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
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
基金supported by the China Agriculture Research System(CARS-11,Sweetpotato)the National Natural Science Foundation of China(31461143017)
文摘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.
文摘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。