An mTRAN-mRNA interaction mediates mitochondrial translation initiation in plants.

Plant mitochondria represent the largest group of respiring organelles on the planet. Plant mitochondrial messenger RNAs (mRNAs) lack Shine-Dalgarno-like ribosome-binding sites, so it is unknown how plant mitoribosomes recognize mRNA. We show that "mitochondrial translation factors" mTRAN1 and mTRAN2 are land plant-specific proteins, required for normal mitochondrial respiration chain biogenesis. Our studies suggest that mTRANs are noncanonical pentatricopeptide repeat (PPR)-like RNA binding proteins of the mitoribosomal "small" subunit. We identified conserved Adenosine (A)/Uridine (U)-rich motifs in the 5' regions of plant mitochondrial mRNAs. mTRAN1 binds this motif, suggesting that it is a mitoribosome homing factor to identify mRNAs. We demonstrate that mTRANs are likely required for translation of all plant mitochondrial mRNAs. Plant mitochondrial translation initiation thus appears to use a protein-mRNA interaction that is divergent from bacteria or mammalian mitochondria.

Impacts of heat, drought, and combined heat-drought stress on yield, phenotypic traits, and gluten protein traits: capturing stability of spring wheat in excessive environments.

Wheat production and end-use quality are severely threatened by drought and heat stresses. This study evaluated stress impacts on phenotypic and gluten protein characteristics of eight spring wheat genotypes (Diskett, Happy, Bumble, SW1, SW2, SW3, SW4, and SW5) grown to maturity under controlled conditions (Biotron) using RGB imaging and size-exclusion high-performance liquid chromatography (SE-HPLC). Among the stress treatments compared, combined heat-drought stress had the most severe negative impacts on biomass (real and digital), grain yield, and thousand kernel weight. Conversely, it had a positive effect on most gluten parameters evaluated by SE-HPLC and resulted in a positive correlation between spike traits and gluten strength, expressed as unextractable gluten polymer (%UPP) and large monomeric protein (%LUMP). The best performing genotypes in terms of stability were Happy, Diskett, SW1, and SW2, which should be further explored as attractive breeding material for developing climate-resistant genotypes with improved bread-making quality. RGB imaging in combination with gluten protein screening by SE-HPLC could thus be a valuable approach for identifying climate stress-tolerant wheat genotypes.

Striving for Stability in the Dough Mixing Quality of Spring Wheat under the Influence of Prolonged Heat and Drought.

The effects of prolonged heat and drought stress and cool growing conditions on dough mixing quality traits of spring wheat (Triticum aestivum L.) were studied in fifty-six genotypes grown in 2017 and 2018 in southern Sweden. The mixing parameters evaluated by mixograph and the gluten protein characteristics studied by size exclusion high-performance liquid chromatography (SE-HPLC) in dough were compared between the two growing seasons which were very different in length, temperature and precipitation. The genotypes varying in gluten strength between the growing seasons (≤5%, ≤12%, and ≤17%) from three groups (stable (S), moderately stable (MS), and of varying stability (VS)) were studied. The results indicate that most of the mixing parameters were more strongly impacted by the interaction between the group, genotype, and year than by their individual contribution. The excessive prolonged heat and drought did not impact the buildup and mixing time expressed as peak time and time 1-2. The gluten polymeric proteins (unextractable, %UPP; total unextractable, TOTU) and large unextractable monomeric proteins (%LUMP) were closely associated with buildup and water absorption in dough. Major significant differences were found in the dough mixing parameters between the years within each group. In Groups S and MS, the majority of genotypes showed the smallest variation in the dough mixing parameters responsible for the gluten strength and dough development between the years. The mixing parameters such as time 1-2, buildup, and peak time (which were not affected by prolonged heat and drought stress) together with the selected gluten protein parameters (%UPP, TOTU, and %LUMP) are essential components to be used in future screening of dough mixing quality in wheat in severe growing environments.

A MYC2/MYC3/MYC4-dependent transcription factor network regulates water spray-responsive gene expression and jasmonate levels.

Mechanical stimuli, such as wind, rain, and touch affect plant development, growth, pest resistance, and ultimately reproductive success. Using water spray to simulate rain, we demonstrate that jasmonic acid (JA) signaling plays a key role in early gene-expression changes, well before it leads to developmental changes in flowering and plant architecture. The JA-activated transcription factors MYC2/MYC3/MYC4 modulate transiently induced expression of 266 genes, most of which peak within 30 min, and control 52% of genes induced >100-fold. Chromatin immunoprecipitation-sequencing analysis indicates that MYC2 dynamically binds >1,300 promoters and trans-activation assays show that MYC2 activates these promoters. By mining our multiomic datasets, we identified a core MYC2/MYC3/MYC4-dependent "regulon" of 82 genes containing many previously unknown MYC2 targets, including transcription factors bHLH19 and ERF109 bHLH19 can in turn directly activate the ORA47 promoter, indicating that MYC2/MYC3/MYC4 initiate a hierarchical network of downstream transcription factors. Finally, we also reveal that rapid water spray-induced accumulation of JA and JA-isoleucine is directly controlled by MYC2/MYC3/MYC4 through a positive amplification loop that regulates JA-biosynthesis genes.

Neofunctionalization of Mitochondrial Proteins and Incorporation into Signaling Networks in Plants.

Because of their symbiotic origin, many mitochondrial proteins are well conserved across eukaryotic kingdoms. It is however less obvious how specific lineages have obtained novel nuclear-encoded mitochondrial proteins. Here, we report a case of mitochondrial neofunctionalization in plants. Phylogenetic analysis of genes containing the Domain of Unknown Function 295 (DUF295) revealed that the domain likely originated in Angiosperms. The C-terminal DUF295 domain is usually accompanied by an N-terminal F-box domain, involved in ubiquitin ligation via binding with ASK1/SKP1-type proteins. Due to gene duplication, the gene family has expanded rapidly, with 94 DUF295-related genes in Arabidopsis thaliana alone. Two DUF295 family subgroups have uniquely evolved and quickly expanded within Brassicaceae. One of these subgroups has completely lost the F-box, but instead obtained strongly predicted mitochondrial targeting peptides. We show that several representatives of this DUF295 Organellar group are effectively targeted to plant mitochondria and chloroplasts. Furthermore, many DUF295 Organellar genes are induced by mitochondrial dysfunction, whereas F-Box DUF295 genes are not. In agreement, several Brassicaceae-specific DUF295 Organellar genes were incorporated in the evolutionary much older ANAC017-dependent mitochondrial retrograde signaling pathway. Finally, a representative set of DUF295 T-DNA insertion mutants was created. No obvious aberrant phenotypes during normal growth and mitochondrial dysfunction were observed, most likely due to the large extent of gene duplication and redundancy. Overall, this study provides insight into how novel mitochondrial proteins can be created via "intercompartmental" gene duplication events. Moreover, our analysis shows that these newly evolved genes can then be specifically integrated into relevant, pre-existing coexpression networks.