臺灣博碩士論文加值系統

本研究將綠球體(green globular body, GGB)繁殖系統應用在馬達加斯加鹿角蕨(Platycerium madagascariense Baker)之培養以大量繁殖孢子體。在添加5.37 μM α-naphthalene acetic acid (NAA)的MS培養基中，可以誘導孢子體幼葉產生綠球體。將其製作成石蠟切片在光學顯微鏡下觀察，可見到多數分生組織。在添加6-benzylaminopurine (BAP)及NAA的MS培養基中，綠球體(鮮重)每4週增殖2.88倍。在添加不同濃度的活性碳(activated charcoal, AC)及聚乙烯吡咯烷酮(polyvinylpolypyrrolidone, PVPP)的1/2 MS固態培養基上，能有效增加綠球體再生孢子體數量。在250 ml震盪瓶中懸浮培養均質化的綠球體10週後，以初始接種量每瓶0.9 g能再生最多正常形態發育的幼孢子體，，含大型者(根莖長(≥ 0.5 cm) 99株，小型者(< 0.5 cm) 780 株。為使這些幼孢子體早期即進入馴化並加速生長，以澆灌不同氮源比例溶液或市售花寶(Hyponex)水溶性肥料肥培。試驗結果以NH4+/NO3- = 0/100與花寶1號在大、小兩型幼孢子體分別能得到最大長、寬幅之葉片生長。本研究建立之懸浮培養系統，能在22週內以接種0.9 g均質化綠球體組織產出約840棵盆苗(營養葉覆蓋範圍約3 cm)。

The method of in vitro culturing green globular body (GGB) was applied in this study to mass propagate sporophytes in Platycerium madagascariense Baker. The GGB was induced from juvenile leaf explants incubating on MS medium supplemented with 5.37 μM α-naphthalene acetic acid (NAA). It was observed, under the light microscope, that the GGB tissue developed multiple meristems. The proliferation of GGB was achieved on a MS medium containing 6-benzylaminopurine (BAP) and NAA, and the fresh weight increase was recorded at 2.88 fold during a 4-week period. The sporophyte regeneration was effectively increased when GGB was transferred to 1/2 MS medium supplemented with activated charcoal (AC) or polyvinylpolypyrrolidone (PVPP). A suspension culture system further achieved the highest sporophyte production, all with normal morphogenesis, 99 large (rhizome size ≥ 0.5 cm) and 780 small (size < 0.5 cm), by incubating 0.9 g homogenized GGB in a 250 ml flask for 10 weeks. Young sporophyte growth was accelerated at early stage of suspension culture by fertility management at acclimatization applying different nitrogen sources in combination or Hyponex fertilizers. Either large or small sporophytes could achieve the maximum length and width growth in leaves under treatment with NH4+/NO3- = 0/100 or Hyponex No. 1 fertilizer. The procedure using 0.9 g homogenized GGB in suspension could produce 840 pot plants in 22 weeks, with diameter of trophophyll coverage at 3 cm.

I前言 1
II前人研究 4
(I)鹿角蕨無菌播孢培養 4
(II)鹿角蕨的無性繁殖 5
(III)懸浮培養及生物反應器 8
(IV)不同氮源影響與肥培培養 10
III材料方法 17
(I)綠球體誘導 17
(II)綠球體增殖 18
(III)綠球體石蠟切片觀察 18
(IV)固態培養再生孢子體 19
(V)懸浮培養均質化綠球體再生孢子體 20
(VI)幼孢子體馴化與澆灌肥培管理 20
(VII)培養條件 23
(VIII)資料處理與統計分析 23
IV結果 25
(I)綠球體誘導及增殖 25
(II)綠球體石蠟切片觀察 25
(III)固態培養再生孢子體 28
(IV)綠球體均質化及懸浮培養 30
(V) 幼孢子體馴化與肥培管理試驗 31
V討論 43
VI參考文獻 49

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