| 发根农杆菌介导的菠菜毛状根遗传转化体系的建立 |
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| 引用本文： | 徐悦,曹英萍,王玉,付春祥,戴绍军. 发根农杆菌介导的菠菜毛状根遗传转化体系的建立[J]. 植物学报, 2019, 54(4): 515-521. DOI: 10.11983/CBB18257 |
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| 作者姓名： | 徐悦 曹英萍 王玉 付春祥 戴绍军 |
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| 作者单位： | 1 东北林业大学盐碱地生物资源环境研究中心, 东北盐碱植被恢复与重建教育部重点实验室, 哈尔滨 1500402 中国科学院青岛生物能源与过程研究所, 中国科学院生物燃料重点实验室, 山东省能源生物遗传资源重点实验室, 山东省合成生物技术创新中心(筹), 青岛 2660003 上海师范大学生命科学学院, 植物种质资源开发协同创新中心, 上海 200234; |
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| 基金项目： | 青岛市创业创新领军人才(16-8-3-15-zhc);中央高校基本科研业务费专项资金(2572017ET01);上海市科学技术委员会科研计划(17391900600) |
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| 摘 要： | 发根农杆菌(Agrobacterium rhizogenes)侵染植物后可诱导植物产生毛状根。菠菜(Spinacia oleracea)是常见的食用蔬菜, 目前尚未见菠菜毛状根的研究报道。经筛选得到适合诱导菠菜毛状根的发根农杆菌菌株LBA9402, LBA9402侵染菠菜外植体茎后, 毛状根的诱导率最高可达16%。菠菜毛状根呈白色, 具有丰富的根毛, 能在无外源激素的固体培养基上快速增殖生长。通过诱导菠菜毛状根产生愈伤组织并进行分化, 获得了菠菜毛状根的再生植株, 再生率为8%。此外, LBA9402可将含有Ri质粒的T-DNA和携带外源GFP基因的Ti质粒T-DNA共同导入外植体中。PCR检测和荧光显微观察结果显示, rolB及GFP基因在菠菜毛状根基因组中稳定表达, 共转化频率为50%。
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| 关 键 词： | 共转化 毛状根 植物再生体系 菠菜 |
| 收稿时间： | 2018-11-30 |
Agrobacterium rhizogenes-mediated Transformation System of Spinacia oleracea |
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| Yue Xu,Yingping Cao,Yu Wang,Chunxiang Fu,Shaojun Dai. Agrobacterium rhizogenes-mediated Transformation System of Spinacia oleracea[J]. Bulletin of Botany, 2019, 54(4): 515-521. DOI: 10.11983/CBB18257 |
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| Authors: | Yue Xu Yingping Cao Yu Wang Chunxiang Fu Shaojun Dai |
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| Abstract: | The plant can produce hairy roots after infection by Agrobacterium rhizogenes harboring Ri plasmids. Spinach (Spinacia oleracea) is a common edible vegetable, and its hairy root system has yet to be reported. In this study, A. rhizogenes LBA9402 was screened to be suitable for hairy root induction of spinach. The highest induction efficiency, 16%, was reached when the spinach stem was infected by LBA9402. Hairy roots grew vigorously on SH solid medium without exogenous hormones. The hairy roots of spinach were white and had enormous root hairs. The regeneration plant obtained from the callus redifferentiated from spinach hair roots; the rate of regeneration was 8%. In addition, Ri T-DNA and Ti T-DNA carrying rolB genes and GFP gene, respectively, were co-transferred into cells of spinach and produced transgenic hairy roots. PCR analysis and fluorescence microscopy assays showed rolB gene and GFP gene in the hairy roots of spinach and expressed stably; the rate of co-transformation was 50%. |
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| Keywords: | co-transformation hairy root plant regeneration system Spinacia oleracea |
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