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Plant & Cell Physiology

Hiroki Tsutsui, Tetsuya Higashiyama
The CRISPR/Cas9 system is widely used as a tool for genome engineering in various organisms. A complex consisting of Cas9 and single guide RNA (sgRNA) induces a DNA double-strand break in a sequence-specific manner, resulting in knockout. Some binary vectors for CRISPR/Cas9 in plants have been reported, but there is a problem with low efficiency. Here, we present a newly developed, highly efficient CRISPR/Cas9 vector for Arabidopsis thaliana, pKAMA-ITACHI Red (pKIR), harboring the RIBOSOMAL PROTEIN S5 A (RPS5A) promoter to drive Cas9 The RPS5A promoter maintains high constitutive expression at all developmental stages starting from the egg cell and including meristematic cells...
November 17, 2016: Plant & Cell Physiology
Moshiul Islam, Wenxiu Ye, Daiki Matsushima, Shintaro Munemasa, Eiji Okuma, Yoshimasa Nakamura, Sanaullah Biswas, Jun'ichi Mano, Yoshiyuki Murata
Drought is responsible for massive reduction in crop yields. In response to drought, plants synthesize the hormone abscisic acid (ABA), which induces stomatal closure, thus reducing water loss. In guard cells, ABA triggers production of reactive oxygen species (ROS), which is mediated by NAD(P)H oxidases. The production of ROS is a key factor for ABA-induced stomatal closure, but it remains to be clarified how the production of ROS is transduced into downstream signaling components in guard cells. We investigated roles of reactive carbonyl species (RCS) in ABA-induced stomatal closure using transgenic tobacco (Nicotiana tabacum) overexpressing Arabidopsis 2-alkenal reductase (AER-OE), which scavenges RCS...
November 12, 2016: Plant & Cell Physiology
Setsuko Fukushima, Masaki Mori, Shoji Sugano, Hiroshi Takatsuji
WRKY62 is a transcriptional repressor regulated downstream of WRKY45, a central transcription factor of the salicylic acid signaling pathway in rice. Previously, WRKY62 was reported to negatively regulate defence. However, our expressional analysis using WRKY62-knockdown rice indicated that WRKY62 positively regulates defence genes, including diterpenoid phytoalexin biosynthetic genes and their transcriptional regulator DPF Blast and leaf blight resistance tests also showed that WRKY62 is a positive defence regulator...
November 10, 2016: Plant & Cell Physiology
Xiaoyang Zhu, Eugénie Robe, Lucile Jomat, Didier Aldon, Christian Mazars, Jean-Philippe Galaud
Calcium is a universal second messenger involved in various cellular processes including plant development and stress responses. Its conversion into biological responses requires the presence of calcium sensor relays such as calmodulin (CaM) and calmodulin-like proteins (CMLs). While the role of CaM is well described, CML functions remain largely uncharacterized. Here, we show that the Arabidopsis CML8 expression is strongly and transiently induced by Pseudomonas syringae and reverse genetic approaches indicated that the over-expression of CML8 confers on plants a better resistance to pathogenic bacteria compared to wild type, knock-down and knock-out lines indicating that CML8 participates as a positive regulator in plant immunity...
November 10, 2016: Plant & Cell Physiology
Amit Srivastava, Klaus Brilisauer, Ashutosh K Rai, Anand Ballal, Karl Forchhammer, Anil K Tripathi
Alternative sigma factors belonging to the Group 3 are thought to play an important role in the adaptation of cyanobacteria to environmental challenges by altering expression of genes needed for coping with such stresses. In this study, the role of an alternative sigma factor, SigJ, was analyzed in the filamentous nitrogen-fixing cyanobacterium, Anabaena sp. PCC 7120 by knocking down the expression of the sigJ gene (alr0277) employing an anti-sense RNA-mediated approach. In the absence of any stress, the knock-down (KD0277) or the wild-type strain both grew similarly...
November 10, 2016: Plant & Cell Physiology
Whitney L Dolan, Clint Chapple
The Mediator complex is a large, multi-subunit, transcription co-regulator that is conserved across eukaryotes. Studies of the Arabidopsis Mediator complex and its subunits have shown that it functions in nearly every aspect of plant development and fitness. In addition to revealing mechanisms of regulation of plant-specific pathways, studies of plant Mediator complexes have the potential to shed light on the conservation and divergence of Mediator structure and function across Kingdoms and plant lineages. The majority of insights into plant Mediator function have come from Arabidopsis because it is the only plant from which Mediator has been purified and from which an array of Mediator mutants have been isolated by forward and reverse genetics...
November 10, 2016: Plant & Cell Physiology
Lotte Caarls, Dieuwertje Van der Does, Richard Hickman, Wouter Jansen, Marcel C Van Verk, Silvia Proietti, Oscar Lorenzo, Roberto Solano, Corné M J Pieterse, Saskia C M Van Wees
Salicylic acid (SA) and jasmonic acid (JA) cross-communicate in the plant immune signaling network to finely regulate induced defenses. In Arabidopsis, SA antagonizes many JA-responsive genes, partly by targeting the ETHYLENE RESPONSE FACTOR (ERF)-type transcriptional activator ORA59. Members of the ERF transcription factor family typically bind to GCC-box motifs in the promoters of JA- and ethylene-responsive genes, thereby positively or negatively regulating their expression. The GCC-box motif is sufficient for SA-mediated suppression of JA-responsive gene expression...
November 10, 2016: Plant & Cell Physiology
Ryudo Ohbayashi, Jun-Ya Yamamoto, Satoru Watanabe, Yu Kanesaki, Taku Chibazakura, Shin-Ya Miyagishima, Hirofumi Yoshikawa
Cyanobacteria exhibit light-dependent cell growth since most of their cellular energy is obtained by photosynthesis. In Synechococcus elongatus PCC 7942, one of the model cyanobacterium, DNA replication depends on the photosynthetic electron transport. However, the critical signal for regulatory mechanism of DNA replication has not been identified. In addition, conservation of this regulatory mechanism has not been investigated among cyanobacteria. To understand this regulatory signal and its dependence on light, we examined the regulation of DNA replication under both light and dark condition among three model cyanobacteria, S...
November 10, 2016: Plant & Cell Physiology
Tomoko Hirano, Teun Munnik, Masa H Sato
Phosphoinositides play an important role in various membrane trafficking events in eukaryotes. One of them, however, phosphatidylinositol 3,5-bisphosphate [PI(3,5)P2], has not been studied widely in plants. Using a combination of fluorescent reporter proteins and the PI(3,5)P2-specific inhibitor YM202636, here we demonstrated that in Arabidopsis thaliana, PI(3,5)P2 affects various membrane trafficking events, mostly in the post-Golgi routes. We found that YM201636 treatment effectively reduced PI(3,5)P2 concentration not only in wild-type but also in FAB1A-overexpressing Arabidopsis plants...
November 1, 2016: Plant & Cell Physiology
Kai Eggenberger, Papia Sanyal, Svenja Hundt, Parvesh Wadhwani, Anne S Ulrich, Peter Nick
Actin filaments are essential for the integrity of the cell membrane. In addition to this structural role, actin can modulate signaling by altering polar auxin flow. On the other hand, the organization of actin filaments is modulated by auxin constituting a self-referring signaling hub. Although the function of this auxin-actin oscillator is not clear, there is evidence for a functional link with stress signaling activated by the NADPH oxidase Respiratory burst oxidase Homolog (RboH). In the current work, we used the cell-penetrating peptide BP100 to induce a mild and transient perturbation of membrane integrity...
October 24, 2016: Plant & Cell Physiology
Menglong Wang, Lan Jiang, Qingen Da, Jun Liu, Dongru Feng, Jinfa Wang, Hong-Bin Wang, Hong-Lei Jin
Chloroplast development is an essential process for plant growth that is regulated by numerous proteins. Plastid-encoded plastid RNA polymerase (PEP) is a large complex that regulates plastid gene transcription and chloroplast development. However, many proteins in this complex remain to be identified. Here, through large-scale screening of Arabidopsis mutants by Chl fluorescence imaging, we identified a novel protein, DELAYED GREENING 238 (DG238), which is involved in regulating chloroplast development and plastid gene expression...
October 22, 2016: Plant & Cell Physiology
Daniel Hey, Patricia Ortega-Rodes, Tingting Fan, Florian Schnurrer, Lea Brings, Boris Hedtke, Bernhard Grimm
In plants, two genes encode ferrochelatase (FC), which catalyzes iron chelation into protoporphyrin IX at the final step of heme biosynthesis. FERROCHELATASE1 (FC1) is continuously, but weakly expressed in roots and leaves, while FC2 is dominantly active in leaves. As a continuation of previous studies on the physiological consequences of FC2 inactivation in tobacco, we aimed to assign FC1 function in plant organs. While reduced FC2 expression leads to protoporphyrin IX accumulation in leaves, FC1 down-regulation and overproduction caused reduced and elevated FC activity in root tissue, respectively, but were not associated with changes in macroscopic phenotype, plant development or leaf pigmentation...
October 22, 2016: Plant & Cell Physiology
Zhenle Yang, Hongying Ji, Dantong Liu
Cyperus esculentus is unique in that it can accumulate rich oil in tuber. However, the underlying mechanism of tuber oil biosynthesis is still unclear. Our transcriptional analyses of the pathways from pyruvate production up to triacylglycerol (TAG) accumulation in tuber revealed many distinct species-specific lipid expression patterns from oil seeds and fruits, indicating that in C. esculentus tuber, (1) carbon flux from sucrose toward plastid pyruvate could be produced mostly through cytosolic glycolytic pathway, (2) acetyl-CoA synthetase might be an important contributor to acetyl-CoA formation for plastid fatty acid biosynthesis, (3) expression pattern for stearoyl-ACP desaturase was associated with high oleic acid composition, (4) it was most likely that endoplasmic reticulum-associated acyl-CoA synthetase played a significant role in the export of fatty acids between the plastid and ER, (5) lipid phosphate phosphatase (LPP)-δ was most probably related to the formation of DAG pool in Kennedy pathway, (6) diacylglyceroltransacylase 2 (DGAT2) and phospholipid:diacylglycerolacyltransferase 1 (PDAT1) might play crucial roles in tuber oil biosynthesis...
October 13, 2016: Plant & Cell Physiology
Maryam Nourimand, Christopher D Todd
Plants apply various molecular, physiological, and morphological strategies in response to undesirable environmental conditions. One of the possible responses which may contribute to surviving stressful conditions is the accumulation of ureides. Ureides are recognized as important nitrogen-rich compounds involved in recycling nitrogen in plants to support growth and reproduction. Amongst them, allantoin not only serves as a transportable nitrogen-rich compound, but has also been suggested to protect plants from abiotic stresses via minimizing oxidative damage...
October 13, 2016: Plant & Cell Physiology
Namiki Mitani-Ueno, Naoki Yamaji, Jian Feng Ma
Rice requires high silicon (Si) for its high and sustainable yield. The efficient uptake of Si in rice is mediated by two transporters OsLsi1 and OsLsi2, which function as influx and efflux transporters, respectively. Our previous studies showed that the mRNA expression levels of these transporter genes were down-regulated by Si. Herein we investigated the mechanism underlying regulation of OsLsi1 and OsLsi2 expression. There was a negative correlation between the expression level of OsLsi1 and OsLsi2 and shoot Si accumulation when the rice seedlings were exposed to different Si supply conditions...
October 13, 2016: Plant & Cell Physiology
Bradley W Abramson, Benjamin Kachel, David M Kramer, Daniel C Ducat
In plants, a limited capacity to utilize or export the end products of the Calvin Benson cycle (CB) from photosynthetically-active source cells to non-photosynthetic sink cells can result in reduced carbon capture, photosynthetic electron transport (PET) and lowered photochemical efficiency. The downregulation of photosynthesis caused by reduced capacity to utilize photosynthate has been termed "sink limitation". Recently, several cyanobacterial and algal strains engineered to over-produce target metabolites have exhibited increased photochemistry suggesting possible source-sink regulatory mechanisms may be involved...
October 13, 2016: Plant & Cell Physiology
Jan Mertens, Alex Van Moerkercke, Robin Vanden Bossche, Jacob Pollier, Alain Goossens
Plants produce many bioactive, specialized metabolites to defend themselves when facing various stress situations. Their biosynthesis is directed by a tightly controlled regulatory circuit that is elicited by phytohormones such as jasmonate (JA). The basic helix-loop-helix (bHLH) transcription factors (TFs) bHLH iridoid synthesis 1 (BIS1) and Triterpene Saponin Activating Regulator (TSAR) 1 and 2, from Catharanthus roseus and Medicago truncatula, respectively, all belong to clade IVa of the bHLH protein family and activate distinct terpenoid pathways, thereby mediating monoterpenoid indole alkaloid (MIA) and triterpene saponin (TS) accumulation, respectively, in these two species...
October 1, 2016: Plant & Cell Physiology
Tanja Kühnlenz, Christian Hofmann, Shimpei Uraguchi, Holger Schmidt, Stefanie Schempp, Michael Weber, Brett Lahner, David E Salt, Stephan Clemens
Phytochelatin (PC) synthesis is essential for the detoxification of non-essential metals such as cadmium (Cd). In vitro experiments with Arabidopsis thaliana seedlings had indicated a contribution to zinc (Zn) tolerance as well. We addressed the physiological role of PC synthesis in Zn homeostasis of plants under more natural conditions. Growth responses, PC accumulation and leaf ionomes of wild-type and AtPCS1 mutant plants cultivated in different soils representing adequate Zn supply, Zn deficiency and Zn excess were analyzed...
October 1, 2016: Plant & Cell Physiology
Huajian Zhang, Tongyao Zhao, Peitong Zhuang, Zhiqiang Song, Hui Du, Zhaozhao Tang, Zhimou Gao
SsCut, which functions as an elicitor, can induce plant immunity. In this study, we utilized Nicotiana benthamiana and virus-induced gene silencing to individually decrease the expression of over 2,500 genes. Using this forward genetics approach, several genes were identified that, when silenced, compromised SsCut-triggered cell death based on a cell death assay. A C2H2-type zinc finger gene was isolated from N. benthamiana Sequence analysis indicated that the gene encodes a 27-kDa protein with 253 amino acids containing two typical C2H2-type zinc finger domains; this gene was named NbCZF1 We found that SsCut-induced cell death could be inhibited by virus-induced gene silencing of NbCZF1 in N...
September 20, 2016: Plant & Cell Physiology
Etsuko Watanabe, Shoji Mano, Mika Nomoto, Yasuomi Tada, Ikuko Hara-Nishimura, Mikio Nishimura, Kenji Yamada
Heat shock protein 90 (HSP90) is a molecular chaperone that is required for the function of various substrate proteins, also known as client proteins. It is proposed that HSP90 buffers or hides phenotypic variations in animals and plants by masking mutations in some of its client proteins. However, none of the client proteins with cryptic mutations has been identified to date. Here, we identify the first client protein example by which HSP90 buffers a mutation: the auxin receptor transport inhibitor response 1 (TIR1)...
November 2016: Plant & Cell Physiology
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