山核桃间接体细胞胚发生和植株再生  

Indirect Somatic Embryogenesis and Plant Regeneration of Carya cathayensis

作　　者：张启香[1,2] 胡恒康[1,2,3] 王正加[1,2] 袁佳[1,2] 万俊丽[1,2] 黄坚钦[1,2] 

机构地区：[1]浙江农林大学林业与生物技术学院,浙江临安311300 [2]浙江农林大学亚热带森林培育国家重点实验室培育基地,浙江临安311300 [3]江西农业大学园林与艺术学院,南昌330045

出　　处：《园艺学报》2011年第6期1063-1070,共8页Acta Horticulturae Sinica

基　　金：浙江省自然科学基金重点项目(Z307534);浙江省自然科学基金项目(Y3090091);浙江农林大学B类创新团队项目;浙江省科技厅面上项目(2009C32027);浙江省大学生科技创新计划项目(2009)

摘　　要：以山核桃(Carya cathayensis Sarg.)自然授粉后10周的幼胚为外植体,对影响胚性愈伤组织和体胚发生的主导因子(基本培养基、植物生长调节物质等)进行了比较分析;对影响体胚萌发的脱水处理时间进行了比较;并采用石蜡切片法对幼胚脱分化产生胚性愈伤组织及体胚发生发育过程进行了组织细胞学观察。结果表明,幼胚胚轴和子叶接种在基本培养基1/2MS上,胚性愈伤组织诱导率显著高于其它处理,达45.3%。接种于基本培养基DW中的幼胚体细胞胚诱导率显著高于其它培养基处理,达16.7%。显微镜下可观察到球形胚、心形胚、鱼雷胚和子叶胚。培养基添加1.0mg.L-16-BA和0.01mg.L-1picloram(氨氯吡啶酸)组合时,胚性愈伤组织诱导率最高,为48.6%;而1.0mg.L-16-BA与0.001mg.L-1picloram组合,体胚诱导率最高,为23.6%。体细胞胚发生方式属于间接体胚发生。用饱和Ca(NO3)2.4H2O对体胚进行脱水处理,脱水处理3d后萌发率为39.03%,显著高于对照及其余处理。将脱水处理后的体胚接种至WPM基本培养基中,光照条件下培养,10周后可长成具3～4片真叶的完整植株。Immature embryos of 10 weeks after pollination of Carya cathayensis were selected as explants.The factors which affect embryogenic calli formation and somatic embryogenesis including basal medium,plant growth regulators（PGRs）and desiccation length of somatic embryos were tested.Cytohistological observations of anatomical structure change during embryogenic callus formation and somatic embryos production were also made.Embryogenic callus percentage was significantly higher onthe medium 1/2 MS than other treatments,reached 45.3%.Somatic embryogenesis percentage was significantly higher on DW medium than other media,reached 16.7%.Global embryo,heart-shaped embryo,embryos and cotyledonary embryos were easily observed under microscope.The highest embryogenic callus percentage was obtained on the medium with 1.0 mg ？ L-1 6-BA and 0.01 mg ？ L-1 picloram,reached 48.6%.The highest somatic embryogenesis percentage was obtained on the medium with 1.0 mg ？ L-1 6-BA and 0.001 mg ？ L-1 picloram,reached 23.6%.The formation and development of somatic embryogenesis is indirect somatic embryogenesis which goes through callus phase.Subsequent desiccation for 3 d with saturated Ca（NO3）2 ？ 4H2O resulted in 39.03% germination.Non-desiccated somatic embryos failed to germinate.Germinated somatic embryos developed into plantlets with 3 to 4 leaves 10 weeks after they were transferred to WPM basal medium under light condition.

关 键 词：山核桃 体细胞胚 间接发生 植株再生 组织细胞学观察 

分 类 号：S664.1[农业科学—果树学]