Microalgae are of substantial biotechnological interest due their polyphyletic nature which grants them access to a wide array of high-value metabolites. The inherent genetic diversity of microalgae combined with their metabolic plasticity when grown using different trophic and illumination strategies necessitate the establishment of a reference knowledge base. In the present study we present a detailed characterisation of the combined effects of wavelength selection and trophic strategy on the growth kinetics and gene expression profile of the model microalgae Chlamydomonas reinhardtii grown under moderate to high light intensity (400 μmol_ph m⁻²·s⁻¹). The aim is twofold: (a) to establish a list of reliable housekeeping genes valid for quantitative comparisons across several combinations of different wavelengths and trophic strategies and (b) to enable the investigation of the response of central carbon metabolic pathways under these process conditions. White, blue and red light emitting diodes (LEDs) were used to grow pH controlled photo- and mixo-trophic cultures over a period of 136 h in batch mode. A panel of 10 candidate genes, along with biomass growth rate and pigment content were dynamically monitored across all conditions. Statistical analysis identified genes (acetyl-CoA carboxylase subunit α and photosystem I reaction centre subunit II) with less variability observed in their expression levels across the entirety of conditions evaluated compared to housekeeping genes often referred to in literature (receptor of activated protein kinase C and ribosomal protein (large subunit) 19). Further analysis of gene expression profiles revealed substantial differences in response to changes in both wavelength selection (upregulation of ribulose bisphosphate carboxylase small subunit under red phototrophic growth) and trophic strategy (upregulation of malate synthase from phototrophic to mixotrophic conditions). The systematic approach used to establish reliable reference genes presented herein enables robust comparisons of cellular responses across different conditions to better understand algal metabolism and improve process performance.

微藻具有广泛的生物多样性，因此具有广泛的高价值代谢产物，因此具有重要的生物技术意义。当使用不同的营养和光照策略进行生长时，微藻固有的遗传多样性及其代谢可塑性需要建立参考知识库。在本研究中，我们提出的波长选择和营养策略对模型微藻的生长动力学和基因表达谱的组合效应的详细表征莱茵衣藻在中度生长到高光强度（400微摩尔_的pH 米^-2 ·秒^{- 1个}）。目的是双重的：（a）建立可靠的管家基因列表，可有效地对不同波长和营养策略的几种组合进行定量比较；以及（b）能够研究这些过程条件下中央碳代谢途径的响应。白色，蓝色和红色发光二极管（LED）用于在136小时内以分批模式生长受pH控制的光养和混养营养培养物。在所有条件下均动态监控一组10个候选基因，以及生物量生长速率和色素含量。统计分析确定的基因（乙酰辅酶A羧化酶亚基α和光系统I反应中心亚基II）与文献中经常提及的管家基因（活化蛋白激酶C和C受体）相比，在整个评估条件下其表达水平差异较小。核糖体蛋白（大亚基）19）。基因表达谱的进一步分析显示，在波长选择（红色光养生长下核糖二磷酸羧化酶小亚基的上调）和营养策略（苹果酸合酶从光养到营养养分的上调）变化方面存在显着差异。