Development of a novel micropropagation system in Lilium longiflorum by film culture vessel

利用一种新型的一次性包膜状培养容器——由氟碳聚合物薄膜（Neofron （] SY.encircledR.[）薄膜）制成的“培养袋”，研究了长花百合鳞片体外培养的球茎生产。本文研究了用改良的Murashige和Skoog （MS）琼脂培养基（宏微量营养成分各占一半，蔗糖含量为4.5%）在烧瓶和用12.5、25、50、100 mm厚的Neoflon FEP或PFA薄膜制成的培养袋（90 × 40 mm）中的液体培养基在光或暗条件下培养鳞片的球形成情况。在两种培养皿中，光照条件下获得的小球粒比黑暗条件下的大，每个鳞片的小球粒数也比黑暗条件下的多。事实上，在任何一个培养袋中，每个鳞片的子弹数量和子弹的新鲜重量都比在烧瓶中要高。FEP膜培养袋中获得的球芽上形成的叶鳞数高于培养袋中的，实际上高于烧瓶中的。FEP膜培养袋中获得的小球茎上形成的叶鳞数量高于PFA膜培养袋。进一步的研究表明，在由PFA膜制成的培养袋中，MS（全强度营养物，4.5%蔗糖）液体培养基中获得的小球比在烧瓶中的琼脂培养基中获得的大得多。还尝试使用比较小尺寸的培养袋大9倍（270 x 120毫米）和较大尺寸的4室培养袋来开发称重系统。在百合的微繁繁殖中，使用培养袋系统可能具有以下优点；(1)与传统的微繁方法相比，该方法可以获得更大的鳞茎，且鳞茎叶和根都很短；(2)将外植体接种到大尺寸的培养袋中可以节省这些鳞茎；(4)与发酵罐和罐式培养相比，该方法可以大大降低鳞茎的生产成本。少

The bulblet production from Lilium longiflorum scales cultured in vitro was studied by using a novel disposable envelope-shaped culture vessel, the "Culture Bag" made of fluorocarbon polymer films (Neofron (〕SY.encircledR.〔) films). This paper dealt with the bulblet formation in scales cultured in vitro on modified Murashige and Skoog (MS) agarmedium (half macro-and micro-nutrients, 4.5% sucrose) in flask and in the liquid medium in the Culture Bag (90 x 40 mm) made of Neoflon FEP or PFA film of 12.5, 25, 50, and 100 mum thickness under light or dark condition. In both culture vessels, larger bulblets and higher number of bulblets per scale were obtained under light than dark. The number of bulblets per scale and fresh weight of bulbrets in any of the Culuture Bag were in fact higher when compared to those in flask. The number of foliage scales formed on bulbrets obtained in the Culuture Bag made of FEP film was higher than those in the Culuture Bag were in fact higher when compared to … More those in flask. The number of foliage scales formed on bulblets obtained in the Culture Bag made of FEP film was higher than those in the Culuture Bag made of PFA film. Further work showed that much larger bulblets were obtained in MS (full-strength nutrients, 4.5% sucrose) liquid medium in the Culture Bag made of PFA film compared to those on the agar medium in flask. An attempt was also made to develop the svale-up systems by using the Culuture Bag which is nine times larger (270 x 120 mm) than the smaller size and the 4-chambered Culuture Bag of larger size.the possible advatages to using the Culuture Bag system in the micropropagation of Lilium are as follows ; (1) bigger bulblets with very short foliage leaves and roots could be obtained than those in the conventional micropropagation, (2) these bulblets could be saved when inoculating explants to the large size Culuture Bag, and (4) the cost for the production of bulblets is expected to be much reduced by this novel system when vompared to those by the fermentor system and the tank culture system. Less