山核桃间接体细胞胚发生和植株再生

园艺学报 ›› 2011, Vol. 38 ›› Issue (6): 1063-1070.

山核桃间接体细胞胚发生和植株再生

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,*

（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）

收稿日期:2010-11-17 修回日期:2011-06-08 出版日期:2011-06-25 发布日期:2011-06-25
通讯作者: 黄坚钦

Indirect Somatic Embryogenesis and Plant Regeneration of Carya cathayensis

ZHANG Qi-xiang1,2，HU Heng-kang1,2,3，WANG Zheng-jia1,2，YUAN Jia1,2，WAN Jun-li1,2，and HUANG Jian-qin1,2,*

（1School of Forestry and Biotechnology，Zhejiang Agriculture and Forestry University，Lin’an，Zhejiang 311300，China;2The Nurturing Station for the State Key Laboratory of Subtropical Silviculture，Zhejiang Agriculture and Forestry University，Lin’an，Zhejiang 311300，China；3College of Landscape Architecture and Art，Jiangxi Agriculture of University，Nanchang 330045，China）

Received:2010-11-17 Revised:2011-06-08 Online:2011-06-25 Published:2011-06-25
Contact: HUANG Jian-qin

摘要/Abstract

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1/2 MS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1.0 mg · L-1 6-BA和0.01 mg · L-1 picloram（氨氯吡啶酸）组合时，胚性愈伤组织诱导率最高，为48.6%；而1.0 mg · L-1 6-BA与0.001 mg · L-1 picloram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3 d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。　　

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

Abstract: 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 on the 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. 　

Key words: Carya cathayensis, somatic embryo, indirect somatic embryogenesis, plant regeneration, cytohistological observation

中图分类号:

S 664.1

张启香;胡恒康;;王正加;袁佳;万俊丽;黄坚钦;. 山核桃间接体细胞胚发生和植株再生 [J]. 园艺学报, 2011, 38(6): 1063-1070.

ZHANG Qi-xiang;HU Heng-kang;;WANG Zheng-jia;YUAN Jia;WAN Jun-li;and HUANG Jian-qin;. Indirect Somatic Embryogenesis and Plant Regeneration of Carya cathayensis [J]. ACTA HORTICULTURAE SINICA, 2011, 38(6): 1063-1070.

[1]	季琳琳, 陈素传, 吴志辉, 常君, 韩文妍, 陶汝鹏. 早花山核桃新品种‘宁国山核桃2号’[J]. 园艺学报, 2022, 49(S2): 53-54.
[2]	蒋思思, 袁军, 周文君, 钮根花, 周俊琴. 薄壳山核桃（Carya illinoinensis）叶绿体基因组及其特征分析[J]. 园艺学报, 2022, 49(8): 1772-1784.
[3]	陈素传, 季琳琳, 吴志辉, 常君, 陶汝鹏, 汪小进 . 大果山核桃新品种‘宁国山核桃3号’[J]. 园艺学报, 2021, 48(S2): 2803-2804.
[4]	常君, 姚小华, 王开良, 滕建华, 邵慰忠, 张建忠, 李雪涛, 任华东, . 薄壳山核桃新品种‘亚优YLC21号’[J]. 园艺学报, 2021, 48(S2): 2805-2806.
[5]	陈素传, 吴志辉, 季琳琳, 常君, 陶汝鹏, 汪小进. 山核桃新品种‘宁国山核桃1号’[J]. 园艺学报, 2020, 47(S2): 2925-2926.
[6]	王开良, 姚小华, 邵慰忠, 任华东, 常君, 方新高, 洪友君. 薄壳山核桃新品种‘亚优YLC28号’[J]. 园艺学报, 2020, 47(S2): 2927-2928.
[7]	雅蓉1，徐伟荣2,3,4,5,*，张莹1，夏思琪2，张宁波2,3,5. 褪黑素对‘无核白’葡萄体细胞胚的诱导作用[J]. 园艺学报, 2020, 47(5): 953-962.
[8]	刘星月, 朱强龙, 李慧英, 孙静宇, 李必聪, 张宏玉, 黄英金, 方加军, 周庆红, . 红芽芋脱毒试管芋诱导及植株再生[J]. 园艺学报, 2020, 47(12): 2427-2438.
[9]	栾爱萍1，贺军虎1,，刘佳柔2，龚雪2，方少秋2，夏靖娴2，林文秋3，许文天3，何业华2,. 矮凤梨短营养生长期突变材料‘14-1’的性状鉴定[J]. 园艺学报, 2019, 46(1): 143-148.
[10]	朱先富1，何国庆2，黄久奎3，徐步青4，夏国华5,*. 大别山山核桃新品种‘皖金1号’[J]. 园艺学报, 2018, 45(S2): 2731-2732.
[11]	孙红梅1,2,*，付麟岚1，王志平1，盖美竹1，王春夏1,2. 基于体细胞胚发生的细叶百合和兰州百合多倍体诱导及鉴定[J]. 园艺学报, 2018, 45(6): 1136-1146.
[12]	夏国华1，王正加1，丁立忠2，张秋月3，黄坚钦1,*. 山核桃新品种‘浙林山3号’[J]. 园艺学报, 2018, 45(3): 603-604.
[13]	黄春颖，黄有军，吴建峰，黄仁，栾雨濛，张深梅，王正加，张启香，黄坚钦. SAD和FAD*家族基因调控山核桃不饱和脂肪酸组分配比[J]. 园艺学报, 2018, 45(2): 250-260.
[14]	姚小华1,*，任华东1，邵慰忠2，王开良1，尹润富3，王年金4，常君1. 山核桃新品种‘亚优7号’[J]. 园艺学报, 2018, 45(2): 401-402.
[15]	常君1，姚小华1，郎学军2，王开良1，滕建华3，王年金4，任华东1,*. 山核桃新品种‘亚优8号’[J]. 园艺学报, 2018, 45(1): 195-196.