Establishment of a chloroplast transformation system in Tisochrysis lutea
- PDF / 509,117 Bytes
- 7 Pages / 595.276 x 790.866 pts Page_size
- 63 Downloads / 178 Views
Establishment of a chloroplast transformation system in Tisochrysis lutea Yuntai Bo 1 & Kang Wang 2 & Yuanyuan Wu 1 & Haiyang Cao 1 & Yulin Cui 3 & Lan Wang 4,5 Received: 2 January 2020 / Revised and accepted: 18 May 2020 # Springer Nature B.V. 2020
Abstract Tisochrysis lutea is a haptophyte microalga commonly used as a commercial feed for juvenile fish and shellfish larvae. Genetic modification is of considerable importance for developing the potential economic value of T. lutea. However, the genetic transformation system of T. lutea has not yet been established, which limits functional genomic studies and strain improvement of this important microalgal species. In the current study, a chloroplast transformation vector harboring the phosphinothricin resistance gene (bar) as a selectable marker was established, and two short peptide-encoding genes (ant1 and ant2) driven by the endogenous psbA and rbcL promoters were cloned in this vector. The recombinant plasmid was transformed using a biolistic method into the trnI/trnA locus of the chloroplast genome via homologous recombination. After continuous selection on phosphinothricin, the integration of foreign genes and the expression of specific products in the transformants were detected using polymerase chain reaction (PCR), Southern blotting, and western blot analysis. This is the first report of establishment of a stable transformation system in T. lutea, which is a prerequisite for functional genomics and applied research on this species. Keywords Tisochrysis lutea . Genetic transformation . Chloroplast . Biolistic method
Introduction Microalgae are unicellular photoautotrophic microorganisms which are widely used for the production of valuable compounds, especially in the biotechnology industry. Microalgal cultivation plays a crucial role in addressing some of the
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10811-020-02159-4) contains supplementary material, which is available to authorized users. * Lan Wang [email protected] 1
Yantai Marine Environmental Monitoring Center Station of SOA, Yantai 264006, China
2
School of Life Sciences, Shandong University of Technology, Zibo 255049, Shandong, China
3
Key Laboratory of Coastal Biology and Biological Resource Utilization, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, Shandong, China
4
Shandong Provincial Key Laboratory of Marine Ecology and Environment & Disaster Prevention and Mitigation, Qingdao 266000, China
5
North China Sea Marine Forecasting Center of SOA, No. 27, Yunling road, Laoshan District, Qingdao 266000, Shandong, China
limitations of traditional biomass production (Zhan et al. 2017). The polysaccharides, proteins, pigments, and lipids present in microalgal cells can be prospectively utilized in the fields of food, health care products, medicine, and biofuels. Tisochrysis lutea is a marine microalga, previously named Isochrysis aff. galbana (Shi et al. 2015; Méndez-Leyva et al. 2019); however, recently
Data Loading...
