日本女貞之微體繁殖

Abstract

本試驗嘗試探討並建立日本女貞其組織培養之微體繁殖體系。初步結果如下： 本試驗以日本女貞之成熟種子為培植體進行培養。將成熟種子先以流水處理24小時，先以1％安其消毒液浸泡20分鐘做表面清潔，再以70％之酒精浸泡5分鐘，最後在5％ NaOCl水溶液（含約0.1％（v/v）Tween20展著劑）中超音波震盪30分鐘，種子可達100％之無污染率。 在器官形成方面：種子與成熟胚植於WPM、1/2MS、MS三種基礎培養基中以WPM對於無菌苗之誘導效果最好。以胚軸為培植體，以WPM為基礎培養基在光照環境下，添加0.5 ppm 之2,4-D培養，可獲得大量淡黃色、鬆軟之癒合組織。以無菌苗之頂芽為培植體，以WPM為基礎培養基在光照環境下，可順利發根。無菌苗之頂芽，在含有0.01 ppm IBA及1 ppm BA 之WPM培養基中，光照環境下可誘導出多芽體。誘導出之多芽體以WPM空白培養基在光度20±5μmol m-2s-1時，抽長效果較好。日本女貞細胞液態培養以0.1 ppm 2,4-D配合10 g/l 蔗糖之WPM培養基最為適當。

This study describes the development conditions and the propagation technique by in vitro culture for the Ligustrum japonicum Thunb. The primary results are as follow: Results of mature seeds culture of Ligustrum japonicum were as follows: mature seeds were first treated with running water for 24 hours, soaked in 1﹪anticeptol solution（Benzel thyonium chloride U.S.P 10﹪(w/v), Allcyl arylpolyther alcohol 10﹪(w/v)）for 20 minutes, then soaked in 70﹪ethanol solution for 5 minutes, then soaked in 5﹪NaOCl（supplemented with 0.1﹪(v/v) Tween 20）and treated with ultrasonic shaker for 30 minutes and the seeds could get the 100﹪non- contamination. The medium of WPM get the best results to establish sterile plants. Light yellow and soft calli were induced after hypocotyls were cultured on WPM medium containing 0.5 ppm 2,4-D under light containing. Rooting was induced after terminal buds were cultured on WPM medium under light containing. Multiple shoots were induced after terminal buds were cultured on WPM medium containing 0.01 ppm IBA and 1 ppm BA under light containing. Multiple buds when transferred to WPM medium lacking plant growth regulators were promoted to elongate their length of internodes under light intensity of 20±5μmol m-2s-1 light environment. The optimal medium for suspension culture was WPM added with 0.1 ppm 2,4-D and 10g/l sucrose of WPM medium.