摘要
为明确黑枸杞LrAN2基因在花青素合成代谢中的调控机制,在与枸杞同属的番茄中做了进一步验证。黑枸杞LrAN2基因参与调控黑色果实花青素生物合成代谢,同时诱导烟草产生紫色表型,受限于枸杞转基因组培技术,选择枸杞亲缘物种番茄进行LrAN2转基因功能分析及其调控机制。基于前期分离的LrAN2开放阅读框序列,通过表达载体构建,荧光定量分析,花青素含量的测定以及液相色谱分析方式对转基因番茄进行检测和分析。结果表明,在番茄材料Maker中过量表达LrAN2基因,得到深绿色番茄植株。转基因番茄的各个组织部位均有不同程度的花青素积累,花、叶和根细胞中都存在颜色较深色素沉着,叶片中花青素含量和LrAN2基因的表达量都是最高。LrAN2的过表达上调DFR和F3′5′H的表达水平,同时激活飞燕草色素和矢车菊色素的积累。说明LrAN2基因能够调控花青素的合成。
In order to clarify the regulation mechanism of LrAN2 gene in anthocyanin biosynthesis of Lycium ruthenicum Murr.,the tomato which belongs to the same Solanaceae as Lycium barbarum wasfurtherverifiedin this study.LrAN2 gene was involved in the regulation of anthocyanin biosynthesis and metabolism in Lycium ruthenicum Murr.,at the same time,LrAN2 gene could induce tobacco to produce purple phenotype.Due to limit of the technology in transgenic tissue culture of Lycium barbarum,tomatoas a relative species of Lycium barbarumwas selected for functional analysis and regulation mechanism of LrAN2.Based on the LrAN2 open reading frame sequence isolated in the previous stage,genetically modified tomato was detected and analyzed byexpression vector construction,fluorescence quantitative analysis,determination of anthocyanin content and liquid chromatography.The LrAN2 gene was overexpressed in the tomato material maker to obtain dark green tomato plants.In genetically modified tomato,the anthocyanin content in each tissue part had different degrees of pigmentation,darker pigmentation in flowers,leaves and root cells.The anthocyanin content and LrAN2 gene expression in leaves were the highest.The overexpression of LrAN2 upregulated the expression of DFR and F3′5′H,and activated the accumulation of delphinidin and cyanidin.
作者
罗香怡
李云
曹东
魏乐
宗渊
刘宝龙
LUO Xiangyi;LI Yun;CAO Dong;WEI Le;ZONG Yuan;LIU Baolong(Qinghai Normal University,Xining 810008,China;Key Laboratory of Crop Molecular Breeding of Qinghai Province,Chinese Academy of Sciences,Xining 810001,China)
出处
《西北农业学报》
CAS
CSCD
北大核心
2021年第9期1374-1381,共8页
Acta Agriculturae Boreali-occidentalis Sinica
基金
青海省应用基础研究项目(2020-SF-132)
中科院“西部之光”青年学者项目(2029051)
中科院特别研究助理项目(Y92961)
青海省应用基础研究项目(2018-ZJ-762)。
