| 紫花苜蓿高频体胚发生及萌发成苗技术与转基因应用 |
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| 引用本文： | 温之雨,董福双,张华宁,杨瑞娟,张艳敏.紫花苜蓿高频体胚发生及萌发成苗技术与转基因应用[J].西北农业学报,2017,26(6):882-889. |
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| 作者姓名： | 温之雨 董福双 张华宁 杨瑞娟 张艳敏 |
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| 作者单位： | （1.河北省农林科学院 遗传生理研究所，河北省植物转基因中心，石家庄 050051；2.河北师范大学 生命科学学院，石家庄 050024） |
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| 基金项目： | 河北省自然科学基金（C2013301033）。 |
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| 摘 要： | 以紫花苜蓿的子叶或叶片为外植体,建立高频体胚发生及成苗技术体系,为苜蓿耐逆性状的遗传改良提供技术支撑。在愈伤诱导、体胚诱导、体胚成熟与萌发等几个关键环节,分析比较培养基组分对培养效果的影响。建立在SHDK培养基上诱导愈伤、MB(-/+)+ABA 0.4mg/L+PEG-6000 50g/L+蔗糖50g/L培养基上诱导体胚、Bio2Y培养基上促体胚发育成熟、1/2 MS(或SH)培养基上体胚萌发成苗的高频再生技术体系。最适条件下,每克胚性愈伤组织可产生77.9个正常萌发成苗的健康体胚。再生植株在生长箱内经过2周20℃、16h/d的光照培养后移栽温室,成活率达90%以上。利用该离体再生技术成功地将凝集素基因PPA导入苜蓿中,获得抗蚜转基因苜蓿。
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| 关 键 词： | 紫花苜蓿 体胚发生 离体再生 转基因 |
High Frequency of Embryogenesis and Germination to Seedling in Alfalfa and Its Application in Transgenic Research |
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| WEN Zhiyu,DONG Fushuang,ZHANG Huaning,YANG Ruijuan and ZHANG Yanmin.High Frequency of Embryogenesis and Germination to Seedling in Alfalfa and Its Application in Transgenic Research[J].Acta Agriculturae Boreali-occidentalis Sinica,2017,26(6):882-889. |
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| Authors: | WEN Zhiyu DONG Fushuang ZHANG Huaning YANG Ruijuan and ZHANG Yanmin |
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| Abstract: | The objective of this study was to establish the technique for high efficient embryogenesis and seedling development using cotyledons or leaves from aseptic seedlings as explants,which will provide a technical support for genetic improvement of tolerance to abiotic stresses in alfalfa(Medicago sativa).The effects of different medium ingredients on callus induction,embryogenetic induction,maturation,and germination were compared.The technique for high-frequency of embryogenesis included induction of callus on SHDK medium,induction of embyros on MB(-/+) + 0.4 mg/L ABA+50 g/L PEG-6000+50 g/L sucrose,growth of embryos on Bio2Y medium,and development of seedling on 1/2 MS(or SH) medium.Under the optimal conditions,about 77.9 healthy embryos that formed seedlings were produced per gram of callus.The seedlings regenerated were grown in a growth chamber at 20 °C with a photoperiod of 16 h light and 8 h dark for two weeks prior to transplanting to a greenhouse,and the survival rate was over 90%.This technique was successfully used in the transformation of PPA gene into alfalfa and the transgenic alfalfa with resistance to aphid was obtained. |
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| Keywords: | Medicago sativa L Somatic embryogenesis In vitro regeneration Transgene |
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