影响文心兰叶片体胚诱导的条件研究  

Somatic embryogenesis induced from leaf explants of Oncidium flexuosum ‘Million Dollar’

作　　者：潘銮银[1,2] 宋红改[2,3] 蒋晶[2] 乔桂荣[2] 李海营[2] 王扬[1] 林永生 卓仁英[2] 

机构地区：[1]云南农业大学植物保护学院,云南昆明650201 [2]中国林业科学研究院亚热带林业研究所,浙江富阳311400 [3]三峡大学生物技术研究中心,天然产物研究与利用湖北省重点实验室,湖北宜昌443002 [4]福建省莆田市江口农业服务中心,福建莆田351115

出　　处：《浙江林学院学报》2010年第4期620-625,共6页Journal of Zhejiang Forestry College

基　　金：国家自然科学基金资助项目(30972340);浙江省自然科学基金杰出青年团队项目(R3090070)

摘　　要：以文心兰Oncidium flexuosum无菌苗叶片为外植体,1/2MS(Murashige and Skoog)为基本培养基,对不同叶位、叶片大小(叶龄)、不同植物生长调节物质处理诱导体胚的效果进行了研究。结果表明:①幼苗的叶尖、叶基部切口、叶面切口、叶面和叶缘均能诱导出体胚,绝大部分体胚能形成二次体胚并能分化成多个类原球茎(PLBs)。②诱导体胚的最佳外植体材料是取自带根幼苗15mm长叶片。③诱导体胚的适宜培养基组成如下:1/2MS培养基(其中微量元素和有机添加物为全量,铁盐减半),磷酸二氢钠170.00mg·L-1,谷胺酰胺1000.00mg·L-1,蛋白胨1000.00mg·L-1,噻苯隆(TDZ)0.50mg·L-1,聚乙烯聚吡咯烷酮(PVPP)250.00mg·L-1,蔗糖20000.00mg·L-1,固化剂(gelrite)2800.00mg·L-1,pH5.2。叶片体胚诱导率为90.00%以上,叶片基部切口和中切口体胚诱导率分别达74.56%和66.91%。Young leaves of Oncidium flexuosum ‘Million Dollar’ were used as explants and cultured in a 1/ 2 Murashige and Skoog（MS） basic medium with three plant growth regulators for somatic embryogenesis induction. A 60 d in vitro culture in darkness of direct somatic embryogenesis for different leaf positions, seedling characteristics, and leaf size （leaf age） was studied along with the influence of different combinations and concentrations of thidiazuron （TDZ）, 6-benzyladenine （6-BA）, and kinetin （KT） on direct somatic embryogenesis. Results after 60 d showed that leaf tips, adaxial sides, margins, cut ends, and cut central parts formed many somatic embryos and repetitive somatic embryos. From these somatic embryos, a number of protocorm-like-bodies eventually formed. The best leaf explant for somatic embryogenesis induction was a 15 mm leaf from seeding with roots. The best embryogenic responses were with 1/2 strength macro-and micro-elements of MS supplemented with 100.00 mg·L-1 myo-inositol, 0.50 mg·L-1 niacin, 0.50 mg·L-1 pyridoxine HCl, 0.10 mg·L-1 thiamine HCl, 2.00 mg·L-1 glycine, 170.00 mg·L-1 NaH2PO4, 13.90mg·L -1 FeSO4·7H2O, 1 000.00 mg·L -1 peptone, 0.50 mg·L -1 TDZ, 250.00 mg·L -1 polyvinylpolypyrrolidone （PVPP）, 20 000.00 mg·L -1 sucrose, and 2 800.0 mg·L -1 GelriteTM at a pH of 5.2. Additionally, 90% of the leaf explants induced somatic embryos, and 74.6% of the cut ends of leaves as well as 66.9% of the cut central parts of leaf explants induced somatic embryos.

关 键 词：植物学 文心兰 叶片 体胚诱导 

分 类 号：S682.31[农业科学—观赏园艺]