Carbon allocation in aspen trees

Trees allocate assimilated carbon between growth and storage. In this PhD thesis, Iinvestigated the regulation of carbon allocation during tree growth both attranscriptional as well as whole-tree level, and with a focus on wood formation.I performed a large-scale DNA affinity purification sequencing (DAP-seq)screen on transcription factor proteins that regulate gene expression in developingwood of aspen (Populus tremula). Together with bioinformaticians, I identified bothnovel and previously reported interactions. The results were integrated into apublicly available database, providing a novel resource for wood biology. We alsopresent a practical guide for the analysis of DAP-seq data to facilitate similar studies.Next, I investigated carbon partitioning between growth and storage in aspen,focusing on the role of starch as the major storage compound. We report that aspengrowth is not limited by starch reserves and suggest a passive starch storagemechanism where sink tissues are the growth-limiting factor.In a study on Arabidopsis (Arabidopsis thaliana), I address the debate on whethersucrose synthase (SUS) enzymes are required in the biosynthesis of cellulose, themost abundant component of wood. As mutants lacking all SUS isoforms grewnormally and their cellulose content was comparable to that of wild-type, I concludethat SUS activity is not required for cellulose biosynthesis in Arabidopsis.Taken together, the results of this PhD study fill key knowledge gaps in the fieldand provide new starting points for future research projects on carbon allocation intrees

Aspen growth is not limited by starch reserves

All photosynthetic organisms balance CO2 assimilation with growth and carbon storage. Stored carbon is used for growth at night and when demand exceeds assimilation. Gaining a mechanistic understanding of carbon partitioning between storage and growth in trees is important for biological studies and for estimating the potential of terrestrial photosynthesis to sequester anthropogenic CO2 emissions.(1,2) Starch represents the main carbon storage in plants.(3,4) To examine the carbon storage mechanism and role of starch during tree growth, we generated and characterized low-starch hybrid aspen (Populus tremula x tremuloides) trees using CRISPR-Cas9-mediated gene editing of two PHOSPHOGLUCOMUTASE (PGM) genes coding for plastidial PGM isoforms essential for starch biosynthesis. We demonstrate that starch deficiency does not reduce tree growth even in short days, showing that starch is not a critical carbon reserve during diel growth of aspen. The low-starch trees assimilated up to similar to 30% less CO2 compared to the wild type under a range of irradiance levels, but this did not reduce growth or wood density. This implies that aspen growth is not limited by carbon assimilation under benign growth conditions. Moreover, the timing of bud set and bud flush in the low-starch trees was not altered, implying that starch reserves are not critical for the seasonal growth-dormancy cycle. The findings are consistent with a passive starch storage mechanism that contrasts with the annual Arabidopsis and indicate that the capacity of the aspen to absorb CO2 is limited by the rate of sink tissue growth

Sucrose synthase activity is not required for cellulose biosynthesis in Arabidopsis

Biosynthesis of plant cell walls requires UDP‐glucose as the substrate for cellulose biosynthesis, and as an intermediate for the synthesis of other matrix polysaccharides. The sucrose cleaving enzyme sucrose synthase (SUS) is thought to have a central role in UDP‐glucose biosynthesis, and a long‐held and much debated hypothesis postulates that SUS is required to supply UDP‐glucose to cellulose biosynthesis. To investigate the role of SUS in cellulose biosynthesis of Arabidopsis thaliana we characterized mutants in which four or all six Arabidopsis SUS genes were disrupted. These sus mutants showed no growth phenotypes, vascular tissue cell wall defects, or changes in cellulose content. Moreover, the UDP‐glucose content of rosette leaves of the sextuple sus mutants was increased by approximately 20% compared with wild type. It can thus be concluded that cellulose biosynthesis is able to employ alternative UDP‐glucose biosynthesis pathway(s), and thereby the model of SUS requirements for cellulose biosynthesis in Arabidopsis can be refuted

Cryopreservation of the Norway spruce tissue culture line able to produce extracellular lignin

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