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SUMMARY: Chloroplast pre‐ribosomal RNA (rRNA) undergoes maturation, which is critical for ribosome assembly. While the central and auxiliary factors in rRNA maturation have been elucidated in bacteria, their mode of action remains largely unexplored in chloroplasts. We now reveal chloroplast‐specific factors involved in 16S rRNA maturation, Arabidopsis thaliana orthologs of bacterial RsmD methyltransferase (AtRsmD) and ribosome maturation factor RimM (AtRimM). A forward genetic screen aimed to find suppressors of the Arabidopsis yellow variegated 2 (var2) mutant defective in photosystem II quality control found a causal nonsense mutation in AtRsmD. The substantially impaired 16S rRNA maturation and translation due to the mutation rescued the leaf variegation phenotype by lowering the levels of chloroplast‐encoded proteins, including photosystem II core proteins, in var2. The subsequent co‐immunoprecipitation coupled with mass spectrometry analyses and bimolecular fluorescence complementation assay found that AtRsmD interacts with AtRimM. Consistent with their interaction, loss of AtRimM also considerably impairs 16S rRNA maturation with decelerated m2G915 modification in 16S rRNA catalyzed by AtRsmD. The atrimM mutation also rescued var2 mutant phenotypes, corroborating the functional interplay between AtRsmD and AtRimM towards modification and maturation of 16S rRNA and chloroplast proteostasis. The maturation and post‐transcriptional modifications of rRNA are critical to assembling ribosomes responsible for protein translation. Here, we revealed that the cooperative regulation of 16S rRNA m2G915 modifications by AtRsmD methyltransferase and ribosome assembly factor AtRimM contributes to 16S rRNA maturation, ribosome assembly, and proteostasis in chloroplasts. Significance Statement: The maturation and post‐transcriptional modifications of ribosomal RNA (rRNA) are critical to assembling ribosomes responsible for protein translation. Here, we revealed that the cooperative regulation of 16S rRNA m2G915 modifications by AtRsmD methyltransferase and ribosome assembly factor AtRimM contributes to 16S rRNA maturation, ribosome assembly, and proteostasis in chloroplasts. [ABSTRACT FROM AUTHOR]

Oxidative post-translational modifications of specific chloroplast proteins contribute to the initiation of retrograde signaling. The Arabidopsis thaliana EXECUTER1 (EX1) protein, a chloroplast-localized singlet oxygen (1O 2) sensor, undergoes tryptophan (Trp) 643 oxidation by 1O 2 , a chloroplast-derived and light-dependent reactive oxygen species. The indole side chain of Trp is vulnerable to 1O 2 , leading to the generation of oxidized Trp variants and priming EX1 for degradation by a membrane-bound FtsH protease. The perception of 1O 2 via Trp643 oxidation and subsequent EX1 proteolysis facilitate chloroplast-to-nucleus retrograde signaling. In this study, we discovered that the EX1-like protein EX2 also undergoes 1O 2 -dependent Trp530 oxidation and FtsH-dependent turnover, which attenuates 1O 2 signaling by decelerating EX1-Trp643 oxidation and subsequent EX1 degradation. Consistent with this finding, the loss of EX2 function reinforces EX1-dependent retrograde signaling by accelerating EX1-Trp643 oxidation and subsequent EX1 proteolysis, whereas overexpression of EX2 produces molecular phenotypes opposite to those observed in the loss–of- function mutants of EX2. Intriguingly, phylogenetic analysis suggests that EX2 may have emerged evolutionarily to attenuate the sensitivity of EX1 toward 1O 2. Collectively, these results suggest that EX2 functions as a negative regulator of the EX1 signalosome through its own 1O 2 -dependent oxidation, providing a new mechanistic insight into the regulation of EX1-mediated 1O 2 signaling. EXECUTER1 (EX1) and EX2 proteins have long been considered major players in mediating singlet oxygen (1O 2)-triggered chloroplast-to-nucleus retrograde signaling, but the specific mechanisms by which they regulate 1O 2 signaling have been unclear. This study demonstrates that the 1O 2 -driven oxidative modification of EX2 modulates EX1-mediated 1O 2 signaling by hindering the oxidation of EX1 that is essential for the initiation of 1O 2 signaling. [ABSTRACT FROM AUTHOR]

In plants, RNA-directed DNA methylation (RdDM) is a well-known de novo DNA methylation pathway that involves two plant-specific RNA polymerases, Pol IV and Pol V. In this study, we discovered and characterized an RdDM factor, RDM15. Through DNA methylome and genome-wide siRNA analyses, we show that RDM15 is required for RdDM-dependent DNA methylation and siRNA accumulation at a subset of RdDM target loci. We show that RDM15 contributes to Pol V-dependent downstream siRNA accumulation and interacts with NRPE3B, a subunit specific to Pol V. We also show that the C-terminal tudor domain of RDM15 specifically recognizes the histone 3 lysine 4 monomethylation (H3K4me1) mark. Structure analysis of RDM15 in complex with the H3K4me1 peptide showed that the RDM15 tudor domain specifically recognizes the monomethyllysine through an aromatic cage and a specific hydrogen bonding network; this chemical feature-based recognition mechanism differs from all previously reported monomethyllysine recognition mechanisms. RDM15 and H3K4me1 have similar genome-wide distribution patterns at RDM15-dependent RdDM target loci, establishing a link between H3K4me1 and RDM15-mediated RdDM in vivo. In summary, we have identified and characterized a histone H3K4me1-specific binding protein as an RdDM component, and structural analysis of RDM15 revealed a chemical feature-based lower methyllysine recognition mechanism. In plants, RNA-directed DNA methylation (RdDM) is a de novo DNA methylation pathway that is responsible for transcriptional silencing of repetitive elements. Here, the authors characterized a new RdDM factor, RDM15, and show that it is required for RdDM-dependent DNA methylation and siRNA accumulation at a subset of RdDM target loci. [ABSTRACT FROM AUTHOR]

Summary: The photosynthetic bacterial phycobiliprotein lyases, also called CpcT lyases, catalyze the biogenesis of phycobilisome, a light‐harvesting antenna complex, through the covalent attachment of chromophores to the antenna proteins. The Arabidopsis CRUMPLED LEAF (CRL) protein is a homolog of the cyanobacterial CpcT lyase. Loss of CRL leads to multiple lesions, including localized foliar cell death, constitutive expression of stress‐related nuclear genes, abnormal cell cycle, and impaired plastid division. Notwithstanding the apparent phenotypes, the function of CRL still remains elusive. To gain insight into the function of CRL, we examined whether CRL still retains the capacity to bind with the bacterial chromophore phycocyanobilin (PCB) and its plant analog phytochromobilin (PΦB). The revealed structure of the CpcT domain of CRL is comparable to that of the CpcT lyase, despite the low sequence identity. The subsequent in vitro biochemical assays found, as shown for the CpcT lyase, that PCB/PΦB binds to the CRL dimer. However, some mutant forms of CRL, substantially compromised in their bilin‐binding ability, still restore the crl‐induced multiple lesions. These results suggest that although CRL retains the bilin‐binding pocket, it seems not functionally associated with the crl‐induced multiple lesions. Significance Statement: Our study revealed that the Arabidopsis CRUMPLED LEAF (CRL) protein retains a bilin‐binding pocket similar to its homolog cyanobacterial bilin lyase. The subsequent reverse genetic approach establishes that the retained bilin‐binding aptitude is not essential for CRL function in Arabidopsis, assuring that CRL evolved with a different function. [ABSTRACT FROM AUTHOR]

As a light-harvesting organelle, the chloroplast inevitably produces a substantial amount of reactive oxygen species (ROS) primarily through the photosystems. These ROS, such as superoxide anion, hydrogen peroxide, hydroxyl radical, and singlet oxygen, are potent oxidizing agents, thereby damaging the photosynthetic apparatus. On the other hand, it became increasingly clear that ROS act as beneficial tools under photo-oxidative stress conditions by stimulating chloroplast-nucleus communication, a process called retrograde signaling (RS). These ROS-mediated RS cascades appear to participate in a broad spectrum of plant physiology, such as acclimation, resistance, programmed cell death (PCD), and growth. Recent reports imply that ROS-driven oxidation of RS-associated components is essential in sensing and responding to an increase in ROS contents. ROS appear to activate RS pathways via reversible or irreversible oxidation of sensor molecules. This review provides an overview of the emerging perspective on the topic of "oxidative modification-associated retrograde signaling." [ABSTRACT FROM AUTHOR]

Singlet oxygen (1O2) is an excited state of molecular oxygen with an electron spin shift in the molecular orbitals, which is extremely unstable and highly reactive. In plants, 1O2 is primarily generated as a byproduct of photosynthesis in the photosystem II reaction center (PSII RC) and the light-harvesting antenna complex (LHC) in the grana core (GC). This occurs upon the absorption of light energy when the excited chlorophyll molecules in the PSII transfer the excess energy to molecular oxygen, thereby generating 1O2. As a potent oxidant, 1O2 promotes oxidative damage. However, at sub-lethal levels, it initiates chloroplast-to-nucleus retrograde signaling to contribute to plant stress responses, including acclimation and cell death. The thylakoid membranes comprise two spatially separated 1O2 sensors: β-carotene localized in the PSII RC in the GC and the nuclear-encoded chloroplast protein EXECUTER1 (EX1) residing in the non-appressed grana margin (GM). Finding EX1 in the GM suggests the existence of an additional source of 1O2 in the GM and the presence of two distinct 1O2-signaling pathways. In this review, we mainly discuss the genesis and impact of 1O2 in plant physiology. [ABSTRACT FROM AUTHOR]

Chloroplast–nucleus communication takes place via processes called anterograde and retrograde signaling pathways. Discovery of the retrograde signaling pathways from the chloroplasts to the nucleus also raised an intriguing proposition that chloroplasts may serve as environmental sensors since multitudes of environmental factors disturb chloroplastic homeostasis. Certain chloroplastic perturbations, mostly impairing transcription/translation, are coupled to the repression of photosynthesis-associated nuclear genes (PhANGs), thus finely coordinating photosynthetic and chloroplastic homeostasis. The unbiased forward genetic screen in Arabidopsis leads to the identification of six independent loci called GENOMES UNCOUPLED (GUN), whose inactivation was found to de-repress the expression of PhANGs under certain conditions promoting retrograde signaling. Of the six GUNs, five encode proteins associated with tetrapyrrole biosynthesis and one, namely GUN1, encodes a member of the pentatricopeptide repeat protein family. Despite the fact that GUN1 plays a role as a central signaling mediator for retrograde communication, the molecular details of GUN1 protein still remain to be elucidated. Here, we recapitulate our current understanding of the GUN1-mediated retrograde signaling pathway and propose a possible mode of action of GUN1 in the chloroplasts together with different aspects of GUN1 protein activity that deserve further investigation. [ABSTRACT FROM AUTHOR]

Cellular protein homeostasis (proteostasis) is maintained through the balance between de novo synthesis and proteolysis. The unfolded/misfolded protein response (UPR) that is triggered by stressed endoplasmic reticulum (ER) also plays an important role in proteostasis in both plants and animals. Although ER-triggered UPR has been extensively studied in plants, the molecular mechanisms underlying mitochondrial and chloroplastic UPRs are largely uncharacterized despite the fact that these organelles are sites of production of harmful reactive oxygen species (ROS), which damage proteins. In this study, we demonstrate that chloroplasts of the Arabidopsis yellow leaf variegation 2 (var2) mutant, which lacks the metalloprotease FtsH2, accumulate damaged chloroplast proteins and trigger a UPR-like response, namely the accumulation of a suite of chloroplast proteins involved in protein quality control (PQC). These PQC proteins include heat-shock proteins, chaperones, proteases, and ROS detoxifiers. Given that FtsH2 functions primarily in photosystem II proteostasis, the accumulation of PQC-related proteins may balance the FtsH2 deficiency. Moreover, the apparent up-regulation of the cognate transcripts indicates that the accumulation of PQC-related proteins in var2 is probably mediated by retrograde signaling, indicating the occurrence of a UPR-like response in var2. [ABSTRACT FROM AUTHOR]

In this paper, the problem of reducing the peak-to-average power ratio (PAPR) of a discrete Fourier transform spread orthogonal frequency-division multiplexing signal is considered for the transmission of BPSK symbols. In particular, a joint optimization of the constellation-rotation angle and the pulse-shaping vector is conducted with or without the allocation of additional sub-carriers. To avoid the exponential complexity required to evaluate the PAPR, a computationally efficient upper bound on the worst-case PAPR is derived and adopted as the objective function. Unlike conventional upper bounds, the proposed upper bound exploits the structure in the relative phase among symbol waveforms and incorporates the constellation-rotation effect. The signal-to-interference-plus-noise ratio (SINR) at the output of a constellation-derotating matched filter is also derived along with a generalized Nyquist criterion and adopted as the inequality constraint function. Then, joint optimizations are conducted through numerical search for various signal parameters and target SINRs. Surprisingly, the optimal rotation angle is neither 0 nor $\pi /2$. It is shown that the proposed combinations significantly outperform the existing combinations of the rotation angle and the shaping vector when the signal is amplified by a nonlinear power amplifier. [ABSTRACT FROM AUTHOR]

Clathrin-mediated vesicular formation and trafficking are responsible for molecular cargo transport and signal transduction among organelles. Our previous study shows that CHLOROPLAST VESICULATION (CV)-containing vesicles (CVVs) are generated from chloroplasts for chloroplast degradation under abiotic stress. Here, we show that CV interacts with the clathrin heavy chain (CHC) and induces vesicle budding toward the cytosol from the chloroplast inner envelope membrane. In the defective mutants of CHC2 and the dynamin-encoding DRP1A , CVV budding and releasing from chloroplast are impeded. The mutations of CHC2 inhibit CV-induced chloroplast degradation and hypersensitivity to water stress. Moreover, CV-CHC2 interaction is impaired by the oxidized GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE (GAPC). GAPC1 overexpression suppresses CV-mediated chloroplast degradation and hypersensitivity to water stress, while CV silencing alleviates the hypersensitivity of the gapc1gapc2 plant to water stress. Together, our work identifies a pathway of clathrin-assisted CVV budding outward from chloroplast, which is involved in chloroplast degradation and stress response. [Display omitted] • CV-CHC interaction induces vesicle budding outward from chloroplast inner envelope membrane • CVV formation is impeded in the chc2 and drp1a mutants • CV-CHC2 interaction is impaired by the oxidized GAPC • GAPC suppresses CV-mediated chloroplast degradation and stress response CV-containing vesicles (CVVs) are involved in stress-induced chloroplast degradation. Pan et al. show that CV interacts with CHC2 to induce vesicle budding outward from the chloroplast inner envelope membrane. DRP1A releases CVVs from chloroplast through membrane scission. The oxidized GAPC inhibits CV-CHC2 interaction and CV-induced chloroplast degradation. [ABSTRACT FROM AUTHOR]

We present here a Tribute to Klaus Apel (1942-2017), a photosynthesis pioneer—an authority on plant molecular genetics—in five parts. The first section is a prologue. The second section deals with a chronological discussion of Apel’s research life, prepared by the editor Govindjee; it is based on a website article at the Boyce Thompson Institute (BTI) by Patricia Waldron (https://btiscience.org/explore-bti/news/post/bti-says-goodbye-klaus-apel/), as approved for use here by Keith C. Hannon and David Stern of BTI. The third section, which focuses on Apel’s pioneering work on singlet oxygen-mediated EXECUTER-dependent signaling in plants, is written by two of us (J-DR and CK). The fourth section includes three selected reminiscences, one from BTI and two from ETH (Eidgenössische Technische Hochschule). This tribute ends with section five, which is a very brief presentation of Klaus Apel’s personal life, by Wiebke Apel. [ABSTRACT FROM AUTHOR]

Abstract: Singlet oxygen (1O2) is a prime cause of photo‐damage of the photosynthetic apparatus. The chlorophyll molecules in the photosystem II reaction center and in the light‐harvesting antenna complex are major sources of 1O2 generation. It has been thought that the generation of 1O2 mainly takes place in the appressed regions of the thylakoid membranes, namely, the grana core, where most of the active photosystem II complexes are localized. Apart from being a toxic molecule, new evidence suggests that 1O2 significantly contributes to chloroplast‐to‐nucleus retrograde signalling that primes acclimation and cell death responses. Interestingly, recent studies reveal that chloroplasts operate two distinct 1O2‐triggered retrograde signalling pathways in which β‐carotene and a nuclear‐encoded chloroplast protein EXECUTER1 play essential roles as signalling mediators. The coexistence of these mediators raises several questions: their crosstalk, source(s) of 1O2, downstream signalling components, and the perception and reaction mechanism of these mediators towards 1O2. In this review, we mainly discuss the molecular genetic basis of the mode of action of these two putative 1O2 sensors and their corresponding retrograde signalling pathways. In addition, we also propose the possible existence of an alternative source of 1O2, which is spatially and functionally separated from the grana core. [ABSTRACT FROM AUTHOR]

The article explains fundamental trade-offs in waveform design and shows how they can be optimized for quadrature amplitude modulation- filter-bank multi-carrier (QAM-FBMC). Topics include efforts to improve spectral efficiency; pros and cons of multicarrier waveforms and performance comparison between cyclic prefix orthogonal frequency-division multiplexing (CP-OFDM) and QAM-FBMC. It mentions that asynchronous heterogeneous network scenario as key features for 5G mobile communications.

In this paper, we propose a new sequence generation methodology to make a constant amplitude zero autocorrelation (CAZAC) preamble available in offset Quadrature Amplitude Modulation-based filter bank multicarrier (FBMC) synchronization. The role of the new sequence is to compensate the filtering and symbol overlap of the FBMC structure so as to finally be able to make a CAZAC preamble in time domain. To derive the new sequence, we formulate a symbol overlap model that shows the relation between the new sequence and a CAZAC preamble. On the other hand, the offset modulation format is a constraint that needs to be considered in the design of the new sequence. By using a property of this offset modulation constraint, we derive a closed-form expression of the new sequence from the symbol overlap model. In addition, we extend an advanced model to create a pseudo-CAZAC preamble that is not data-dependent, thus enabling convenience of implementation. Simulation results show the superiority of the proposed model with respect to synchronization accuracy, out-of-band radiation, and peak to average power ratio. Moreover, we verify that the proposed model is well-suited for use in the cellular environment through complexity and resource efficiency comparisons. [ABSTRACT FROM PUBLISHER]

A filter-bank multicarrier–quadrature amplitude modulation (FBMC-QAM) system with two prototype filters for transmitting QAM symbol is proposed, and the orthogonality conditions for the FBMC-QAM system without the intrinsic interference is derived. The proposed transmitter performs the individual filtering for the even and odd-numbered sub-carrier symbols, respectively. In order to satisfy the derived orthogonality conditions, we perform a sub-block wise reverse ordering procedure for the outputs of the odd-numbered sub-carrier filter. Also, this paper shows that the proposed FBMC-QAM system satisfies the suggested orthogonality conditions and can use the multiple-input multiple-output transmission schemes as the conventional orthogonal frequency division multiplexing (OFDM) systems. The signal-to-interference power ratio and the bit error rate (BER) for the proposed FBMC-QAM system are evaluated. Numerical results show that the proposed FBMC-QAM system has almost the same BER performance compared with the FBMC-OQAM and OFDM systems. [ABSTRACT FROM PUBLISHER]

Necrotrophic fungal pathogens produce toxic compounds that induce cell death in infected plants. Often, the primary targets of these toxins and the way a plant responds to them are not known. In the present work, the effect of tenuazonic acid ( TeA), a non-host-specific toxin of A lternaria alternata, on A rabidopsis thaliana has been analysed. TeA blocks the QB-binding site at the acceptor side of photosystem II ( PSII). As a result, charge recombination at the reaction centre ( RC) of PSII is expected to enhance the formation of the excited triplet state of the RC chlorophyll that promotes generation of singlet oxygen (1 O 2). 1 O 2 activates a signalling pathway that depends on the two EXECUTER ( EX) proteins EX1 and EX2 and triggers a programmed cell death response. In seedlings treated with TeA at half-inhibition concentration 1 O 2-mediated and EX-dependent signalling is activated as indicated by the rapid and transient up-regulation of 1 O 2-responsive genes in wild type, and its suppression in ex 1/ ex 2 mutants. Lesion formation occurs when seedlings are exposed to higher concentrations of TeA for a longer period of time. Under these conditions, the programmed cell death response triggered by 1 O 2-mediated and EX-dependent signalling is superimposed by other events that also contribute to lesion formation. [ABSTRACT FROM AUTHOR]

Singlet oxygen (O)-mediated signaling has been established in the conditional fluorescent ( flu) mutant of Arabidopsis. In the dark, the flu mutant accumulates free protochlorophyllide (Pchlide), a photosensitizer that in the light generates O. The release of O leads to growth inhibition of mature plants and bleaching of seedlings. These O-mediated responses depend on two plastid proteins, EXECUTER (EX) 1 and 2. An ex1/ ex2/ flu mutant accumulates in the dark Pchlide and upon illumination generates similar amounts of O as flu, but O-mediated responses are abrogated in the triple mutant. The O- and EX-dependent signaling pathway operates also in wild type placed under light stress. However, it does not act alone as in flu, but interacts with other signaling pathways that modulate O-mediated responses. Depending on how severe the light stress is, O- and EX-dependent signaling may be superimposed by O-mediated signaling that does not depend on EX and is associated with photo-oxidative damage. Because of its high reactivity and short half-life, O is unlikely to be a signal that is translocated across the chloroplast envelope, but is likely to interact with other plastid components close to its site of production and to generate more stable signaling molecules during this interaction. Depending on the site of O production and the severity of stress, different signaling molecules may be expected that give rise to different O-mediated responses. [ABSTRACT FROM AUTHOR]

Seedlings of Arabidopsis have been exposed to norflurazon. Depending on the developmental stage at which seedlings were first exposed to the inhibitor, enhanced production of reactive oxygen species occurred and, among others, 1O2-mediated and EXECUTER-dependent retrograde signaling was induced.Chloroplast development depends on the synthesis and import of a large number of nuclear-encoded proteins. The synthesis of some of these proteins is affected by the functional state of the plastid via a process known as retrograde signaling. Retrograde plastid-to-nucleus signaling has been often characterized in seedlings of Arabidopsis thaliana exposed to norflurazon (NF), an inhibitor of carotenoid biosynthesis. Results of this work suggested that, throughout seedling development, a factor is released from the plastid to the cytoplasm that indicates a perturbation of plastid homeostasis and represses nuclear genes required for normal chloroplast development. The identity of this factor is still under debate. Reactive oxygen species (ROS) were among the candidates discussed as possible retrograde signals in NF-treated plants. In the present work, this proposed role of ROS has been analyzed. In seedlings grown from the very beginning in the presence of NF, ROS-dependent signaling was not detectable, whereas, in seedlings first exposed to NF after light-dependent chloroplast formation had been completed, enhanced ROS production occurred and, among others, 1O2-mediated and EXECUTER-dependent retrograde signaling was induced. Hence, depending on the developmental stage at which plants are exposed to NF, different retrograde signaling pathways may be activated, some of which are also active in non-treated plants under light stress. [ABSTRACT FROM AUTHOR]

In this paper, we derive the exact probability distribution of post-scheduling signal-to-interference-plus-noise ratio (SINR) considering both user feedback and scheduling. We also develop an optimized adaptive modulation scheme in orthogonal random beamforming systems with M transmit antennas and K single-antenna users. The exact probability distributions of each user's feedback SINR and the exact post-scheduling SINR are derived rigorously by direct integration and multinomial distribution. It is also shown that the derived cumulative distribution function (CDF) of the post-scheduling SINR happens to be identical to the the existing approximate CDF for SINR higher than 0 dB. The closed form expressions of system performance, such as average spectral efficiency (ASE) and average bit error ratio (A-BER), are derived using the CDF of the post-scheduling SINR. The optimal SINR thresholds that maximize the ASE with a target A-BER constraint are solved using the derived closed form CDF and a Lagrange multiplier. Key contributions of this paper include the derivation of the exact CDF of post-scheduling SINR by direct integration, and its application to an optimized adaptive modulation based on a Lagrange multiplier. Simulations show the correspondence between theoretical and empirical CDF's, and the performance improvement of the proposed adaptive modulation method in terms of ASE. [ABSTRACT FROM PUBLISHER]

Recently the porA- 1 null mutant of Arabidopsis thaliana has been identified, which contains an insertion of the Dissociation ( Ds) element in the PORA gene (Paddock et al. in Plant Mol Biol 78:447-460, ). Light-grown porA- 1 seedlings suffer from a drastically reduced chlorophyll content and a developmental arrest beyond the cotyledon stage, suggesting that PORA is not only transiently involved in initiating chlorophyll synthesis during illumination of etiolated seedlings but is also essential for normal growth and plant development. Here we report the presence of a second Ds element in this porA- 1 mutant line that inactivates the Speechless gene required for stomata formation. Similar to porA- 1, speechless seedlings are severely impaired in their development. Our results suggest that the lack of stomata in porA- 1 may contribute to the dwarfed phenotype of the mutant and thus emphasizes the need to re-address the proposed role of PORA during plant development by studying a porA mutant that retains its stomata formation. [ABSTRACT FROM AUTHOR]

Enhanced levels of singlet oxygen (1O2) in chloroplasts trigger programmed cell death. The impact of 1O2 production in chloroplasts was monitored first in the conditional fluorescent (flu) mutant of Arabidopsis thaliana that accumulates 1O2 upon a dark/light shift. The onset of 1O2 production is rapidly followed by a loss of chloroplast integrity that precedes the rupture of the central vacuole and the final collapse of the cell. Inactivation of the two plastid proteins EXECUTER (EX1) and EX2 in the flu mutant abrogates these responses, indicating that disintegration of chloroplasts is due to EX-dependent signaling rather than 1O2 directly. In flu seedlings, 1O2 -mediated cell death signaling operates as a default pathway that results in seedlings committing suicide. By contrast, EX-dependent signaling in the wild type induces the formation of microlesions without decreasing the viability of seedlings. 1O2 -mediated and EX-dependent loss of plastid integrity and cell death in these plants occurs only in cells containing fully developed chloroplasts. Our findings support an as yet unreported signaling role of 1O2 in the wild type exposed to mild light stress that invokes photoinhibition of photosystem II without causing photooxidative damage of the plant. [ABSTRACT FROM AUTHOR]

Summary Retrograde plastid-to-nucleus signaling tightly controls and coordinates the nuclear and plastid gene expression that is required for plastid biogenesis and chloroplast activity. As chloroplasts act as sensors of environmental changes, plastid-derived signaling also modulates stress responses of plants by transferring stress-related signals and altering nuclear gene expression. Various mutant screens have been undertaken to identify constituents of plastid signaling pathways. Almost all mutations identified in these screens target plastid-specific but not extraplastidic functions. They have been suggested to define either genuine constituents of retrograde signaling pathways or components required for the synthesis of plastid signals. Here we report the characterization of the constitutive activator of AAA-ATPase ( caa33) mutant, which reveals another way of how mutations that affect plastid functions may modulate retrograde plastid signaling. caa33 disturbs a plastid-specific function by impeding plastid division, and thereby perturbing plastid homeostasis. This results in preconditioning plants by activating the expression of stress genes, enhancing pathogen resistance and attenuating the capacity of the plant to respond to plastid signals. Our study reveals an intimate link between chloroplast activity and the susceptibility of the plant to stress, and emphasizes the need to consider the possible impact of preconditioning on retrograde plastid-to-nucleus signaling. [ABSTRACT FROM AUTHOR]

The conditional flu mutant of Arabidopsis thaliana generates singlet oxygen (1O2) in plastids during a dark-to-light shift. Seedlings of flu bleach and die, whereas mature plants stop growing and develop macroscopic necrotic lesions. Several suppressor mutants, dubbed singlet oxygen-linked death activator ( soldat), were identified that abrogate 1O2-mediated cell death of flu seedlings. One of the soldat mutations, soldat10, affects a gene encoding a plastid-localized protein related to the human mitochondrial transcription termination factor mTERF. As a consequence of this mutation, plastid-specific rRNA levels decrease and protein synthesis in plastids of soldat10 is attenuated. This disruption of chloroplast homeostasis in soldat10 seedlings affects communication between chloroplasts and the nucleus and leads to changes in the steady-state concentration of nuclear gene transcripts. The soldat10 seedlings suffer from mild photo-oxidative stress, as indicated by the constitutive up-regulation of stress-related genes. Even though soldat10/ flu seedlings overaccumulate the photosensitizer protochlorophyllide in the dark and activate the expression of 1O2-responsive genes after a dark-to-light shift they do not show a 1O2-dependent cell death response. Disturbance of chloroplast homeostasis in emerging soldat10/ flu seedlings seems to antagonize a subsequent 1O2-mediated cell death response without suppressing 1O2-dependent retrograde signaling. The results of this work reveal the unexpected complexity of what is commonly referred to as ‘plastid signaling’. [ABSTRACT FROM AUTHOR]

Low and high temperatures are life-threatening stress factors, diminishing plant productivity. One of the earliest responses of plants to stress is a rapid burst of reactive oxygen species (ROS) in chloroplasts. Widespread efforts over the past decade shed new light on the chloroplast as an environmental sensor, translating the environmental fluctuation into varying physiological responses by utilizing distinct retrograde (chloroplast-to-nucleus) signals. Recent studies have unveiled that chloroplasts mediate a similar unfolded/misfolded/damaged protein response (cpUPR) as observed in the endoplasmic reticulum and mitochondria. Although observing cpUPR is not surprising since the chloroplast is a prime organelle producing harmful ROS, the intertwined relationship among ROS, protein damage, and chloroplast protein quality controls (cpPQCs) with retrograde signaling has recently been reported. This finding also gives rise to critical attention on chloroplast proteins involved in cpPQCs, ROS detoxifiers, transcription/translation, import of precursor proteins, and assembly/maturation, the deficiency of which compromises chloroplast protein homeostasis (proteostasis). Any perturbation in the protein may require readjustment of proteostasis by transmitting retrograde signal(s) to the nucleus, whose genome encodes most of the chloroplast proteins involved in proteostasis. This review focuses on recent findings on cpUPR and chloroplast-targeted FILAMENTOUS TEMPERATURE-SENSITIVE H proteases involved in cpPQC and retrograde signaling and their impacts on plant responses to temperature stress. [ABSTRACT FROM AUTHOR]

When plant cells are under environmental stress, several chemically distinct reactive oxygen species (ROS) are generated simultaneously in various intracellular compartments and these can cause oxidative damage or act as signals. The conditional flu mutant of Arabidopsis, which generates singlet oxygen in plastids during a dark-to-light transition, has allowed the biological activity of singlet oxygen to be determined, and the criteria to distinguish between cytotoxicity and signalling of this particular ROS to be defined. The genetic basis of singlet-oxygen-mediated signalling has been revealed by the mutation of two nuclear genes encoding the plastid proteins EXECUTER (EX)1 and EX2, which are sufficient to abrogate singlet-oxygen-dependent stress responses. Conversely, responses due to higher cytotoxic levels of singlet oxygen are not suppressed in the ex1/ex2 background. Whether singlet oxygen levels lower than those that trigger genetically controlled cell death activate acclimation is now under investigation. [ABSTRACT FROM AUTHOR]

The NADPH-dependent protochlorophyllide (Pchlide) oxidoreductase (POR) is unique because it is a photoenzyme that requires light for its catalytic activity and uses Pchlide itself as a photoreceptor. In Arabidopsis, there are three structurally related PORs, denoted PORA, PORB, and PORC. The import of one of them, PORA, into plastids of cotyledons is substrate dependent. This substrate dependence is demonstrated in intact seedlings of wild-type Arabidopsis and two mutants, xantha2, which is devoid of Pchlide, and flu, which upon redarkening rapidly accumulates Pchlide. In true leaves, PORA up, take does not require the presence of Pchlide. The organ specificity of the substrate-dependent import of PORA reveals a means of controlling plastid protein translocation that is closely associated with a key step in plant development, the light-dependent transformation of cotyledons from a storage organ to a photosynthetically active leaf. [ABSTRACT FROM AUTHOR]

Regulation of tetrapyrrole biosynthesis in higher plants has been attributed to negative feedback control of steps prior to δ-aminolevulinic acid (ALA) formation. One of the first mutants with a defect in this control had been identified in barley. The tigrina (tig) d mutant accumulates 10–15-fold higher amounts of protochlorophyllide than wild type, when grown in the dark. The identity of the TIGRINA d protein and its mode of action are not known yet. Initially this protein had been proposed to act as a repressor of genes that encode enzymes involved in early steps of ALA formation, but subsequent attempts to confirm this experimentally failed. Here we demonstrate that the TIGRINA d gene of barley is an ortholog of the FLU gene of Arabidopsis thaliana. The FLU protein is a nuclear-encoded plastid protein that plays a key role in negative feedback control of chlorophyll biosynthesis in higher plants. Sequencing of the FLU gene of barley revealed a frame shift mutation in the FLU gene of the tig d mutant that results in the loss of two tetratricopeptide repeats that in the FLU protein of Arabidopsis are essential for its biological activity. This mutation cosegregates strictly with the tigrina phenotype within the F1 population of a heterozygous tig d mutant, thus providing additional support for the flu gene being responsible for the tigrina phenotype of barley. [Copyright &y& Elsevier]

Environmental information perceived by chloroplasts can be translated into retrograde signals that alter the expression of nuclear genes. Singlet oxygen (1O2) generated by photosystem II (PSII) can cause photo-oxidative damage of PSII but has also been implicated in retrograde signaling. We previously reported that a nuclear-encoded chloroplast FtsH2 metalloprotease coordinates 1O2-triggered retrograde signaling by promoting the degradation of the EXECUTER1 (EX1) protein, a putative 1O2 sensor. Here, we show that a 1O2-mediated oxidative post-translational modification of EX1 is essential for initiating 1O2-derived signaling. Specifically, the Trp643 residue in DUF3506 domain of EX1 is prone to oxidation by 1O2. Both the substitution of Trp643 with 1O2-insensitive amino acids and the deletion of the DUF3506 domain abolish the EX1-mediated 1O2 signaling. We thus provide mechanistic insight into how EX1 senses 1O2 via Trp643 located in the DUF3506 domain. Singlet oxygen generated by photosynthesis under photo-oxidative stress conditions triggers retrograde signaling from plastids to nuclei. Here, the authors show that singlet oxygen perception and subsequent signaling events require oxidative post-translational modification of the EXECUTER1 protein. [ABSTRACT FROM AUTHOR]

Derived are the exact cumulative distribution functions (CDFs) of each user's feedback signal-to-interference-plus-noise ratio (SINR), and the post-scheduling SINR in orthogonal random beamforming systems with M transmit antennas and K single-antenna users considering both user feedback and scheduling. A key contribution is to derive the exact CDF of the post-scheduling SINR by direct integration, and it is verified by Monte-Carlo simulations. It is also shown that the existing approximate CDF is different from the exact distribution for SINR smaller than 0 dB. [ABSTRACT FROM AUTHOR]

\boldmath In this letter, we analyze the post-scheduling SINR mismatch of a multiuser orthogonal random beamforming (ORBF) system. The mismatch probability is derived for a Gaussian broadcast channel with M transmit antennas and K single-antenna users. Numerical results are shown to be identical to theoretical results. The probability analysis enables us to calculate the minimum number of users to satisfy a target mismatch probability. Another way to use the mismatch probability is to control CQI mismatch handling mechanisms. [ABSTRACT FROM AUTHOR]