| 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 |
| DOI:10.7606/j.issn.1004-1389.2017.06.011 |
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| 基金项目:河北省自然科学基金（C2013301033）。 |
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| 中文摘要: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 0.4 mg/L+PEG-6000 50 g/L+蔗糖50 g/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 ℃、16 h/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。 |
| 中文关键词:紫花苜蓿 体胚发生 离体再生 转基因 |
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| High Frequency of Embryogenesis and Germination to Seedling in Alfalfa and Its Application in Transgenic Research |
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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. |
| keywords:Medicago sativa L Somatic embryogenesis In vitro regeneration Transgene |
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