Summary: Plants undergo several developmental transitions during their life cycle. One of these, the differentiation of the young embryo from a meristem‐like structure into a highly specialized storage organ, is believed to be controlled by local connections between sugars and hormonal response systems. However, we know little about the regulatory networks underpinning the sugar–hormone interactions in developing seeds. By modulating the trehalose 6‐phosphate (T6P) content in growing embryos of garden pea (Pisum sativum), we investigate here the role of this signaling sugar during the seed‐filling process. Seeds deficient in T6P are compromised in size and starch production, resembling the wrinkled seeds studied by Gregor Mendel. We show also that T6P exerts these effects by stimulating the biosynthesis of the pivotal plant hormone, auxin. We found that T6P promotes the expression of the auxin biosynthesis gene TRYPTOPHAN AMINOTRANSFERASE RELATED2 (TAR2), and the resulting effect on auxin concentrations is required to mediate the T6P‐induced activation of storage processes. Our results suggest that auxin acts downstream of T6P to facilitate seed filling, thereby providing a salient example of how a metabolic signal governs the hormonal control of an integral phase transition in a crop plant.

A previously unrecognized interaction between light and NADPH-dependent thiol-redox systems affects photosynthetic metabolism and growth in response to varying

Heterotrophic plastids of seeds perform many biosynthetic reactions. Understanding their function in crop plants is crucial for seed production. Physiological functions depend on the uptake of precursors by a range of different metabolite translocators. The 2‐oxoglutarate/malate translocator gene (PsOMT), which is highly expressed during pea (Pisum sativum) embryo maturation, has an important role during seed storage. PsOMT functions have been studied by antisense repression in maturing pea embryos, and were found to reduce mRNA levels and transport rates of 2‐oxoglutarate and malate by 50–70%. Combined metabolite and transcript profiling revealed that OMT repression affects the conversion of carbohydrates from sucrose into amino acids and proteins, decreases seed weight and delays maturation. OMT‐repressed pea embryos have increased levels of organic acids, ammonia, and higher ratios of Asn : Asp and Gln : Glu. Decreased levels of most other amino acids indicate the reduced usage of organic acids and ammonia for amino acid biosynthesis in plastids, possibly caused by substrate limitation of the plastidial glutamine synthetase/glutamine–2‐oxoglutarate aminotransferase cycle. Expression of storage proteins is delayed, and mature seeds have reduced protein content. Downregulated gene expression of starch biosynthesis and plastidial glucose‐6‐phosphate transport in asOMT embryos reveals that decreased 2‐oxoglutarate/malate transport capacity affects other pathways of central carbon metabolism. Gene expression analysis related to plastid physiology revealed that OMT repression delays differentiation of storage plastids, thereby maintaining gene expression associated with green chloroplasts. We conclude that OMT is important for protein‐storing crop seeds, and is necessary for amino acid biosynthesis in pea seeds. In addition, carbon supply as mediated by OMT controls plastid differentiation during seed maturation.

The large and highly repetitive genomes of the cultivated species Hordeum vulgare (barley), Triticum aestivum (wheat), and Secale cereale (rye) belonging to the Triticeae tribe of grasses appear to be particularly rich in gene-like sequences including partial duplicates. Most of them have been classified as putative pseudogenes. In this study we employ transient and stable gene silencing- and over-expression systems in barley to study the function of HvARM1 (for H. vulgare Armadillo 1), a partial gene duplicate of the U-box/armadillo-repeat E3 ligase HvPUB15 (for H. vulgare Plant U-Box 15). The partial ARM1 gene is derived from a gene-duplication event in a common ancestor of the Triticeae and contributes to quantitative host as well as nonhost resistance to the biotrophic powdery mildew fungus Blumeria graminis. In barley, allelic variants of HvARM1 but not of HvPUB15 are significantly associated with levels of powdery mildew infection. Both HvPUB15 and HvARM1 proteins interact in yeast and plant cells with the susceptibility-related, plastid-localized barley homologs of THF1 (for Thylakoid formation 1) and of ClpS1 (for Clp-protease adaptor S1) of Arabidopsis thaliana. A genome-wide scan for partial gene duplicates reveals further events in barley resulting in stress-regulated, potentially neo-functionalized, genes. The results suggest neo-functionalization of the partial gene copy HvARM1 increases resistance against powdery mildew infection. It further links plastid function with susceptibility to biotrophic pathogen attack. These findings shed new light on a novel mechanism to employ partial duplication of protein-protein interaction domains to facilitate the expansion of immune signaling networks.

NADPH-dependent thioredoxin reductase C (NTRC) is a chloroplast redox regulator in algae and plants. Here, we used site-specific mutation analyses of the thioredoxin domain active site of NTRC in the green alga Chlamydomonas reinhardtii to show that NTRC mediates cold tolerance in a redox-dependent manner. By means of coimmunoprecipitation and mass spectrometry, a redox- and cold-dependent binding of the Calvin–Benson Cycle Protein 12 (CP12) to NTRC was identified. NTRC was subsequently demonstrated to directly reduce CP12 of C. reinhardtii as well as that of the vascular plant Arabidopsis thaliana in vitro. As a scaffold protein, CP12 joins the Calvin–Benson cycle enzymes phosphoribulokinase (PRK) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) to form an autoinhibitory supracomplex. Using size-exclusion chromatography, NTRC from both organisms was shown to control the integrity of this complex in vitro and thereby PRK and GAPDH activities in the cold. Thus, NTRC apparently reduces CP12, hence triggering the dissociation of the PRK/CP12/GAPDH complex in the cold. Like the ntrc::aphVIII mutant, CRISPR-based cp12::emx1 mutants also exhibited a redox-dependent cold phenotype. In addition, CP12 deletion resulted in robust decreases in both PRK and GAPDH protein levels implying a protein protection effect of CP12. Both CP12 functions are critical for preparing a repertoire of enzymes for rapid activation in response to environmental changes. This provides a crucial mechanism for cold acclimation.