苋菜原生质体和瞬时转化体系的建立及应用  

Establishment and Application of Protoplast and Transient Transformation System of Edible Amaranth (Amaranthus spp.)

作　　者：仝亚楠 刘雪 刘秀洁 李大勇 张德春 Tong Ya'nan;Liu Xue;Liu Xiujie;Li Dayong;Zhang Dechun(Biotechnology Research Center,China Three Gorges University,Yichang,443002;Vegetable Research Center,Beijing Academy of Agricultural and Forestry Sciences,Beijing,100097;College of Life Sciences,Yantai University,Yantai,264005)

机构地区：[1]三峡大学,生物技术研究中心,宜昌443002 [2]北京市农林科学院蔬菜研究中心,北京100097 [3]烟台大学生命科学学院,烟台264005

出　　处：《分子植物育种》2021年第19期6476-6481,共6页Molecular Plant Breeding

基　　金：北京市农林科学院科技创新能力建设专项(KJCX20200113)资助。

摘　　要：为了建立稳定、高效的苋菜原生质体制备和瞬时转化体系,本研究以栽培苋菜(Amaranthus tricolor)品种‘京苋三号’无菌组培苗叶片为材料,通过反复试验确立了苋菜原生质体分离制备和瞬时转化体系,并利用该体系对一个苋菜甜菜红素合成调控相关的MYB转录因子进行了亚细胞定位分析。结果表明,以1.0%纤维素酶+0.25%离析酶组合,酶解3~4 h,能得到活力较高的叶肉细胞原生质体,可直接用于植物蛋白亚细胞定位等实验;苋菜甜菜红素合成调控相关的MYB转录因子定位在细胞核中。苋菜原生质体制备和瞬时转化体系的建立,为苋菜基因功能研究提供了重要的技术支撑。In order to estab lish a stable and efficient system of protoplast preparation and instantaneous trans formation of amaranth(Amaranthus spp.), in this study, the leaves of sterile tissue culture seedlings of cultivated amaranth cultivar ’Jingxian 3’ were used as materials to establish a system of protoplast separation and preparation and instantaneous transformation of Amaranth through repeated experiments. The subcellular localization of MYB transcription factor related to the synthesis of betacarotene in amaranth was analyzed by using this system. The results showed that a combination of 1.0% cellulase and 0.25% macerozyme for 3 to 4 hours could produce mesophyll cell protoplasts with high activity, which could be directly used for subcellular localization of plant proteins. MYB transcription factors related to the regulation of betacarotene synthesis in Amaranth are located in the nucleus. The preparation of protoplasts and establishment of transient transformation system of Amaranth provide important technical support for the study of gene function of Amaranth.

关 键 词：苋菜(Amaranthus spp.) 原生质体制备 瞬时表达 MYB转录因子 

分 类 号：S636.4[农业科学—蔬菜学]