Search Results (37)

The plastid stroma-localized chaperone HSP90C is essential for maintaining chloroplast proteostasis and facilitating protein translocation. Prior research has established HSP90C’s imperative role in the SEC translocase-dependent transport of the photosystem II subunit PsbO1 and its interaction with the SEC1 translocase motor protein SecA1. However, the exact mechanism of this interaction remains to be explored. In this study, we delineated the interactional mode of HSP90C and SecA1 with the model client protein. Yeast two-hybrid and in vitro ATPase activity analyses with purified proteins revealed PsbO1 may bind to HSP90C at multiple sites, including the DPW motif within the C-terminal extension (CTE) region, suggesting a possible client-loading mechanism unique to plastid orthologs. We also confirmed that glycine-646 is important in mediating substrate interaction, though it conferred a much weaker binding than the CTE region, thereby elucidating a critical role for the amino acid whose mutation resulted in visible plant phenotypes. Our in vitro biochemical assays also demonstrated that the stromal intermediate form of PsbO1 with the thylakoid signal peptide (tSP) significantly enhanced SecA1 ATPase activity, suggesting a preferential binding to the motor protein. On the other hand, the mature domain of the PsbO1, excluding the tSP sequence, inhibited HSP90C ATPase activity. We also observed the HSP90C-PsbO1-SecA1 ternary complex was stabilized by the presence of the client tSP. This work therefore provides new insights into the functional mechanisms of HSP90C and its contribution to chloroplast stromal protein stabilization and thylakoid protein transport. Full article

Plant architecture-related traits are key agronomic traits affecting crop growth and yield. To unravel the genetic architecture of plant height (PH), ear height (EH), tassel length (TL), and tassel primary branch number (TPBN), 379 DH lines derived from 21 maize hybrids were used for genome-wide association study (GWAS) and genomic selection (GS) analyses. Although plant architecture-related traits were significantly influenced by genotype and genotype-by-environment interactions, moderate to high broad-sense heritability was observed for PH (81.3%), EH (79.6%), TL (86.4%), and TPBN (82.5%). Using six different models for GWAS, seven unique SNPs on chromosomes 1, 2, and 3 were identified for PH, 92 unique SNPs located on chromosomes 1 to 9 were identified for EH, three unique SNPs on chromosome 6 were detected for TL, and 18 unique SNPs located on chromosomes 1, 4, 5, 8, and 10 were identified for TPBN at the p-value threshold of 7.42 × 10⁻⁶. A few hotspot genomic regions conferring plant architecture-related traits were identified, located in bins 2.07, 4.07, 8.03, 6.01, and 10.00. A total of 144 putative candidate genes were identified, which were enriched in endocytosis and lipid biosynthetic process, electron carrier activity, chloroplast stroma, and plastid stroma. The prediction accuracy evaluated through 5-fold cross-validation was 0.44 for PH, 0.43 for EH, 0.31 for TL, and 0.30 for TPBN. When the training population size (TPS) reached 60–70% or marker density (MD) reached 3000, the prediction accuracy tends to stabilize, indicating that the optimum size of TPS and MD were 60–70% and 3000 for GS, respectively. The highest prediction accuracy evaluated by using 30–5000 significant SNPs corresponding to the lowest p-value was 0.70 for PH, 0.85 for EH, 0.58 for TL, and 0.75 for TPBN, with an increase in accuracy of 59.1% to 150.0%. These results demonstrate that integrating GS with a subset of highly significant SNPs can substantially enhance prediction efficiency, thereby facilitating the selection of superior genotypes and accelerating the breeding of maize varieties with optimized plant architecture. This study has further elucidated the genetic basis of maize architecture-related traits and provided valuable information on how to implement GS to breed novel maize varieties with optimized plant types. Full article

Shallot (Allium hirtifolium Boiss.) is of considerable nutritional and medical significance due to its strong antioxidant properties; however, no nanophytotoxicity studies have assessed whether the use of nanofertilizers would improve shallot performance, micronutrient iron (Fe) enrichment, and yield in semi-arid regions. Herein, we evaluated the effects of magnetite nanoparticles (nFe₃O₄) on shallot grown for a full lifecycle in two semi-arid regions through bulb-priming followed by foliar application and compared them with conventional ferrous sulfate (FeSO₄) fertilizer and untreated control. Our results showed remarkable cellular adaptations to semi-arid climate upon nFe₃O₄ treatment as leaves displayed thickened cell walls, distinct chloroplasts featuring organized thylakoid grana and stroma, normal mitochondria, abundant starch grains, and plastoglobuli around chloroplasts compared to FeSO₄ or untreated control. At 900 mg/L nFe₃O₄, chlorophyll-a, chlorophyll-b, and carotenoid increased by 27–55%, 108–126%, and 77–97%, respectively, compared to FeSO₄ applied at recommended field rate (1800 mg/L). Significant increments in bulb diameter (38–39%) and sister bulb number (300–500%) were observed upon 900 mg/L nFe₃O₄ treatment compared to FeSO₄ (1800 mg/L) and control. Furthermore, with 900 mg/L nFe₃O₄ treatment, total phenol, flavonoids, and Fe in bulbs increased by 27–46%, 29–73%, and 486–549%, respectively, compared to FeSO₄ (1800 mg/L). These findings demonstrate that bulb-priming followed by foliar application of 900 mg/L of nFe₃O₄ could significantly promote cellular adaptation, thereby improving photosynthetic efficiency, bulb yield, antioxidant activities, and Fe biofortification in shallot, and may serve as a novel approach for improving shallot production in semi-arid regions. Full article

The Kinesin superfamily members are ATP-dependent microtubule-based motor proteins that are conserved among all eukaryotic organisms and play vital roles in diverse cellular processes, such as vesicle trafficking, mitosis and meiosis, and cytoskeletal dynamics. Here, OsKIF14.3, a kinesin-14 subfamily protein, was map-based cloned and functionally analyzed. The OsKIF14.3 gene exhibited a constitutive expression pattern. OsKIF14.3 protein localized on the microtubule and formed homodimer via the conserved Coiled Coil 1 (CC1) domain. Mutation of OsKIF14.3 altered OsSWEET11′s subcellular location from the plasma membrane into both the plasma membrane and the cytoplasm, leading to abnormal starch metabolism, excess starch accumulation in the chloroplast, broken stroma lamella and yellowish leaves in oskif14.3 mutant. These results enriched our understanding of the kinesin superfamily and leaf color regulation mechanism. Full article

The stromules are tubular extensions of chloroplasts, or broader plastids, formed by the organelle envelope and filled with the stroma, the internal content of organelle. The formation of stromules is related to the cytoskeleton. Stromules occur in photosynthetic tissues under illumination and are therefore proposed to be important for retrograde signaling, which is essential for adaptation to stress factors. Stromules are also observed after pathogen attack. Some groups propose that stromules are a resemblance to endoplasmic reticulum dynamics, without having actual significance in molecules transport within a cell. However, there is no consensus among researchers regarding the actual function and significance of stromules, which can be the result of different models used to study stromules, and the necessity of using fluorescent labels, with all advantages and limitations of fluorescence-based methodology. Here I briefly discuss current knowledge on the subject from a perspective of stromule origin—the chloroplast envelope, and the potential advantages of having a conduit within organelles instead of relying on diffusion through cytosol. Full article

Background: The proper development of grana and stroma within chloroplasts is critical for plant vitality and crop yield in rice and other cereals. While the molecular mechanisms underpinning these processes are known, the genetic networks governing them require further exploration. Methods and Results: In this study, we characterize a novel rice mutant termed yellow leaf and dwarf 7 (yld7), which presents with yellow, lesion-like leaves and a dwarf growth habit. The yld7 mutant shows reduced photosynthetic activity, lower chlorophyll content, and abnormal chloroplast structure. Transmission electron microscopy (TEM) analysis revealed defective grana stacking in yld7 chloroplasts. Additionally, yld7 plants accumulate high levels of hydrogen peroxide (H₂O₂) and exhibit an up-regulation of senescence-associated genes, leading to accelerated cell death. Map-based cloning identified a C-to-T mutation in the LOC_Os07g33660 gene, encoding the YLD7 protein, which is a novel ankyrin domain-containing protein localized to the chloroplast. Immunoblot analysis of four LHCI proteins indicated that the YLD7 protein plays an important role in the normal biogenesis of chloroplast stroma and grana, directly affecting leaf senescence and overall plant stature. Conclusions: This study emphasizes the significance of YLD7 in the intricate molecular mechanisms that regulate the structural integrity of chloroplasts and the senescence of leaves, thus providing valuable implications for the enhancement of rice breeding strategies and cultivation. Full article

Water deficit is the major stress factor magnified by climate change that causes the most reductions in plant productivity. Knowledge of photosystem II (PSII) response mechanisms underlying crop vulnerability to drought is critical to better understanding the consequences of climate change on crop plants. Salicylic acid (SA) application under drought stress may stimulate PSII function, although the exact mechanism remains essentially unclear. To reveal the PSII response mechanism of celery plants sprayed with water (WA) or SA, we employed chlorophyll fluorescence imaging analysis at 48 h, 96 h, and 192 h after watering. The results showed that up to 96 h after watering, the stroma lamellae of SA-sprayed leaves appeared dilated, and the efficiency of PSII declined, compared to WA-sprayed plants, which displayed a better PSII function. However, 192 h after watering, the stroma lamellae of SA-sprayed leaves was restored, while SA boosted chlorophyll synthesis, and by ameliorating the osmotic potential of celery plants, it resulted in higher relative leaf water content compared to WA-sprayed plants. SA, by acting as an antioxidant under drought stress, suppressed phototoxicity, thereby offering PSII photoprotection, together with enhanced effective quantum yield of PSII photochemistry (Φ_PSII) and decreased quantity of singlet oxygen (¹O₂) generation compared to WA-sprayed plants. The PSII photoprotection mechanism induced by SA under drought stress was triggered by non-photochemical quenching (NPQ), which is a strategy to protect the chloroplast from photo-oxidative damage by dissipating the excess light energy as heat. This photoprotective mechanism, triggered by NPQ under drought stress, was adequate in keeping, especially in high-light conditions, an equal fraction of open PSII reaction centers (qp) as of non-stress conditions. Thus, under water deficit stress, SA activates a regulatory network of stress and light energy partitioning signaling that can mitigate, to an extent, the water deficit stress on PSII functioning. Full article

The plastid stroma-localized chaperone HSP90C plays a crucial role in maintaining optimal proteostasis within chloroplasts and participates in protein translocation processes. While existing studies have revealed HSP90C’s direct interaction with the Sec translocase-dependent client pre-protein PsbO1 and the SecY1 subunit of the thylakoid membrane-bound Sec1 translocase channel system, its direct involvement with the extrinsic homodimeric Sec translocase subunit, SecA1, remains elusive. Employing bimolecular fluorescence complementation (BiFC) assay and other in vitro analyses, we unraveled potential interactions between HSP90C and SecA1. Our investigation revealed dynamic interactions between HSP90C and SecA1 at the thylakoid membrane and stroma. The thylakoid membrane localization of this interaction was contingent upon active HSP90C ATPase activity, whereas their stromal interaction was associated with active SecA1 ATPase activity. Furthermore, we observed a direct interaction between these two proteins by analyzing their ATP hydrolysis activities, and their interaction likely impacts their respective functional cycles. Additionally, using PsbO1, a model Sec translocase client pre-protein, we studied the intricacies of HSP90C’s possible involvement in pre-protein translocation via the Sec1 system in chloroplasts. The results suggest a complex nature of the HSP90C-SecA1 interaction, possibly mediated by the Sec client protein. Our studies shed light on the nuanced aspects of HSP90C’s engagement in orchestrating pre-protein translocation, and we propose a potential collaborative role of HSP90C with SecA1 in actively facilitating pre-protein transport across the thylakoid membrane. Full article

Leaves are the main site of photosynthesis in plants, and leaf color plays a major role in crop quality, yield, resistance, as well as other aspects. Although the genes related to photosynthesis have been well characterized in plants in general, yellow-green leaf mutants have not yet been fully studied in tomatoes. In the present study, a dark green leaf (GL) mutant was isolated from yellow-leaf tomato (wild-type). The dark GL displays a distinct yellow-green phenotype, and has a greater chlorophyll content and higher photosynthetic rate. Furthermore, the lamellae were clear, and the stroma and grana were orderly, with more stacking and larger starch grains according to the ultrastructure analysis of chloroplasts in GL leaves. Comparative transcriptome analysis of GL and wild-type plants was performed to identify the pathways and genes related to photosynthesis. In this work, a total of 292 differentially expressed genes (DEGs) between GL plants and WT plants were identified, of which 131 genes were upregulated and 161 genes were downregulated. The diterpenoid biosynthesis and photosynthesis antenna proteins were the two most significantly enriched in the first 20 pathways according to KEGG analysis. Most of the DEGs involved in diterpenoid biosynthesis and photosynthesis were antenna proteins. The photosynthesis antenna protein Solyc02g071030 (LHCB1) and the diterpenoid biosynthesis-related genes, Solyc08g005710 and Solyc09g059240, were significantly upregulated in GL leaves compared with WT leaves. The expression patterns of the DEGs were similar to those determined by qRT-PCR. Overall, our research not only revealed the diterpenoid biosynthesis and photosynthesis pathways involving in leaf color variation, but also identified the putative target genes for genetic manipulation in the future. Full article

Trichomes, epidermal protrusions in terrestrial plants, play diverse roles in plant defense, metabolism, and development. Arabidopsis thaliana, a model plant with single-celled and non-glandular trichomes, is a valuable system for studying cell differentiation in plants. However, organelle biology in Arabidopsis trichomes is relatively underexplored. Using light and transmission electron microscopy, we investigated the phenotypes of intracellular structures in Arabidopsis trichomes caused by tgd5 mutations, which are known to disrupt lipid transfer from the endoplasmic reticulum to plastids and have a large impact on chloroplast morphology in pavement and guard cells. Significant phenotypic changes in the plastid structure were observed in tgd5 trichome cells, including the absence of plastoglobuli, the emergence of clusters of electron-dense particles in the stroma, and the possibly cup-shaped morphology of plastids. Additionally, the tgd5 mutations triggered the formation of giant, up to 15 µm in diameter, neutral lipid-containing droplets in the trichome cells, as revealed using histochemical staining with lipophilic dyes. These lipid droplets were substantially larger and more frequent in trichome cells than in other types of cells in tgd5. These findings highlight the role of TGD5 in maintaining plastid structure and implicate the unique activity of lipid metabolism in Arabidopsis trichomes. Full article

In vascular plants, the final photosynthetic electron transfer from ferredoxin (Fd) to NADP⁺ is catalyzed by the flavoenzyme ferredoxin:NADP⁺ oxidoreductase (FNR). FNR is recruited to thylakoid membranes via an integral membrane protein TROL (thylakoid rhodanese-like protein) and the membrane associated protein Tic62. We have previously demonstrated that the absence of TROL triggers a very efficient superoxide (O₂^•−) removal mechanism. The dynamic TROL–FNR interaction has been shown to be an apparently overlooked mechanism that maintains linear electron flow before alternative pathway(s) is(are) activated. In this work, we aimed to further test our hypothesis that the FNR–TROL pair could be the source element that triggers various downstream networks of chloroplast ROS scavenging. Tandem affinity purification followed by the MS analysis confirmed the TROL–FNR interaction and revealed possible interaction of TROL with the thylakoid form of the enzyme ascorbate peroxidase (tAPX), which catalyzes the H₂O₂-dependent oxidation of ascorbate and is, therefore, the crucial component of the redox homeostasis system in plants. Further, EPR analyses using superoxide spin trap DMPO showed that, in comparison with the wild type, plants overexpressing TROL (TROL OX) propagate more O₂^•− when exposed to high light stress. This indicates an increased sensitivity to oxidative stress in conditions when there is an excess of membrane-bound FNR and less free FNR is found in the stroma. Finally, immunohistochemical analyses of glutathione in different Arabidopsis leaf cell compartments showed highly elevated glutathione levels in TROL OX, indicating an increased demand for this ROS scavenger in these plants, likely needed to prevent the damage of important cellular components caused by reactive oxygen species. Full article

Waterlogging and heavy mental (e.g., cadmium) stress are two primary threats to crop growth. The combination of abiotic stresses was common and frequent, especially in the field condition. Even though the effects of individual waterlogging and cadmium on tomato plants have been widely investigated, the response of tomatoes under combined waterlogging and cadmium stress remains unclear. This study aimed to clarify and compare physiological, biochemical characteristics and plant growth of two tomato genotypes under individual and combined stress. Two tomato genotypes (‘MIX-002’ and ‘LA4440’) were treated under control, waterlogging, cadmium stress and their combination. The results showed that chloroplast ultrastructure of tomatoes under individual and combined stress was damaged with disordered stroma and grana lamellae. The H₂O₂ (hydrogen peroxide) content and O₂^·− (superoxide anion radical) production rate of plants under all the three stresses was not significantly higher than the control except for ‘LA4440’ under the combined stress. Antioxidant enzymes actively responded in the two tomato genotypes, as shown by significant increase in SOD activity from ‘MIX-002’ under waterlogging and combined stress and from ‘LA4440’ under cadmium. Meanwhile, CAT activity of ‘MIX-002’ under waterlogging and ‘LA4440′ under combined stress significantly decreased, and the POD activity of ‘MIX-002’ under combined stress significantly increased as compared with the respective control. The APX activity of ‘MIX-002’ and ‘LA4440’ under combined stress was significantly lower and higher than the respective controls. This indicated that tomato plants were able to secure redox homeostasis and protect plants from oxidative damage through the synergetic regulation of antioxidant enzymes. Plant height and biomass of the two genotypes under individual and combined stress significantly decreased, which could be a direct result from the chloroplast alteration and resource re-allocation. Overall, the effects of combined waterlogging and cadmium stress were not simply the sum of individual effects on two tomato genotypes. Distinct ROS (reactive oxygen species) scavenging systems of two tomato genotypes under stresses suggest a genotype-dependent antioxidant enzymes regulation. Full article

Chloroplasts play crucial roles in biotic and abiotic stress responses, regulated by nuclear gene expression through changes in the cellular redox state. Despite lacking the N-terminal chloroplast transit peptide (cTP), nonexpressor of pathogenesis-related genes 1 (NPR1), a redox-sensitive transcriptional coactivator was consistently found in the tobacco chloroplasts. Under salt stress and after exogenous application of H₂O₂ or aminocyclopropane-1-carboxylic acid, an ethylene precursor, transgenic tobacco plants expressing green fluorescent protein (GFP)-tagged NPR1 (NPR1-GFP) showed significant accumulation of monomeric nuclear NPR1, irrespective of the presence of cTP. Immunoblotting and fluorescence image analyses indicated that NPR1-GFP, with and without cTP, had similar molecular weights, suggesting that the chloroplast-targeted NPR1-GFP is likely translocated from the chloroplasts to the nucleus after processing in the stroma. Translation in the chloroplast is essential for nuclear NPR1 accumulation and stress-related expression of nuclear genes. An overexpression of chloroplast-targeted NPR1 enhanced stress tolerance and photosynthetic capacity. In addition, compared to the wild-type lines, several genes encoding retrograde signaling-related proteins were severely impaired in the Arabidopsis npr1-1 mutant, but were enhanced in NPR1 overexpression (NPR1-Ox) transgenic tobacco line. Taken together, chloroplast NPR1 acts as a retrograding signal that enhances the adaptability of plants to adverse environments. Full article

Psb28 is a soluble protein in the photosystem II (PSII) complex, but its role in the drought stress response of wheat remains unclear. Here, we functionally characterized the TaPsb28 gene, which positively regulates drought tolerance in wheat. When the full-length 546-bp TaPsb28 cDNA was transferred into Arabidopsis thaliana, it was located in the guard cell chloroplast around the stroma. Overexpression of TaPsb28 conferred drought tolerance, as exhibited by the increases in the survival rate. Transgenic plants maintained lower MDA content and higher chlorophyll content by inducing chlorophyll synthase (ChlG) gene transcription. The content of abscisic acid (ABA) and zeatin increased significantly in wild-type (WT) plants under drought stress, and the transcriptional expression levels of RD22, dihydroflavonol 4-reductase (DFR) and anthocyanin reductase (ANR) genes were induced, thus enhancing the contents of endogenous cyanidin, delphinidin, and proanthocyanidins. However, in transgenic plants, although anthocyanins were further aggregated, the ABA increase was inhibited, zeatin was restored to the control level under drought stress, and stomatal closure was promoted. These findings indicate ABA and zeatin have opposite synergistic effects in the process of drought tolerance caused by TaPsb28 because only after the effect of zeatin is alleviated can ABA better play its role in promoting anthocyanin accumulation and stomatal closure, thus enhancing the drought tolerance of transgenic plants. The results suggest that overexpression of TaPsb28 exerts a positive role in the drought response by influencing the functional metabolism of endogenous hormones. The understanding acquired through the research laid a foundation for further in-depth investigation of the function of TaPsb28 in drought resistance in wheat, especially its relationship with anthocyanidin accumulation. Full article

Ornamental plant species may vary substantially in their tolerance response to heavy metals. The aim of this research was to check chrysanthemum cultivars, namely Donglin Ruixue (C), Yellow (F), Red pocket (G), and New 9714 (I), which are commonly used as landscape plants to determine their levels of cadmium (Cd) tolerance at different cadmium concentrations through hydroponic cultures. Chrysanthemum cultivars were treated with five different Cd concentrations (0, 10, 20, 50, and 100 mg L⁻¹) and different physiological, enzymatic, and ultra-structure traits were taken under consideration in vitro. The results showed that cadmium concentration significantly inhibited the total chlorophyll content, chlorophyll a, chlorophyll b, and carotenoid content. Chlorophyll contents were significantly reduced at higher Cd concentrations in all cultivars, but the reduction rates were higher in cultivar F (59.49%), G (40.41%), I (44.97%), and C (33.86%). Similarly, the chlorophyll b reduction was higher than that of chlorophyll a in I (73.33%), followed by G (58.06%), F (61.66%), and C (32.43%), under Cd stress conditions. Additionally, the relative conductivity was recorded in cultivars C (146.48%), F (223.66%), G (165.96%), and I (154.92%), respectively, at 100 mg L⁻¹ Cd concentrations. Likewise, MDA was significantly increased with high Cd stress, at 155.56, 325.27, 173.91, and 322.18%, in C, F, G, and I cultivars at 100 mg L⁻¹, but it was promoted with a greater increase in F and I cultivars. Similarly, SOD and CAT activities were increased with the increase in Cd stress, but reduced in F and I cultivars at higher stress levels of 100 mg L⁻¹. In the same way, POD activity was significantly higher in the C and G cultivars. Additionally, ultrastructure changes also occurred with the increase in the Cd stress, i.e., 20 mg L⁻¹ to 100 mg L⁻¹, and these changes caused alterations in cell organelles, including in the chloroplast, grana, lamella, thylakoid, and stroma. They also caused noticeable damage to mitochondria at higher Cd concentrations. It was concluded that the higher levels of antioxidative defense of the C and G cultivars of chrysanthemum indicated their ability to tolerate high Cd stress conditions. These could, therefore, be used for their phytoremediation potential in Cd-contaminated areas. Full article