香蕉胚性悬浮细胞系建立过程中非胚性成分的去除  

Removal of Non-embryogenic Components during Establishment of Embryogenic Cell Suspension of Banana

作　　者：常胜合[1,2,3] 徐立[3] 李敬阳[1,2] 孙威[1,2] 王甲水[1,2] 许桂莺[1,2] 孙佩光[1,2] 吴琼[1,2] 金志强[1,2] 舒海燕[1,2] 

机构地区：[1]中国热带农业科学院海口实验站,海南海口570102 [2]海南省香蕉遗传改良重点实验室,海南海口570102 [3]中国热带农业科学院热带作物品种资源研究所,海南儋州571737

出　　处：《安徽农业科学》2013年第20期8477-8479,共3页Journal of Anhui Agricultural Sciences

基　　金：中央级公益性科研院所基本科研业务费专项资金(中国热带农业科学院热带作物品种资源研究所)项目(1630032013022);海南省自然科学基金项目(313065)

摘　　要：[目的]研究香蕉胚性悬浮细胞系建立过程中非胚性成分的去除方法.[方法]以巴西蕉的花蕾进行诱导培养后,运用孔径在250～ 500 μm的不锈铜筛网进行过滤,并用倒置显微镜对培养物进行实时观测.[结果]运用孔径在250～500 μm的不锈钢筛网能够有效去除悬浮细胞系中的褐化坏死组织和分生组织小球,但如果在悬浮细胞系诱导后3d立即进行去除,则易使之后胚性细胞的增殖变得困难;如果在悬浮细胞系诱导后14 d左右去除,则褐化坏死组织释放的酚类物质很容易损伤并杀死正在增殖的胚性细胞.悬浮细胞系诱发后7d是一个比较恰当的过滤去除时间点.[结论]在培养的过程中,将悬浮细胞系定期放置在倒置显微镜下进行观察,用移液器能将液泡化的细胞和胚去除.在操作前,需将倒置显微镜放置在超净工作台内,用紫外线长时间照射进行表面灭菌.这样即可以将悬浮细胞系中的非胚性成分过滤去除,又能预防感染.[ Objective ] The aim was to study the removal of non-embryogenie components during the establishment of the embryogenie cell sus- pension （ECS） of banana. [ Method] The buds of Musa acuminate AAA Cavendish cv. Brazil were induced and cultured, and the culture was fil- tered by meshes With aperture of 250 -500 μm. Then the culture was observed in real time by inverted microscope. [ Result ] The used of meshes with aperture of 250 - 500 μm could effectively remove the necrotic tissues and meristematic globules. But if the meristematic globules were re- moved after 3 days of ECS induction, the following proliferation of embryogenie cells would become difficult. If the meshes were used after 14 days of ECS induction,the embryogenic cells would be injured by phenolic compounds that were released from the necrotic tissues. It was appropriate to sieve the fractions after 7 days of ECS induction. [ Conclusion] ECS was put under inverted microscope and pipette could be used to remove embryos and highly vacuolated cells. The inverted microscope should be put in the superclean bench and surface-sterilized with UV light,that could guarantee that microbes would not be introduced.

关 键 词：香蕉 胚性悬浮细胞系 非胚性成分 去除 

分 类 号：S668.1[农业科学—果树学]