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1
Gong, Pan, Michiel Bontinck, Kirin Demuynck, Jolien De Block, Kris Gevaert, Dominique Eeckhout, Geert Persiau et al. « SAMBA controls cell division rate during maize development ». Plant Physiology 188, no 1 (13 novembre 2021) : 411–24. http://dx.doi.org/10.1093/plphys/kiab514.
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Abstract SAMBA has been identified as a plant-specific regulator of the anaphase-promoting complex/cyclosome (APC/C) that controls unidirectional cell cycle progression in Arabidopsis (Arabidopsis thaliana), but so far its role has not been studied in monocots. Here, we show the association of SAMBA with the APC/C is conserved in maize (Zea mays). Two samba genome edited mutants showed growth defects, such as reduced internode length, shortened upper leaves with erect leaf architecture, and reduced leaf size due to an altered cell division rate and cell expansion, which aggravated with plant age. The two mutants differed in the severity and developmental onset of the phenotypes, because samba-1 represented a knockout allele, while translation re-initiation in samba-3 resulted in a truncated protein that was still able to interact with the APC/C and regulate its function, albeit with altered APC/C activity and efficiency. Our data are consistent with a dosage-dependent role for SAMBA to control developmental processes for which a change in growth rate is pivotal.
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Huang, X., P. S. Springer et I. Kaloshian. « Expression of the Arabidopsis MCM Gene PROLIFERA During Root-Knot and Cyst Nematode Infection ». Phytopathology® 93, no 1 (janvier 2003) : 35–41. http://dx.doi.org/10.1094/phyto.2003.93.1.35.
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Expression of the Arabidopsis thaliana gene PROLIFERA (PRL) was examined during development of root-knot and cyst nematode feeding sites. These obligate plant parasites establish specialized feeding structures in roots that allow them to withdraw nutrients from the host. In the process of establishing feeding sites, nematodes alter cell cycle regulation. PRL is normally expressed specifically in dividing cells at all stages of plant development and was used here as a marker for cell division. PRL expression, reported from a PRL∷GUS fusion protein, was detected in nematode feeding sites of both root-knot and cyst nematodes from the earliest stages of infection in both giant cells and syncytia. However, unlike other cell cycle genes, expression of PRL was detected only occasionally in cells surrounding the feeding sites. PRL∷GUS activity persisted until late in the infection cycle, past the time when other cell cycle genes are expressed. These data indicate that some aspects of the PRL expression pattern during nematode infection differ from that of other cell cycle genes.
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Shimizu, Takayuki, Rintaro Yasuda, Yui Mukai, Ryo Tanoue, Tomohiro Shimada, Sousuke Imamura, Kan Tanaka, Satoru Watanabe et Tatsuru Masuda. « Proteomic analysis of haem-binding protein from Arabidopsis thaliana and Cyanidioschyzon merolae ». Philosophical Transactions of the Royal Society B : Biological Sciences 375, no 1801 (4 mai 2020) : 20190488. http://dx.doi.org/10.1098/rstb.2019.0488.
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Chloroplast biogenesis involves the coordinated expression of the plastid and nuclear genomes, requiring information to be sent from the nucleus to the developing chloroplasts and vice versa. Although it is well known how the nucleus controls chloroplast development, it is still poorly understood how the plastid communicates with the nucleus. Currently, haem is proposed as a plastid-to-nucleus (retrograde) signal that is involved in various physiological regulations, such as photosynthesis-associated nuclear genes expression and cell cycle in plants and algae. However, components that transduce haem-dependent signalling are still unidentified. In this study, by using haem-immobilized high-performance affinity beads, we performed proteomic analysis of haem-binding proteins from Arabidopsis thaliana and Cyanidioschyzon merolae . Most of the identified proteins were non-canonical haemoproteins localized in various organelles. Interestingly, half of the identified proteins were nucleus proteins, some of them have a similar function or localization in either or both organisms. Following biochemical analysis of selective proteins demonstrated haem binding. This study firstly demonstrates that nucleus proteins in plant and algae show haem-binding properties. This article is part of the theme issue ‘Retrograde signalling from endosymbiotic organelles’.
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Pedroza-Garcia, Jose Antonio, Thomas Eekhout, Ignacio Achon, Maher-Un Nisa, Griet Coussens, Ilse Vercauteren, Hilde Van den Daele et al. « Maize ATR safeguards genome stability during kernel development to prevent early endosperm endocycle onset and cell death ». Plant Cell 33, no 8 (4 juin 2021) : 2662–84. http://dx.doi.org/10.1093/plcell/koab158.
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Abstract The ataxia-telangiectasia mutated (ATM) and ATM and Rad3-related (ATR) kinases coordinate the DNA damage response. The roles described for Arabidopsis thaliana ATR and ATM are assumed to be conserved over other plant species, but molecular evidence is scarce. Here, we demonstrate that the functions of ATR and ATM are only partially conserved between Arabidopsis and maize (Zea mays). In both species, ATR and ATM play a key role in DNA repair and cell cycle checkpoint activation, but whereas Arabidopsis plants do not suffer from the absence of ATR under control growth conditions, maize mutant plants accumulate replication defects, likely due to their large genome size. Moreover, contrarily to Arabidopsis, maize ATM deficiency does not trigger meiotic defects, whereas the ATR kinase appears to be crucial for the maternal fertility. Strikingly, ATR is required to repress premature endocycle onset and cell death in the maize endosperm. Its absence results in a reduction of kernel size, protein and starch content, and a stochastic death of kernels, a process being counteracted by ATM. Additionally, while Arabidopsis atr atm double mutants are viable, no such mutants could be obtained for maize. Therefore, our data highlight that the mechanisms maintaining genome integrity may be more important for vegetative and reproductive development than previously anticipated.
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Ikram, Aziz, Yong Ding et Yanhua Su. « OsARP6 Is Involved in Internode Elongation by Regulating Cell-Cycle-Related Genes ». Biomolecules 11, no 8 (26 juillet 2021) : 1100. http://dx.doi.org/10.3390/biom11081100.
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The SWR1 complex (SWR1-C) is important for the deposition of histone variant H2A.Z into chromatin to regulate gene expression. Characterization of SWR1-C subunits in Arabidopsis thaliana has revealed their role in variety of developmental processes. Oryza sativa actin related protein 6 (OsARP6) is a subunit of rice SWR1-C. Its role in rice plant development is unknown. Here, we examined the subcellular localization, expression patterns, and loss of function phenotypes for this protein and found that OsARP6 is a nuclear localized protein, and is broadly expressed. OsARP6 interacted with OsPIE1, a central ATPase subunit of rice SWR1-C. The osarp6 knockout mutants displayed pleiotropic phenotypic alterations in vegetative and reproductive traits, including semi-dwarf phenotype, lower tillers number, short leaf length, changes in spikelet morphology, and seed abortion. Microscopic thin sectioning of the top internode revealed that the dwarf phenotype of osarp6 was due to reduced number of cells rather than reduced cell length. The altered transcript level of genes involved in cell division suggested that OsARP6 affects cell cycle regulation. In addition, H2A.Z levels were reduced at the promoters and transcription start sites (TSS) of the regulated genes in osarp6 plants. Together, these results suggest that OsARP6 is involved in rice plant development, and H2A.Z deposition.
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Carrillo-Flores, Elizabeth, Jonanci Arreola Rivera, Denni Mariana Pazos-Solis, Moises Bocanegra-Mondragon, Grisel Fierros Romero, Maria Elena Mellado-Rojas et Elda Beltran-Pena. « TOR participation on the root system changes of Arabidopsis during its interaction with Azospirillum ». Journal of Applied Biotechnology & ; Bioengineering 9, no 2 (7 mars 2022) : 18–23. http://dx.doi.org/10.15406/jabb.2022.09.00280.
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The root system of the plant is essential for taking up water and nutrients, serves as an anchor and is the organ where plant-microorganism interaction takes place. When the Plant Growth Promoting Rhizobacteria (PGPR) Azospirillum brasilense Sp245 colonizes the root of the plants, it halts the growth of the primary root and stimulates the development of the lateral roots and root hairs which support vegetative, green biomass. Target of Rapamycin (TOR) is a highly conserved protein in all eukaryotes, and it controls anabolic processes, such as cell cycle, ribosome biogenesis, protein synthesis, cell wall changes and photosynthesis among others. TOR in plants forms part of the TORC1 complex, which when is activated by auxins and light, activates anabolic processes and represses autophagy. TOR regulates the growth of the primary root of Arabidopsis through cell proliferation and elongation. In the present investigation, the participation of TOR during the Arabidopsis-Azospirillum interaction was determined using two approaches, a pharmacology and other genetic. The results showed that TOR is involved in the development of the lateral roots of A. thaliana seedlings inoculated with A. brasilense.
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Boudonck, K., L. Dolan et P. J. Shaw. « Coiled body numbers in the Arabidopsis root epidermis are regulated by cell type, developmental stage and cell cycle parameters ». Journal of Cell Science 111, no 24 (18 décembre 1998) : 3687–94. http://dx.doi.org/10.1242/jcs.111.24.3687.
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We have used whole mount immunofluorescence labelling with the antibody 4G3, raised against the human snRNP-specific protein U2B″, and whole mount in situ hybridization with an anti-sense probe to a conserved region of U2 snRNA, in combination with confocal microscopy, to examine the organization of spliceosomal components throughout the development of the Arabidopsis thaliana root epidermis. We show that the number of coiled bodies, nuclear organelles in which splicing snRNPs and snRNAs concentrate, is developmentally regulated in the Arabidopsis root epidermis. Firstly, there is a progression from a small number of coiled bodies in the quiescent centre and initial cells, to a larger number in the cell division zone, returning to a lower number in the cell elongation and differentiation zone. Secondly, trichoblasts (root-hair forming epidermal cells) have on average 1.5 times more and often smaller coiled bodies than atrichoblasts (hairless epidermal cells). Moreover, we have shown that these differences in coiled body numbers are related to differences in cell cycle stage, cell type and developmental stage, but are not due to differences in nucleolar or general metabolic activity per se. We discuss possible explanations, including a model in which coiled bodies coalesce during interphase, for the developmental dynamics of coiled bodies.
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Johnstone, Aaron D., Robert T. Mullen et Dev Mangroo. « Arabidopsis At2g40730 encodes a cytoplasmic protein involved in nuclear tRNA export ». Botany 89, no 3 (mars 2011) : 175–90. http://dx.doi.org/10.1139/b10-090.
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Nuclear tRNA export plays an essential role in several key cellular processes, such as regulation of protein synthesis, cell cycle progression, response to nutrient availability and DNA damage, and development. While the overall mechanism of nuclear tRNA export is, in general, poorly understood, the details of specific steps are emerging from studies conducted in different organisms aimed at identifying and characterizing components involved in the process. Here, we report that Arabidopsis thaliana (L.) Heynh At2g40730 encodes CTEXP, a cytoplasmic protein component of the nuclear tRNA export process. CTEXP bound tRNA directly and saturably, and like the nuclear tRNA export receptor PAUSED, overexpression of CTEXP restored export of a nuclear export-defective lysine amber suppressor tRNA in tobacco cells. CTEXP was also found to associate with nucleoporins of the nuclear pore complex (NPC), PAUSED, and the GTPase Ran in vivo. CTEXP interacted directly with PAUSED in vitro and RanGTP, but not RanGDP. Furthermore, a portion of CTEXP appeared to associate with the NPC. Taken together, the data suggest that CTEXP assists with unloading of tRNAs from PAUSED at the cytoplasmic side of the NPC in plant cells.
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Odgerel, Khongorzul, et Zsófia Bánfalvi. « In silico promoter analysis and expression of the BIG BROTHER gene in different organs of potato ». Columella : Journal of Agricultural and Environmental Sciences 9, no 1 (8 juillet 2022) : 31–41. http://dx.doi.org/10.18380/szie.colum.2022.9.1.31.
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The ubiquitin E3 ligase BIG BROTHER/ENHANCER OF DA1 (BB) gene encoding a RING finger protein was identified as a central growth regulator in Arabidopsis thaliana. It was found that BB restricts cell proliferation and promotes leaf senescence. Besides of Arabidopsis, however, the role and regulation of BB in other plant species is only sparsely known. Supposing that the BB gene, like in Arabidopsis, has an important role in the development of potato we aimed to analyse a 3.0-kb promoter sequence of the potato BB gene, StBB, in silico and study the level of StBB expression by quantitative reverse transcription PCR in different organs. A total of 48 binding sites for 15 transcription factor (TF) families were predicted. Most of them were located in the -1.5-kb promoter region. The dominating family of TFs was DOF. It was found that 20 out of the 24 TFs with known functions are involved in developmental processes such as for example, the flower-, leaf-, stem- and root development or cell cycle regulation. In line with this finding, the StBB mRNA was detected in each organ tested with the largest amounts in petal and stamen. These results suggest a similar function of StBB in potato than that is of BB in Arabidopsis, i.e., restriction of organ overgrowth during development and limitation of the plant growth.
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Paraskevopoulou, Dafni, Nikolaos Anezakis, Eleni Giannoutsou, Penelope Sotiriou et Ioannis-Dimosthenis S. Adamakis. « The Stomata of the Katanin Mutants, fra2, lue1 and bot1 ». Biology and Life Sciences Forum 4, no 1 (1 décembre 2020) : 30. http://dx.doi.org/10.3390/iecps2020-08730.
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Katanin, is a microtubule severing protein that orchestrates microtubule organization throughout the plant cell cycle. Taking into consideration the role of the microtubule cytoskeleton in the stomatal development, the Arabidopsis thaliana katanin mutants, fra2, lue1 and bot1 were studied to observe how the absence of function of/malfunction of katanin affects stomatal development. Katanin mutants are characterised by less mature stomata and more young stomata and meristemoids forming clusters. The size of the mature stomata differed from col-0, with the katanin mutants having shorter guard cells and pores as well as smaller pore aperture. In addition, a unique type of cell was observed in the fra2 mutant, the persistent guard mother cells (GMC’s), where the GMC persisted and did not divide symmetrically to form a stoma. Another rather significant observation was that the cell walls of some epidermal cells in the mutants appeared to be incomplete. As far as the cell wall matrix components distribution is concerned, callose did not display significant differences compared to col-0 while pectins and hemicelluloses were differentially dispersed. Microtubules in cytokinetic GMCs were long, bended and connected to the nuclei, while microtubule arrays in katanin mutant leaf epidermis were aberrant and stomatal complexes had astral microtubule arrays as it was observed in the wild type. In conclusion, the malfunction of katanin appears to affect the development of stomata in the epidermis of young leaves in Arabidopsis thaliana, affecting not only stomatal patterning, since the one-cell spacing rule was compromised, but also the morphology of the stomatal complexes. The cell wall-matrix appears altered in the katanin mutants, possibly affecting the function of the stomata since katanin mutant stomata had a reduced pore aperture.
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Machida, Yasunori, Takanori Suzuki, Michiko Sasabe, Hidekazu Iwakawa, Shoko Kojima et Chiyoko Machida. « Arabidopsis ASYMMETRIC LEAVES2 (AS2) : roles in plant morphogenesis, cell division, and pathogenesis ». Journal of Plant Research 135, no 1 (19 octobre 2021) : 3–14. http://dx.doi.org/10.1007/s10265-021-01349-6.
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AbstractThe ASYMMETRIC LEAVES2 (AS2) gene in Arabidopsis thaliana is responsible for the development of flat, symmetric, and extended leaf laminae and their vein systems. AS2 protein is a member of the plant-specific AS2/LOB protein family, which includes 42 members comprising the conserved amino-terminal domain referred to as the AS2/LOB domain, and the variable carboxyl-terminal region. Among the members, AS2 has been most intensively investigated on both genetic and molecular levels. AS2 forms a complex with the myb protein AS1, and is involved in epigenetic repression of the abaxial genes ETTIN/AUXIN RESPONSE FACTOR3 (ETT/ARF3), ARF4, and class 1 KNOX homeobox genes. The repressed expression of these genes by AS2 is markedly enhanced by the cooperative action of various modifier genes, some of which encode nucleolar proteins. Further downstream, progression of the cell division cycle in the developing organs is stimulated; meristematic states are suppressed in determinate leaf primordia; and the extension of leaf primordia is induced. AS2 binds the specific sequence in exon 1 of ETT/ARF3 and maintains methylated CpGs in several exons of ETT/ARF3. AS2 forms bodies (designated as AS2 bodies) at nucleolar peripheries. AS2 bodies partially overlap chromocenters, including inactive 45S ribosomal DNA repeats, suggesting the presence of molecular and functional links among AS2, the 45S rDNAs, and the nucleolus to exert the repressive regulation of ETT/ARF3. The AS2/LOB domain is characterized by three subdomains, the zinc finger (ZF) motif, the internally conserved-glycine containing (ICG) region, and the leucine-zipper-like (LZL) region. Each of these subdomains is essential for the formation of AS2 bodies. ICG to LZL are required for nuclear localization, but ZF is not. LZL intrinsically has the potential to be exported to the cytoplasm. In addition to its nuclear function, it has been reported that AS2 plays a positive role in geminivirus infection: its protein BV1 stimulates the expression of AS2 and recruits AS2 to the cytoplasm, which enhances virus infectivity by suppression of cytoplasmic post transcriptional gene silencing.
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Wang, Chao, Mu Li, Guorui Li, Xinsen Liu, Wensheng Zhao, Bin Yu, Junfeng Liu, Jun Yang et You-Liang Peng. « Two distinct nucleic acid binding surfaces of Cdc5 regulate development ». Biochemical Journal 476, no 21 (15 novembre 2019) : 3355–68. http://dx.doi.org/10.1042/bcj20190502.
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Cell division cycle 5 (Cdc5) is a highly conserved nucleic acid binding protein among eukaryotes and plays critical roles in development. Cdc5 can simultaneously bind to DNA and RNA by its N-terminal DNA-binding domain (DBD), but molecular mechanisms describing its nucleic acid recognition and the regulation of development through its nucleic acid binding remain unclear. Herein, we present a crystal structure of the N-terminal DBD of MoCdc5 (MoCdc5-DBD) from the rice blast fungus Magnaporthe oryzae. Residue K100 of MoCdc5 is on the periphery of a positively charged groove that is formed by K42, K45, R47, and N92 and is evolutionally conserved. Mutation of K100 significantly reduces the affinity of MoCdc5-DBD to a Cdc5-binding element but not to a conventional myeloblastosis (Myb) domain-binding element, suggesting that K100 is a key residue of the high binding affinity to Cdc5-binding element. Another conserved residue (R31) is located close to the U6 RNA in the structure of the spliceosome, and its mutation dramatically reduces the binding capacity of MoCdc5-DBD for U6 RNA. Importantly, mutations in these key residues, including R31, K42, and K100 in AtCDC5, an Arabidopsis thaliana ortholog of MoCdc5, greatly impair the functions of AtCDC5, resulting in pleiotropic development defects and reduced levels of primary microRNA transcripts. Taken together, our findings suggest that Cdc5-DBD binds nucleic acids with two distinct binding surfaces, one for DNA and another for RNA, which together contribute to establishing the regulation mechanism of Cdc5 on development through nucleic acid binding.
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Bernacki, Maciej Jerzy, Weronika Czarnocka, Magdalena Zaborowska, Elżbieta Różańska, Mateusz Labudda, Anna Rusaczonek, Damian Witoń et Stanisław Karpiński. « EDS1-Dependent Cell Death and the Antioxidant System in Arabidopsis Leaves is Deregulated by the Mammalian Bax ». Cells 9, no 11 (10 novembre 2020) : 2454. http://dx.doi.org/10.3390/cells9112454.
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Cell death is the ultimate end of a cell cycle that occurs in all living organisms during development or responses to biotic and abiotic stresses. In the course of evolution, plants and animals evolve various molecular mechanisms to regulate cell death; however, some of them are conserved among both these kingdoms. It was found that mammalian proapoptotic BCL-2 associated X (Bax) protein, when expressed in plants, induces cell death, similar to hypersensitive response (HR). It was also shown that changes in the expression level of genes encoding proteins involved in stress response or oxidative status regulation mitigate Bax-induced plant cell death. In our study, we focused on the evolutional compatibility of animal and plant cell death molecular mechanisms. Therefore, we studied the deregulation of reactive oxygen species burst and HR-like propagation in Arabidopsis thaliana expressing mammalian Bax. We were able to diminish Bax-induced oxidative stress and HR progression through the genetic cross with plants mutated in ENHANCED DISEASE SUSCEPTIBILITY 1 (EDS1), which is a plant-positive HR regulator. Plants expressing the mouse Bax gene in eds1-1 null mutant background demonstrated less pronounced cell death and exhibited higher antioxidant system efficiency compared to Bax-expressing plants. Moreover, eds1/Bax plants did not show HR marker genes induction, as in the case of the Bax-expressing line. The present study indicates some common molecular features between animal and plant cell death regulation and can be useful to better understand the evolution of cell death mechanisms in plants and animals.
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Pan, Ting, Shan Gao, Xiaoyu Cui, Lili Wang et Shunping Yan. « APC/CCDC20 targets SCFFBL17 to activate replication stress responses in Arabidopsis ». Plant Cell, 12 décembre 2022. http://dx.doi.org/10.1093/plcell/koac360.
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Abstract DNA replication stress threatens genome stability and affects plant growth and development. How plants resolve replication stress is poorly understood. The protein kinase WEE1-mediated cell cycle arrest is required for replication stress responses. The E3 ubiquitin ligases Anaphase-promoting complex/cyclosome (APC/C) and Skp1/Cullin 1/F-box (SCF) are essential regulators of the cell cycle. Here, we show that APC/CCDC20 mediates the degradation of SCFFBL17 during replication stress responses in Arabidopsis thaliana. Biochemically, WEE1 interacts with and phosphorylates the APC/C co-activator APC10, which enhances the interaction between F BOX-LIKE17 (FBL17) and CELL DIVISION CYCLE 20 (CDC20), an activator of APC/C. Both APC10 and CDC20 are required for the polyubiquitination and degradation of FBL17. Genetically, silencing CDC20 or APC10 confers plant hypersensitivity to replication stress, which is suppressed by loss of FBL17. Collectively, our study suggests that WEE1 activates APC/C to inhibit FBL17, providing insight into replication stress responses in plants.
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Mergner, Julia, Martin Frejno, Maxim Messerer, Daniel Lang, Patroklos Samaras, Mathias Wilhelm, Klaus F. X. Mayer, Claus Schwechheimer et Bernhard Kuster. « Proteomic and transcriptomic profiling of aerial organ development in Arabidopsis ». Scientific Data 7, no 1 (9 octobre 2020). http://dx.doi.org/10.1038/s41597-020-00678-w.
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Abstract Plant growth and development are regulated by a tightly controlled interplay between cell division, cell expansion and cell differentiation during the entire plant life cycle from seed germination to maturity and seed propagation. To explore some of the underlying molecular mechanisms in more detail, we selected different aerial tissue types of the model plant Arabidopsis thaliana, namely rosette leaf, flower and silique/seed and performed proteomic, phosphoproteomic and transcriptomic analyses of sequential growth stages using tandem mass tag-based mass spectrometry and RNA sequencing. With this exploratory multi-omics dataset, development dynamics of photosynthetic tissues can be investigated from different angles. As expected, we found progressive global expression changes between growth stages for all three omics types and often but not always corresponding expression patterns for individual genes on transcript, protein and phosphorylation site level. The biggest difference between proteomic- and transcriptomic-based expression information could be observed for seed samples. Proteomic and transcriptomic data is available via ProteomeXchange and ArrayExpress with the respective identifiers PXD018814 and E-MTAB-7978.
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Cabral, Danila, Helkin Forero Ballesteros, Bruno Paes de Melo, Isabela Tristan Lourenço-Tessutti, Kércya Maria Simões de Siqueira, Luciana Obicci, Maria Fatima Grossi-de-Sa, Adriana S. Hemerly et Janice de Almeida Engler. « The Armadillo BTB Protein ABAP1 Is a Crucial Player in DNA Replication and Transcription of Nematode-Induced Galls ». Frontiers in Plant Science 12 (30 avril 2021). http://dx.doi.org/10.3389/fpls.2021.636663.
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The biogenesis of root-knot nematode (Meloidogyne spp.)-induced galls requires the hyperactivation of the cell cycle with controlled balance of mitotic and endocycle programs to keep its homeostasis. To better understand gall functioning and to develop new control strategies for this pest, it is essential to find out how the plant host cell cycle programs are responding and integrated during the nematode-induced gall formation. This work investigated the spatial localization of a number of gene transcripts involved in the pre-replication complex during DNA replication in galls and report their akin colocation with the cell cycle S-phase regulator Armadillo BTB Arabidopsis Protein 1 (ABAP1). ABAP1 is a negative regulator of pre-replication complex controlling DNA replication of genes involved in control of cell division and proliferation; therefore, its function has been investigated during gall ontogenesis. Functional analysis was performed upon ABAP1 knockdown and overexpression in Arabidopsis thaliana. We detected ABAP1 promoter activity and localized ABAP1 protein in galls during development, and its overexpression displayed significantly reduced gall sizes containing atypical giant cells. Profuse ABAP1 expression also impaired gall induction and hindered nematode reproduction. Remarkably, ABAP1 knockdown likewise negatively affected gall and nematode development, suggesting its involvement in the feeding site homeostasis. Microscopy analysis of cleared and nuclei-stained whole galls revealed that ABAP1 accumulation resulted in aberrant giant cells displaying interconnected nuclei filled with enlarged heterochromatic regions. Also, imbalanced ABAP1 expression caused changes in expression patterns of genes involved in the cell division control as demonstrated by qRT-PCR. CDT1a, CDT1b, CDKA;1, and CYCB1;1 mRNA levels were significantly increased in galls upon ABAP1 overexpression, possibly contributing to the structural changes in galls during nematode infection. Overall, data obtained in galls reinforced the role of ABAP1 controlling DNA replication and mitosis and, consequently, cell proliferation. ABAP1 expression might likely take part of a highly ordered mechanism balancing of cell cycle control to prevent gall expansion. ABAP1 expression might prevent galls to further expand, limiting excessive mitotic activity. Our data strongly suggest that ABAP1 as a unique plant gene is an essential component for cell cycle regulation throughout gall development during nematode infection and is required for feeding site homeostasis.
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Guo, Binhui, Lin Chen, Lu Dong, Chunhong Yang, Jianhua Zhang, Xiaoyan Geng, Lijuan Zhou et Li Song. « Characterization of the soybean KRP gene family reveals a key role for GmKRP2a in root development ». Frontiers in Plant Science 14 (27 janvier 2023). http://dx.doi.org/10.3389/fpls.2023.1096467.
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Kip-related proteins (KRPs), as inhibitory proteins of cyclin-dependent kinases, are involved in the growth and development of plants by regulating the activity of the CYC-CDK complex to control cell cycle progression. The KRP gene family has been identified in several plants, and several KRP proteins from Arabidopsis thaliana have been functionally characterized. However, there is little research on KRP genes in soybean, which is an economically important crop. In this study, we identified nine GmKRP genes in the Glycine max genome using HMM modeling and BLASTP searches. Protein subcellular localization and conserved motif analysis showed soybean KRP proteins located in the nucleus, and the C-terminal protein sequence was highly conserved. By investigating the expression patterns in various tissues, we found that all GmKRPs exhibited transcript abundance, while several showed tissue-specific expression patterns. By analyzing the promoter region, we found that light, low temperature, an anaerobic environment, and hormones-related cis-elements were abundant. In addition, we performed a co-expression analysis of the GmKRP gene family, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) set enrichment analysis. The co-expressing genes were mainly involved in RNA synthesis and modification and energy metabolism. Furthermore, the GmKRP2a gene, a member of the soybean KRP family, was cloned for further functional analysis. GmKRP2a is located in the nucleus and participates in root development by regulating cell cycle progression. RNA-seq results indicated that GmKRP2a is involved in cell cycle regulation through ribosome regulation, cell expansion, hormone response, stress response, and plant pathogen response pathways. To our knowledge, this is the first study to identify and characterize the KRP gene family in soybean.
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Takasaki, Hironori, Miho Ikeda, Reika Hasegawa, Zhang Yilin, Shingo Sakamoto, Daisuke Maruyama, Nobutaka Mitsuda, Tetsu Kinoshita et Masaru Ohme-Takagi. « Elongation of Siliques Without Pollination 3 Regulates Nutrient Flow Necessary for Embryogenesis ». Plant and Cell Physiology, 20 octobre 2022. http://dx.doi.org/10.1093/pcp/pcac151.
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Abstract Apomixis, defined as the transfer of maternal germplasm to offspring without fertilization, enables the fixation of F1-useful traits, providing advantages in crop breeding. However, most apomictic plants require pollination to produce endosperm. Endosperm is essential for embryogenesis and its development is suppressed until fertilization. We show that expression of a chimeric repressor of the Elongation of Siliques without Pollination 3 (ESP3) gene (Pro35S:ESP3-SRDX) induces ovule enlargement without fertilization in Arabidopsis thaliana. The ESP3 gene encodes a protein similar to the FWA homeodomain transcription factor containing a StAR-related lipid-transfer (START) domain. However, ESP3 lacks the homeobox-encoding region. Genes related to the cell cycle and sugar metabolism were upregulated in unfertilized Pro35S:ESP3-SRDX ovules similar as in fertilized seeds, while those related to autophagy were downregulated similar to fertilized seeds. Unfertilized Pro35S:ESP3-SRDX ovules partially nourished embryos when only the egg was fertilized, accumulating hexoses without central cell proliferation. ESP3 may regulate nutrient flow during seed development, and ESP3-SRDX could be a useful tool for complete apomixis that does not require pseudo-fertilization.
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Zheng, Guanghui, Shaoqiang Hu, Simin Cheng, Liyang Wang, Lijun Kan, Zhengming Wang, Qiang Xu, Zhongchi Liu et Chunying Kang. « Factor of DNA Methylation 1 affects woodland strawberry plant stature and organ size via DNA methylation ». Plant Physiology, 6 octobre 2022. http://dx.doi.org/10.1093/plphys/kiac462.
Texte intégralRésumé :
Abstract RNA-directed DNA methylation (RdDM) is an epigenetic process that directs silencing to specific genomic regions and loci. The biological functions of RdDM are not well studied in horticultural plants. Here, we isolated the EMS mutant reduced organ size (ros) producing small leaves, flowers, and fruits in woodland strawberry (Fragaria vesca) due to reduced cell numbers compared with that in the wild type. The candidate mutation causes a premature stop codon in FvH4_6g28780, which shares high similarity to Arabidopsis (Arabidopsis thaliana) FACTOR OF DNA METHYLATION1 (FDM1) encoding an RdDM pathway component and was named FveFDM1. Consistently, the fvefdm1CR mutants generated by CRISPR/Cas9 also produced smaller organs. Overexpressing FveFDM1 in an Arabidopsis fdm1-1 fdm2-1 double mutant restored DNA methylation at the RdDM target loci. FveFDM1 acts in a protein complex with its homologue INVOLVED IN DE NOVO 2 (FveIDN2). Furthermore, whole genome bisulfite sequencing revealed that DNA methylation, especially in the CHH context, was remarkably reduced throughout the genome in fvefdm1. Common and specific differentially expressed genes were identified in different tissues of fvefdm1 compared to in wild-type tissues. DNA methylation and expression levels of several gibberellic acid (GA) biosynthesis and cell cycle genes were validated. Moreover, the contents of GA and auxin were substantially reduced in the young leaves of fvefdm1 compared to in the wild type. However, exogenous application of GA and auxin could not recover the organ size of fvefdm1. In addition, expression levels of FveFDM1, FveIDN2, NUCLEAR RNA POLYMERASE D1 (FveNRPD1), DOMAINS REARRANGED METHYLASE 2 (FveDRM2), and cell cycle genes were greatly induced by GA treatment. Overall, our work demonstrated the critical roles of FveFDM1 in plant growth and development via RdDM-mediated DNA methylation in horticultural crops.
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Xie, Lulu, Jianfan Tian, Lixin Peng, Qingqing Cui, Yang Liu, Jiyang Liu, Fu Li, Siyuan Zhang et Jianchang Gao. « Conserved Regulatory Pathways for Stock-Scion Healing Revealed by Comparative Analysis of Arabidopsis and Tomato Grafting Transcriptomes ». Frontiers in Plant Science 12 (24 février 2022). http://dx.doi.org/10.3389/fpls.2021.810465.
Texte intégralRésumé :
Many plants can successfully join root and shoot sections at cut surfaces when severed at the stem. Graft healing is complex and conserved in diverse taxonomic groups with different vascular structures. Herein, we compared transcriptome data from autografted and separated stem sections of Arabidopsis thaliana and tomato (Solanum lycopersicum) to explore changes related to graft healing. Using orthologous gene pairs identified between the two species, temperal expression patterns of evolutionary associated genes in grafted top and bottom, separated top and bottom, and intact stems were exhibited. Genes with expression preference indicate functional diversification of genes related to anatomical structure and cellular development in the two species. Expression profiles of the variable genes revealed common pathways operating during graft healing, including phenylpropanoid metabolism, response to oxygen-containing compounds, xylan, and cell wall biogenesis, mitosis and the cell cycle, carboxylic acid catabolism, and meristem structural organization. In addition, vascular differentiation related NAC domain transcription factors and genome-wide members in Arabidopsis and tomato were used for phylogenetic and expression analysis. Expression differences were largely consistent with sequence differences, reflecting high similarity for protein-coding and regulatory regions of individual clades. NAC proteins mainly clustered in accordance with their reported functions in xylem differentiation or cambium formation. The putative conserved mechanisms suggested by conserved genes and functions could help to expand graft healing theory to a wider range of species, and temporal fluctuations in common pathways imply conserved biological processes during graft healing.
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Perrotta, Lara, Roberta Giordo, Dennis Francis, Hilary J. Rogers et Diego Albani. « Molecular Analysis of the E2F/DP Gene Family of Daucus carota and Involvement of the DcE2F1 Factor in Cell Proliferation ». Frontiers in Plant Science 12 (12 mars 2021). http://dx.doi.org/10.3389/fpls.2021.652570.
Texte intégralRésumé :
E2F transcription factors are key components of the RB/E2F pathway that, through the action of cyclin-dependent kinases, regulates cell cycle progression in both plants and animals. Moreover, plant and animal E2Fs have also been shown to regulate other cellular functions in addition to cell proliferation. Based on structural and functional features, they can be divided into different classes that have been shown to act as activators or repressors of E2F-dependent genes. Among the first plant E2F factors to be reported, we previously described DcE2F1, an activating E2F which is expressed in cycling carrot (Daucus carota) cells. In this study, we describe the identification of the additional members of the E2F/DP family of D. carota, which includes four typical E2Fs, three atypical E2F/DEL genes, and three related DP genes. Expression analyses of the carrot E2F and DP genes reveal distinctive patterns and suggest that the functions of some of them are not necessarily linked to cell proliferation. DcE2F1 was previously shown to transactivate an E2F-responsive promoter in transient assays but the functional role of this protein in planta was not defined. Sequence comparisons indicate that DcE2F1 could be an ortholog of the AtE2FA factor of Arabidopsis thaliana. Moreover, ectopic expression of the DcE2F1 cDNA in transgenic Arabidopsis plants is able to upregulate AtE2FB and promotes cell proliferation, giving rise to polycotyly with low frequency, effects that are highly similar to those observed when over-expressing AtE2FA. These results indicate that DcE2F1 is involved in the control of cell proliferation and plays important roles in the regulation of embryo and plant development.
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Wu, Yajie, Lei Zhang, Jinglong Zhou, Xiaojian Zhang, Zili Feng, Feng Wei, Lihong Zhao, Yalin Zhang, Hongjie Feng et Heqin Zhu. « Calcium-Dependent Protein Kinase GhCDPK28 Was Dentified and Involved in Verticillium Wilt Resistance in Cotton ». Frontiers in Plant Science 12 (15 décembre 2021). http://dx.doi.org/10.3389/fpls.2021.772649.
Texte intégralRésumé :
Verticillium dahliae is a soil-borne fungus that causes vascular wilt through the roots of plants. Verticillium wilt caused by V. dahliae is one of the main diseases in cotton producing areas of the world, resulting in huge economic losses. Breeding resistant varieties is the most economical and effective method to control Verticillium wilt. Calcium-dependent protein kinases (CDPKs) play a pivotal role in plant innate immunity, including regulation of oxidative burst, gene expression as well as hormone signal transduction. However, the function of cotton CDPKs in response to V. dahliae stress remains unexplored. In this study, 96, 44 and 57 CDPKs were identified from Gossypium hirsutum, Gossypium raimondii and Gossypium arboretum, respectively. Phylogenetic analysis showed that these CDPKs could be divided into four branches. All GhCDPKs of the same clade are generally similar in gene structure and conserved domain arrangement. Cis-acting elements related to hormones, stress response, cell cycle and development were predicted in the promoter region. The expression of GhCDPKs could be regulated by various stresses. Gh_D11G188500.1 and Gh_A11G186100.1 was up-regulated under Vd0738 and Vd991 stress. Further phosphoproteomics analysis showed that Gh_A11G186100.1 (named as GhCDPK28-6) was phosphorylated under the stress of V. dahliae. Knockdown of GhCDPK28-6 expression, the content of reactive oxygen species was increased, a series of defense responses were enhanced, and the sensitivity of cotton to V. dahliae was reduced. Moreover, overexpression of GhCDPK28-6 in Arabidopsis thaliana weakened the resistance of plants to this pathogen. Subcellular localization revealed that GhCDPK28-6 was localized in the cell membrane. We also found that GhPBL9 and GhRPL12C may interact with GhCDPK28-6. These results indicate that GhCDPK28-6 is a potential molecular target for improving resistance to Verticillium wilt in cotton. This lays a foundation for breeding disease-resistant varieties.
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He, Chengcheng, Jinghui Liang, Zhaoqun Wu, Xianglin Zhuge, Nan Xu et Hailing Yang. « Study on the interaction preference between CYCD subclass and CDK family members at the poplar genome level ». Scientific Reports 12, no 1 (7 octobre 2022). http://dx.doi.org/10.1038/s41598-022-20800-9.
Texte intégralRésumé :
AbstractCyclin-dependent kinases (CDKs) control the progression of the cell cycle. D-type cyclin (CYCD) is generally believed to form a complex with CDK and control the G1/S transition. In plants, CYCD and CDK gene families can be divided into 6 (D1–D7) and 7 (CDKA–CDKG) subclasses, respectively. Different subclasses in the CYCD and CDK families have different numbers, structures and functions. In some heterologous woody plants, the functions of these subclass family members remain unclear. In this study, 43 CYCD and 27 CDK gene family members were identified in the allodiploid Populus tomentosa Carr. Phylogenetic analysis suggested that these CYCDs and CDKs were divided into 6 and 7 subclasses, respectively, which were the same as other species. The analysis of protein properties, gene structure, motifs, domains, cis-acting elements and tissue-specific expression of all members of these CYCDs and CDKs showed that the differences between members of different subclasses varied widely, but members of the same subclass especially in the CDK gene family were very similar. These findings also demonstrated a strong correlation between CYCD and CDK gene family members in response to hormones and specific expression. The collinear analysis of P. tomentosa, Populus trichocarpa and Arabidopsis thaliana showed that the expansion patterns of CYCD and CDK gene families were predominantly whole genome duplications (WGD). The protein interaction prediction results of different subclasses of CYCD and CDKs showed that the interaction between different subclasses of CYCD and CDKs was significantly different. Our previous study found that transgenic PtoCYCD2;1 and PtoCYCD3;3 poplars exhibited opposite phenotypes. Y2H and BIFC results showed that the interaction between PtoCYCD2;1 and PtoCYCD3;3 was significantly different with CDKs. This finding might suggest that the functional differences of different CYCD subclasses in plant growth and development were closely related to the different interactions between CYCD and CDK. Our results provide a good idea and direction for the functional study of CYCD and CDK proteins in woody plants.