2014, Gruber, Ansgar, Kroth, Peter G.

In the recent years, a large number of genomes from a variety of different organisms have been sequenced. Most of the sequence data has been publicly released and can be assessed by interested users. However, this wealth of information is currently underexploited by scientists not directly involved in genome annotation. This is partially because sequencing, assembly, and automated annotation can be done much faster than the identification, classification, and prediction of the intracellular localization of the gene products. This part of the annotation process still largely relies on manual curation and addition of contextual information. Users of genome databases who are unfamiliar with the types of data available from (whole) genomes might therefore find themselves either overwhelmed by the vast amount and multiple layers of data or dissatisfied with less-than-meaningful analyses of the data.
In this chapter we present procedures and approaches to identify and characterize gene models of enzymes involved in metabolic pathways based on their similarity to known sequences. Furthermore we describe how to predict the subcellular location of the proteins using publicly available prediction servers and how to interpret the obtained results. The strategies we describe are generally applicable to organisms with primary plastids such as land plants or green algae. Additionally, we describe strategies suitable for those groups of algae with secondary plastids (for instance diatoms), which are characterized by a different cellular topology and a larger number of intracellular compartments compared to plants.

2014, Matsuda, Yusuke, Kroth, Peter G.

Diatoms are unicellular photoautotrophic algae and very successful primary producers in the oceans. Their high primary productivity is probably sustained by their high adaptability and a uniquely arranged metabolism. Diatom belongs to the Chromista, a large eukaryotic group, which has evolved by multiple endosymbiotic steps. As a result, diatoms possess a plastids with four membranes together with complicated translocation systems to transport proteins and metabolites including inorganic substances into and out of the plastids. In addition to the occurrence of potential plasma-membrane transporters, there are numerous carbonic anhydrases (CAs) within the matrix of the layered plastidic membranes, strongly suggesting large interconversion activity between CO₂ and HCO₃ − within the chloroplast envelope as a part of a CO2-concentrating mechanism (CCM). In diatoms also the Calvin cycle and its adjacent metabolism reveal unique characteristics as, for instance, ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) activase, the plastidic sedoheptulose-1,7-bisphosphatase (SBPase), and the plastidic oxidative pentose phosphate pathway (OPP) are absent. Furthermore, the Calvin cycle metabolism in diatoms is not under the strict redox control by the thioredoxin (Trx) system. Instead, a CO₂-supplying system in the pyrenoid shows CA activities which are probably regulated by chloroplastic Trxs. Pyrenoidal CAs are also regulated on the transcriptional level by CO₂ concentrations via cAMP as a second messenger, suggesting an intense control system of CO₂ acquisition in response to CO₂ availability. The photorespiratory carbon oxidation cycle (PCOC) is the major pathway to recycle phosphoglycolate in diatoms although this process might not be involved in recycling of 3-phosphoglycerate but instead produces glycine and serine. In this review we focus on recent experimental data together with supportive genome information of CO₂ acquisition and fixation systems primarily in two marine diatoms, Phaeodactylum tricornutum and Thalassiosira pseudonana.