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.

黄褐线虫（Tisochrysis lutea）是一种触藻微藻，通常用作幼鱼和贝类幼虫的商业饲料。基因改造对于开发黄茶的潜在经济价值具有重要意义。但是，尚未建立T. lutea的遗传转化系统，这限制了功能基因组研究和该重要微藻物种的菌株改良。在当前的研究中，建立了一个以膦丝菌素抗性基因（bar）为选择标记的叶绿体转化载体，并由内源性psbA和rbcL驱动的两个短肽编码基因（ant 1和ant 2）启动子被克隆到该载体中。使用同源枪法，通过同源重组将重组质粒转化到叶绿体基因组的trnI / trnA基因座中。在膦丝菌素上连续选择后，使用聚合酶链反应（PCR），Southern印迹和Western印迹分析检测外源基因的整合和特定产物在转化体中的表达。这是在黄茶中建立稳定转化系统的第一份报告，这是功能基因组学和对该物种进行应用研究的前提。