Chloroplast Breakdown Research Articles

Exogenous application of calcium ameliorates salinity stress of mungbean

An experiment was conducted to investigate the impact of exogenous Ca in alleviating salinity stress of mungbean. Two mungbean genotypes, BD 6895 and BD 6905, were used with four different Ca levels (5, 10, 15 and 20 mM) under 10 dSm-1 salinity conditions in a hydroponic culture. The application of exogenous Ca positively influenced on overall plant growth and development. Although both genotypes exhibited similar plant heights, they gave the peak (56.65 and 48.6 cm for BD 6895 and BD 6905, respectively) at 10 mM Ca. Higher levels of exogenous Ca were associated with increased chlorophyll content. Maximum Chlorophyll (1.6 mg g-1) was observed at 15 mM Ca treatment. Furthermore, exogenous Ca application reduced Malondialdehyde levels (at 15 mM Ca: 1.14 and 1.69 μmole/g for BD 6895 and BD 6905, respectively). Both the genotypes exhibited a proline content pattern, demonstrating proline content upsurge with increased Ca application. Microscopic analysis revealed larger vascular areas with exogenous Ca (BD 6895: 402 µm, BD 6905: 398.3 µm) compared to smaller areas under salinity stress (BD 6895: 258.7 µm, BD 6905: 248.4 µm). Salinity stress induced changes in upper epidermis thickness, leaf tissue compactness, chloroplast breakdown and chlorosis in mungbean plants. However, exogenous Ca application counteracted these detrimental effects, enabling mungbean plants to thrive in saline conditions. In conclusion, the study highlights the positive influence of exogenous Ca (10-15 mM) in promoting mungbean growth and managing salt stress. Ann. Bangladesh Agric. (2023) 27 (2): 91-104

The kiwifruit amyloplast proteome (kfALP): a resource to better understand the mechanisms underlying amyloplast biogenesis and differentiation.

The biogenesis and differentiation (B&D) of amyloplasts contributes to fruit flavor and color. Here, remodeling of starch granules, thylakoids and plastoglobules was observed during development and ripening in two kiwifruit (Actinidia spp.) cultivars - yellow-fleshed 'Hort16A' and green-fleshed 'Hayward'. A protocol was developed to purify starch-containing plastids with a high degree of intactness, and amyloplast B&D was studied using label-free-based quantitative proteomic analyses in both cultivars. Over 3000 amyloplast-localized proteins were identified, of which >98% were quantified and defined as the kfALP (kiwifruit amyloplast proteome). The kfALP data were validated by Tandem-Mass-Tag (TMT) labeled proteomics in 'Hort16A'. Analysis of the proteomic data across development and ripening revealed: 1) a conserved increase in the abundance of proteins participating in starch synthesis/degradation during both amyloplast B&D; 2) up-regulation of proteins for chlorophyll degradation and of plastoglobule-localized proteins associated with chloroplast breakdown and plastoglobule formation during amyloplast differentiation; 3) constitutive expression of proteins involved in ATP supply and protein import during amyloplast B&D. Interestingly, two different pathways of amyloplast B&D were observed in the two cultivars. In 'Hayward', significant increases in abundance of photosynthetic- and tetrapyrrole metabolism-related proteins were observed, but the opposite trend was observed in 'Hort16A'. In conclusion, analysis of the kfALP provides new insights into the potential mechanisms underlying amyloplast B&D with relevance to key fruit quality traits in contrasting kiwifruit cultivars.

The interplay between autophagy and chloroplast vesiculation pathways under dark-induced senescence.

In cellular circumstances where carbohydrates are scarce, plants can use alternative substrates for cellular energetic maintenance. In plants, the main protein reserve is present in the chloroplast, which contains most of the total leaf proteins and represents a rich source of nitrogen and amino acids. Autophagy plays a key role in chloroplast breakdown, a well-recognised symptom of both natural and stress-induced plant senescence. Remarkably, an autophagic-independent route of chloroplast degradation associated with chloroplast vesiculation (CV) gene was previously demonstrated. During extended darkness, CV is highly induced in the absence of autophagy, contributing to the early senescence phenotype of atg mutants. To further investigate the role of CV under dark-induced senescence conditions, mutants with low expression of CV (amircv) and double mutants amircv1xatg5 were characterised. Following darkness treatment, no aberrant phenotypes were observed in amircv single mutants; however, amircv1xatg5 double mutants displayed early senescence and altered dismantling of chloroplast and membrane structures under these conditions. Metabolic characterisation revealed that the functional lack of both CV and autophagy leads to higher impairment of amino acid release and differential organic acid accumulation during starvation conditions. The data obtained are discussed in the context of the role of CV and autophagy, both in terms of cellular metabolism and the regulation of chloroplast degradation.

Transcription factor LcNAC002 coregulates chlorophyll degradation and anthocyanin biosynthesis in litchi.

Chlorophyll degradation and anthocyanin biosynthesis, which often occur almost synchronously during fruit ripening, are crucial for vibrant coloration of fruits. However, the interlink point between their regulatory pathways remains largely unknown. Here, 2 litchi (Litchi chinensis Sonn.) cultivars with distinctively different coloration patterns during ripening, i.e. slow-reddening/stay-green "Feizixiao" (FZX) vs rapid-reddening/degreening "Nuomici" (NMC), were selected as the materials to study the key factors determining coloration. Litchi chinensis STAY-GREEN (LcSGR) was confirmed as the critical gene in pericarp chlorophyll loss and chloroplast breakdown during fruit ripening, as LcSGR directly interacted with pheophorbide a oxygenase (PAO), a key enzyme in chlorophyll degradation via the PAO pathway. Litchi chinensis no apical meristem (NAM), Arabidopsis transcription activation factor 1/2, and cup-shaped cotyledon 2 (LcNAC002) was identified as a positive regulator in the coloration of litchi pericarp. The expression of LcNAC002 was significantly higher in NMC than in FZX. Virus-induced gene silencing of LcNAC002 significantly decreased the expression of LcSGR as well as L. chinensis MYELOBLASTOSIS1 (LcMYB1), and inhibited chlorophyll loss and anthocyanin accumulation. A dual-luciferase reporter assay revealed that LcNAC002 significantly activates the expression of both LcSGR and LcMYB1. Furthermore, yeast-one-hybrid and electrophoretic mobility shift assay results showed that LcNAC002 directly binds to the promoters of LcSGR and LcMYB1. These findings suggest that LcNAC002 is an important ripening-related transcription factor that interlinks chlorophyll degradation and anthocyanin biosynthesis by coactivating the expression of both LcSGR and LcMYB1.

Melatonin delays yellowing of broccoli during storage by regulating chlorophyll catabolism and maintaining chloroplast ultrastructure

• Exogenous melatonin markedly extended the shelf life of broccoli. • Melatonin delayed the decline of chlorophyll content and chloroplast breakdown. • Melatonin regulated activities and gene expressions of chlorophyll catabolic enzymes. • Melatonin regulated the expressions of BoSGR1 and BoSGR2 . The effect of melatonin treatment on color, chlorophyll degradation and chloroplast structure in broccoli during storage was evaluated. One hundred μmmol L −1 melatonin markedly extended the shelf life of broccoli. Compared with control florets, melatonin treatment delayed the decline of chlorophyll content, and maintained more intact chloroplasts. Meanwhile, melatonin also suppressed the activities of chlorophyll catabolic enzymes such as chlorophyllase (CLH), pheophytinase (PPH), pheophorbide a oxygenase (PAO) and red chlorophyll catabolite reductase (RCCR), and down-regulated the expressions of BoNYC1 , BoNOL , BoCLH , BoPPH , BoPAO , BoRCCR and BoSGR1 involved in chlorophyll catabolism. Taken together, our results indicated that melatonin could be a good candidate for delaying yellowing in broccoli during storage at 20 ℃.

PDM4, a Pentatricopeptide Repeat Protein, Affects Chloroplast Gene Expression and Chloroplast Development in Arabidopsis thaliana.

Extensive studies have been carried out on chloroplast gene expression and chloroplast development; however, the regulatory mechanism is still largely unknown. Here, we characterized Pigment-Defective Mutant4 (PDM4), a P-type PPR protein localized in chloroplast. The pdm4 mutant showed seedling-lethal and albino phenotype under heterotrophic growth conditions. Transmission electron microscopic analysis revealed that thylakoid structure was totally disrupted in pdm4 mutant and eventually led to the breakdown of chloroplasts. The levels of several chloroplast- and nuclear-encoded proteins are strongly reduced in pdm4 mutant. Besides, transcript profile analysis detected that, in pdm4 mutant, the expression of plastid-encoded RNA polymerase-dependent genes was markedly affected, and deviant chloroplast rRNA processing was also observed. In addition, we found that PDM4 functions in the splicing of group II introns and may also be involved in the assembly of the 50S ribosomal particle. Our results demonstrate that PDM4 plays an important role in chloroplast gene expression and chloroplast development in Arabidopsis.

Postharvest 1-methylcyclopropene application delays leaf yellowing of pak choi (Brassica rapa subsp. chinensis) by improving chloroplast antioxidant capacity and maintaining chloroplast structural integrity during storage at 20 °C

The aim of this study was to uncover the regulatory mechanism underlying the leaf yellowing of pak choi after 1-methylcyclopropene (1-MCP) treatment at (20 ± 0.5) °C. The ultrastructural, physiological and biochemical metabolism changes of the chloroplast in pak choi were investigated. Chlorophyll degradation was effectively alleviated by 1-MCP treatment, and the market life of pak choi was extended by 2 days compared with that of control. Electron microscopy confirmed that the chloroplast breakdown was reduced by 1-MCP. This treatment not only preserved the high activities of chloroplast antioxidant enzymes (superoxide dismutase SOD and ascorbic peroxidase APX) and non-enzymatic antioxidants (ascorbate-glutathione (AsA-GSH)) for scavenging reactive oxygen species (ROS) (superoxide anion O2·− and hydrogen peroxide H2O2), but also decreased the chloroplast chlorophyll-degrading peroxidase (Chl-POX) activity, thus retarding chlorophyll degradation in the peroxidase-H2O2 system. Abundant O2·- and H2O2 may function as major toxic molecules to enhance the activities of chloroplast phospholipase (PLD), lipoxygenase (LOX) and lipase. The loss of the chloroplast structural integrity and function caused by the high activities of PLD, lipase and LOX aggravated the enzymatic degradation of chlorophyll in pak choi, which was rescued by using 1-MCP that retarded the degradation of the chloroplast membrane, as well as maintained high unsaturated fatty acid levels and low malondialdehyde (MDA) level. In conclusion, 1-MCP fumigation may augment the chloroplast antioxidant capacity for scavenging ROS (O2·- and H2O2), which attenuated the activities of chloroplast PLD, lipase and LOX, inhibited the degradation of the chloroplast membrane and kept the membrane integrity and normal function, ultimately relieving the yellowing of pak choi.

Photosystem II 22kDa protein level - a prerequisite for excess light-inducible memory, cross-tolerance to UV-C and regulation of electrical signalling.

It is well known that PsbS is a key protein for the proper management of excessive energy in plants. Plants without PsbS cannot trigger non-photochemical quenching, which is crucial for optimal photosynthesis under variable conditions. Our studies showed wild-type plants had enhanced tolerance to UV-C-induced cell death (CD) upon induction of light memory by a blue or red light. However, npq4-1 plants, which lack PsbS, as well as plants overexpressing this protein (oePsbS), responded differently. Untreated oePsbS appeared more tolerant to UV-C exposure, whereas npq4-1 was unable to adequately induce cross-tolerance to UV-C. Similarly, light memory induced by episodic blue or red light was differently deregulated in npq-4 and oePsbS, as indicated by transcriptomic analyses, measurements of the trans-thylakoid pH gradient, chlorophyll a fluorescence parameters, and measurements of foliar surface electrical potential. The mechanism of the foliar CD development seemed to be unaffected in the analysed plants and is associated with chloroplast breakdown. Our results suggest a novel, substantial role for PsbS as a regulator of chloroplast retrograde signalling for light memory, light acclimation, CD, and cross-tolerance to UV radiation.

Functional inactivation of OsGCNT induces enhanced disease resistance to Xanthomonas oryzae pv. oryzae in rice

BackgroundSpotted-leaf mutants are important to reveal programmed cell death and defense-related pathways in rice. We previously characterized the phenotype performance of a rice spotted-leaf mutant spl21 and narrowed down the causal gene locus spl21(t) to an 87-kb region in chromosome 12 by map-based cloning.ResultWe showed that a single base substitution from A to G at position 836 in the coding sequence of Oryza sativa beta-1,6-N-acetylglucosaminyl transferase (OsGCNT), effectively mutating Tyr to Cys at position 279 in the translated protein sequence, was responsible for the spotted-leaf phenotype as it could be rescued by functional complementation. Compared to the wild type IR64, the spotted-leaf mutant spl21 exhibited loss of chlorophyll, breakdown of chloroplasts, down-regulation of photosynthesis-related genes, and up-regulation of senescence associated genes, which indicated that OsGCNT regulates premature leaf senescence. Moreover, the enhanced resistance to the bacterial leaf blight pathogen Xanthomonas oryzae pv. oryzae, up-regulation of pathogenesis-related genes and increased level of jasmonate which suggested that OsGCNT is a negative regulator of defense response in rice. OsGCNT was expressed constitutively in the leaves, sheaths, stems, roots, and panicles, and OsGCNT-GFP was localized to the Golgi apparatus. High throughput RNA sequencing analysis provided further evidence for the biological effects of loss of OsGCNT function on cell death, premature leaf senescence and enhanced disease resistance in rice. Thus, we demonstrated that the novel OsGCNT regulated rice innate immunity and immunity-associated leaf senescence probably by changing the jasmonate metabolic pathway.ConclusionsThese results reveal that a novel gene Oryza sativa beta-1,6-N-acetylglucosaminyl transferase (OsGCNT) is responsible for the spotted-leaf mutant spl21, and OsGCNT acts as a negative-regulator mediating defense response and immunity-associated premature leaf senescence probably by activating jasmonate signaling pathway.

Chloroplast breakdown during dehydration of a homoiochlorophyllous resurrection plant proceeds via senescence-like processes

The vegetative tissues of resurrection plants are able to withstand severe protoplasmic dehydration and revive quickly upon rehydration. Resurrection species defined as ‘homoiochlorophyllous’ retain most or part of their chlorophyll and photosynthetic complement in the dry state, and rely on various mechanisms to protect themselves against photo-damage. In this study, we investigated the changes in chlorophyll distribution, light absorption gradients as well as the alterations in ultrastructure that take place during dehydration of the homoiochlorophyllous species Craterostigma pumilum. Chlorophyll fluorescence profiles show that light absorption is attenuated in dry leaves, likely minimizing generation of reactive oxygen species. These are accompanied by changes that take place in the supramolecular organization of the photosynthetic protein complexes, and ordered functional adjustments of the photosynthetic apparatus, further lessening the excitation and electron pressures. Albeit these, the ultrastructural studies reveal that chloroplasts in dehydrated leaf tissues exhibit features indicative of oxidative stress, which are also reminiscent of senescing chloroplasts. These include mass proliferation of plastoglobules, variable degrees of thylakoid dismantling, as well as chloroplast fragmentation and seemingly vacuolar degradation of such fragments. In addition, unique vesicular structures between the two chloroplast envelope membranes were visualized, some of which appeared to detach from chloroplasts, likely en route to degradation. Together, the data indicate that homoiochlorophyllous resurrection species handle photo-induced damage during dehydration on two levels. Minimization of photo-damage is achieved by attenuation of light absorption and other photo-protective mechanisms. When this is insufficient and significant damage does occur, elimination of damaged components takes place via processes resembling senescence. Nevertheless, these processes are reversible, enabling the plants to avoid the terminal steps of senescence and, hence, to recover.

PDM3, a pentatricopeptide repeat-containing protein, affects chloroplast development.

The chloroplast, as the photosynthetic organelle of plants, plays a crucial role in plant development. Extensive studies have been conducted on chloroplast development; however, the related regulatory mechanism still remains elusive. Here, we characterized a mutant with defective chloroplasts in Arabidopsis, termed pigment-defective mutant3 (pdm3), which exhibits a distinct albino phenotype in leaves, eventually leading to pdm3 seedling lethality under autotrophic growth conditions. Electron microscopy demonstrated that the number of thylakoids was reduced and the structure of those thylakoids was disrupted in the pdm3 mutant, which eventually led to the breakdown of chloroplasts. Sequence analysis showed that PDM3 encodes a chloroplast protein consisting of 12 pentratricopeptide repeat domains that belongs to the P subgroup. Both confocal microscopic analysis and immunoblotting in the chloroplast protein fraction showed that PDM3 was located in the stroma. Furthermore, analysis of the transcript profiles of chloroplast genes revealed that plastid-encoded polymerase-dependent transcript levels were markedly reduced, while nuclear-encoded polymerase-dependent transcript levels were increased in pdm3 mutants. In addition, we found that the splicing of introns in trnA, ndhB, and clpP-1 is also affected in pdm3. Taken together, we propose that PDM3 plays an essential role in chloroplast development in Arabidopsis.

SPL33, encoding an eEF1A-like protein, negatively regulates cell death and defense responses in rice.

Lesion-mimic mutants are useful to dissect programmed cell death and defense-related pathways in plants. Here we identified a new rice lesion-mimic mutant, spotted leaf 33 (spl33) and cloned the causal gene by a map-based cloning strategy. SPL33 encodes a eukaryotic translation elongation factor 1 alpha (eEF1A)-like protein consisting of a non-functional zinc finger domain and three functional EF-Tu domains. spl33 exhibited programmed cell death-mediated cell death and early leaf senescence, as evidenced by analyses of four histochemical markers, namely H2O2 accumulation, cell death, callose accumulation and TUNEL-positive nuclei, and by four indicators, namely loss of chlorophyll, breakdown of chloroplasts, down-regulation of photosynthesis-related genes, and up-regulation of senescence-associated genes. Defense responses were induced in the spl33 mutant, as shown by enhanced resistance to both the fungal pathogen Magnaporthe oryzae and the bacterial pathogen Xanthomonas oryzae pv. oryzae and by up-regulation of defense response genes. Transcriptome analysis of the spl33 mutant and its wild type provided further evidence for the biological effects of loss of SPL33 function in cell death, leaf senescence and defense responses in rice. Detailed analyses showed that reactive oxygen species accumulation may be the cause of cell death in the spl33 mutant, whereas uncontrolled activation of multiple innate immunity-related receptor genes and signaling molecules may be responsible for the enhanced disease resistance observed in spl33. Thus, we have demonstrated involvement of an eEF1A-like protein in programmed cell death and provided a link to defense responses in rice.

Characterization of a novel β-barrel protein (AtOM47) from the mitochondrial outer membrane of Arabidopsis thaliana.

In plant cells, mitochondria are major providers of energy and building blocks for growth and development as well as abiotic and biotic stress responses. They are encircled by two lipid membranes containing proteins that control mitochondrial function through the import of macromolecules and metabolites. Characterization of a novel β-barrel protein, OUTER MEMBRANE PROTEIN 47 (OM47), unique to the green lineage and related to the voltage-dependent anion channel (VDAC) protein family, showed that OM47 can complement a VDAC mutant in yeast. Mutation of OM47 in Arabidopsis thaliana by T-DNA insertion had no effect on the import of proteins, such as the β-barrel proteins translocase of the outer membrane 40 (TOM40) or sorting and assembly machinery 50 (SAM50), into mitochondria. Molecular and physiological analyses revealed a delay in chlorophyll breakdown, higher levels of starch, and a delay in the induction of senescence marker genes in the mutant lines. While there was a reduction of >90% in OM47 protein in mitochondria isolated from 3-week-old om47 mutants, in mitochondria isolated from 8-week-old plants OM47 levels were similar to that of the wild type. This recovery was achieved by an up-regulation of OM47 transcript abundance in the mutants. Combined, these results highlight a role in leaf senescence for this plant-specific β-barrel protein, probably mediating the recovery and recycling of chloroplast breakdown products by transporting metabolic intermediates into and out of mitochondria.

Infection strategy of Ramularia collo‐cygni and development of ramularia leaf spot on barley and alternative graminaceous hosts

Ramularia leaf spot (RLS) is a newly important disease of barley across temperate regions worldwide. Despite this recent change in importance, the infection biology of the causal agent Ramularia collo‐cygni (Rcc) remains poorly understood. Confocal microscopy of the infection process of two transgenic Rcc isolates, expressing either GFP or DsRed reporter markers, was combined with light microscopy during field infection to track the progression of Rcc in planta. Infection of stomata, including the development of a previously unreported stomatopodium structure, results in symptomless development and intercellular colonization of the mesophyll tissue. Transition to necrotrophy is associated with breakdown of host chloroplasts and the formation of aggregates of conidiophores. In addition to barley, Rcc forms a compatible interaction with winter wheat and a number of perennial grass species. An incompatible reaction was observed with two dicotyledonous species. These results provide further insights into the host interactions of this fungus and suggest that RLS could be a potential threat to other agriculturally important crops.

An Ancient Bacterial Signaling Pathway Regulates Chloroplast Function to Influence Growth and Development in Arabidopsis.

The chloroplast originated from the endosymbiosis of an ancient photosynthetic bacterium by a eukaryotic cell. Remarkably, the chloroplast has retained elements of a bacterial stress response pathway that is mediated by the signaling nucleotides guanosine penta- and tetraphosphate (ppGpp). However, an understanding of the mechanism and outcomes of ppGpp signaling in the photosynthetic eukaryotes has remained elusive. Using the model plant Arabidopsis thaliana, we show that ppGpp is a potent regulator of chloroplast gene expression in vivo that directly reduces the quantity of chloroplast transcripts and chloroplast-encoded proteins. We then go on to demonstrate that the antagonistic functions of different plant RelA SpoT homologs together modulate ppGpp levels to regulate chloroplast function and show that they are required for optimal plant growth, chloroplast volume, and chloroplast breakdown during dark-induced and developmental senescence. Therefore, our results show that ppGpp signaling is not only linked to stress responses in plants but is also an important mediator of cooperation between the chloroplast and the nucleocytoplasmic compartment during plant growth and development.

Photosynthesis in the seeds of chloroembryophytes

Depending on the presence or absence of chlorophylls in the embryo, angiosperms are divided into chloroembryophytes and leucoembryophytes. Synthesis of chlorophylls (Chl) in the chloroembryos starts in the globular stage, rises as the embryo is formed, and stops in the late phase of seed maturation. The seeds also contain carotenoids that participate in photosynthesis and act as ABA precursors. The chloroembryos contain photochemically active chloroplasts that contain all the main photosynthetic complexes at a necessary stoichiometric ratio. Dark reactions of photosynthesis in developing seeds are notable for the fact that the main source of carbon therein is sucrose arriving from the maternal plant. Therefore, function of chloroplasts mainly aims at production of NADPH and ATP that are spent on conversion of sucrose into acetyl-CoA and, subsequently, to fatty acids. The CO2 fixation system involving Rubisco and/or phosphoenolpyruvate carboxylase operates in the chloroembryos. In the course of photosynthesis, oxygen is released, which prevents hypoxia and maintains seed respiration. In late stages of ripening, the seeds enter the state of dormancy, which is associated with dehydration, disintegration of photosynthetic apparatus, Chl breakdown, and transformation of chloroplasts into plastids filled with reserve nutrient substances. At the same time, very often Chl are not destroyed completely and their residues are present in mature seeds of numerous plant species.

Hydrogen sulfide delays leaf yellowing of stored water spinach (Ipomoea aquatica) during dark-induced senescence by delaying chlorophyll breakdown, maintaining energy status and increasing antioxidative capacity

During dark-induced senescence at 12–14°C, changes of endogenous hydrogen sulfide (H2S) metabolism, chlorophyll degradation, energy status, and antioxidant capacity of Ipomoea aquatica fumigated with or without exogenously applied H2S donor sodium hydrosulfide (NaHS) were assessed. Compared to the control, NaHS treatment brought about higher contents of endogenous H2S and chlorophyll, both of which were matched with increased activities of H2S synthetic enzymes and decreased activities of chlorophyll-degrading enzyme. Electron microscopy confirmed that the breakdown of chloroplasts was reduced by NaHS. H2S not only sustained the energy status, but also decreased the respiration rate and increased antioxidant capacity. The inhibition of H2S production caused by the application of its synthetic inhibitor (dl-propargylglucine) aggravated the yellowing of I. aquatica, and this response could be rescued by exogenously applied NaHS. Nevertheless, no obvious differences were observed between treatment of hypotaurine (a scavenger of H2S) and control. These results suggest that NaHS fumigation-induced endogenous H2S might improve the energy status, leading to enhancement of antioxidant capacity and inhibition of the respiration rate, and consequently, reducing the senescence of I. aquatica.

Senescence-Associated Vacuoles, a Specific Lytic Compartment for Degradation of Chloroplast Proteins?

Degradation of chloroplasts and chloroplast components is a distinctive feature of leaf senescence. In spite of its importance in the nutrient economy of plants, knowledge about the mechanism(s) involved in the breakdown of chloroplast proteins is incomplete. A novel class of vacuoles, “senescence-associated vacuoles” (SAVs), characterized by intense proteolytic activity appear during senescence in chloroplast-containing cells of leaves. Since SAVs contain some chloroplast proteins, they are candidate organelles to participate in chloroplast breakdown. In this review we discuss the characteristics of SAVs, and their possible involvement in the degradation of Rubisco, the most abundant chloroplast protein. Finally, SAVs are compared with other extra-plastidial protein degradation pathways operating in senescing leaves.

Leaf Morphology and Ultrastructure Responses to Elevated O3 in Transgenic Bt (cry1Ab/cry1Ac) Rice and Conventional Rice under Fully Open-Air Field Conditions

BackgroundElevated tropospheric ozone severely affects not only yield but also the morphology, structure and physiological functions of plants. Because of concerns regarding the potential environmental risk of transgenic crops, it is important to monitor changes in transgenic insect-resistant rice under the projected high tropospheric ozone before its commercial release.Methodology/Principal FindingsUsing a free-air concentration enrichment (FACE) system, we investigated the changes in leaf morphology and leaf ultrastructure of two rice varieties grown in plastic pots, transgenic Bt Shanyou 63 (Bt-SY63, carrying a fusion gene of cry1Ab and cry1Ac) and its non-transgenic counterpart (SY63), in elevated O3 (E-O3) versus ambient O3 (A-O3) after 64-DAS (Days after seeding), 85-DAS and 102-DAS. Our results indicated that E-O3 had no significant effects on leaf length, leaf width, leaf area, stomatal length and stomatal density for both Bt-SY63 and SY63. E-O3 increased the leaf thickness of Bt-SY63, but decreased that of SY63. O3 stress caused early swelling of the thylakoids of chloroplasts, a significant increase in the proportion of total plastoglobule area in the entire cell area (PCAP) and a significant decrease in the proportion of total starch grain area in the entire cell area (SCAP), suggesting that E-O3 accelerated the leaf senescence of the two rice genotypes. Compared with SY63, E-O3 caused early swelling of the thylakoids of chloroplasts and more substantial breakdown of chloroplasts in Bt-SY63.Conclusions/SignificanceOur results suggest that the incorporation of cry1Ab/Ac into SY63 could induce unintentional changes in some parts of plant morphology and that O3 stress results in greater leaf damage to Bt-SY63 than to SY63, with the former coupled with higher O3 sensitivity in CCAP (the proportions of total chloroplast area in the entire cell area), PCAP and SCAP. This study provides valuable baseline information for the prospective commercial release of transgenic crops under the projected future climate.

Knockdown of GDCH gene reveals reactive oxygen species‐induced leaf senescence in rice

Glycine decarboxylase complex (GDC) is a multi-protein complex, comprising P-, H-, T- and L-protein subunits, which plays a major role in photorespiration in plants. While structural analysis has demonstrated that the H subunit of GDC (GDCH) plays a pivotal role in GDC, research on the role of GDCH in biological processes in plants is seldom reported. Here, the function of GDCH, stresses resulting from GDCH-knockdown and the interactions of these stresses with other cellular processes were studied in rice plants. Under high CO(2), the OsGDCH RNA interference (OsGDCH-RNAi) plants grew normally, but under ambient CO(2), severely suppressed OsGDCH-RNAi plants (SSPs) were non-viable, which displayed a photorespiration-deficient phenotype. Under ambient CO(2), chlorophyll loss, protein degradation, lipid peroxidation and photosynthesis decline occurred in SSPs. Electron microscopy studies showed that chloroplast breakdown and autophagy took place in these plants. Reactive oxygen species (ROS), including O2(-) and H(2)O(2), accumulated and the antioxidant enzyme activities decreased in the leaves of SSPs under ambient CO(2). The expression of transcription factors and senescence-associated genes (SAGs), which was up-regulated in SSPs after transfer to ambient CO(2), was enhanced in wild-type plants treated with H(2)O(2). Evidences demonstrate ROS induce senescence in SSPs, and transcription factors OsWRKY72 may mediate the ROS-induced senescence.