文献类型：期刊文献

英文题名：Isolation of leaf mesophyll protoplasts optimized by orthogonal design for transient gene expression in Catalpa bungei

作者：Ma, Wenjun[1] Yi, Fei[1,2] Xiao, Yao[1] Yang, Guijuan[1] Chen, Faju[2] Wang, Junhui[1]

第一作者：麻文俊

通信作者：Yi, F[1];Wang, JH[1];Yi, F[2]

机构：[1]Chinese Acad Forestry, Res Inst Forestry, Natl Innovat Alliance Catalpa Bungei,State Key La, Natl Forestry & Grassland Adm,Key Lab Tree Breedi, Beijing 100091, Peoples R China;[2]Three Gorges Univ, Coll Biol & Pharmaceut Sci, Key Lab Gorges Reg Plant Genet & Germplasm Enhanc, Yichang 443002, Peoples R China

年份：2020

卷号：274

外文期刊名：SCIENTIA HORTICULTURAE

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000572433900002)】；

基金：This work was supported by the Open Foundation of Key Laboratory of Three Gorges Regional Plant Genetics and Germplasm Enhancement (China Three Gorges University) (2016KBC11).

语种：英文

外文关键词：Catalpa bungei; Protoplast; Transient gene expression; PEG-mediated transformation

摘要：Catalpa bungei is an important ornamental plant and timber species that originated in China. With the forth-coming C. bungei genome sequence, an efficient and convenient transformation system should be developed. The transient gene expression system using protoplasts is fast and efficient and can be applied to study the functions of genes in C. bungei. In this study, the main factors influencing the isolation of protoplasts from C. bungei were optimized by orthogonal design. The results showed that when the enzymolysis solution contained 3% Cellulase RS (w/v), 2% Macemzyme R-10 (w/v), and 0.4 M mannitol and the enzymolysis time was 8 h, the protoplast yield was 1 x 10(6) protoplasts/g fresh weight [FM]. The viability of protoplasts was approximately 90 % under optimal enzymolysis conditions. Furthermore, using GFP as the reporter gene, we verified the feasibility of PEG-mediated protoplast transformation of C. bungei. We established an efficient protoplast isolation procedure and feasible transient gene expression system. This methodology, when combined with genetics and omics techniques, will allow further study of the functions of the genes in C. bungei.