臺灣博碩士論文加值系統

仙履蘭(Paphiopedilum)在組織培養的過程中，常因培植體褐化影響其再生增殖，使用100 mg∙L-1抗壞血酸(ascorbic acid, AA)或抗壞血酸配合激動素(ascorbic acid and kinetin, AA+KT)在消毒殺菌前處理仙履蘭Maudiae Type雜交種Paph. Hsinying Web ‘Giant’ × Paph. Pulsar ‘Hsinying Flame’花梗芽培植體，以AA+KT的效果優於單獨施用AA，花梗芽培植體取自初期發育的花苞較成熟花朵的褐化率低，且再生芽體誘導率高。在培養基中添加半胱胺酸(cysteine, Cys)，能夠有效降低仙履蘭培植體在培養初期之多酚氧化酶(polyphenol oxidase, PPO)、苯丙胺酸脫氨酶(phenylalanine ammonia lyase, PAL)等活性，培植體添加50-100 mg∙L-1 Cys組的PAL活性都較對照組低，顯著降低仙履蘭培植體PAL合成酚類化合物如香豆酸(coumaric acid)的含量，亦減少和PPO反應的受質供源，因此降低酵素型褐化的生成，並能誘導芽體增殖與生根。高氮鹽培養基會加劇仙履蘭培植體之褐化，修改培養基NO3--N/NH4+-N的氮源比例和型態，以除去硝酸銨的MS培養基能有效減輕仙履蘭培植體的褐化，利於仙履蘭分生苗之增殖。此外，以硝酸鉀為唯一無機態氮源之培養基最能降低仙履蘭分生苗培植體褐化且提高芽體的增殖。

Abstract
Explant browning was frequently found to impede plant regeneration and multiplication during in vitro propagation of Paphiopedilum (Paph.) orchids. To reduce explant browning, lateral flower buds (LFBs) of Maudiae Type Paph. Hsinying Web ‘Giant’ × Paph. Pulsar ‘Hsinying Flame’ were presoaked with 100 mg∙L-1 ascorbic acid (AA) solution or ascorbic acid +kinetin (AA+KT) solution before surface sterilization. AA+KT was superior to AA alone in controlling explant browning of LFBs. After AA+KT pretreatment, LFB explants excised from young floral buds grew better than those from mature flowers with lower explant browning rate and higher shoot induction rate. Supplement of cysteine (Cys) in culture medium effectively decreased polyphenol oxidase (PPO) and phenylalanine ammonia lyase (PAL) activities of Paph. explants during early incubation. Particularly, the presence of 50 -100 mg∙L-1 Cys depressed Paph. explant PAL activities compared with the control groups, and thus greatly reduced phenolic compounds, such as coumaric acid. The reduction of phenolic substrates for PPO oxidization significantly prevented enzymatic browning in Cys-containing media and subsequently induced shoot multiplication and rooting in Paph. explants. Medium with high nitrogen salts exacerbated the incidence of Paph. explant browning, therefore alteration of NO3--N/NH4+-N ratio and nitrogen form in media were investigated. Removal of ammonia nitrate in MS medium positively reduced explant browning and beneficially improved the development of Paph. mericlonal propagation. In addition, potassium nitrate as a sole inorganic nitrogen source lowered the occurrence of Paph. explant browning and increased shoot multiplication.

中文摘要 i
英文摘要 ii
目次 iii
表目次 v
圖目次 vi
縮寫與全名對照表 viii
第一章 緒論 1
第一節 前言 1
第二節 前人研究 3
第二章 仙履蘭培植體前處理抗氧化物與細胞分裂素對再生芽體之影響 12
摘要 12
英文摘要 13
第一節 前言 14
第二節 材料與方法 17
第三節 結果 19
第四節 討論 23
第三章 半胱胺酸對仙履蘭分生苗培植體多酚氧化酶、苯丙胺酸脫氨酶活 性影響與芽體增殖 43
摘要 43
英文摘要 44
第一節 前言 45
第二節 材料與方法 48
第三節 結果 51
第四節 討論 54
第四章 培養基氮源比例及型態對仙履蘭分生苗發育之影響 65
摘要 65
英文摘要 66
第一節 前言 67
第二節 材料與方法 69
第三節 結果 74
第四節 討論 77
第五章 綜合討論 98
參考文獻 101
附錄 115

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