海州骨碎補試管內培養及體胚發生

In vitro culture and somatic embryogenesis of Davallia mariesii Moore ex Bak

蔡羽婷(Yung-Ting Tsai)

指導教授 ： 何錦玟

大同大學/工程學院/生物工程學系所/碩士(2009年)

摘要

海州骨碎補(Davallia mariesii Moore ex Bak)是一種多年附生性蕨類，又稱台灣骨碎補，具有觀賞以及藥用價值。海州骨碎補的孢子消毒後播種於改良1/2 MS基礎鹽類固態培養基，萌芽後以原葉體為培植體，添加3 mg l-1 GA3對孢子體形成有些微促進。
將幼小孢子體劃傷培養於改良1/2 MS液態培養基，可產生癒合組織並形成體胚，在液態培養下部份體胚可轉成幼小孢子體。為了使孢子體適應試管外培養，將含體胚癒合組織轉至1/2 MS基礎鹽類減糖固態培養，分別添加2,4-D、kinetin、picloram、BA四種生長調節劑，濃度為1、5、10、15 mg l-1，結果顯示15 mg l-1 kinetin可以使芽抽長並且癒合組織不褐化，同時促進癒合組織中芽分化。將2,4-D及kinetin以1、5、10、15 mg l-1搭配測試，以5 mg l-1 2,4-D及1 mg l-1 kinetin組合結果最佳，可使芽抽長同時新增芽數最多。孢子體出瓶培養馴化後型態與原本試管外的母株相同。

關鍵字

體胚發生 ； 骨碎補 ； 蕨類

並列摘要

Davallia mariesii Moore ex Bak is a kind of perennial fern. It is an ornamental plant and also medicinal plant. It is a traditional Chinese herbal medicine to be used to treat about injure and hurt in bones.
The immature and mature spores were seeded on 1/2 modified MS medium after sterilization, and germinated spores( prothallus ), used as explant for the formation of sporophyte. Explant was cultured on modified 1/2 MS medium with different concentration of GA3. The medium supplemented with 3 mg l-1 GA3 had the best effect of the formation of sporophyte .
The wounded young sporophyte was inoculation in modified 1/2 MS liquid medium, and lots of embryogenic callus are proliferated. Some of the somatic embryos can regeneration into young sporophytes. In order to the formation of frond and root, the embryogenic callus were transfer to 1/2 MS medium, supplement with different concentration( 1, 5, 10,and 15 mg l-1 ) of 2,4-D, kinetin, picloram and BA. Compared to the medium without PGRs, more buds differentiation from the sporophyte when supplement with 15 mg l-1 kinetin. Further test showed that the combination of 5 mg l-1 2,4-D and 1 mg l-1 kinetin have the best effect on buds differentiation The sporophyte can survive in vivo and the same phenotype with mother plant.

並列關鍵字

embryogenesis ； Gu-sui-bu ； fern

參考文獻

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