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Interpreting gene co-regulation and metabolic profiling in the unique starch metabolism of Arabidopsis dpe2/phs1 mutant reveals insights into control mechanism of the starch granule number

ABSTRACT: An Arabidopsis double mutant lacking both the cytosolic Disproportionating enzyme 2 (DPE2) and the plastidial Glucan phosphorylase (PHS1) revealed a unique starch metabolism. Dpe2/phs1 was reported to have a dwarf growth phenotype, an uneven starch distribution in the rosettes, and a strongly reduced starch granule number per chloroplast when grown under diurnal rhythm. Here we analyzed dpe2/phs1 in more detail and found that it showed three distinct growth periods. In young plants the starch granule number was similar to Col-0, then the starch granule number decreased massively down to one or no granule per chloroplast followed by an increase of the granule number. Thus, in dpe2/phs1 the control over the starch granule number is impaired and it is not defective in starch granule initiation. The data also show that the granule number is not fixed and is regulated over the entire plant growth. Further, also the chloroplasts revealed alterations during these three periods with a partially strong aberrant morphology in the middle phase. Interestingly, the unique metabolism perpetuated if starch degradation is further impaired by additionally lack of Isoamylase 3 and Starch excess 4. Transcriptomic studies and metabolic profiling of dpe2/phs1 revealed a gene co-regulation of most starch metabolism related genes and a clear metabolic separation. Further most senescence-induced genes were found to up-regulated more than 2-fold in the starch-less mature leave. Thus, dpe2/phs1 is a unique source to understand especially the starch granule number regulation in detail. We performed gene expression profiling analysis using data obtained from RNA-seq of 3 stages from both Col-0 and dpe2/phs1.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE201804 | GEO | 2022/05/23

REPOSITORIES: GEO

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Project description:Cotton (Gossypium hirsutum L) is an important crop world wide that provides fiber for the textile industry. Cotton is a perennial plant that stores starch in stems and roots to provide carbohydrates for growth in subsequent seasons. These reserves are not available to produce seed and fiber when cotton is usually grown as an annual crop. Analysis of developing cotton plants indicated that starch levels peaked about the time of first anthesis then began to decline. An earlier peak of levels of starch was occasionally observed and in some greenhouse-grown samples starch increased 2 week after first bloom. Microarray analyses compared gene expression in tissues containing low levels of starch with tissues rapidly accumulating starch. Statistical analysis of differentially expressed genes indicated increased expression among genes associated with carbohydrate metabolism, transcription activity and the proteasome. Genes associated with starch synthesis, starch degradation, sucrose metabolism, hexose metabolism, raffinose synthesis and trehalose synthesis increased in expression in starch accumulating tissues. The anticipated changes in these sugars were largely confirmed by measuring soluble sugars in relevant tissues. We propose that altering expressions of genes and pathways identified in this work could be used to more efficiently mobilize stored carbohydrate to fiber production. Keywords: starch accumulating, stem, root Genes expression was compared between cotton stems that were low in starch and accumulating starch. Gene expression was also compared between cotton roots that were low in starch and accumulating starch. A total of three microarrays were used. One dye swap was used. Material from the field were harvested 2 weeks apart. Greenhouse grown material were planted at two week intervals and harvested at the same time. NOTE that the channel representing the low starch material only gave about half of the total signal than the high starch samples. QPCR of 9 genes confirmed differential expression of 8 of them. QPCR also confirmed similar expression of two genes not predicted to be differentially expressed by the microarray analysis. Therefore no correction was made for the apparent difference in the hybridization of high and low starch samples.

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Interpreting gene co-regulation and metabolic profiling in the unique starch metabolism of Arabidopsis dpe2/phs1 mutant reveals insights into control mechanism of the starch granule number

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