commonly infected by the Bamboo mosaic virus (BaMV) which has severely hindered the development of bamboo production. Furthermore, common breeding methods of bamboo have many limitations which pose challenges for the industry of ma bamboo.
Many successful ma bamboo tissue culture systems have been established. In tissue culture system, explants should be cultured in aseptic condition. Therefore, having a good sterilization method is the very first and an important step of system establishment. However, complete sterilization of microorganisms and its endophytes remain inefficient using common disinfectants. Research shows that HgCl2 is the most widely used disinfectant, yet it causes heavy environmental pollution. This study aims to find the optimal sterilization method that use disinfectants with less environmental impact and seek the best condition for ma bamboo callus induction.
Using the nodal branch of ma bamboo as material, results show that sterilization at high concentrations of disinfectant for a short time combined with antibiotic-supplemented mediums can lower contamination rates by more than 80% without affecting nodal growth. Ma bamboo shoots were also used to induce callus. Results show that there was successful callus induction when ma bamboo shoots were treated with 1% NaOCl for 45 min, washed thoroughly with sterilized distilled water, and cultured in 1/2 MSp680 medium that was supplemented with 2.41 mg/L Picloram in the absence of light. It was also found that adding 100μg/ml Cefotaxime, and 100μg/ml Timetin to the medium reduced contamination rates. The lowest contamination rate was observed when the ma bamboo budding branch was treated with 2% NaOCl for 30 min, sonication for 15 min, three thorough washes in sterilized distilled water, and cultured in a medium containing 50 μg/ml Cefotaxime and 50 μg/ml Timetin. The best callus induction rate (82.8%) occurred when the budding branch was cultured on 1/2 MSp680 medium supplemented with 2.21 mg/L 2,4-D in the absence of light. Despite these observations, the growth of callus still depended on explants and could not survive alone.
These results can be used as basis for further study of ma bamboo regeneration system, callus induction and reproduction. And be applied to understand the changes and mechanisms of cell growth, providing the cornerstone of future research and application of bamboo growth in the foreseeable future.2016-01-01T00:00:00Z麻竹筍生長之細胞壁特性及基因表現分析Wan-Jung Chang張菀蓉http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/582182021-06-16T08:08:33Z2014-01-01T00:00:00Z標題: 麻竹筍生長之細胞壁特性及基因表現分析; Cell Wall Characteristics and Gene Expression of Dendrocalamus latiflorus Shoot during Different Growth Stages
作者: Wan-Jung Chang; 張菀蓉
摘要: 麻竹(Dendrocalamus latiflorus)為臺灣常見竹種之中,分布最廣且適應力最佳之竹種。麻竹具有生長快速及竹種高大等優勢,為良好之木質纖維材料應用來源。本試驗以四周齡大之麻竹筍為材料,依生長部位之型態區分為頂部、中部及基部,分析其不同生長階段之細胞壁組成特性、代謝成分差異及基因表現,期能藉由上述分析,瞭解麻竹的生長特性,以利未來進一步推廣利用。
由組織切片之免疫標定及染色觀察結果得知,麻竹筍於頂部初生長階段,導管即開始有明顯的分化,此時之細胞壁結構以初級細胞壁為主,尚未有明顯的木質素及纖維素的累積。麻竹筍基部可觀察到完整的維管束組織,且木質素及纖維素大量累積於維管束組織。維管束的發育會先由原生木質部開始,並伴隨著導管的生成;接著為韌皮部的成熟,最後才為木質部導管及週邊組織的發育及纖維鞘的累積。細胞壁成分分析結果顯示,麻竹筍木質素含量約占6 – 9%,且於麻竹筍生長初期,酸可溶木質素含量會高於酸不溶木質素。麻竹筍細胞壁醣類組成主要為葡萄糖及木糖,並在生長過程有明顯的累積,葡萄糖的含量由頂部至基部分別為34.5、43.8及47.9%;木糖的含量則分別為14.8、27.0及29.2%。
代謝成分分析結果顯示,葡萄糖、呋喃木糖、果糖及蔗糖為極性層主要代謝成分。蔗糖於麻竹筍頂部有較高代謝,而葡萄糖、呋喃木糖及果糖此3種糖類於基部代謝較為旺盛。脂質層則以Pentadecanoic acid及8,11-Octadecadienoic acid為主要代謝成分,且此2種成分於麻竹筍頂部代謝較為旺盛。
基因差異表現結果顯示,於麻竹筍頂部主要為細胞活動及產生能量相關之基因,進一步以墨點分析及即時定量聚合酶鏈鎖反應結果可知,麻竹筍頂部之myo-Inositol phosphate synthase、Auxin response factor、Signal recognition particle 14 kDa protein及Glutathione S-transferase的表現量較高;麻竹筍基部表現量較高的基因則與光合作用香觀,分別為Chloroplast oxygen-evolving enhancer protein 及Succinate dehydrogenase。
本研究結合了物理、化學性質、代謝及基因表現分析,瞭解單株麻竹筍於不同生長階段之特性。由試驗結果可知,麻竹筍頂部之維管束發育尚未完全,此時細胞壁結構以初級細胞壁為主,基因表現及代謝以細胞分化及能量產生較為相關;麻竹筍基部具有成形之維管束,此時細胞壁已有明顯之二次生長,基因表現及代謝則以醣類生成及光合作用為主。; Ma bamboo (Dendrocalamus latiflous) is a common bamboo specie that is the most distributed and well adapted in Taiwan. Having the advantages of rapid growth and relatively large bulk, it provides a great source of ligno-cellulosic material. One four-week-old ma bamboo shoot was used for this experiment and was divided into top, middle and base portions. Each portion was analyzed according to different growth phases for differences in cell wall properties, metabolites and gene expression. A better understanding of the growth of ma bamboo shoot provides opportunities for further utilization in the future.
From immunolocalization and staining results, the vessels in the top portion of ma bamboo shoot began to show apparent cell division in primary growth phases. No clear accumulation of cellulose and lignin were observed in this primary growth phase. However, complete vascular bundles were observed in the base portion along with vast accumulation of cellulose and lignin. Initiation of vascular bundles starts in the region of the protoxylem accompanied by the formation of vessels, and follows the maturation of the phloem. Finally, the fiber sheath and surrounding tissue develop around the xylem vessel. Cell wall component analysis shows that lignin takes up 6 - 9% of the cell wall and that during early stages of growth, the contents of acid soluble lignin are higher than that of acid insoluble lignin. Glucose and xylose are the main sugar components of ma bamboo shoot cell wall and show significant deposition during growth. Glucose contents from top to base portion are 34.5%, 43.8%, and 47.9% respectively; while xylose contents from top to base portion are 14.8%, 27.0%, and 29.2% respectively.
Metabolite profiling shows that glucose, xylofuranose, fructose, and sucrose are the major compounds of the polar phase. Of these compounds, the metabolism of sucrose in the top portion of ma bamboo shoot is greater than that of the base portion, whereas the metabolism of glucose, xylofuranose, and fructose is greater in the base portion. In the lipid phase, pentadecanoic acid and 8, 11-octadecadienoic acid are the main compounds and they both present higher metabolism in the top portion.
According to gene differential expression results, the genes expressed in the top portion of ma bamboo shoot were mostly found to be related to cell activity and energy generation. Using dot blot and qRT-PCR methods, the main genes were recognized as myo-inositol phosphate synthase, auxin response factor, signal recognition particle 14 kDa protein, and glutathione S-transferase. In the base portion, the most expressive genes were chloroplast oxygen-evolving enhancer protein and succinate dehydrogenase which deals with photosynthesis.
Current research is an integrated analysis of physical, chemical, metabolic, and gene expressional properties, which allows us to understand the growth characteristics of ma bamboo shoot. In the top portion of the bamboo shoot, vascular bundles are immature. Functions of gene expression and metabolic processes involve cell differentiation and energy generation. On the other hand, the base portion contains fully developed vascular bundles, gene expression is related to sugar metabolism and photosynthesis, and cell wall exhibits noticeable secondary growth.2014-01-01T00:00:00Z