Translation initiation factor 3 families: what are their roles in regulating cyanobacterial and chloroplast gene expression?

Initiation is a key control point for the regulation of translation in prokaryotes and prokaryotic-like translation systems such as those in plant chloroplasts. Genome sequencing and biochemical studies are increasingly demonstrating differences in many aspects of translation between well-studied microbes such as Escherichia coli and lesser studied groups such as cyanobacteria. Analyses of chloroplast translation have revealed its prokaryotic origin but also uncovered many unique aspects that do not exist in E. coli. Recently, a novel form of posttranscriptional regulation by light color was discovered in the filamentous cyanobacterium Fremyella diplosiphon that requires a putative stem-loop and involves the use of two different prokaryotic translation initiation factor 3s (IF3s). Multiple (up to five) putative IF3s have now been found to be encoded in 22 % of sequenced cyanobacterial genomes and 26 % of plant nuclear genomes. The lack of similar light-color regulation of gene expression in most of these species suggests that IF3s play roles in regulating gene expression in response to other environmental and developmental cues. In the plant Arabidopsis, two nuclear-encoded IF3s have been shown to localize to the chloroplasts, and the mRNA levels encoding these vary significantly in certain organ and tissue types and during several phases of development. Collectively, the accumulated data suggest that in about one quarter of photosynthetic prokaryotes and eukaryotes, IF3 gene families are used to regulate gene expression in addition to their traditional roles in translation initiation. Models for how this might be accomplished in prokaryotes versus eukaryotic plastids are presented.

The role of a 14-3-3 protein in stomatal opening mediated by PHOT2 in Arabidopsis.

The 14-3-3 λ isoform is required for normal stomatal opening mediated by PHOT2 in Arabidopsis thaliana. Arabidopsis phototropin2 (PHOT2) interacts with the λ-isoform 14-3-3 protein both in yeast two-hybrid screening and in an in vitro pull-down assay. Further yeast two-hybrid analysis also showed that the PHOT2 C-terminal kinase domain was required for the interaction. Site-directed mutagenesis indicated that PHOT2 Ser-747 is essential for the yeast interaction. Phenotypic characterization of a loss-of-function 14-3-3 λ mutant in a phot1 mutant background showed that the 14-3-3 λ protein was necessary for normal PHOT2-mediated blue light-induced stomatal opening. PHOT2 Ser-747 was necessary for complementation of the blue light-activated stomatal response in a phot1 phot2 double mutant. The 14-3-3 λ mutant in the phot1 mutant background allowed normal phototropism and normal chloroplast accumulation and avoidance responses. It also showed normal stomatal opening mediated by PHOT1 in a phot2 mutant background. The 14-3-3 κ mutant had no effect on stomatal opening in response to blue light. Although the 14-3-3 λ mutant had no chloroplast movement phenotype, the 14-3-3 κ mutation caused a weaker avoidance response at an intermediate blue light intensity by altering the balance between the avoidance and accumulation responses. The results highlight the strict specificity of phototropin-mediated signal transduction pathways.

Cover crop inclusion and residue retention improves soybean production and physiology in drought conditions.

Soybean (Glycine max (L.) Merr.) planting has increased in central and western North Dakota despite frequent drought occurrences that limit productivity. Soybean plants need high photosynthetic and transpiration rates to be productive, but they also need high water use efficiency when water is limited. Crop residues and cover crops in crop rotations may improve soybean drought tolerance in northern Great Plains. We aimed to examine how a management practice that included cover crops and residue retention impacts agronomic, ecosystem water and carbon dioxide flux, and canopy-scale physiological attributes of soybeans in the northern Great Plains under drought conditions. The experiment consisted of two soybean fields over two years with business-as-usual (no-cover crops and spring wheat residue removal) and aspirational management (cover crops and spring wheat residue retention) during a drought year. We compared yield; aboveground biomass; green chromatic coordinates, and CO2 and H2O fluxes from eddy covariance, Phenocam images, and ancillary micrometeorological measurements. These measurements were used to derive ecosystem-scale physical, and physiological attributes with the 'big leaf' framework to diagnose underlying processes. Soybean yields were 29 % higher under drought conditions in the field managed in a system that included cover crops and residue retention. This yield increase was associated with a 5 day increase in the green-chromatic-coordinate defined maturity phenophase, increasing agronomic and intrinsic water use efficiency by 27 % and 33 %, respectively, increasing water uptake, and increasing the rubisco-limited photosynthetic capacity (Vcmax25) by 42 %. The inclusion of cover crops and residue retention into a cropping system improved soybean productivity because of differences in water use, phenology timing, and photosynthetic capacity. These results suggest that farmers can improve soybean productivity and yield stability by incorporating cover crops and residue retention into their management suite because these practices to facilitate more aggressive water uptake.

The Long-Term Agroecosystem Research cropland common experiment at Northern Plains.

Cropland agriculture in the northern Great Plains is challenged by variable weather, agricultural intensification, and competing use for energy development. Innovative cropland practices that address these challenges are needed to ensure regional agriculture can sustainably meet future food, fuel, and fiber demand. In response to this need, the Northern Plains Long-Term Agroecosystem Research Network site established a cropland experiment in 2019 that contrasts prevailing and alternative practices at plot and field scales over a proposed 30-year time frame. The experimental site is located on the Area IV Soil Conservation Districts Cooperative Research Farm near Mandan, ND. Cropping practices for the first 6 years of the experiment were developed with input from stakeholders and include a 3-year crop rotation of spring wheat (Triticum aestivum L.), corn (Zea mays L.), and soybean (Glycine max L.) with cover crops (alternative practice) and without (prevailing practice). The prevailing practice also involves the removal of crop residue, while a second alternative practice of perennial forages is included in the plot-scale experiment. Biophysical measurements are made at both spatial scales at frequencies aligned with approved methods for each agronomic and environmental metric. Findings from the first 6 years of the experiment will help identify tradeoffs associated with cover crop use and residue removal in dryland cropping systems. In the future, the experiment will adopt a knowledge co-production approach whereby researchers and stakeholders will work collaboratively to identify problems, implement research, and interpret results.

A mutation in Arabidopsis seedling plastid development1 affects plastid differentiation in embryo-derived tissues during seedling growth.

Oilseed plants like Arabidopsis (Arabidopsis thaliana) develop green photosynthetically active embryos. Upon seed maturation, the embryonic chloroplasts degenerate into a highly reduced plastid type called the eoplast. Upon germination, eoplasts redifferentiate into chloroplasts and other plastid types. Here, we describe seedling plastid development1 (spd1), an Arabidopsis seedling albino mutant capable of producing normal green vegetative tissues. Mutant seedlings also display defects in etioplast and amyloplast development. Precocious germination of spd1 embryos showed that the albino seedling phenotype of spd1 was dependent on the passage of developing embryos through the degreening and dehydration stages of seed maturation, suggesting that SPD1 is critical during eoplast development or early stages of eoplast redifferentiation. The SPD1 gene was found to encode a protein containing a putative chloroplast-targeting sequence in its amino terminus and also domains common to P-loop ATPases. Chloroplast localization of the SPD1 protein was confirmed by targeting assays in vivo and in vitro. Although the exact function of SPD1 remains to be defined, our findings reveal aspects of plastid development unique to embryo-derived cells.

Changes in leaf optical properties associated with light-dependent chloroplast movements.

We surveyed 24 plant species to examine how leaf anatomy influenced chloroplast movement and how the optical properties of leaves change with chloroplast position. All species examined exhibited light-dependent chloroplast movements but the associated changes in leaf absorptance varied considerably in magnitude. Chloroplast movement-dependent changes in leaf absorptance were greatest in shade species, in which absorptance changes of >10% were observed between high- and low-light treatments. Using the Kubelka-Munk theory, we found that changes in the absorption (k) and chlorophyll a absorption efficiency (k*) associated with chloroplast movement correlated with cell diameter, such that the narrower, more columnar cells found in sun leaves restricted the ability of chloroplasts to move. The broader, more spherical cells of shade leaves allowed greater chloroplast rearrangements and in low-light conditions allowed efficient light capture. Across the species tested, light-dependent chloroplast movements modulated leaf optical properties and light absorption efficiency by manipulating the package (sieve or flattening) effect but not the detour (path lengthening) effect.

Bird and invertebrate communities appear unaffected by fracking traffic along rural roads despite dust emissions.

The development of shale petroleum resources has industrialized rural landscapes. We investigated how traffic from energy development expands and intensifies the road-effect zone through increased dust exposure, and how birds and invertebrates inhabiting the road-effect zone in agricultural areas of the Bakken region might be affected by dust exposure. We used dust collectors, trail cameras, and sweep-netting at increasing distances from unpaved roads to determine dust deposition, relative bird abundance, and invertebrate abundance, respectively. We found that traffic associated with fracking along unpaved roads emitted substantial dust 180 m into adjacent crop fields. But neither bird abundance or behavior, nor invertebrate abundance or community composition, appeared to be affected by dust or traffic. These findings suggest that wildlife in previously intensified agricultural landscapes like crop fields are resilient to intensification from energy development, but the same might not be true for wildlife in previously undisturbed habitat.

PLASTID MOVEMENT IMPAIRED1 mediates ABA sensitivity during germination and implicates ABA in light-mediated Chloroplast movements.

The plant hormone abscisic acid (ABA) controls many aspects of plant growth and development, including seed development, germination and responses to water-deficit stress. A complex ABA signaling network integrates environmental signals including water availability and light intensity and quality to fine-tune the response to a changing environment. To further define the regulatory pathways that control water-deficit and ABA responses, we carried out a gene-trap tagging screen for water-deficit-regulated genes in Arabidopsis thaliana. This screen identified PLASTID MOVEMENT IMPAIRED1 (PMI1), a gene involved in blue-light-induced chloroplast movement, as functioning in ABA-response pathways. We provide evidence that PMI1 is involved in the regulation of seed germination by ABA, acting upstream of the intersection between ABA and low-glucose signaling pathways. Furthermore, PMI1 participates in the regulation of ABA accumulation during periods of water deficit at the seedling stage. The combined phenotypes of pmi1 mutants in chloroplast movement and ABA responses indicate that ABA signaling may modulate chloroplast motility. This result was further supported by the detection of altered chloroplast movements in the ABA mutants aba1-6, aba2-1 and abi1-1.

THRUMIN1 is a light-regulated actin-bundling protein involved in chloroplast motility.

Chloroplast movement in response to changing light conditions optimizes photosynthetic light absorption. This repositioning is stimulated by blue light perceived via the phototropin photoreceptors and is transduced to the actin cytoskeleton. Some actin-based motility systems use filament reorganizations rather than myosin-based translocations. Recent research favors the hypothesis that chloroplast movement is driven by actin reorganization at the plasma membrane, but no proteins affecting chloroplast movements have been shown to associate with both the plasma membrane and actin filaments in vivo. Here we identified THRUMIN1 as a critical link between phototropin photoreceptor activity at the plasma membrane and actin-dependent chloroplast movements. THRUMIN1 bundles filamentous actin in vitro, and it localizes to the plasma membrane and displays light- and phototropin-dependent localization to microfilaments in vivo. These results suggest that phototropin-induced actin bundling via THRUMIN1 is important for chloroplast movement. A mammalian homolog of THRUMIN1, GRXCR1, has been implicated in auditory responses and hair cell stereocilla development as a regulator of actin architecture. Studies of THRUMIN1 will help elucidate the function of this family of eukaryotic proteins.

The "sensational" power of movement in plants: A Darwinian system for studying the evolution of behavior.

Darwin's research on botany and plant physiology was a landmark attempt to integrate plant movements into a biological perspective of behavior. Since antiquity, people have sought to explain plant movements via mechanical or physiological forces, and yet they also constructed analogies between plant and animal behavior. During the Renaissance and Enlightenment, thinkers began to see that physiochemical explanations of plant movements could equally apply to animal behavior and even human thought. Darwin saw his research on plant movements as a strategic front against those who argued that his theory of evolution could not account for the acquisition of new behavioral traits. He believed that his research explained how the different forms of plant movement evolved as modified habits of circumnutation, and he presented evidence that plants might have a brain-like organ, which could have acquired various types of plant sensitivity during evolution. Upon publication of The Power of Movement in Plants, his ideas were overwhelmingly rejected by plant physiologists. Subsequently, plant biologists came to view the work as an important contribution to plant physiology and biology, but its intended contribution to the field of evolution and behavior has been largely overlooked.

A brassinosteroid-hypersensitive mutant of BAK1 indicates that a convergence of photomorphogenic and hormonal signaling modulates phototropism.

The phototropic response of Arabidopsis (Arabidopsis thaliana) is induced by the phototropin photoreceptors and modulated by the cryptochrome and phytochrome photoreceptors. Downstream of these photoreceptors, asymmetric lateral redistribution of auxin underlies the differential growth, which results in phototropism. Historical physiological evidence and recent analysis of hormone-induced gene expression demonstrate that auxin and brassinosteroid signaling function interdependently. Similarly, in this study we report evidence that interactions between brassinosteroids and auxin signaling modulate phototropic responsiveness. We found that elongated, a previously identified photomorphogenesis mutant, enhances high-light phototropism and represents a unique allele of BAK1/SERK3, a receptor kinase implicated in brassinosteroid perception. Altogether, our results support the hypothesis that phototropic responsiveness is modulated by inputs that influence control of auxin response factor-mediated transcription.

Second positive phototropism results from coordinated co-action of the phototropins and cryptochromes.

Phototropism and hypocotyl growth inhibition are modulated by the coaction of different blue-light photoreceptors and their signaling pathways. How seedlings integrate the activities of the different blue-light photoreceptors to coordinate these hypocotyl growth responses is still unclear. We have used time-lapse imaging and a nontraditional mathematical approach to conduct a detailed examination of phototropism in wild-type Arabidopsis and various blue-light photoreceptor mutants. Our results indicate that high fluence rates of blue light (100 micro mol m(-)(2) s(-)(1)) attenuate phototropism through the coaction of the phototropin and cryptochrome blue-light photoreceptors. In contrast, we also demonstrate that phototropins and cryptochromes function together to enhance phototropism under low fluence rates (<1.0 micro mol m(-)(2) s(-)(1)) of blue light. Based on our results, we hypothesize that phototropins and cryptochromes regulate phototropism by coordinating the balance between stimulation and inhibition of growth of the hypocotyl depending on the fluence rate of blue light.