臺灣博碩士論文加值系統

摘 要
台灣金線連為台灣之名貴中藥草，且具廣泛藥效。除根腐病、莖腐病與蟎類等危害嚴重外，生長緩慢也是金線連無法快速大量生產之原因。因此，本論文的研究目的如下：
1.具促進金線連生長之蘭菌菌種之篩選；
2.改良蘭菌接種源生產之形式與速率；
3.有效菌種的大量生產；
4.促進金線連生長的生長速率。
在春季，非套袋栽培以單一菌種R09菌株與混合菌種R01.7.8.9(即R.01+R,07+R.08+R.09)接種較佳，套袋栽培金線連以單一菌種R07菌株與混合菌種R02.7.8.9(即R.02+R,07+R.08+R.09)接種具顯著生長促進效果。接種時應考慮氣溫與菌株間之競爭性，並建議慎選栽培期與菌種是共生栽培成敗之關鍵。
接種後混合菌種不如預期效果好，推測是受到低溫和介質的影響。經兩次重新接種有效單一菌種R01、R02與R04，然後以SSR(simple sequence repeat)進行分析，僅能分離出R01。因此建議單獨以R01或與其它分離菌株混合作為單一或混合菌種。
絲核菌屬代號R02菌株為蘭科植物良好之蘭共生真菌，並且可以顯著的促進金線連、彩葉蘭、蝴蝶蘭及拖鞋蘭等蘭科作物的生長。以GY培養基(glucose 2%、yeast extract 1%)來培養R02菌株，結果顯示出，R02菌株喜黑暗環境，在溫度25~30℃，培養基pH5.5~6.5，葡萄糖濃度為2~4%，酵母抽出物濃度為1~2%等條件下培養10天，即能生產出具有高活力且高產量之菌絲。
泥炭土 (含有20% V8蔬菜汁) 添加「安佳」脫脂與「克寧」全脂奶粉後，對菌絲之直線生長促進效果並不顯著，但菌絲密度顯著較高，整體以添加25 g/L脫脂奶粉效果最佳。水苔菌種為新式菌種，以蘭花瓶 (450mL容量) 生產時，水苔 (體積300mL) 以2.5%安佳脫脂奶粉浸泡，並適當調整溶液含量，滅菌後在無菌操作台下，每瓶植入蘭菌菌種 (5×5mm2) 三塊，培養6天可提升俱活力菌絲20~70倍；以MPS+1/5CM培養時，僅需四天即可生產出高發育率與高活力之菌絲。

Abstract
Anoectochilus formosanus Hayata is a valuable herb in Taiwan using as a multi-purposes of medicine. But they suffered from severe infection of root-rot, stem-rot and mites. Also, slow growth of A. formosanus hindered the production of this plant. Thus, this thesis aimed as follows：
1.To search for the effective inoculum ;
2.To improved the production method and speed of orchid mycorrhiza fungi (OMF) inoculum ;
3.To produce the effective OMF inoculum in large scale ;
4.To enhance the growth rate of A. formosanus.
In spring, the growth of A. formosanus could be enhanced by the single inoculum inoculation of R09 or by the mixed inocula of R01+R07+R08+R09 ( R01.7.8.9 ) . While A. formosanus plants were cultivated by plastic bag cultivation method ( PBCM ), R07 or R02+R07+R08+R09 ( R02.7.8.9 ) could stimulate their growth. The effect of the temperature and the competition among OMF inocula should be considered. So, it was suggested that right timing for inoculation with proper OMF was primary important.
The result of mixed inocula was not as good as expected. It was postulated that low temperature and growth medium might had some effects. Twice replication with the inoculation of effective inoculum such as R01、R02 and R04, then analyzed by SSR ( simple sequence repeat ) technique, the results showed that only R01 could be isolated. Therefore, it was recommended to use R01 isolate alone or to mix with other inoculum as mixed inoculum.
Isolate of Rhizoctonia spp. encoded as R02 was an effective OMF for orchidaceae , and could highly improve the growth of A. formosanus、Hamaria discolor (Ker.) Lindl.、Phalaenopsis spp. and Paphiopedilum spp., using GY medium ( glucose 2% + yeast extract 1% ) to grow R02 isolate, and the result showed that this isolate grew better in darkness、25~30℃ temperature、growth medium pH 5.5~6.5 in glucose 2~4%, and yeast extract 1~2% medium for 10 days, then high-viability mycelia in large scale could be produced.
Adding defatted ‘ Anchor ’ and full-cream ‘ Klim ’ milk powder to peat ( contaning 20% of V8 juice ) resulted in non-significant growth of OMF mycelial linear growth rate, but significantly increased the thickness of mycelia. It was found that by adding 25 g/l defatted ‘ Anchor ‘ dry milk to peat showed the best result. Sphagnum OMF inoculum was newly produced in an orchid flask (450ml in vol.). Three hundred mL in volume of wet sphagnum pre-soaked with 2.5% defatted ‘ Anchor ‘ milk powder solution ( MPS ), and adjusted the water content properly. After sterilization, then 3 pieces of OMF inoculum ( 5×5 mm2 ) were put into each orchid flask under laminard flow , then 20~70 times of viabile hyphae could be produced by incubation for 6 days. If the mycelia were grown in MPS+1/5 CM ( glucose, yeast extract and malt extract were 1 g/1, separately) , in only 4 days, high germinability and viability of mycelia could be produced.

論文中文摘要
論文英文摘要
前 言
第一章前人研究
第二章蘭菌之單一與混合菌種對台灣金線連幼苗生長與發育之影響 第三章利用分菌及SSR技術探討絲核菌對臺灣金線連根部之感染 獻第四章蘭菌生長特性之探
第五章蘭菌固體接種源繁殖速率之改

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