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表达bar基因的抗除草剂转基因甘薯的获得 被引量:13
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作者 臧宁 翟红 +3 位作者 王玉萍 于波 何绍贞 刘庆昌 《分子植物育种》 CAS CSCD 2007年第4期475-479,共5页
用农杆菌介导法将bar基因导入甘薯主栽品种徐薯18的胚性悬浮细胞中,获得了抗除草剂的转基因植株。农杆菌菌株EHA105携带的双元载体pCAMBIA3300上含有bar基因。来自于徐薯18胚性悬浮细胞的直径为0.7~1.3mm的280个胚性细胞团用于遗传转... 用农杆菌介导法将bar基因导入甘薯主栽品种徐薯18的胚性悬浮细胞中,获得了抗除草剂的转基因植株。农杆菌菌株EHA105携带的双元载体pCAMBIA3300上含有bar基因。来自于徐薯18胚性悬浮细胞的直径为0.7~1.3mm的280个胚性细胞团用于遗传转化。共培养3d后,首先在含有2mg/L2,4-D、100mg/LCarb的液体MS培养基中培养1周,然后将胚性细胞团转移到添加2mg/L2,4-D、100mg/LCarb和0.3mg/LPPT的固体MS培养基上进行选择培养。选择8周后,将获得的37个PPT抗性愈伤组织转移到添加1mg/LABA、100mg/LCarb和0.3mg/LPPT的固体MS培养基上,其中的34个愈伤组织诱导得到体细胞胚并发芽形成小植株,共获得了164株拟转基因植株。PCR分析表明,其中的123株为转基因植株。Southernblot和Northernblot分析表明,bar基因稳定整合到转基因植株的基因组中并正确表达。除草剂喷洒试验结果表明,转基因植株具有高度除草剂抗性。 展开更多
关键词 甘薯(ipomoea batatas(l.)lam.) 农杆菌介导转化 胚性悬浮细胞 除草剂 转基因植株
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Analysis of drought tolerance and genetic and epigenetic variations in a somatic hybrid between Ipomoea batatas (L.) Lam. and I. triloba L. 被引量:3
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作者 JIA Li-cong ZHAI Hong +2 位作者 HE Shao-zhen YANG Yu-feng LIU Qing-chang 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2017年第1期36-46,共11页
The somatic hybrid KT1 was previously obtained from protoplast fusion between sweetpotato (Ipomoea batatas (L.) Lam.) cv. Kokei No. 14 and its wild relative I. triloba L. However, its genetic and epigenetic variat... The somatic hybrid KT1 was previously obtained from protoplast fusion between sweetpotato (Ipomoea batatas (L.) Lam.) cv. Kokei No. 14 and its wild relative I. triloba L. However, its genetic and epigenetic variations have not been investigated. This study showed that KT1 exhibited significantly higher drought tolerance compared to the cultivated parent Kokei No. 14. The content of proline and activities of superoxide dismutase (SOD) and photosynthesis were significantly increased, while malonaldehyde (MDA) content was significantly decreased compared to Kokei No. 14 under drought stress. KT1 also showed higher expression level of well-known drought stress-responsive genes compared to Kokei No. 14 under drought stress. Amplified fragment length polymorphism (AFLP) and methylation-sensitive amplified polymorphism (MSAP) analyses indicated that KT1 had AFLP and MSAP band patterns consisting of both parent specific bands and changed bands. Fur- ther analysis demonstrated that in KT1. the proportions of Kokei No. 14 specific genome components and methylation sites were much greater than those of I. triloba. KT1 had the same chloroplast and mitochondrial genomes as Kokei No. 14. These results will aid in developing the useful genes ofI. triloba and understanding the evolution and phylogeny of the cultivated sweetpotato. 展开更多
关键词 drought tolerance epigenetic variation genetic variation ipomoea batatas l.) lain. I. triloba l. somatic hybrid
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甘薯高类胡萝卜素突变体农大辐14块根cDNA文库的构建及鉴定 被引量:3
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作者 杨元军 王玉萍 +3 位作者 翟红 陈伟 何绍贞 刘庆昌 《分子植物育种》 CAS CSCD 2007年第6期879-882,共4页
为分析和克隆甘薯块根中类胡萝卜素生物合成的相关基因,以甘薯品种高系14号经^(60)Coγ射线慢照射结合茎尖培养获得的高类胡萝卜素含量突变体农大辐14的块根mRNA为材料,利用Clontech SMART cD- NA Library Construction Kit构建了其cDN... 为分析和克隆甘薯块根中类胡萝卜素生物合成的相关基因,以甘薯品种高系14号经^(60)Coγ射线慢照射结合茎尖培养获得的高类胡萝卜素含量突变体农大辐14的块根mRNA为材料,利用Clontech SMART cD- NA Library Construction Kit构建了其cDNA文库。初始文库的独立克隆为0.57×10~6pfu,滴度为4.9×10~6pfu/mL,重组率为88%,插入片段大小为0.3~2.5kb,大于0.4kb的克隆比例为70%,独立克隆数为理论值的3倍。扩增文库的滴度为5×10~9 pfu/mL。鉴定结果表明该文库可用于基因克隆与分析。 展开更多
关键词 甘薯(tpomoea batatas(l.)lam.) 类胡萝卜素 突变体 块根 cDNA文库
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