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作 者:乔利仙[1] 隋炯明[1] 谭玲玲[1] 郭宝太[1] 孙世孟[1] 王晶珊[1]
机构地区:[1]青岛农业大学生命科学学院/山东省高校植物生物技术重点实验室,山东青岛266109
出 处:《农学学报》2012年第10期9-13,共5页Journal of Agriculture
基 金:山东省自然基金"花生离体诱变及创造抗旱耐盐种质资源研究"(ZR2012CM014);山东省自然基金青年基金"花生维生素E合成关键基因的克隆及其抗氧化机制研究"(ZR2011CQ026);国家自然基金青年基金"花生几丁质酶基因;β-1;3-葡聚糖酶基因启动子的克隆与功能分析"(31101178);山东省中青年科学家奖励基金"花生Chi基因及Glu基因的克隆;双价表达载体的构建及遗传转化研究"(BS2009NY028)
摘 要:以成熟种子胚小叶为外植体,对花生体胚诱导及植株再生进行了研究,旨在为花生体细胞杂交、离体诱变及遗传转化提供培养方法。将花生5大类型17个品种的胚小叶外植体分别培养在添加10mg/L2,4-D的培养基上诱导体胚形成。培养4周后将形成体胚的外植体转移到添加4mg/LBAP的培养基上进行培养,促使体胚萌发成苗。结果表明,不同类型间体胚诱导率和植株再生率存在明显差异,中间型供试品种获得了较高的胚诱导率(84.1%~91.2%)和植株再生率(81.1%~97.0%);珍珠豆型和普通型内供试品种间体胚诱导率差异较大,分别为43.9%~85.0%和42.2%~82.2%,而植株再生率均较高,分别为94.3%~96.5%和88.1%~98.7%;多粒型品种体胚诱导率(32.2%~50.0%)和植株再生率(41.2%~55.1%)均最低;而龙生型体胚诱导率(61.1%~87.8%)和植株再生率(63.7%~87.6%)均表现为中间。再生苗经嫁接驯化后移栽于田间,植株正常生长和结实。本研究结果说明,花生不同类型间再生能力存在显著差异,多粒型品种再生能力最低。Efficient somatic embryogenesis and plant regeneration in embryonic leaflet culture of peanut (Arachis hypogaea L.) were studied, for providing the methods of somatic hybridization, In vitro mutagenesis and genetic transformation. The embryonic leaflets of 17 peanut cultivars in 5 types were cultured on MSB5 medium (Murashige and Skoog salts plus B5 vitamins) supplemented with 10.0 mg/L 2,4-D to induce somatic embryos. After 4 weeks culture, the explants with somatic embryos were transferred to the medium supplemented with 4 mg/L BAP for somatic embryo germination and plant regeneration. The result showed that the frequencies of somatic embryo induction and plant regeneration were different significantly in 5 peanut types. Intermediate type gave a higher frequency (84.1%-91.2%) of somatic embryo induction and a higher frequency (81.1%-97.0%) of plant regeneration. The frequencies of somatic embryo induction were significantly different among the cuhivars in Arachis hypogaea var. vulgaris with a range from 43.9% to 85.0% and Arachis hypogaea var. hypogaea with a range from 42.2%-82.2%, respectively, but the higher frequencies of plant regeneration were obtained in them, sharing a range from 94.3%-96.5% and 88.1%-98.7%,respectively. Araehis hypogaea var. fastigiata showed the lowest frequencies of somatic embryo induction (32.2%-50.0%) and plant regeneration (41.2%-55.1%). The frequencies of somatic embryo induction (61.1%-87.8%) and plant regeneration (63.7%-87.6%) were obtained in Araehis hypogaea vat. hirsute. When the plantlets were grafted onto germinated seedlings and transplanted to the field, they grew to maturity and produced viable seeds. The result showed that the regeneration capacity was different among various types of peanut, and the regeneration capacity of Araehis hypogaea var. fastigiata was the lowest.
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