Chloroplast Structure Research Articles

Animal-derived plant biostimulant alleviates drought stress by regulating photosynthesis, osmotic adjustment, and antioxidant systems in tomato plants

• Pig blood-derived protein hydrolysate alleviated negative effects of drought stress on tomato growth. • Pig blood-derived protein hydrolysate improved chlorophyll content and photosynthetic capacity. • Pig blood-derived protein hydrolysate enhanced accumulation of proline, soluble proteins, soluble sugars, and inorganic ions. • Pig blood-derived protein hydrolysate reduced drought-induced oxidative damage by stimulating antioxidant systems. Drought stress is one of the major abiotic factors limiting crop growth and yield production. Protein hydrolysates have been used as plant biostimulants in agriculture due to their positive impacts on plant productivity under abiotic stress; however, little is known about their roles in inducing drought tolerance and the underlying mechanisms. Therefore, we investigated the effects of a new pig blood-derived protein hydrolysate (PP) in increasing tomato tolerance to drought stress. We found that foliar PP application reduced the inhibited impacts of drought stress (10% PEG-6000) on tomato growth, as indicated by improved plant growth parameters. Exogenous PP application inhibited the degradation of chlorophyll, maintained chloroplast structures, increased stomatal aperture, and thereby improved photosynthetic rate under drought stress. The higher accumulation of proline, soluble sugars, soluble proteins, inorganic ions including K + , Ca 2+ , and Mg 2+ , and subsequently higher relative water content were observed in PP-treated tomato leaves and roots. Moreover, PP application mitigated oxidative damages of drought affected-tomato plants by increasing the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) and the accumulation of total phenolic, total flavonoid, ascorbic acid, and glutathione. These findings indicated that foliar PP application could obviously alleviate drought stress by regulating stomatal aperture, chloroplast ultrastructure, osmotic alteration, and antioxidant systems. Therefore, protein hydrolysate derived from animal protein is an effective, economical, and environmental plant biostimulant for improving plant performances under drought stress.

Response and Function of Solanum lycopersicum L. SlSGR2 Gene under Cadmium Stress

Cd stress can cause severe damage to the chloroplast structure and plant function. The chloroplast stay-green gene (SGR2) is senescence-inducible and has several important functions such as regulating senescence, responding to abiotic stress, and regulating seed and fruit ripening. The SlSGR2 gene was cloned from the tomato “Hongluocheng” (full-length of 819 bp encoding 272 amino acids). Subcellular localization prediction and laser confocal observation revealed that the SlSGR2 protein was localized to the chloroplast of A. thaliana protoplasts. A qRT-PCR analysis suggested that the SlSGR2 gene was upregulated by different Cd concentrations and that the gene expression was significantly upregulated in tomato roots and leaves under 50 μM CdCl2 treatment. The pCAMBIA2300-SlSGR2 overexpression constructs were introduced into wild-type Arabidopsis plants via Agrobacterium-mediated transformation. Homozygous mutant lines of sgr2 were identified using the three-primer method. The growth of SlSGR2-overexpressed lines under Cd stress was significantly better than that of WT and sgr2. The chlorophyll, SOD, POD, and CAT contents of OE4-1 and OE4-2 were significantly higher than those of WT and sgr2 while the MDA content was significantly lower than other lines under cadmium stress. This study showed that the SlSGR2 gene is involved in Cd-induced plant senescence and is a negative regulator of plant chlorophyll degradation.

The tobacco chloroplast YCF4 gene is essential for transcriptional gene regulation and plants photoautotrophic growth.

A tobacco chloroplast hypothetical open reading frame 4 (YCF4) has been reported as a non-essential assembly factor for photosynthesis based on an incomplete knockout of YCF4, just 93 of 184 amino acids from the N-terminus were knocked out. On the other hand, we removed the complete sequence of YCF4 from tobacco chloroplasts and observed that ΔYCF4 plants were unable to survive photoautotrophically as their growth was hampered in the absence of an external carbon supply, clearly showing that the YCF4 is essential for photosynthesis. Initially, the aadA gene was introduced into the tobacco plastome replacing the complete YCF4 gene through homologous recombination events. The replacement of YCF4 with aadA was confirmed by PCR and Southern blot analysis in ΔYCF4 plants. Homoplasmic ΔYCF4 plants had a light green phenotype, and the leaves became pale yellow as the plants grew older. The structure of chloroplasts of ΔYCF4 mutants of light green phenotype was studied using a transmission electron microscope (TEM), and the micrographs demonstrated structural anomalies in the chloroplasts; including shape, size, and grana stacking compared to the wild-type plants. Further, transcriptome analysis revealed that the expression of PSI, PSII, and ribosomal genes remained unchanged in ∆YCF4 plants. On the other hand, transcriptome levels of rbcL (Ribulose 1,5-bisphosphatecarboxylase/oxygenaselarge subunit), LHC (Light-Harvesting Complex), and ATP Synthase (atpB and atpL) decreased, indicating that the YCF4 has the function(s) in addition to assembling the photosynthetic complex. This was confirmed by in-silico protein-protein interactions of full-length YCF4 as well as 93 and 91 of 184 amino acids from N- and C-termini of the full-length protein, which revealed that the C-terminus (91 aa) of YCF4 is important in interacting with other chloroplast proteins. These findings provide genetic support for the plastid YCF4 gene's critical role in regulating the plastid gene expression and assembling the photosynthetic complex.

Cytological, genetic and transcriptomic characterization of a cucumber albino mutant.

Photosynthesis, a fundamental process for plant growth and development, is dependent on chloroplast formation and chlorophyll synthesis. Severe disruption of chloroplast structure results in albinism of higher plants. In the present study, we report a cucumber albino alc mutant that presented white cotyledons under normal light conditions and was unable to produce first true leaf. Meanwhile, alc mutant could grow creamy green cotyledons under dim light conditions but died after exposure to normal light irradiation. No chlorophyll and carotenoid were detected in the alc mutant grown under normal light conditions. Using transmission electron microscopy, impaired chloroplasts were observed in this mutant. The genetic analysis indicated that the albino phenotype was recessively controlled by a single locus. Comparative transcriptomic analysis between the alc mutant and wild type revealed that genes involved in chlorophyll metabolism and the methylerythritol 4-phosphate pathway were affected in the alc mutant. In addition, three genes involved in chloroplast development, including two FtsH genes and one PPR gene, were found to have negligible expression in this mutant. The quality of RNA sequencing results was further confirmed by real-time quantitative PCR analysis. We also examined 12 homologous genes from alc mutant in other plant species, but no genetic variation in the coding sequences of these genes was found between alc mutant and wild type. Taken together, we characterized a cucumber albino mutant with albinism phenotype caused by chloroplast development deficiency and this mutant can pave way for future studies on plastid development.

Lead exposure dose-dependently affects oxidative stress, AsA-GSH, photosynthesis, and mineral content in pakchoi (Brassica chinensis L.).

Lead (Pb) is a heavy metal pollutant and negatively affects agriculture and ecosystems. Pb can cause oxidative stress and abnormal plant growth. The ascorbic acid-glutathione (AsA-GSH) cycle mainly exists in chloroplasts and resists oxidative stress, scavenges reactive oxygen radicals, and maintains normal photosynthesis. However, the dosage related effects of Pb on pakchoi photosynthesis, via oxidative stress and the AsA-GSH system, remains unclear. In this study, various Pb dosage stress models were tested (low: 300 mg/kg; medium: 600 mg/kg; high: 900 mg/kg). Pb stress induced a dose-dependent increase in Pb content in pakchoi leaves (P < 0.05). Principal component analysis showed that Se, B, and Pb were significantly and negatively correlated. Pb stress also increased MDA content and decreased antioxidant enzymes SOD, GSH-Px, and T-AOC activities (P < 0.05). We also found that Vc content, as well as the GSH/GSSG ratio, decreased. Additionally, Pb stress destroyed chloroplast structure, decreased photosynthesis indicators Pn, Tr, Gs, Ci and VPD, and attenuated Fv/Fm and Fv/Fo (P < 0.05). In the high-dose group, the contents of chlorophyll a, chlorophyll b, and carotenoids decreased significantly, while the expression of chloroplast development genes (GLK, GLN2) decreased (P < 0.05). Our data suggest that Pb stress leads to dosage-dependent, aberrant photosynthesis by inhibiting the AsA-GSH system in pakchoi. This study expands the Pb toxicology research field and provides indications for screening antagonists.

Abscisic acid involved in trehalose improved melon photosynthesis via regulating oxidative stress tolerance and cell morphology structure under cold stress

Melon is a thermophilic vegetable crop with high economic benefits. Cold stress is often encountered in the growth period of melon seedlings in early spring and limits the normal growth and development of seedlings. Exogenous trehalose (Tre) is a green and healthy sugar, known as the ‘sugar of life’, and has a significant role in regulating crop cold tolerance. In this study, we explored whether exogenous Tre induced abscisic acid (ABA) signal involved in regulating melon photosynthesis under cold tolerance and clarified the associated regulatory mechanism. Melon seedlings were pre-treated with nordihydroguaiaretic acid (NDGA), a synthetic inhibitor of the key enzyme 9-cis-epoxycarotenoid dioxygenase for ABA synthesis, followed by spraying with exogenous Tre and then treated with normal temperature or cold stress. Results showed that exogenous Tre induced the up-regulation of CmNCED3 gene expression and increase in ABA level under normal temperature conditions. Cold stress treatment significantly caused leaf membrane lipid peroxidation damage; reduced relative water content, net photosynthetic rate, transpiration rate, stomatal conductance, water use efficiency and the spongy ratio of leaves; caused chloroplast structure damage. The use of NDGA in inhibiting the synthesis of endogenous ABA further aggravated the damages compared with cold treatment alone. Exogenous Tre treatment alleviated the inhibitory effects of cold stress and NDGA and maintained relatively normal cell morphology structure and photosynthesis. Therefore, we speculate that exogenous Tre pretreatment increases the level of endogenous ABA by up-regulating the expression of the CmNCED3 gene and triggers the defence system of melon plants. Under cold stress, cell membrane lipid peroxidation damage in the melon seedlings was significantly reduced by Tre treatment. Meanwhile, the absorption and utilisation of water maintained the relative integrity of the structure and function of mesophyll cells and chloroplasts, thereby reducing the inhibitory effect of cold stress on plant photosynthesis. In conclusion, ABA participates in exogenous Tre-enhanced melon oxidative stress tolerance and alleviation of cell morphology structure damage, which combined to improve melon cold tolerance and photosynthesis. • Exogenous trehalose triggers endogenous ABA levels. • ABA are involved in exogenous Tre-alleviated melon oxidative stress damage. • ABA participate in the Tre-regulated maintenance of cell integrity and function under cold stress.

Identification and function analysis of yellow-leaf mutant (YX-yl) of broomcorn millet

BackgroundBroomcorn millet is highly tolerant to drought and barren soil. Changes in chlorophyll content directly affect leaf color, which subsequently leadsleading to poor photosynthetic performance and reduced crop yield. Herein, we isolated a yellow leaf mutant (YX-yl) using a forward genetics approach and evaluated its agronomic traits, photosynthetic pigment content, chloroplast ultrastructure, and chlorophyll precursors. Furthermore, the molecular mechanism of yellowing was explored using transcriptome sequencing.ResultsThe YX-yl mutant showed significantly decreased plant height and low yield. The leaves exhibited a yellow-green phenotype and poor photosynthetic capacity during the entire growth period. The content of chlorophyll a, chlorophyll b, and carotenoids in YX-yl leaves was lower than that in wild-type leaves. Chlorophyll precursor analysis results showed that chlorophyll biosynthesis in YX-yl was hindered by the conversion of porphobilinogen to protoporphyrin IX. Examination of chloroplast ultrastructure in the leaves revealed that the chloroplasts of YX-yl accumulated on one side of the cell. Moreover, the chloroplast structure of YX-yl was degraded. The inner and outer membranes of the chloroplasts could not be distinguished well. The numbers of grana and grana thylakoids in the chloroplasts were low. The transcriptome of the yellowing mutant YX-yl was sequenced and compared with that of the wild type. Nine chlorophyll-related genes with significantly different expression profiles were identified: PmUROD, PmCPO, PmGSAM, PmPBDG, PmLHCP, PmCAO, PmVDE, PmGluTR, and PmPNPT. The proteins encoded by these genes were located in the chloroplast, chloroplast membrane, chloroplast thylakoid membrane, and chloroplast matrix and were mainly involved in chlorophyll biosynthesis and redox-related enzyme regulation.ConclusionsYX-yl is an ideal material for studying pigment metabolism mechanisms. Changes in the expression patterns of some genes between YX-yl and the wild type led to differences in chloroplast structures and enzyme activities in the chlorophyll biosynthesis pathway, ultimately resulting in a yellowing phenotype in the YX-yl mutant. Our findings provide an insight to the molecular mechanisms of leaf color formation and chloroplast development in broomcorn millet.

Plastome structure, phylogenomic analyses and molecular dating of Arecaceae.

Arecaceae is a species-rich clade of Arecales, while also being regarded as a morphologically diverse angiosperm family with numerous species having significant economic, medicinal, and ornamental value. Although in-depth studies focused on the chloroplast structure of Arecaceae, as well as inferring phylogenetic relationships using gene fragments, have been reported in recent years, a comprehensive analysis of the chloroplast structure of Arecaceae is still needed. Here we perform a comprehensive analysis of the structural features of the chloroplast genome of Arecaceae, compare the variability of gene sequences, infer phylogenetic relationships, estimate species divergence times, and reconstruct ancestral morphological traits. In this study, 74 chloroplast genomes of Arecaceae were obtained, covering five subfamilies. The results show that all chloroplast genomes possess a typical tetrad structure ranging in size between 153,806-160,122 bp, with a total of 130-137 genes, including 76-82 protein-coding genes, 29-32 tRNA genes, and 4 rRNA genes. Additionally, the total GC content was between 36.9-37.7%. Analysis of the SC/IR boundary indicated that the IR region underwent expansion or contraction. Phylogenetic relationships indicate that all five subfamilies in Arecaceae are monophyletic and that Ceroxyloideae and Arecoideae are sister groups (BS/PP = 100/1). The results of molecular dating indicate that the age of the crown group of Arecaceae is likely to be 96.60 [84.90-107.60] Ma, while the age of the stem group is 102.40 [93.44-111.17] Ma. Reconstruction of ancestral traits indicate that the ancestral characteristics of the family include monoecious plants, one seed, six stamens, and a smooth pericarp.

Manufactured Nano-Objects Confer Viral Protection against Cucurbit Chlorotic Yellows Virus (CCYV) Infecting Nicotiana benthamiana.

Nanotechnology has emerged as a new tool to combat phytopathogens in agricultural crops. Cucurbit chlorotic yellows virus (CCYV) mainly infects Solanaceae crops and causes significant crop losses. Nanomaterials (NMs) may have efficacy against plant viruses, but the mechanisms underlying complex nanomaterials-plant-virus interactions remain elusive. We challenged Nicotiana benthamiana plants with GFP-tagged CCYV and observed morphological, physiological, and molecular changes in response to 21-d foliar exposure to nanoscale Fe and Zn and C60 fullerenes at 100 mg/L concentration for 21 days. We observed that in response to C60 (100 mg/L) treatment, plants displayed a normal phenotype while the viral infection was not seen until 5 days post-inoculation. On the contrary, Fe and Zn were unable to suppress viral progression. The mRNA transcriptional analysis for GFP and viral coat protein revealed that the transcripts of both genes were 5-fold reduced in response to C60 treatment. Evaluation of the chloroplast ultrastructure showed that NMs treatment maintained the normal chloroplast structure in the plants as compared to untreated plants. C60 upregulated the defense-related phytohormones (abscisic acid and salicylic acid) by 42–43%. Our results demonstrate the protective function of carbon-based NMs, with suppression of CCYV symptoms via inhibition of viral replication and systemic movement.

A chloroplast-localized pentatricopeptide repeat protein involved in RNA editing and splicing and its effects on chloroplast development in rice

BackgroundThe chloroplast is the organelle responsible for photosynthesis in higher plants. The generation of functional chloroplasts depends on the precise coordination of gene expression in the nucleus and chloroplasts and is essential for the development of plants. However, little is known about nuclear-plastid regulatory mechanisms at the early stage of chloroplast generation in rice.ResultsIn this study, we identified a rice (Oryza sativa) mutant that exhibited albino and seedling-lethal phenotypes and named it ssa1(seedling stage albino1). Transmission electron microscopy (TEM) analysis indicated that the chloroplasts of ssa1 did not have organized thylakoid lamellae and that the chloroplast structure was destroyed. Genetic analysis revealed that the albino phenotypes of ssa1 were controlled by a pair of recessive nuclear genes. Map-based cloning experiments found that SSA1 encoded a pentapeptide repeat (PPR) protein that was allelic to OSOTP51,which was previously reported to participate in Photosystem I (PSI) assembly. The albino phenotype was reversed to the wild type (WT) phenotype when the normal SSA1 sequence was expressed in ssa1 under the drive of the actin promoter. Knockout experiments further created mutants ssa1–2/1–9, which had a phenotype similar to that of ssa1. SSA1 consisted of 7 pentatricopeptide repeat domains and two C-terminal LAGLIDADG tandem sequence motifs and was located in the chloroplast. GUS staining and qRT–PCR analysis showed that SSA1 was mainly expressed in young leaves and stems. In the ssa1 mutants, plastid genes transcribed by plastid-encoded RNA polymerase decreased, while those transcribed by nuclear-encoded RNA polymerase increased at the mRNA level. Loss-of-function SSA1 destroys RNA editing of ndhB-737 and intron splicing of atpF and ycf3–2 in the plastid genome. Yeast two-hybrid and BiFC assays revealed that SSA1 physically interacted with two new RNA editing partners, OsMORF8 and OsTRXz, which have potential functions in RNA editing and chloroplast biogenesis.ConclusionsRice SSA1 encodes a pentatricopeptide repeat protein, which is targeted to the chloroplast. SSA1 regulates early chloroplast development and plays a critical role in RNA editing and intron splicing in rice. These data will facilitate efforts to further elucidate the molecular mechanism of chloroplast biogenesis.

Comparative transcriptome analysis identified ChlH and POLGAMMA2 in regulating yellow-leaf coloration in Forsythia.

Leaf color is one of the most important features for plants used for landscape and ornamental purposes. However, the regulatory mechanism of yellow leaf coloration still remains elusive in many plant species. To understand the complex genetic mechanism of yellow-leaf Forsythia, we first compared the pigment content and leaf anatomical structure of yellow-leaf and green-leaf accessions derived from a hybrid population. The physiological and cytological analyses demonstrated that yellow-leaf progenies were chlorophyll deficient with defected chloroplast structure. With comparative transcriptome analysis, we identified a number of candidate genes differentially expressed between yellow-leaf and green-leaf Forsythia plants. Among these genes, we further screened out two candidates, ChlH (magnesium chelatase Subunit H) and POLGAMMA2 (POLYMERASE GAMMA 2), with consistent relative-expression pattern between different colored plants. To verify the gene function, we performed virus-induced gene silencing assays and observed yellow-leaf phenotype with total chlorophyll content reduced by approximately 66 and 83% in ChlH-silenced and POLGAMMA2-silenced plants, respectively. We also observed defected chloroplast structure in both ChlH-silenced and POLGAMMA2-silenced Forsythia. Transient over-expression of ChlH and POLGAMMA2 led to increased chlorophyll content and restored thylakoid architecture in yellow-leaf Forsythia. With transcriptome sequencing, we detected a number of genes related to chlorophyll biosynthesis and chloroplast development that were responsive to the silencing of ChlH and POLGAMMA2. To summarize, ChlH and POLGAMMA2 are two key genes that possibly related to yellow-leaf coloration in Forsythia through modulating chlorophyll synthesis and chloroplast ultrastructure. Our study provided insights into the molecular aspects of yellow-leaf Forsythia and expanded the knowledge of foliage color regulation in woody ornamental plants.

Cotton (Gossypium hirsutum) VIRMA as an N6-Methyladenosine RNA Methylation Regulator Participates in Controlling Chloroplast-Dependent and Independent Leaf Development.

N6-methyladenosine (m6A) is one of the most abundant internal modifications of mRNA, which plays important roles in gene expression regulation, and plant growth and development. Vir-like m6A methyltransferase associated (VIRMA) serves as a scaffold for bridging the catalytic core components of the m6A methyltransferase complex. The role of VIRMA in regulating leaf development and its related mechanisms have not been reported. Here, we identified and characterized two upland cotton (Gossypium hirsutum) VIRMA genes, named as GhVIR-A and GhVIR-D, which share 98.5% identity with each other. GhVIR-A and GhVIR-D were ubiquitously expressed in different tissues and relatively higher expressed in leaves and main stem apexes (MSA). Knocking down the expression of GhVIR genes by the virus-induced gene silencing (VIGS) system influences leaf cell size, cell shape, and total cell numbers, thereby determining cotton leaf morphogenesis. The dot-blot assay and colorimetric experiment showed the ratio of m6A to A in mRNA is lower in leaves of GhVIR-VIGS plants compared with control plants. Messenger RNA (mRNA) high-throughput sequencing (RNA-seq) and a qRT-PCR experiment showed that GhVIRs regulate leaf development through influencing expression of some transcription factor genes, tubulin genes, and chloroplast genes including photosystem, carbon fixation, and ribosome assembly. Chloroplast structure, chlorophyll content, and photosynthetic efficiency were changed and unsuitable for leaf growth and development in GhVIR-VIGS plants compared with control plants. Taken together, our results demonstrate GhVIRs function in cotton leaf development by chloroplast dependent and independent pathways.

Cadmium contributes to heat tolerance of a hyperaccumulator plant species Sedum alfredii

Hyperaccumulators are plant species that tolerate and accumulate very high concentrations of toxic metals, including Cd. Hyperaccumulation of heavy metals is reported to benefit plant biotic resistance; however, no prior study has examined the possible role of toxic metals on abiotic stress resistance in hyperaccumulators. A preliminary experiment found that Cd significantly improved plant growth of a hyperaccumulator, Sedum alfredii Hance, under heat stress. This study investigated the possible role of Cd in S. alfredii’s heat resistance, using infrared thermography, transmission electron microscopy (TEM), real-time quantitative polymerase chain reaction (RTqPCR), and high-throughput sequencing. The results showed that high temperatures irreversibly damaged stomatal function, chloroplast structure, photosynthesis in S. alfredii, and lowered survival rates to 25%. However, Cd application significantly decreased the leaf temperature of S. alfredii and increased the survival rate to 75%. Cd penetrated the guard cells, restored stomatal function, and mitigated excessive water loss from S. alfredii under heat stress. Moreover, it activated antioxidant enzymes, promoted phytohormone biosynthesis, and upregulated a series of unigenes, thereby augmenting heat resistance in S. alfredii. These results indicate that Cd effectively improved thermotolerance in S. alfredii by regulating stomatal movement and antioxidant systems via upregulation of phytohormones and heat shock proteins.

Parental salt priming improves the low temperature tolerance in wheat offspring via modulating the seed proteome

Low temperature is one of the main abiotic stresses that inhibit wheat growth and development. To understand the physiological mechanism of salt priming induced low temperature tolerance and its transgenerational effects, the chlorophyl b-deficient mutant (ANK) and its wild type (WT) wheat were subjected to low temperature stress after parental salt priming. Salt priming significantly decreased the levels of superoxide anions, hydrogen peroxide and malondialdehyde in both parental and offspring plants under low temperature. The catalase activity in parental wheat and activities of dehydroascorbate reductase and glutathione reductase in the offspring were significantly increased by salt priming under low temperature. Meanwhile, salt priming contributed to mantaining the integrity of chloroplast structure and relatively higher net photosynthetic rate (Pn) in both generations under low temperature. Salt priming also improved the carbohydrate metabolism enzyme activities of parental and offspring plants, such as phosphoglucomutase, fructokinase and sucrose synthase. In addition, ANK plants had significantly higher carbohydrate metabolism enzyme activities than WT plants. The differential expressed proteins (DEP) in seeds of two genotypes under salt priming were mainly related to homeostasis, electron transfer activity, photosynthesis and carbohydrate metabolism. Correlation network analysis showed that the expression of DEP under salt priming was significantly correlated to sucrose concentration and cytoplasmic peroxidase (POX) activity in WT, while that was correlated to various carbohydrate metabolism enzyme activities in ANK plants. These results indicated that the parental salt priming induced modulations of seed proteome regulated the ROS metabolism, photosynthetic carbon assimilation and carbohydrate metabolism, hence enhancing the low temperature tolerance in offspring wheat.

Acidification of seawater attenuates the allelopathic effects of Ulva pertusa on Karenia mikimotoi.

Acidification of seawater resulting from absorption of excessive carbon dioxide from the atmosphere is posing a serious threat to marine ecosystem. In this study, we hypothesized that acidified seawater attenuates allelopathic effects of macroalgae on red tide algae because the increase of dissolved carbon dioxide benefits algal growth, and investigated the allelopathic effects of Ulva pertusa on Karenia mikimotoi in response to seawater acidification by determining cell density, photosynthetic pigment content, chlorophyll fluorescence parameters, and chloroplast structure of K. mikimotoi under U. pertusa stress in original (pH=8.2) and acidified (pH=7.8) seawater. U. pertusa inhibited the growth of K. mikimotoi in the original and acidizing seawater, and the inhibition rate was positively correlated with treatment time and concentration of U. pertusa. However, acidizing condition significantly weakened the inhibition degree of U. pertusa on K. mikimotoi (P < 0.05), with the inhibition rates decreased from 51.85 to 43.16% at 10 gFW/L U. pertusa for 96 h. U. pertusa reduced contents of chlorophyll a, chlorophyll c, and carotenoid, maximum photochemical quantum yield (Fv/Fm), actual quantum yield, maximum relative electron transfer efficiency (rETRmax) of PSII, real-time fluorescence value (F), and maximum fluorescence value (Fm') of PSII of K. mikimotoi under original and acidified conditions. And, the inhibition degree of U. pertusa under acidizing condition was significantly lower than that of original seawater group. Furthermore, the damage degree of chloroplast structure of K. mikimotoi under U. pertusa stress was more serious under original seawater condition. These results indicate that acidification of seawater attenuates the allelopathic effects of U. pertusa on K. mikimotoi.

Amaranth calcium oxalate crystals are associated with chloroplast structures and proteins

Calcium oxalate (CaOx) crystals in plants are formed in crystal idioblasts cells and have specific geometric shapes. Their proposed functions include calcium homeostasis and carbon source, among others. Amaranth is a plant that presents high tolerance to abiotic stresses and accumulates considerable amounts of CaOx crystals; however, few studies have focused on characterizing the crystals ultrastructure and none is related to identifying proteins bound to them. This information is of great interest to understand the mechanisms related to CaOx crystal formation and to support their proposed functions. Thus, this work aimed to characterize CaOx crystals in amaranth leaves. Crystals were purified and the proteins bound to them were isolated and identified by nLC-MS/MS. Leaf sections were analyzed by light and electron microscopy. The identified proteins were related to the chloroplast such as ATPb synthase, RuBisCO large subunit, and cell wall-related proteins, which were validated by immunohistochemistry and immunogold labeling. In addition, it was observed that CaOx crystal idioblasts were formed from parenchyma cells associated with mesophyll and veins, in which the thylakoid membranes of degraded chloroplasts turned into crystal chambers. These results significantly advance our understanding of the mechanisms of CaOx crystal formation and the potential function as an alternative carbon source in leaves.

Trehalose alleviates salt tolerance by improving photosynthetic performance and maintaining mineral ion homeostasis in tomato plants

Trehalose (Tre), which was an osmoprotective or stabilizing molecule, played a protective role against different abiotic stresses in plants and showed remarkable perspectives in salt stress. In this study, the potential role of Tre in improving the resistance to salt stress in tomato plants was investigated. Tomato plants (Micro Tom) were treated with Hoagland nutrient solution (CK), 10 mM Tre (T), 150 mM sodium chloride (NaCl, S), and 10 mM Tre+150 mM NaCl (S+T) for 5 days. Our results showed that foliar application of Tre alleviated the inhibition of tomato plant growth under salt stress. In addition, salt stress decreased the values of net photosynthetic rate (Pn, 85.99%), stomata conductance (gs, 57.3%), and transpiration rate (Tr, 47.97%), but increased that of intercellular carbon dioxide concentration (Ci, 26.25%). However, exogenous application of Tre significantly increased photosynthetic efficiency, increased the activity of Calvin cycle enzymes [ribulose diphosphate carboxylase/oxygenase (Rubisco), fructose-1,6-bisphosphate aldolase (FBA), fructose-1, 6-bisphosphatase (FBPase), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and transketolase (TK)], up-regulated the expression of genes encoding enzymes, induced stomatal opening, and alleviated salt-induced damage to the chloroplast membrane and structure. In the saline environment, photosynthetic electron transport was restricted, resulting the J-I-P phase to decrease. At the same time, the absorption, capture, and transport energies per excited cross-section and per active reaction center decreased, and the dissipation energy increased. Conversely, Tre reversed these values and enhanced the photosystem response to salt stress by protecting the photosynthetic electron transport system. In addition, foliage application with Tre significantly increased the potassium to sodium transport selectivity ratio (SK–Na) by 16.08%, and increased the levels of other ions to varying degrees. Principal component analysis (PCA) analysis showed that exogenous Tre could change the distribution of elements in different organs and affect the expressions of SlSOS1, SlNHX, SlHKT1.1, SlVHA, and SlHA-A at the transcriptional level under salt stress, thereby maintaining ion homeostasis. This study demonstrated that Tre was involved in the process of mitigating salt stress toxicity in tomato plants and provided specific insights into the effectiveness of Tre in mediating salt tolerance.

LSL1 controls cell death and grain production by stabilizing chloroplast in rice.

Lesion mutants can be valuable tools to reveal the interactions between genetic factors and environmental signals and to improve grain production. Here we identified a rice (Oryza sativa) mutant, lesion spotleaf1 (lsl1), which produces necrotic leaf lesions throughout its life cycle. LSL1 encodes a protein of unknown function and belongs to a grass-specific clade. The lesion phenotype of the lsl1 mutant was sharply induced by shading, and its detached leaves incubated in 6-benzylamino purine similarly formed lesions in the dark. In addition, the lsl1 mutant exhibited reactive oxygen species accumulation and cell death. The terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) and comet assays revealed that the lsl1 mutant contained severe DNA damage, resulting in reduced grain yield and quality. RNA sequencing, gene expression, and protein activity analyses indicate that LSL1 is required for chloroplast function. Furthermore, LSL1 interacts with PsaD and PAP10 to form a regulatory module that functions in chlorophyll synthesis and chloroplast development to maintain redox balance. Our results reveal that LSL1 maintains chloroplast structure, redox homeostasis, and DNA stability, and plays important roles in the interaction between genetic factors and environmental signals and in regulating grain size and quality.

Melatonin delays leaf senescence and improves cucumber yield by modulating chlorophyll degradation and photoinhibition of PSII and PSI

Decreases in chlorophyll and photosynthetic efficiency co-occur during aging and leaf senescence. Here, we report the positive functions of melatonin (MT) in delaying leaf senescence in cucumber plants. The results showed that natural leaf senescence induced the accumulation of endogenous MT and that spraying MT delayed leaf senescence in a concentration-dependent manner. Moreover, application of MT and transient overexpression of CsASMT, the key gene in MT synthesis, both suppressed leaf senescence simulated by darkness. This was evidenced by a decrease in expression of the senescence-related gene SAG12 and the cell death-related gene PDCD, an increase in chlorophyll content in terms of downregulation of chlorophyll-degradation genes and normal chloroplast structure, and higher activity of PSI and PSII reaction centers, electron transfer efficiency and photosynthesis-associated protein levels, compared with these measures in the H2O-treated seedlings. In addition, the data from field spraying experiments demonstrated that MT indeed alleviated the decline in chlorophyll content and increased photosynthetic efficiency in both functional and senescent leaves, which ultimately promoted the growth and yield of cucumber in the solar greenhouse. Together, these results indicate that MT acts as a positive regulator to downregulate SAG12 and PDCD mRNA abundances and alleviate damage to the photosynthetic apparatus by suppressing chloroplast disintegration and photosynthesis-associated protein degradation during leaf senescence.

The strigolactone receptor SlDWARF14 plays a role in photosynthetic pigment accumulation and photosynthesis in tomato.

Tomato DWARF14 regulates the development of roots, shoot branches and leaves, and also plays a role in photosynthetic pigment accumulation and photosynthetic capacity. Strigolactones (SLs) are a novel class of plant hormones. DWARF14 (D14) is the only SL receptor identified to date, but it is not functionally analyzed in tomato (Solanum lycopersicum). In the present study, we identified the potential SL receptor in tomato by bioinformatic analysis, which was designated as SlD14. SlD14 was expressed in roots, stems, flowers and developing fruits, with the highest expression level in leaves. sld14 mutant plants produced by the CRISPR/Cas9 system displayed reduced plant height and root biomass, increased shoot branching and altered leaf shape comparing with WT plants. The cytokinin biosynthetic gene ISOPENTENYLTRANSFERASE 3 (SlIPT3), auxin biosynthetic genes FLOOZY (SlFZY) and TRYPTOPHAN AMINOTRANSFERASE RELATED 1 (SlTAR1) and several auxin transport genes SlPINs, which are involved in branch formation, showed higher expression levels in the sld14 plant stem. In addition, sld14 plants exhibited light-green leaves, reduced chlorophyll and carotenoid contents, abnormal chloroplast structure and reduced photosynthetic capacity. Transcriptomic analysis showed that the transcript levels of six chlorophyll biosynthetic genes, three carotenoid biosynthetic genes and numerous chlorophyll a/b-binding protein genes were decreased in sld14 plants. These results suggest that tomato SL receptor gene SlD14 not only regulates the development of roots, shoot branches and leaves, but also plays a role in regulating photosynthetic pigment accumulation and photosynthetic capacity.