文献类型：期刊文献

英文题名：Agrobacterium-Mediated Genetic Transformation of Larix kaempferi (Lamb.) Carr. Embryogenic Cell Suspension Cultures and Expression Analysis of Exogenous Genes

作者：Dang, Shaofei[1,2] Zhang, Lifeng[1,2] Han, Suying[3] Qi, Liwang[1,2]

第一作者：Dang, Shaofei

通信作者：Zhang, LF[1];Zhang, LF[2]

机构：[1]Chinese Acad Forestry, Res Inst Forestry, State Key Lab Tree Genet & Breeding, Beijing 100091, Peoples R China;[2]Chinese Acad Forestry, Res Inst Forestry, Key Lab Tree Breeding & Cultivat, Natl Forestry & Grassland Adm, Beijing 100091, Peoples R China;[3]Chinese Acad Forestry, Ecol & Nat Conservat Inst, Beijing 100091, Peoples R China

年份：2022

卷号：13

期号：9

外文期刊名：FORESTS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000859612600001)】；

基金：This research was funded by the Fundamental Research Funds for the Central Non-profit Research Institution of CAF (CAFYBB2019SY011), the National Natural Science Foundation of China (32171811 and 31600544), and the National Transgenic Major Program of China (2018ZX08020-003).

语种：英文

外文关键词：larch; genetic transformation; somatic embryogenesis; Agrobacterium tumefaciens

摘要：A simple and efficient protocol for the genetic transformation of Japanese larch (Larix kaempferi (Lamb.) Carr.) was developed by altering the infection method and duration and the bacterial removal process. More than 600 hygromycin-resistant embryonal masses with the vector pCAMBIA1301 were obtained, with an average of 20 transgenic lines per gram of fresh weight. Nine hygromycin-resistant transformation events (designated P1-P9) were analyzed using PCR, quantitative real-time PCR, and histochemical beta-glucuronidase (GUS) assays. The GUS transcript abundance in each transformed cell line ranged from 10(1) to 10(3) magnitudes, with a maximum abundance of 2.89 x 10(3). In addition, the pLaTCTP::GUS vector, which contains GUS under the control of the L. kaempferi LaTCTP promoter, led to constitutive expression of GUS in embryonal-suspensor mass and somatic embryos. The transcript abundance of the exogenous genes HPT and GUS, driven by the CaMV 35S or LaTCTP promoter, ranged from 10(1) to 10(4), which was equivalent to genes with moderate and low abundances in Japanese larch. The relatively low expression levels of exogenous genes in transformants might reflect the large genome of Japanese larch. Additional transgenic cell lines need to be screened to obtain transformants with higher expression levels of foreign genes for further functional research in Japanese larch.