Lie Alle A Y, Liu Zhenfeng, Terrado Ramon, Tatters Avery O, Heidelberg Karla B, Caron David A
Department of Biological Sciences, University of Southern California, 3616 Trousdale Parkway, Los Angeles, CA, 90089-0371, USA.
BMC Genomics. 2017 Feb 14;18(1):163. doi: 10.1186/s12864-017-3549-1.
Ochromonas is a genus of mixotrophic chrysophytes that is found ubiquitously in many aquatic environments. Species in this genus can be important consumers of bacteria but vary in their ability to perform photosynthesis. We studied the effect of light and bacteria on growth and gene expression of a predominantly phagotrophic Ochromonas species. Axenic cultures of Ochromonas sp. were fed with heat-killed bacteria (HKB) and grown in constant light or darkness. RNA was extracted from cultures in the light or in the dark with HKB present (Light + HKB; Dark + HKB), and in the light after HKB were depleted (Light + depleted HKB).
There were no significant differences in the growth or bacterial ingestion rates between algae grown in light or dark conditions. The availability of light led to a differential expression of only 8% of genes in the transcriptome. A number of genes associated with photosynthesis, phagotrophy, and tetrapyrrole synthesis was upregulated in the Light + HKB treatment compared to Dark + HKB. Conversely, the comparison between the Light + HKB and Light + depleted HKB treatments revealed that the presence of HKB led to differential expression of 59% of genes, including the majority of genes involved in major carbon and nitrogen metabolic pathways. Genes coding for unidirectional enzymes for the utilization of glucose were upregulated in the presence of HKB, implying increased glycolytic activities during phagotrophy. Algae without HKB upregulated their expression of genes coding for ammonium transporters, implying uptake of inorganic nitrogen from the culture medium when prey were unavailable.
Transcriptomic results agreed with previous observations that light had minimal effect on the population growth of Ochromonas sp. However, light led to the upregulation of a number of phototrophy- and phagotrophy-related genes, while the availability of bacterial prey led to prominent changes in major carbon and nitrogen metabolic pathways. Our study demonstrated the potential of transcriptomic approaches to improve our understanding of the trophic physiologies of complex mixotrophs, and revealed responses in Ochromonas sp. not apparent from traditional culture studies.
赭球藻属是一类混合营养型金藻,广泛存在于许多水生环境中。该属物种可能是细菌的重要消费者,但其光合作用能力存在差异。我们研究了光照和细菌对一种主要营吞噬营养的赭球藻属物种生长和基因表达的影响。将无菌培养的赭球藻用热灭活细菌(HKB)投喂,并在持续光照或黑暗条件下培养。从光照或黑暗条件下且存在HKB(光照+HKB;黑暗+HKB)以及HKB耗尽后的光照条件下(光照+耗尽的HKB)的培养物中提取RNA。
在光照或黑暗条件下生长的藻类,其生长速率或细菌摄取率没有显著差异。光照的可利用性导致转录组中仅8%的基因差异表达。与黑暗+HKB处理相比,光照+HKB处理中一些与光合作用、吞噬营养和四吡咯合成相关的基因上调。相反,光照+HKB与光照+耗尽的HKB处理之间的比较表明,HKB的存在导致59%的基因差异表达,包括参与主要碳和氮代谢途径的大多数基因。编码利用葡萄糖的单向酶的基因在HKB存在时上调,这意味着吞噬营养过程中糖酵解活性增加。没有HKB的藻类上调了编码铵转运蛋白的基因表达,这意味着在没有猎物时从培养基中摄取无机氮。
转录组学结果与之前的观察结果一致,即光照对赭球藻属种群生长的影响最小。然而,光照导致一些与光营养和吞噬营养相关的基因上调,而细菌猎物的可利用性导致主要碳和氮代谢途径发生显著变化。我们的研究证明了转录组学方法在增进我们对复杂混合营养生物营养生理学理解方面的潜力,并揭示了赭球藻属中传统培养研究未发现的反应。