| 免预培养高效菊苣遗传转化方法 |
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| 引用本文： | 李小冬,蔡璐,李世歌,莫本田,韩永芬,王小利.免预培养高效菊苣遗传转化方法[J].草业学报,2016,25(10):124-131. |
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| 作者姓名： | 李小冬 蔡璐 李世歌 莫本田 韩永芬 王小利 |
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| 作者单位： | 贵州省草业研究所, 贵州 贵阳 550006 |
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| 基金项目： | 贵州省联合基金“利用 AtmiR156培育高产耐刈菊苣种子新材料”(黔科合 J LKN[2013]04)，贵州省农科院博士启动基金“四倍体菊苣种质创新应用研究”(黔农科院人才启动项目[2013]01)，农科院自主创新科研专项“贵州主要优良牧草种质资源发掘与创新利用研究”(黔农科院自主创新科研专项字[2014]010号)资助。 |
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| 摘 要： | 转基因技术对基因功能研究与农作物种质资源创新具有重要作用,在牧草类作物菊苣中相应的研究还处于初级阶段。本研究改进了菊苣组织培养与遗传转化方法,采用暗光培养的菊苣子叶为外植体,经农杆菌侵染后置于光照条件下进行共培养,利用子叶细胞从暗光转移到光照条件下细胞迅速分裂的特点,实现植物遗传转化。与已报道的转化方法(光照条件下的真叶外植体先进行2~3 d预培养,再进行农杆菌转化与共培养)相比,本方法出苗率与已报道方法相当,达90%以上;比不经预培养方法的出苗率要高80%~90%,能极大节省人力物力。同时本研究发现,在愈伤诱导、植物分化以及生根培养中,依次递减细胞分裂素的强度与浓度,能显著降低愈伤和再生苗的玻璃化程度,与一直采用强细胞分裂素相比,改进方法愈伤玻璃化比率下降约9%,再生苗玻璃化比例下降约16%。
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| 关 键 词： | 菊苣 组织培养 转基因 玻璃化 子叶 |
| 收稿时间： | 2016-03-09 |
An effective transformation method mediated by Agrobacterium in chicory (Cichorium intybus) |
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| LI Xiao-Dong,CAI Lu,LI Shi-Ge,MO Ben-Tian,HAN Yong-Fen,WANG Xiao-Li.An effective transformation method mediated by Agrobacterium in chicory (Cichorium intybus)[J].Acta Prataculturae Sinica,2016,25(10):124-131. |
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| Authors: | LI Xiao-Dong CAI Lu LI Shi-Ge MO Ben-Tian HAN Yong-Fen WANG Xiao-Li |
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| Institution: | Guizhou Institute of Prataculturae, Guiyang 550006, China |
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| Abstract: | Transgenic technology plays an important role in gene function analysis and crop germplasm innova-tion.However,transgenic studies are relatively recent in chicory.In this study we sought to improve tissue culture and genetic transformation methodology in chicory.Cotyledons grown in the dark were collected as ex-plants.After transformation mediated by Agrobacteria,these explants were grown under illuminated condi-tions.The cotyledon cells divided rapidly when they were transferred from dark to light.The emergence rate of regenerated seedlings was greater than 90%,similar to that of previous studies however,the germination rate could be improved by 80%-90% if the explants were not pre-cultured.Our results were time,money and la-bor-saving.We also found that during callus induction,plant differentiation and root growth,decreased inten-sity and concentration of cytokinin significantly reduced the vitrification of callus and regenerated plantlets. Compared with continuously applied cytokinin our method decreased vitrification of callus and regenerated pla-ntlets by about 9% and 16% respectively. |
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| Keywords: | chicory tissue culture transgenic vitrification cotyledon |
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