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21 pages, 8595 KiB  
Article
Genome-Wide Identification of Xyloglucan Endotransglucosylase/Hydrolase Multigene Family in Chinese Jujube (Ziziphus jujuba) and Their Expression Patterns Under Different Environmental Stresses
by Mohamed Refaiy, Muhammad Tahir, Lijun Jiao, Xiuli Zhang, Huicheng Zhang, Yuhan Chen, Yaru Xu, Shuang Song and Xiaoming Pang
Plants 2024, 13(24), 3503; https://doi.org/10.3390/plants13243503 - 15 Dec 2024
Viewed by 317
Abstract
The Xyloglucan endotransglucosylase/hydrolase (XTH) family, a group of cell wall-modifying enzymes, plays crucial roles in plant growth, development, and stress adaptation. The quality and yield of Chinese jujube (Ziziphus jujuba) fruit are significantly impacted by environmental stresses, including excessive salinity, drought, [...] Read more.
The Xyloglucan endotransglucosylase/hydrolase (XTH) family, a group of cell wall-modifying enzymes, plays crucial roles in plant growth, development, and stress adaptation. The quality and yield of Chinese jujube (Ziziphus jujuba) fruit are significantly impacted by environmental stresses, including excessive salinity, drought, freezing, and disease. However, there has been no report of the XTH encoding genes present in the Chinese jujube genome and their response transcription level under various stresses. This study provides an in-depth analysis of ZjXTH genes in the genome of Chinese jujube and elucidates their structural motifs, regulatory networks, and expression patterns under various stresses. A total of 29 ZjXTH genes were identified from the Ziziphus jujuba genome. Phylogenetic analysis classifies ZjXTH genes into four distinct groups, while conserved motifs and domain analyses reveal coordinated xyloglucan modifications, highlighting key shared motifs and domains. Interaction network predictions suggest that ZjXTHs may interact with proteins such as Expansin-B1 (EXPB1) and Pectin Methylesterase 22 (PME22). Additionally, cis-regulatory element analysis enhances our understanding of Chinese jujube plant’s defensive systems, where TCA- and TGACG-motifs process environmental cues and orchestrate stress responses. Expression profiling revealed that ZjXTH1 and ZjXTH5 were significantly upregulated under salt, drought, freezing, and phytoplasma infection, indicating their involvement in biotic and abiotic stress responses. Collectively, these findings deepen our understanding of the functional roles of Chinese jujube XTHs, emphasizing their regulatory function in adaptive responses in Chinese jujube plants. Full article
(This article belongs to the Special Issue Genetic Breeding of Trees)
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<p>Phylogenetic analysis of XTH proteins among 29 <span class="html-italic">ZjXTHs</span> from <span class="html-italic">Ziziphus jujuba</span>, 33 <span class="html-italic">AtXTHs</span> from <span class="html-italic">Arabidopsis thaliana</span>, 29 <span class="html-italic">OsXTHs</span> from <span class="html-italic">Oryza sativa</span>, and 15 <span class="html-italic">MdXTHs</span> from <span class="html-italic">Malus domestica</span>. Whole protein sequences of the <span class="html-italic">XTHs</span> gene family were used for alignment using MEGA X software. The phylogenetic tree was constructed ussssing the IQ-TREE 2 web tool using maximum likelihood with 1000 bootstrap replicates. Different-colored branches correspond to distinct XTH subfamilies, and the XTH IDs of arabidopsis, apple, and rice were assigned based on previous studies.</p>
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<p>Comparative analysis of the phylogenetics, exon–intron structures, and conserved motifs of the XTH family in Chinese jujube (<span class="html-italic">ZjXTHs</span>). (<b>A</b>) Motif composition models of 29 XTH proteins, with different motifs color-coded according to the legend. (<b>B</b>) Two conserved domains were identified and are represented in green and yellow. (<b>C</b>) The gene structures of <span class="html-italic">ZjXTH</span> were analyzed and visualized, including introns (black lines), exons (coding sequences, blue rectangles), and untranslated regions (UTRs, red rectangles).</p>
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<p>Chromosomal localization and synteny analysis of <span class="html-italic">ZjXTH</span> proteins in the Chinese jujube genome. Genes IDs in black indicate an absence of collinearity, genes and lines colored in green indicate dispersed duplication, red indicates whole genome duplication, and blue-colored lines indicate transposed duplicated pairs (<b>A</b>). Protein–protein interaction analyses were performed using the String web tool and visualized using Cytoscape software v3.10.3. The network consists of various proteins represented as nodes, with interactions depicted by edges. Proteins highlighted in yellow form key hubs with multiple interactions, suggesting their significant role in the network. Green nodes represent additional interacting proteins (<b>B</b>). Syntenic relationships of <span class="html-italic">ZjXTH</span> genes between <span class="html-italic">Arabidopsis thaliana</span>, <span class="html-italic">Glycine max</span>, and <span class="html-italic">Oryza sativa</span>. The brown lines in the background represent the collinear blocks within <span class="html-italic">Ziziphus jujuba</span> and other plant genomes, while the red lines highlight the syntenic <span class="html-italic">ZjXTH</span> gene pairs (<b>C</b>).</p>
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<p>Analysis of cis-regulatory elements (CREs) in the putative promoter region of <span class="html-italic">ZjXTH</span> genes using the PlantCARE database. (<b>A</b>) The number of predicted CREs located in the 2k bp upstream of the <span class="html-italic">ZjXTH</span> genes and the distribution of the three categories of CREs among the members of the <span class="html-italic">ZjXTH</span> gene family. (<b>B</b>) Venn diagram plot and pie chart showing the distribution of different functional categories of CREs identified in the <span class="html-italic">ZjXTH</span> promoter region.</p>
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<p>Gene ontology (GO) analysis was conducted on the <span class="html-italic">ZjXTH</span> gene family to assess its functional distribution across the genome. GO annotations were assigned to the <span class="html-italic">ZjXTH</span> gene sequences, categorizing them into three primary domains: (<b>A</b>) biological process, (<b>B</b>) cellular component, and (<b>C</b>) molecular function. The resulting bar graph illustrates the proportional distribution of <span class="html-italic">ZjXTH</span> genes across these categories, providing insights into their potential roles in various biological pathways and cellular functions.</p>
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<p>Heatmaps were generated to examine the expression patterns of <span class="html-italic">ZjXTHs</span> under various cellular compartments, developmental stages, and stress conditions. The heatmaps were constructed and visualized using TBTools software v2.102. (<b>A</b>) The sub-cellular localization of <span class="html-italic">ZjXTH</span> proteins was predicted using the WoLF PSORT web tool. (<b>B</b>) The tissue-specific expression profiles of <span class="html-italic">ZjXTH</span> at different developmental stages of the Chinese jujube plant were analyzed using publicly available transcriptome data and displayed in a heatmap. The normalized fragments per kilobase of transcript per million fragments (FPKM) values. A deeper red indicates higher expression levels, while a deeper green represents lower expression levels.</p>
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<p>Expression patterns of <span class="html-italic">ZjXTHs</span> of 29 differentially expressed genes in <span class="html-italic">Z. jujuba</span>. var. spinosa diploid and tetraploid seedlings, representing sensitive and tolerant types, respectively, were used in a salinity treatment, gradually applied at 50, 100, and 150 mM NaCl. A deeper red indicates higher expression levels, while a deeper green represents lower expression levels.</p>
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<p>Expression patterns of 29 differentially expressed <span class="html-italic">ZjXTH</span> genes were analyzed in diploid and tetraploid <span class="html-italic">Z. jujuba</span>. var. spinosa seedlings, representing sensitive and tolerant types, respectively, under PEG6000 concentrations of 5%, 10%, 15%, and 20% applied over 1-day intervals. The heatmaps represent the average FPKM values of the genes. A deeper red indicates higher expression levels, while a deeper green represents lower expression levels.</p>
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<p>Expression patterns of 29 differentially expressed <span class="html-italic">ZjXTH</span> genes were analyzed in the cold-sensitive cultivar ‘Dongzao’ and the cold-tolerant cultivar ‘Jinsixiaozao’. A deeper red indicates higher expression levels, while a deeper green represents lower expression levels.</p>
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<p>Heatmap of 29 differentially expressed genes in Chinese jujube under biotic stress caused by jujube witches’ broom phytoplasma (JWB). (<b>A</b>) <span class="html-italic">Z. jujuba</span> ‘Huping,’ a sensitive cultivar, and (<b>B</b>) <span class="html-italic">Z. mauritiana</span> ‘Cuiming,’ a tolerant cultivar, were grafted onto the diseased ‘Jinsixiaozao’ (<span class="html-italic">Z. jujuba</span>). Phenotypic observations were conducted 21 weeks after grafting. The heatmaps display the average FPKM values of the genes, where deeper red indicates higher expression levels and deeper green represents lower expression levels.</p>
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13 pages, 2842 KiB  
Article
Fast-Swelling Tamarind Xyloglucan/PVA Hydrogels with Interconnected Macroporous Structures for Biomedical Applications
by Umpornpan Ninjumrat, Piyachat Chuysinuan, Thitirat Inprasit, Sarute Ummartyotin, Kittipong Chainok and Penwisa Pisitsak
Polymers 2024, 16(24), 3457; https://doi.org/10.3390/polym16243457 - 10 Dec 2024
Viewed by 418
Abstract
This work demonstrates the preparation of fast-swelling hydrogels based on poly(vinyl alcohol) (PVA) and tamarind xyloglucan (XG), utilizing freeze-drying to achieve an interconnected macroporous structure. Although XG is non-toxic and abundant, it has poor mechanical properties. Therefore, XG was mixed with PVA and [...] Read more.
This work demonstrates the preparation of fast-swelling hydrogels based on poly(vinyl alcohol) (PVA) and tamarind xyloglucan (XG), utilizing freeze-drying to achieve an interconnected macroporous structure. Although XG is non-toxic and abundant, it has poor mechanical properties. Therefore, XG was mixed with PVA and crosslinked with citric acid (CA). Without XG, the crosslinked PVA sample contained partially aligned channels several hundred microns wide. The addition of XG (25% w/w) reduced the structural order of the hydrogels. However, the addition of XG improved the swelling ratio from 308 ± 19% in crosslinked PVA to 533.33% in crosslinked PVA/XG. XG also increased the porosity, as the porosity of the crosslinked PVA, XG, and PVA/XG samples was 56.09 ± 2.79%, 68.99 ± 2.06%, and 66.49 ± 1.62%, respectively. Resistance to compression was decreased by the incorporation of XG but was increased by CA crosslinking. The determination of the gel fraction revealed that CA crosslinking was more effective for the PVA component than the XG component. The swelling of all hydrogels was very rapid, reaching equilibrium within 10 s, due to the interconnected macroporous structure that allowed for capillary action. In conclusion, the prepared hydrogels are non-cytotoxic and well suited for biomedical applications such as drug delivery, wound dressings, and hygienic products. Full article
(This article belongs to the Special Issue Polymer-Based Materials for Drug Delivery and Biomedical Applications)
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<p>Structural representations of (<b>a</b>) tamarind XG [<a href="#B2-polymers-16-03457" class="html-bibr">2</a>,<a href="#B5-polymers-16-03457" class="html-bibr">5</a>,<a href="#B6-polymers-16-03457" class="html-bibr">6</a>], (<b>b</b>) partially hydrolyzed PVA [<a href="#B20-polymers-16-03457" class="html-bibr">20</a>], (<b>c</b>) CA, and (<b>d</b>) the crosslinking reaction between PVA and XG with CA as the crosslinker.</p>
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<p>FTIR spectra of the samples before and after crosslinking.</p>
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<p>SEM images of the crosslinked samples taken at 100× and 1000× magnifications: (<b>a</b>,<b>b</b>) PVA/CA, (<b>c</b>,<b>d</b>) XG/CA, and (<b>e</b>,<b>f</b>) PVA/XG/CA.</p>
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<p>DSC thermograms of the prepared hydrogels.</p>
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<p>Swelling ratio of the prepared hydrogels.</p>
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<p>Textural analysis results.</p>
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<p>MTT assay results showing cell viability at various extraction medium concentrations.</p>
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12 pages, 4414 KiB  
Communication
Homogalacturonans and Hemicelluloses in the External Glands of Utricularia dichotoma Traps
by Bartosz J. Płachno, Małgorzata Kapusta, Marcin Feldo and Piotr Świątek
Int. J. Mol. Sci. 2024, 25(23), 13124; https://doi.org/10.3390/ijms252313124 - 6 Dec 2024
Viewed by 512
Abstract
The Utricularia (bladderworts) species are carnivorous plants that prey mainly on invertebrates using traps (bladders) of leaf origin. On the outer surfaces of the trap, there are dome-shaped glands (capitate trichomes). Each such trichome consists of a basal cell, a pedestal cell, and [...] Read more.
The Utricularia (bladderworts) species are carnivorous plants that prey mainly on invertebrates using traps (bladders) of leaf origin. On the outer surfaces of the trap, there are dome-shaped glands (capitate trichomes). Each such trichome consists of a basal cell, a pedestal cell, and a terminal cell. During the maturation of these external glands, there are changes in the cell wall of the terminal cell of the gland (deposited layers of secondary wall material). Thus, due to changes in the cell wall, these glands are excellent models for studying the specialization of cell walls. The main aim of this study was to check whether different cell wall layers in terminal gland cells have a different composition in the case of homogalacturonans (low-methylesterified HGs, fully de-esterified HGs, and galactan) and hemicelluloses (galactoxyloglucan, xyloglucan, and xylan). The antibodies were used against cell wall components (anti-pectins JIM5, JIM7, LM19, CCRC-M38, and LM5 and anti-hemicelluloses LM25, LM15, CCRC-M1, and CCRC-M138). The localization of the examined compounds was determined using immunohistochemistry techniques, Carbotrace 680, and Calcofluor White. Our study showed the presence of various components in the cell walls of external gland cells: methylesterified and demethylesterified homogalacturonans, galactan, xylan, galactoxyloglucan, and xyloglucan. In the terminal cell, the primary cell wall contains different pectins in contrast to the secondary wall material, which is rich in cellulose and hemicelluloses. We also found that the basal cell differs from the other gland cells by the presence of galactan in the cell wall, which resembles the epidermal cells and parenchyma of traps. A particularly noteworthy part of the cell wall functions as a Casparian strip in the pedestal cell. Here, we found no labeling with Carbotrace 680, possibly due to cell wall modification or cell wall chemical composition variation. We have shown that the apoplastic space formed by the cell walls of the terminal cell is mainly composed of cellulose and hemicelluloses (galactoxyloglucan and xyloglucan). This composition of the cell walls allows the easy uptake of components from the external environment. Our research supports the external glands’ function as hydropotens. Full article
(This article belongs to the Special Issue Latest Research on Plant Cell Wall)
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<p>External gland distribution and structure. (<b>A</b>) Trap epidermis with external glands (arrows), treated with toluidine blue; the glands absorbed the dye; the bar is 100 µm. (<b>B</b>) The structure of the external gland, terminal cell (Tc), pedestal cell (Pc), and basal cell (Bc); the bar is 10 µm.</p>
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<p>Homogalacturonan distribution in the external gland (intense green color—signal of antibody, blue color—cellulose stained by Calcofluor White), terminal cell (Tc), pedestal cell (Pc), and basal cell (Bc). (<b>A</b>) A section through the external gland, labeled with JIM5; the bar is 10 µm. (<b>B</b>) The same section as in A, labeled with JIM5 and Calcofluor White; the bar is 10 µm. (<b>C</b>) A section through the external gland, labeled with JIM5; the bar is 10 µm. (<b>D</b>) A section through the external gland, labeled with LM19; the bar is 10 µm. (<b>E</b>) The same section as in (<b>D</b>), labeled with LM19 and Calcofluor White; the bar is 10 µm. (<b>F</b>) A section through the external gland, labeled with LM19; the bar is 10 µm. (<b>G</b>) A section through the external gland, labeled with CCRC-M38; the bar is 10 µm. (<b>H</b>) The same section as in (<b>G</b>), labeled with CCRC-M38 and Calcofluor White; the bar is 10 µm. (<b>I</b>) A section through the external gland, labeled with CCRC-M38; the bar is 10 µm.</p>
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<p>Homogalacturonan distribution in the external gland (intense green color—signal of antibody, blue color—cellulose stained by Calcofluor White), terminal cell (Tc), pedestal cell (Pc), and basal cell (Bc). (<b>A</b>) A section through the external gland, labeled with JIM7; the bar is 10 µm. (<b>B</b>) A section through the external gland, labeled with LM5; the bar is 10 µm. (<b>C</b>) A section through the external gland, labeled with LM5; the bar is 10 µm. (<b>D</b>) The same section as in (<b>C</b>), labeled with LM5 and Calcofluor White; the bar is 10 µm.</p>
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<p>Hemicellulose (xyloglucan) distribution in the external gland (intense green color—signal of antibody, blue color—cellulose stained by Calcofluor White), terminal cell (Tc), pedestal cell (Pc), and basal cell (Bc). (<b>A</b>) A section through the external gland, labeled with CCRC-M138; the bar is 10 µm. (<b>B</b>) The same section as in A, labeled with CCRC-M138 and Calcofluor White; the bar is 10 µm. (<b>C</b>) A section through the external gland, labeled with CCRC-M138; the bar is 10 µm. (<b>D</b>) A section through the external gland, labeled with CCRC-M1; the bar is 10 µm. (<b>E</b>,<b>F</b>) A section through the external gland and through the terminal cell, labeled with LM15; the bar is 10 µm.</p>
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<p>Hemicellulose (galactoxyloglucan) distribution in the external gland (intense green color—signal of antibody, blue color—cellulose stained by Calcofluor White), terminal cell (Tc), pedestal cell (Pc), and basal cell (Bc). (<b>A</b>) A section through the external gland, labeled with LM25; the bar is 10 µm. (<b>B</b>) The same section as in A, labeled with LM25 and Calcofluor White; the bar is 10 µm. (<b>C</b>) A section through the external gland, labeled with LM25, noting the cell wall ingrowths in the pedestal cell (arrow); the bar is 10 µm. (<b>D</b>) A section through the external gland, labeled with LM25; the bar is 10 µm.</p>
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<p>Dye staining the external gland, terminal cell (Tc), pedestal cell (Pc), and basal cell (Bc). (<b>A</b>,<b>B</b>) A section through the external gland stained by Carbotrace 680 (red color); the bar is 10 µm. (<b>C</b>) A section through the external gland stained by Calcofluor White (blue color); the bar is 10 µm.</p>
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17 pages, 1493 KiB  
Review
Sources and Methods for the Production of Xyloglucan, a Promising Stimulus-Sensitive Biopolymer: A Review
by Elena O. Bakhrushina, Victor S. Pyzhov, Jana A. Yuntunen, Alexander S. Gulenkov, Shoyad D. Arislanov, Ksenia V. Eremeeva, Anastasiya V. Belyatskaya, Natalia B. Demina, Ivan I. Krasnyuk and Ivan I. Krasnyuk
Polymers 2024, 16(21), 3022; https://doi.org/10.3390/polym16213022 - 28 Oct 2024
Viewed by 1041
Abstract
Xyloglucan is a highly promising ‘green’ polymer that has found its application in the food and pharmaceutical industries. Due to its molecular structure similarity to mucin, it has remarkable mucoadhesion properties, which has led to a high research interest in this excipient for [...] Read more.
Xyloglucan is a highly promising ‘green’ polymer that has found its application in the food and pharmaceutical industries. Due to its molecular structure similarity to mucin, it has remarkable mucoadhesion properties, which has led to a high research interest in this excipient for the development of transmucosal delivery systems. Thermosensitivity is another promising property of xyloglucan derivatives, which is mainly exhibited by synthetic block copolymers such as pluronics and PLGA derivatives. Delivery systems whose mechanism of active ingredient release is based on temperature sensitivity are widely used in many medical fields, ranging from antitumour therapy to intranasal delivery. Thus, conducting research on the possibility of obtaining and using a new mucoadhesive, fully biocompatible and affordable polymer—xyloglucan—is a promising task. Full article
(This article belongs to the Special Issue Natural-Based Biodegradable Polymeric Materials II)
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<p>Structure of xyloglucan molecule [<a href="#B16-polymers-16-03022" class="html-bibr">16</a>].</p>
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<p>Structure of xyloglucan (<b>A</b>) compared to that of mucin (<b>B</b>) [<a href="#B20-polymers-16-03022" class="html-bibr">20</a>].</p>
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<p>Xyloglucan content in plant raw materials [<a href="#B11-polymers-16-03022" class="html-bibr">11</a>,<a href="#B24-polymers-16-03022" class="html-bibr">24</a>,<a href="#B28-polymers-16-03022" class="html-bibr">28</a>,<a href="#B29-polymers-16-03022" class="html-bibr">29</a>,<a href="#B30-polymers-16-03022" class="html-bibr">30</a>,<a href="#B31-polymers-16-03022" class="html-bibr">31</a>,<a href="#B32-polymers-16-03022" class="html-bibr">32</a>,<a href="#B33-polymers-16-03022" class="html-bibr">33</a>].</p>
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<p>Comparison of different methods for extracting xyloglucan from <span class="html-italic">Tamarindus indica</span> seeds [<a href="#B35-polymers-16-03022" class="html-bibr">35</a>,<a href="#B44-polymers-16-03022" class="html-bibr">44</a>,<a href="#B45-polymers-16-03022" class="html-bibr">45</a>,<a href="#B46-polymers-16-03022" class="html-bibr">46</a>,<a href="#B47-polymers-16-03022" class="html-bibr">47</a>,<a href="#B48-polymers-16-03022" class="html-bibr">48</a>,<a href="#B49-polymers-16-03022" class="html-bibr">49</a>].</p>
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<p>Change in research interest in development of xyloglucan-based in situ moulds from 1999 to 2024, according to PubMed database, keywords: ‘xyloglucan “+” in situ forming “+” in situ gelling “+” thermosensitive’.</p>
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14 pages, 2003 KiB  
Article
Proteomic Analysis to Understand the Promotive Effect of Ethanol on Soybean Growth Under Salt Stress
by Setsuko Komatsu and Takumi Nishiuchi
Biology 2024, 13(11), 861; https://doi.org/10.3390/biology13110861 - 24 Oct 2024
Viewed by 678
Abstract
Finding solutions to mitigate the impact of salinity on crops is important for global food security because soil salinity significantly reduces plant growth and grain yield. Ethanol may play an important role in mitigating the negative salt-induced effects on crops. Soybean root growth [...] Read more.
Finding solutions to mitigate the impact of salinity on crops is important for global food security because soil salinity significantly reduces plant growth and grain yield. Ethanol may play an important role in mitigating the negative salt-induced effects on crops. Soybean root growth was significantly reduced under salt stress; however, it was restored and comparable to control values by ethanol application even under stress. To study the positive mechanism of ethanol on soybean growth, a proteomic approach was carried out. The categories with the greatest changes in protein numbers were protein metabolism, transport, and cell organization in biological processes, nucleus and cytosol in cellular components, and nucleic acid binding activity in molecular functions. Proteomic data were confirmed using immunoblot analysis. Reactive oxygen species enzymes increased under salt stress; among them, mitochondrial ascorbate peroxidase was further accumulated by ethanol application. Among the cell wall and membrane-associated proteins, xyloglucan xyloglucosyl transferase and H+-ATPase increased and decreased, respectively, under salt stress; however, they were restored to control levels by ethanol application. These results suggest that soybeans were adversely affected by salt stress and recovered with ethanol application via the regulation of cell wall and membrane functions through the detoxification of reactive oxygen species. Full article
(This article belongs to the Special Issue Adaptation of Living Species to Environmental Stress)
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<p>Design of experiments to elucidate the function of ethanol on soybeans under salt stress. Soybean seedlings treated with salt and ethanol were subjected to morphological parameters, and their proteins were further analyzed by proteomics. Proteins identified by proteomics were validated by immunoblots. For all experiments, 3 independent biological replicates were performed.</p>
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<p>Effect of ethanol on soybean morphology under salt stress. Three-day-old soybean seedlings were treated with or without 300 mM of ethanol as well as with or without 150 mM of NaCl for 2 days. At 5 days after sowing, the hypocotyl length (<b>A</b>), hypocotyl-fresh weight (<b>B</b>), taproot length (<b>C</b>), and total-root fresh weight (<b>D</b>) were measured as morphological parameters. Data are presented as mean ± SD from 3 independent biological replicates. The means of points with different letters are significantly different according to a one-way ANOVA followed by Tukey’s multiple comparison test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Summary of soybean-proteomic data based on principal component analysis. Three-day-old soybean seedlings were treated with or without 300 mM of ethanol as well as with or without 150 mM of NaCl for 2 days. Soybean roots were collected for protein extraction. Proteomic analysis was conducted in 3 independent biological replicates for each treatment. Principal component analysis was performed in Proteome Discoverer using proteins from 6 different kinds of samples from salt/control (<b>A</b>) and salt + ethanol/salt (<b>B</b>).</p>
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<p>Functional classification of proteins with altered accumulation in the roots of soybeans treated with ethanol under salt stress. Sample collection and experimental methods were the same as in <a href="#biology-13-00861-f003" class="html-fig">Figure 3</a>. After proteomics, gene-ontology analysis was used to determine functional categories: biological process (<b>A</b>), cellular component (<b>B</b>), and molecular function (<b>C</b>) (<a href="#app1-biology-13-00861" class="html-app">Tables S1 and S2</a>). The orange and blue columns indicate the number of increased and decreased proteins, respectively.</p>
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<p>Immunoblot analysis of selected proteins from ROS scavenging enzymes in soybeans treated with ethanol under salt stress. Proteins of soybean roots and hypocotyls were extracted and separated using SDS-polyacrylamide gel electrophoresis. A staining pattern with Coomassie-brilliant blue was used as a loading control (<a href="#app1-biology-13-00861" class="html-app">Figure S2</a>). After immuno-reaction, the integrated density of bands was calculated using ImageJ software. As primary antibodies, anti-ascorbate peroxidase (<b>A</b>,<b>B</b>) and peroxiredoxin (<b>C</b>) antibodies were used. Data are presented as the mean ± SD from 3 independent biological replicates (<a href="#app1-biology-13-00861" class="html-app">Figures S3 and S4</a>). The means of points with star marks are significantly different according to a Student’s <span class="html-italic">t</span>-test between 2 groups (**, <span class="html-italic">p</span> &lt; 0.01).</p>
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<p>Immunoblot analysis of selected proteins related to the cell wall and membrane in soybeans treated with ethanol under salt stress. Sample collection and experimental methods were the same as in <a href="#biology-13-00861-f005" class="html-fig">Figure 5</a>. As primary antibodies, anti-xyloglucan xyloglucosyl transferase (<b>A</b>), cellulose synthase (<b>B</b>), and H<sup>+</sup>-ATPase (<b>C</b>) antibodies were used. Data are presented as mean ± SD from 3 independent biological replicates (<a href="#app1-biology-13-00861" class="html-app">Figures S5–S7</a>). Statistical analysis is the same as in <a href="#biology-13-00861-f005" class="html-fig">Figure 5</a> (*, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01).</p>
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14 pages, 4410 KiB  
Article
Identification of Candidate Genes Associated with Flesh Firmness by Combining QTL Mapping and Transcriptome Profiling in Pyrus pyrifolia
by Shuang Jiang, Jiaying Zhang, Xiaoqing Wang, Chunhui Shi and Jun Luo
Int. J. Mol. Sci. 2024, 25(21), 11347; https://doi.org/10.3390/ijms252111347 - 22 Oct 2024
Viewed by 533
Abstract
Flesh firmness is an important quality of pear fruits. Breeding cultivars with suitably low flesh firmness is one of the popular pear breeding goals. At present, SNP markers related to pear flesh firmness and genes affecting flesh firmness are still uncertain. In this [...] Read more.
Flesh firmness is an important quality of pear fruits. Breeding cultivars with suitably low flesh firmness is one of the popular pear breeding goals. At present, SNP markers related to pear flesh firmness and genes affecting flesh firmness are still uncertain. In this study, a QTL analysis was performed, and the result showed that the position of 139.857 cM in lineage group 14 (LG14) had the highest average logarithm of odds (3.41) over two years. This newly discovered locus was identified as a flesh firmness-related QTL (qFirmness-LG14). The ‘C/T’ SNP was found in corresponding Marker1512129. The ‘C’ genotype is the high-firmness genotype, which is a dominant trait. The average firmness of fruits with genotype C is 21.4% higher than genotype without the C genotype. Transcriptome profiling was obtained between ‘Zaoshengxinshui’ and ‘Qiushui’ at five time points. Three candidate genes in the interval of qFirmness-LG14 might affect firmness. A gene of xyloglucan endotransglucosylase 1 (PpXTH1) was upregulated in ‘Qiushui’ at all five time points. Two transcription factors (PpHY5 and PpERF113) were upregulated in ‘Zaoshengxinshui’, which might be negative regulatory genes for high flesh firmness. The transcriptome results also isolated a large number of cell wall-related genes (e.g., Pectate lyase, Pectin acetylesterase, Pectin methylesterase, and 4-coumarate-CoA ligase) and transcription factors (e.g., ERF, WRKY). These genes are all potential upstream and downstream genes related to flesh firmness. In conclusion, this study provides valuable insights into the QTLs and molecular mechanisms associated with fruit firmness in Pyrus pyrifolia. Full article
(This article belongs to the Section Molecular Plant Sciences)
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<p>Flesh firmness of 92 offspring counted at 100 DAB in 2017 and 2018. Each dot represents a hybrid individual.</p>
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<p>The mapping result diagram of firmness traits in 2017 and 2018. The logarithm of odds (LOD) value, marked in blue, and the explained variance (Expl %), marked in red, are shown in the left y-axis and right y-axis, respectively. The red arrows indicate a common peak, and the green arrows indicate the specific peaks in the two years.</p>
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<p>The average LOD value of markers from 2017 and 2018 data in Linkage Group 14. Each point represents a marker. The intervals of qFirmness-LG14 are labeled in peach color.</p>
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<p>The flesh firmness of ‘Zaoshengxinshui’ and ‘Qiushui’ at five time points.</p>
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<p>The Venn diagram of differentially expressed genes between ‘Zaoshengxinshui’ and ‘Qiushui’ at five time points. ‘Qiushui’ was used as the control cultivar; the upregulated and downregulated genes were classified in ‘Zaoshengxinshui’. The number of DEGs are shown in the column charts. Yellow, orange, green, blue, and purple represent 75, 82, 89, 96, and 103 DAB, respectively.</p>
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<p>Heatmap of the expression of DEGs related to cell wall-related genes and transcription factors in fruit development. The letter ‘Z’ represents ‘Zaoshengxinshui’, and ‘Q’ represents ‘Qiushui’. P1 to P5 represent five time points (75, 82, 89, 96, and 103 DAB) for sampling. Green to red represents the gene expression level from low to high.</p>
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<p>Heatmap of the expression of 14 genes related to flesh firmness around the position of qFirmness-LG14. Letter Z represents ‘Zaoshengxinshui’, and Q represents ‘Qiushui’. P1~P5 represent 75, 82, 89, 96, and 103 DAB, respectively. Green to red represents the gene expression level from low to high.</p>
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<p>Potential regulatory pathways in pear cultivars with low and high flesh firmness. The upregulated genes in the low-firmness cultivars are labeled in red. The downregulated genes are labeled in green. ‘T’ and ‘C’ represent the genotype of Marker1512129.</p>
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17 pages, 6397 KiB  
Article
Functional Analysis of Cucumis melo CmXTH11 in Regulating Drought Stress Tolerance in Arabidopsis thaliana
by Shiwen Zhao, Qianqian Cao, Lei Li, Wenqin Zhang, Yongjun Wu and Zhenchao Yang
Int. J. Mol. Sci. 2024, 25(20), 11031; https://doi.org/10.3390/ijms252011031 - 14 Oct 2024
Viewed by 728
Abstract
The CmXTH11 gene, a member of the XTH (xyloglucan endotransglycosylase/hydrolase) family, plays a crucial role in plant responses to environmental stress. In this study, we heterologously expressed the melon gene CmXTH11 in Arabidopsis to generate overexpressing transgenic lines, thereby elucidating the regulatory role [...] Read more.
The CmXTH11 gene, a member of the XTH (xyloglucan endotransglycosylase/hydrolase) family, plays a crucial role in plant responses to environmental stress. In this study, we heterologously expressed the melon gene CmXTH11 in Arabidopsis to generate overexpressing transgenic lines, thereby elucidating the regulatory role of CmXTH11 in water stress tolerance. Using these lines of CmXTH11 (OE1 and OE2) and wild-type (WT) Arabidopsis as experimental materials, we applied water stress treatments (including osmotic stress and soil drought) and rewatering treatments to investigate the response mechanisms of melon CmXTH11 in Arabidopsis under drought stress from a physiological and biochemical perspective. Overexpression of CmXTH11 significantly improved root growth under water stress conditions. The OE lines exhibited longer roots and a higher number of lateral roots compared to WT plants. The enhanced root system contributed to better water uptake and retention. Under osmotic and drought stress, the OE lines showed improved survival rates and less wilting compared to WT plants. Biochemical analyses revealed that CmXTH11 overexpression led to lower levels of malondialdehyde (MDA) and reduced electrolyte leakage, indicating decreased oxidative damage. The activities of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD), were significantly higher in OE lines, suggesting enhanced oxidative stress tolerance. The CmXTH11 gene positively regulates water stress tolerance in Arabidopsis by enhancing root growth, improving water uptake, and reducing oxidative damage. Overexpression of CmXTH11 increases the activities of antioxidant enzymes, thereby mitigating oxidative stress and maintaining cellular integrity under water deficit conditions. These findings suggest that CmXTH11 is a potential candidate for genetic improvement of drought resistance in crops. Full article
(This article belongs to the Section Molecular Plant Sciences)
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<p><span class="html-italic">CmXTH11</span> protein structure and physicochemical properties analysis. (<b>a</b>) Conserved domains of <span class="html-italic">CmXTH11</span> protein; (<b>b</b>) signal peptide domain analysis of <span class="html-italic">CmXTH11</span> protein; (<b>c</b>) transmembrane domain analysis of <span class="html-italic">CmXTH11</span> protein; (<b>d</b>) SWISS modeling of <span class="html-italic">CmXTH11</span>.</p>
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<p><span class="html-italic">CmXTH11</span> expression analysis. (<b>a</b>) Tissue-specific expression analysis of <span class="html-italic">CmXTH11</span>; (<b>b</b>) expression levels of <span class="html-italic">CmXTH11</span> under different hormone treatments. Note: Data in the same column with different letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Analysis of <span class="html-italic">CmXTH11</span> amino acid sequences and phylogenetic tree construction. (<b>a</b>) Amino acid sequence alignment: Comparative analysis of the amino acid sequences of <span class="html-italic">CmXTH11</span> with <span class="html-italic">XTH</span> proteins from six different plant species; (<b>b</b>) phylogenetic tree analysis: Phylogenetic tree showing the evolutionary relationships between <span class="html-italic">CmXTH11</span> and <span class="html-italic">XTH</span> proteins from various plant species.</p>
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<p>Analysis of <span class="html-italic">CmXTH11</span> amino acid sequences and phylogenetic tree construction. (<b>a</b>) Amino acid sequence alignment: Comparative analysis of the amino acid sequences of <span class="html-italic">CmXTH11</span> with <span class="html-italic">XTH</span> proteins from six different plant species; (<b>b</b>) phylogenetic tree analysis: Phylogenetic tree showing the evolutionary relationships between <span class="html-italic">CmXTH11</span> and <span class="html-italic">XTH</span> proteins from various plant species.</p>
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<p>Confirmation of transgenic <span class="html-italic">Arabidopsis</span> lines. (<b>a</b>) Construction of the pCAMBIA1303-CaMV35S-<span class="html-italic">CmXTH11</span> expression vector; (<b>b</b>) expression levels of <span class="html-italic">CmXTH11</span> in 4-week-old <span class="html-italic">Arabidopsis</span> leaves of WT and two transgenic lines (OE1 and OE2); (<b>c</b>) phenotypic comparison of 8-week-old WT and <span class="html-italic">CmXTH11-OE</span> lines under normal growth conditions. Note: Data in the same column with different letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Analysis of root growth in WT and OE <span class="html-italic">Arabidopsis</span> lines. (<b>a</b>,<b>b</b>) Phenotypic images of root growth in WT and OE lines grown on 1/2 MS medium; (<b>c</b>) number of lateral roots in WT and OE lines grown on 1/2 MS medium; (<b>d</b>) root length of WT and OE lines grown on 1/2 MS medium. Note: Data in the same column with different letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The impact of drought stress on seed germination in WT and <span class="html-italic">CmXTH11</span> overexpressing <span class="html-italic">Arabidopsis</span>. (<b>a</b>) Phenotypic images of root lengths of WT and transgenic lines on 1/2 MS medium containing 0, 100, 200, and 250 mM mannitol after 10 days of cultivation. (<b>b</b>) Phenotypic images of seed germination under the same conditions. (<b>c</b>) Seed germination rates under different mannitol concentrations. (<b>d</b>) Number of green leaves. (<b>e</b>) Root lengths of seeds grown under different mannitol concentrations. (<b>f</b>) Fresh weight of seeds grown under different mannitol concentrations. Note: Data in the same column with different letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p><span class="html-italic">CmXTH11</span> gene enhances drought resistance in <span class="html-italic">Arabidopsis</span>. (<b>a</b>) Phenotypes of WT and <span class="html-italic">CmXTH11</span> overexpressing lines (OE1, OE2) under normal growth conditions, after 9 days of drought stress, and after 5 days of rewatering; (<b>b</b>–<b>d</b>): malondialdehyde (MDA) content (<b>b</b>), relative electrolyte conductivity (REC) (<b>c</b>), and relative water content (RWC) (<b>d</b>) in WT and <span class="html-italic">CmXTH11</span>-OE lines under normal growth conditions, after 9 days of drought stress, and after 5 days of rewatering. Note: Data in the same column with different letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Overexpression of <span class="html-italic">CmXTH11</span> gene affects ROS accumulation and antioxidant activity in <span class="html-italic">Arabidopsis</span> leaves under drought stress. (<b>a</b>,<b>b</b>) NBT and DAB staining: The staining of leaves with NBT and DAB reveals the accumulation of O<sup>2−</sup> and H<sub>2</sub>O<sub>2</sub> in WT and <span class="html-italic">CmXTH11</span>-OE <span class="html-italic">Arabidopsis</span> plants under normal conditions, drought stress for 9 days, and after 5 days of rewatering. (<b>c</b>,<b>d</b>) ROS content: The quantification of O<sup>2−</sup> and H<sub>2</sub>O<sub>2</sub>; (<b>e</b>–<b>g</b>) antioxidant enzyme activity: The activities of POD (<b>e</b>), CAT (<b>f</b>), and SOD (<b>g</b>) measured in leaves of WT and transgenic lines. Note: Data in the same column with different letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Impact of overexpressing <span class="html-italic">CmXTH11</span> gene on stomatal characteristics of <span class="html-italic">Arabidopsis</span> under drought stress. (<b>a</b>–<b>c</b>): (<b>a</b>) Stomatal density (<b>b</b>), stomatal aperture, and (<b>c</b>) pore area of WT and <span class="html-italic">CmXTH11</span> transgenic <span class="html-italic">Arabidopsis</span> leaves under normal growth conditions, after 9 days of drought stress, and after 5 days of rewatering. Note: Data in the same column with different letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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18 pages, 22909 KiB  
Article
Integrated Biological Experiments and Proteomic Analyses of Nicotiana tabacum Xylem Sap Revealed the Host Response to Tomato Spotted Wilt Orthotospovirus Infection
by Hongping Feng, Waiwai Mon, Xiaoxia Su, Yu Li, Shaozhi Zhang, Zhongkai Zhang and Kuanyu Zheng
Int. J. Mol. Sci. 2024, 25(20), 10907; https://doi.org/10.3390/ijms252010907 - 10 Oct 2024
Viewed by 726
Abstract
The plant vascular system is not only a transportation system for delivering nutrients but also a highway transport network for spreading viruses. Tomato spotted wilt orthotospovirus (TSWV) is among the most destructive viruses that cause serious losses in economically important crops worldwide. However, [...] Read more.
The plant vascular system is not only a transportation system for delivering nutrients but also a highway transport network for spreading viruses. Tomato spotted wilt orthotospovirus (TSWV) is among the most destructive viruses that cause serious losses in economically important crops worldwide. However, there is minimal information about the long-distance movements of TSWV in the host plant vascular system. In this this study, we confirm that TSWV virions are present in the xylem as observed by transmission electron microscopy (TEM). Further, a quantitative proteomic analysis based on label-free methods was conducted to reveal the uniqueness of protein expression in xylem sap during TSWV infection. Thus, this study identified and quantified 3305 proteins in two groups. Furthermore, TSWV infection induced three viral structural proteins, N, Gn and Gc, and 315 host proteins differentially expressed in xylem (163 up-regulated and 152 down-regulated). GO enrichment analysis showed up-regulated proteins significantly enriched in homeostasis, wounding, defense response, and DNA integration terms, while down-regulated proteins significantly enriched in cell wall biogenesis/xyloglucan metabolic process-related terms. KEGG enrichment analysis showed that the differentially expressed proteins (DEPs) were most strongly associated with plant-pathogen interaction, MAPK signaling pathway, and plant hormone signal transduction. Cluster analysis of DEPs function showed the DEPs can be categorized into cell wall metabolism-related proteins, antioxidant proteins, PCD-related proteins, host defense proteins such as receptor-like kinases (RLKs), salicylic acid binding protein (SABP), pathogenesis related proteins (PR), DNA methylation, and proteinase inhibitor (PI). Finally, parallel reaction monitoring (PRM) validated 20 DEPs, demonstrating that the protein abundances were consistent between label-free and PRM data. Finally, 11 genes were selected for RT-qPCR validation of the DEPs and label-free-based proteomic analysis concordant results. Our results contribute to existing knowledge on the complexity of host plant xylem system response to virus infection and provide a basis for further study of the mechanism underlying TSWV long-distance movement in host plant vascular system. Full article
(This article belongs to the Special Issue Advances in Plant Virus Diseases and Virus-Induced Resistance)
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<p>Symptoms of TSWV in <span class="html-italic">N. tabacum</span> cv. K326 and RT-PCR detection of TSWV. (<b>A</b>) Symptoms of TSWV in <span class="html-italic">N. tabacum</span> cv. K326. The red circle and arrow point indicate the symptoms of TSWV infection. Photos were taken 14 days after inoculation. (<b>B</b>) RT-PCR identification of TSWV. CK-: healthy tobacco leaf, CK+: positive control infected with TSWV. 1–10: tobacco samples inoculated with TSWV.</p>
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<p>(<b>A</b>) The ultrastructure of xylem vessel from TSWV-infected <span class="html-italic">N. tabacum</span> cv. K326 was observed by TEM.The dashed box represents the enlarged details of the local area. (<b>B</b>) Xylem sap from TSWV-infected <span class="html-italic">N. tabacum</span> cv. K326 was collected and observed by TEM following negative staining. VE: vessel, CW: cell wall, V: TSWV virons.</p>
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<p>(<b>A</b>) Differential proteins between TSWV-infected and mock plants determined via SDS-PAGE analysis. Note: Lane Marker, Lane 1–3: TSWV-infected tobacco xylem sap, Lane 4–6: mock tobacco xylem sap. (<b>B</b>,<b>C</b>) Western blots of TSWV N protein and TSWV Gn proteins in xylem sap. Note: Lane Marker, Lane 1: mock tobacco xylem sap, Lane 2: TSWV-infected tobacco xylem sap.</p>
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<p>(<b>A</b>) Number of xylem sap proteins identified by LC-MS/MS. (<b>B</b>) The number of upregulated and downregulated proteins. (<b>C</b>) PCA of the tobacco xylem sap samples.</p>
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<p>GO enrichment and KEGG pathways. BP: biological process; MF: molecular function; CC: cellular component. (<b>A</b>) The upregulated proteins that significantly enriched GO terms. (<b>B</b>) The downregulated proteins significantly enriched GO terms. (<b>C</b>) KEGG pathway analysis.</p>
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<p>Cluster analysis of DEPs.</p>
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<p>PPI network analysis of DEPs.</p>
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<p>Comparison of the quantification results between label-free and PRM of the 20 candidate proteins. The <span class="html-italic">X</span>-axis represents the protein names, and the <span class="html-italic">Y</span>-axis represents fold changes of protein abundances between TSWV-infection and control in tobacco xylem sap.</p>
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<p>qRT-PCR analysis of 11 candidate genes with significant changes in protein abundance. The bars represent the means ± SD (<span class="html-italic">n</span> = 3) of three biological replicates. The asterisks indicate the significance level (* <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001) based on a Student’s <span class="html-italic">t</span>-test.</p>
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16 pages, 4495 KiB  
Article
A Genome-Wide Identification and Expression Analysis of the Xyloglucan Endotransglucosylase/Hydrolase Gene Family in Melon (Cucumis melo L.)
by Shiwen Zhao, Yushi Kang, Yuqin Lin, Xue Zheng, Yongjun Wu and Zhenchao Yang
Horticulturae 2024, 10(10), 1017; https://doi.org/10.3390/horticulturae10101017 - 24 Sep 2024
Cited by 1 | Viewed by 844
Abstract
The xyloglucan endotransglucosylase/hydrolase (XTH) family is an important multigene family in plants that plays a key role in cell wall reconstruction and stress tolerance. However, the specific traits of XTH genes and their expression patterns under different stresses have not been [...] Read more.
The xyloglucan endotransglucosylase/hydrolase (XTH) family is an important multigene family in plants that plays a key role in cell wall reconstruction and stress tolerance. However, the specific traits of XTH genes and their expression patterns under different stresses have not been systematically studied in melon. In this study, based on the genomic data of Cucumis melon, 29 XTH genes were identified; most of these genes contain two conserved domains (Glyco_hydro_16 and XET_C domains). Based on neighbor-joining phylogenetic analysis, the CmXTHs were divided into four subfamilies, I/II, IIIA, and IIIB, which are distributed across nine chromosomes of melon. Collinearity analysis showed that the melon XTH genes have an evolutionary history consistent with three species: Arabidopsis, tomato, and cucumber. The promoter regions of the CmXTH genes contain numerous cis-acting elements, which are associated with plant growth, hormonal response, and stress responses. RNA-Seq analysis indicated that CmXTH genes exhibit different expression patterns under drought and salt stress treatments, suggesting that this gene family plays an important role under abiotic stress. This study provides a theoretical basis for further studies on the molecular function of XTH genes in melon. Full article
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<p>A phylogenetic tree analysis of XTH proteins in melon and <span class="html-italic">Arabidopsis thalian</span>a. (The evolutionary tree was divided into four subfamilies with different colors. The blue square represents Arabidopsis. The red five-pointed star represents the melon).</p>
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<p>Chromosomal localization of <span class="html-italic">XTH</span> gene in melon.</p>
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<p>Phylogenetic tree, gene structure, and conserved motifs of melon <span class="html-italic">XTHs</span>. (<b>a</b>) Phylogenetic relationships of 29 <span class="html-italic">CmXTH</span> genes, (<b>b</b>) protein motifs, and (<b>c</b>) gene structures. Horizontal coordinates denote lengths of genes and amino acid sequences.</p>
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<p>A conserved domain analysis of CmXTH proteins. The green box represents the Glyco_hydro_16 domain, while the yellow box represents the XET_C domain.</p>
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<p>Cis-element analysis of 29 <span class="html-italic">CmXTH</span> gene promoters. Each box is filled with different colors representing different cis-acting elements.</p>
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<p>A fragment duplication analysis of the melon <span class="html-italic">XTHs</span>. The black lines represent the fragmentary duplicated gene pair, and the gray lines represent the synteny blocks of the <span class="html-italic">XTH</span> genes in the melon genome. The line and heat map in the outer circle represent gene density on the chromosome.</p>
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<p>A collinearity analysis of <span class="html-italic">XTH</span> genes between melon and <span class="html-italic">Arabidopsis thaliana</span>, <span class="html-italic">Solanum lycopersicum</span>, and <span class="html-italic">Cucumis sativus</span>. The red lines represent XTH syntenic gene pairs of melon with three other species, and the gray lines represent orthologous genes of melon with three other species.</p>
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<p>Expression pattern of melon <span class="html-italic">XTHs</span> under different abiotic stress. (<b>a</b>) Expression pattern of <span class="html-italic">CmXTHs</span> under drought stress. (<b>b</b>) Expression pattern of <span class="html-italic">CmXTHs</span> under salt stress. Samples for expression profiling were collected from melon at 0, 3, 6, 9, 12, and 24 h post drought and salt stress. Red represents high expression, and blue represents low expression. All ratios are log<sub>2</sub>-transformed so that inductions and repressions of identical magnitude are numerically equal but have opposite signs.</p>
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26 pages, 18602 KiB  
Article
Integration of Phenotypes, Phytohormones, and Transcriptomes to Elucidate the Mechanism Governing Early Physiological Abscission in Coconut Fruits (Cocos nucifera L.)
by Lilan Lu, Zhiguo Dong, Xinxing Yin, Siting Chen and Ambreen Mehvish
Forests 2024, 15(8), 1475; https://doi.org/10.3390/f15081475 - 22 Aug 2024
Viewed by 979
Abstract
The abscission of fruits has a significant impact on yield, which in turn has a corresponding effect on economic benefits. In order to better understand the molecular mechanism of early coconut fruit abscission, the morphological and structural characteristics, cell wall hydrolysis and oxidase [...] Read more.
The abscission of fruits has a significant impact on yield, which in turn has a corresponding effect on economic benefits. In order to better understand the molecular mechanism of early coconut fruit abscission, the morphological and structural characteristics, cell wall hydrolysis and oxidase activities, phytohormones, and transcriptomes were analyzed in the abscission zone (AZ) from early-abscised coconut fruits (AFs) and non-abscised coconut fruits (CFs). These results indicated that the weight and water content of AFs are significantly lower than those of CFs, and the color of AFs is a grayish dark red, with an abnormal AZ structure. Cellulase (CEL), polygalacturonase (PG), pectinesterase (PE), and peroxidase (POD) activities were significantly lower than those of CFs. The levels of auxin (IAA), gibberellin (GA), cytokinins (CKs), and brassinosteroid (BR) in AFs were significantly lower than those in CFs. However, the content of abscisic acid (ABA), ethylene (ETH), jasmonic acid (JA), and salicylic acid (SA) in AFs was significantly higher than in CFs. The transcriptome analysis results showed that 3601 DEGs were functionally annotated, with 1813 DEGs upregulated and 1788 DEGs downregulated. Among these DEGs, many genes were enriched in pathways such as plant hormone signal transduction, carbon metabolism, peroxisome, pentose and gluconate interconversion, MAPK signaling pathway—plant, and starch and sucrose metabolism. Regarding cell wall remodeling-related genes (PG, CEL, PE, POD, xyloglucan endoglucosidase/hydrogenase (XTH), expansin (EXP), endoglucanase, chitinase, and beta-galactosidase) and phytohormone-related genes (IAA, GA, CKs, BR, ABA, JA, SA, and ETH) were significantly differentially expressed in the AZ of AFs. Additionally, BHLH, ERF/AP2, WRKY, bZIP, and NAC transcription factors (TFs) were significantly differently expressed, reflecting their crucial role in regulating the abscission process. This study’s results revealed the molecular mechanism of early fruit abscission in coconuts. This provided a new reference point for further research on coconut organ development and abscission. Full article
(This article belongs to the Section Genetics and Molecular Biology)
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<p>Development of early-abscised coconut fruits (AFs) and non-abscised coconut fruits (CFs). (<b>a</b>) The anatomical structure of fruit morphology and abscission zone (AZ). (<b>b</b>) Fresh weight and water content of coconut fruits. The data represent the mean ± standard deviation (SD) of ten samples, and the significance of fresh weight and water content of coconut fruits between CFs and AFs was determined using Student’s <span class="html-italic">t</span>-tests. * significant at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Enzyme activity and phytohormones in AZ of AFs and CFs. CEL, cellulase; PG, polygalacturonase; PE, pectinesterase; POD, peroxidase; IAA, auxin; GA, gibberellin; CK, cytokinin; BR, brassinosteroid; ABA, abscisic acid; ETH, ethylene; JA, jasmonic acid; SA, salicylic acid. The data represent the mean ± standard deviation (SD) of three biological replicates, and the significance of enzyme activity and plant hormone contents in AZ between CFs and AFs was determined using Student’s <span class="html-italic">t</span>-tests. * significant at <span class="html-italic">p</span> &lt; 0.05. ** significant at <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Differentially expressed genes (DEGs) enriched in the top 20 enriched GO terms in terms of biological processes, molecular functions, and cellular components in AFs and CFs.</p>
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<p>KEGG analysis from CF vs. AF group. (<b>a</b>) KEGG classification chart of DEGs. (<b>b</b>) KEGG enrichment bar chart of all DEGs. (<b>c</b>) KEGG enrichment bar chart of upregulated DEGs. (<b>d</b>) KEGG enrichment bar chart of downregulated DEGs.</p>
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<p>Expression of identified DEGs involved in the main KEGG enrichment pathways in CF vs. AF group.</p>
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<p>Expression of identified DEGs involved in cell wall modification in CF vs. AF group.</p>
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<p>Heatmap of relative changes in expression patterns of 8 phytohormone-related genes in AZ of CF vs. AF group. The color scales on each heatmap display their expression values.</p>
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<p>Transcription factor analysis in CF vs. AF group. (<b>a</b>) Expression of identified DEGs involved in transcription factors in CF vs. AF group. (<b>b</b>) Distribution of overexpression of the regulatory transcription factor family in CF vs. AF group.</p>
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<p>Verification of the expression of 11 coconut fruit abscission-related genes through qRT-PCR analysis. The bar chart represents the value of FPKM. The line graph represents qRT-PCR values. The error bar represents the standard deviation of three biological replicates (<b>a</b>–<b>k</b>). Correlation of expression changes observed through RNA-seq (<span class="html-italic">y</span>-axis) and qRT-PCR (<span class="html-italic">x</span>-axis) (<b>l</b>).</p>
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<p>A hypothetical model for coconut fruit abscission.</p>
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22 pages, 5638 KiB  
Review
Plant Cell Wall Polysaccharide O-Acetyltransferases
by Ruiqin Zhong, Dayong Zhou, Lirong Chen, John P. Rose, Bi-Cheng Wang and Zheng-Hua Ye
Plants 2024, 13(16), 2304; https://doi.org/10.3390/plants13162304 - 19 Aug 2024
Viewed by 1145
Abstract
Plant cell walls are largely composed of polysaccharide polymers, including cellulose, hemicelluloses (xyloglucan, xylan, mannan, and mixed-linkage β-1,3/1,4-glucan), and pectins. Among these cell wall polysaccharides, xyloglucan, xylan, mannan, and pectins are often O-acetylated, and polysaccharide O-acetylation plays important roles in cell [...] Read more.
Plant cell walls are largely composed of polysaccharide polymers, including cellulose, hemicelluloses (xyloglucan, xylan, mannan, and mixed-linkage β-1,3/1,4-glucan), and pectins. Among these cell wall polysaccharides, xyloglucan, xylan, mannan, and pectins are often O-acetylated, and polysaccharide O-acetylation plays important roles in cell wall assembly and disease resistance. Genetic and biochemical analyses have implicated the involvement of three groups of proteins in plant cell wall polysaccharide O-acetylation: trichome birefringence-like (TBL)/domain of unknown function 231 (DUF231), reduced wall acetylation (RWA), and altered xyloglucan 9 (AXY9). Although the exact roles of RWAs and AXY9 are yet to be identified, members of the TBL/DUF231 family have been found to be O-acetyltransferases responsible for the O-acetylation of xyloglucan, xylan, mannan, and pectins. Here, we provide a comprehensive overview of the occurrence of O-acetylated cell wall polysaccharides, the biochemical properties, structural features, and evolution of cell wall polysaccharide O-acetyltransferases, and the potential biotechnological applications of manipulations of cell wall polysaccharide acetylation. Further in-depth studies of the biochemical mechanisms of cell wall polysaccharide O-acetylation will not only enrich our understanding of cell wall biology, but also have important implications in engineering plants with increased disease resistance and reduced recalcitrance for biofuel production. Full article
(This article belongs to the Section Plant Molecular Biology)
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Figure 1
<p>Phylogenetic relationship of TBL members from <span class="html-italic">Arabidopsis</span> (At), <span class="html-italic">Picea abies</span> (Pa/Ma), <span class="html-italic">Selaginella moellendorffii</span> (Sm), <span class="html-italic">Physcomitrium patens</span> (Pp), <span class="html-italic">Marchantia polymorpha</span> (Mp), and <span class="html-italic">Klebsormidium nitens</span> (Kn). The phylogenetic tree was constructed using MEGA11 software with the maximum likelihood method. The numbers at the nodes represent bootstrap values as percentages of 1000 replicates and the 0.1 scale denotes 10% change.</p>
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<p><span class="html-italic">O</span>-acetyltransferases mediating <span class="html-italic">O</span>-acetylation of xyloglucan. (<b>A</b>) Diagram of representative structural motifs of the XXXG-type xyloglucan showing acetyl groups attached to side-chain Gal residues. The letters underneath the diagram denote side chain structures and the underlined letters indicate the presence of <span class="html-italic">O</span>-acetyl groups. Question marks indicate enzymes that have not been identified or biochemically verified. (<b>B</b>) Diagram of representative structural motifs of the tomato XXGG<sub>n</sub>-type xyloglucan showing acetyl groups attached to side-chain Gal and Ara<span class="html-italic">f</span> residues and to backbone Glc residues. The question marks indicate that the corresponding <span class="html-italic">O</span>-acetyltransferases have not yet been identified. (<b>C</b>) Phylogenetic relationship of biochemically characterized XGOATs and XyBATs from <span class="html-italic">Arabidopsis</span> (At), poplar (<span class="html-italic">Populus trichocarpa</span>; Ptr), tomato (<span class="html-italic">Solanum lycopersicum</span>; Sl), rice (<span class="html-italic">Oryza sativa</span>; Os), and <span class="html-italic">Brachypodium distachyon</span> (Bd). The phylogenetic tree was constructed using MEGA11 software with the maximum likelihood method. The numbers at the nodes represent bootstrap values as percentages of 1000 replicates and the 0.1 scale denotes 10% change. (<b>D</b>) <sup>1</sup>H-NMR spectra of the acetyl resonance region of unacetylated xyloglucan oligomers (control) and acetylated xyloglucan oligomers catalyzed by XGOAT2/AXY4L showing resonance signals corresponding to acetyl groups attached to <span class="html-italic">O</span>-3, <span class="html-italic">O</span>-4, and <span class="html-italic">O</span>-6 of side-chain Gal residues. See Zhong et al. (2018) [<a href="#B22-plants-13-02304" class="html-bibr">22</a>] for details. (<b>E</b>) <sup>1</sup>H-NMR spectra of unacetylated cellohexaose (control) and acetylated cellohexaose catalyzed by OsXyBAT6 showing resonance signals corresponding to 6-<span class="html-italic">O</span>-acetylated backbone Glc residues (highlighted in yellow). See Zhong et al. (2020) [<a href="#B26-plants-13-02304" class="html-bibr">26</a>] for details. Abbreviations: Ac, acetyl; Ara<span class="html-italic">f</span>, arabinofuranose; Fuc, fucose; Gal, galactose; Glc, glucose; Xyl, xylose.</p>
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<p><span class="html-italic">O</span>-acetyltransferases mediating <span class="html-italic">O</span>-acetylation of xylan. (<b>A</b>) Diagram of the structural feature of xylan showing acetyl groups attached to <span class="html-italic">O</span>-2 and/or <span class="html-italic">O</span>-3 of Xyl residues. Abbreviations: Ac, acetyl; GlcA, glucuronic acid; Xyl, xylose. (<b>B</b>) Phylogenetic relationship of biochemically characterized XOATs from <span class="html-italic">Arabidopsis</span> (At), poplar (<span class="html-italic">Populus trichocarpa</span>; Ptr), and rice (<span class="html-italic">Oryza sativa</span>; Os). The phylogenetic tree was constructed using MEGA11 software with the maximum likelihood method. The numbers at the nodes represent bootstrap values as percentages of 1000 replicates and the 0.1 scale denotes 10% change. (<b>C</b>) <sup>1</sup>H-NMR spectra of acetyl xylan isolated from <span class="html-italic">Arabidopsis</span>, poplar, and rice. Shown on the left are resonances corresponding to carbohydrate (3.0–5.5 ppm) and acetyl groups (2.0–2.25 ppm). Shown on the right are enlarged acetyl resonances attributed to acetyl groups attached to <span class="html-italic">O</span>-2 (Xyl-2Ac), <span class="html-italic">O</span>-3 (Xyl-3Ac), both <span class="html-italic">O</span>-2 and <span class="html-italic">O</span>-3 (Xyl-2,3Ac) of Xyl residues, and <span class="html-italic">O</span>-3 of 2-<span class="html-italic">O</span>-GlcA-substituted Xyl residues. See Zhong et al. (2017, 2018 and 2018) [<a href="#B35-plants-13-02304" class="html-bibr">35</a>,<a href="#B36-plants-13-02304" class="html-bibr">36</a>,<a href="#B47-plants-13-02304" class="html-bibr">47</a>] for details. DS<sub>AC</sub>, degree of substitutions by acetyl groups.</p>
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<p><span class="html-italic">O</span>-acetyltransferases mediating <span class="html-italic">O</span>-acetylation of mannan. (<b>A</b>) Diagram of the structural feature of glucomannan showing acetyl groups attached to <span class="html-italic">O</span>-2 or <span class="html-italic">O</span>-3 of Man residues. Abbreviations: Ac, acetyl; Glc, glucose; Man, mannose. (<b>B</b>) Phylogenetic relationship of biochemically characterized MOATs from <span class="html-italic">Arabidopsis</span> (At), poplar (<span class="html-italic">Populus trichocarpa</span>; Ptr), rice (<span class="html-italic">Oryza sativa</span>; Os), voodoo lily (<span class="html-italic">Amorphophallus Konjac</span>; Ak), pine (<span class="html-italic">Pinus taeda</span>; Pt), spruce (<span class="html-italic">Picea abies</span>; Pa), <span class="html-italic">Selaginella moellendorffii</span> (Sm), moss (<span class="html-italic">Physcomitrium Patens</span>; Pp), and <span class="html-italic">Marchantia polymorpha</span> (Mp). The phylogenetic tree was constructed using MEGA11 software with the maximum likelihood method. The numbers at the nodes represent bootstrap values as percentages of 1000 replicates and the 0.1 scale denotes 10% change. (<b>C</b>) <sup>1</sup>H-NMR spectra of acetyl mannan isolated from <span class="html-italic">Arabidopsis</span>. The top panel shows resonances corresponding to carbohydrate (3.0–5.5 ppm) and acetyl groups (2.0–2.25 ppm). The bottom panel displays enlarged resonances attributed to Man residues acetylated at <span class="html-italic">O</span>-2 (Man-2Ac) or <span class="html-italic">O</span>-3 (Man-3Ac). Man-2Ac-<span class="underline">Man-2Ac</span> and <span class="underline">Man-2Ac</span>-Man-3Ac refer to resonances corresponding to the 2-O-acetylated Man (underlined) in two consecutive acetylated Man residues. See Zhong et al. (2018) [<a href="#B61-plants-13-02304" class="html-bibr">61</a>] for details. DS<sub>AC</sub>, degree of substitutions by acetyl groups.</p>
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<p><span class="html-italic">O</span>-acetyltransferases mediating <span class="html-italic">O</span>-acetylation of pectins. (<b>A</b>) Diagrams of the structural units of the pectins HG and RG-I and the side chain B of RG-II showing acetyl groups attached to <span class="html-italic">O</span>-2 and/or <span class="html-italic">O</span>-3 of GalA residues in HG and RG-I and to the side-chain MeFuc and Ace<span class="html-italic">f</span> residues in RG-II. The question marks indicate that the corresponding <span class="html-italic">O</span>-acetyltransferases have not yet been identified. Abbreviations: Ac, acetyl; Acef, aceric acid; Api, apiose; Ara<span class="html-italic">p</span>, arabinopyranose; Gal, galactose; GalA, galacturonic acid; MeFuc, 2-<span class="html-italic">O</span>-methylfucose; Rha, rhamnose. (<b>B</b>) Phylogenetic relationship of biochemically characterized POATs from <span class="html-italic">Arabidopsis</span> (At), <span class="html-italic">Marchantia polymorpha</span> (Mp), and <span class="html-italic">Klebsormidium nitens</span> (Kn). The phylogenetic tree was constructed using MEGA11 software with the maximum likelihood method. The numbers at the nodes represent bootstrap values as percentages of 1000 replicates and the 0.1 scale denotes 10% change. (<b>C</b>) <sup>1</sup>H-NMR spectra of the acetyl resonance region of unacetylated HG (control) and acetylated HG catalyzed by AtPOAT8 showing resonances attributed to acetyl groups. (<b>D</b>) <sup>1</sup>H-NMR spectra of the acetyl resonance region of unacetylated RG-I (control) and acetylated RG-I catalyzed by AtPOAT8 showing resonances attributed to acetyl groups. See Zhong et al. (2024) [<a href="#B73-plants-13-02304" class="html-bibr">73</a>] for details.</p>
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<p>Structural similarities of plant cell wall polysaccharide <span class="html-italic">O</span>-acetyltransferases. The structure of the bacterial peptidoglycan <span class="html-italic">O</span>-acetyltransferase SaOatA<sub>c</sub> is shown for comparison. The structural models of the catalytic domains of AtMOAT3 (amino acids 92-456), AtXGOAT1 (amino acids 72-416), OsXyBAT6 (amino acids 61-420), and AtPOAT8 (amino acids 168-533) were predicted using AlphaFold2, and the structures of the catalytic domains of AtXOAT1 (amino acids 133-487) and SaOatA<sub>c</sub> (amino acids 445-601) were obtained from the Protein Data Bank (6CCI and 6VJP, respectively). The top panels display the whole view of the structural models of AtMOAT3 (<b>A</b>), AtXGOAT1 (<b>B</b>), OsXyBAT6 (<b>C</b>), AtPOAT8 (<b>D</b>), AtXOAT1 (<b>E</b>), and SaOatA<sub>c</sub> (<b>F</b>). The bottom panels show close-up views of the active site of each protein from the boxed area in the top panel. The conserved Ser-His-Asp catalytic triad at the active site of each protein is highlighted.</p>
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9 pages, 724 KiB  
Brief Report
Cell Wall Profiling of the Resurrection Plants Craterostigma plantagineum and Lindernia brevidens and Their Desiccation-Sensitive Relative, Lindernia subracemosa
by John P. Moore, Brock Kuhlman, Jeanett Hansen, Leonardo Gomez, Bodil JØrgensen and Dorothea Bartels
Plants 2024, 13(16), 2235; https://doi.org/10.3390/plants13162235 - 12 Aug 2024
Viewed by 1018
Abstract
Vegetative desiccation tolerance has evolved within the genera Craterostigma and Lindernia. A centre of endemism and diversification for these plants appears to occur in ancient tropical montane rainforests of east Africa in Kenya and Tanzania. Lindernia subracemosa, a desiccation-sensitive relative of Craterostigma [...] Read more.
Vegetative desiccation tolerance has evolved within the genera Craterostigma and Lindernia. A centre of endemism and diversification for these plants appears to occur in ancient tropical montane rainforests of east Africa in Kenya and Tanzania. Lindernia subracemosa, a desiccation-sensitive relative of Craterostigma plantagineum, occurs in these rainforests and experiences adequate rainfall and thus does not require desiccation tolerance. However, sharing this inselberg habitat, another species, Lindernia brevidens, does retain vegetative desiccation tolerance and is also related to the resurrection plant C. plantagineum found in South Africa. Leaf material was collected from all three species at different stages of hydration: fully hydrated (ca. 90% relative water content), half-dry (ca. 45% relative water content) and fully desiccated (ca. 5% relative water content). Cell wall monosaccharide datasets were collected from all three species. Comprehensive microarray polymer profiling (CoMPP) was performed using ca. 27 plant cell-wall-specific antibodies and carbohydrate-binding module probes. Some differences in pectin, xyloglucan and extension epitopes were observed between the selected species. Overall, cell wall compositions were similar, suggesting that wall modifications in response to vegetative desiccation involve subtle cell wall remodelling that is not reflected by the compositional analysis and that the plants and their walls are constitutively protected against desiccation. Full article
(This article belongs to the Special Issue New Perspectives on the Plant Cell Wall)
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<p>Monosaccharide compositional analysis of the total AIR isolated from leaf material of <span class="html-italic">Craterostigma plantagineum</span> (<b>a</b>), and <span class="html-italic">Lindernia brevidens</span> (<b>b</b>). White bars represent hydrated leaves, mid-grey shaded bars represent partially hydrated leaves and shaded bars represent desiccated leaves. Monosaccharide codes are for arabinose (Ara), rhamnose (Rha), fucose (Fuc), xylose (Xyl), mannose (Man), galactose (Gal), galacturonic acid (GalUA), glucose (Glc) and glucuronic acid (GlcUA). Error bars represent the standard error (SE) of the mean of four biological samples with two technical replicates per biological sample. Statistically significant differences, based on one-way ANOVA variance testing, are indicated on the bar graphs as an asterisk.</p>
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<p>Comprehensive microarray polymer profiling (CoMPP) analysis of plant-leaf cell wall fractions from CDTA-extractable material (<b>a</b>) and NaOH-extractable material (<b>b</b>) isolated from <span class="html-italic">Craterostigma plantagineum</span>, <span class="html-italic">Lindernia brevidens</span> and <span class="html-italic">Lindernia subracemosa</span> leaves that were hydrated (H), partially hydrated (PD) or desiccated (D). The heatmaps indicate the relative abundance of plant cell wall glycan-associated epitopes present in the AIR, and the colour intensity is correlated to the mean spot signals. The values in the heatmap are the mean spot signals from three experiments. The highest signal in the entire data set was set to 100, and all other data were adjusted accordingly.</p>
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17 pages, 570 KiB  
Article
Comparative Analysis of CAZymes from Trichoderma longibrachiatum LMBC 172 Cultured with Three Different Carbon Sources: Sugarcane Bagasse, Tamarind Seeds, and Hemicellulose Simulation
by Alex Graça Contato, Tiago Cabral Borelli, Ana Karine Furtado de Carvalho, Heitor Buzetti Simões Bento, Marcos Silveira Buckeridge, Janet Rogers, Steven Hartson, Rolf Alexander Prade and Maria de Lourdes Teixeira de Moraes Polizeli
Clean Technol. 2024, 6(3), 994-1010; https://doi.org/10.3390/cleantechnol6030050 - 8 Aug 2024
Viewed by 916
Abstract
The examination of fungal secretomes has garnered attention for its potential to unveil the repertoire of secreted proteins, notably CAZymes (Carbohydrate-Active enzymes), across various microorganisms. This study presents findings on categorizing the secretome profile of CAZymes by their function and family, derived from [...] Read more.
The examination of fungal secretomes has garnered attention for its potential to unveil the repertoire of secreted proteins, notably CAZymes (Carbohydrate-Active enzymes), across various microorganisms. This study presents findings on categorizing the secretome profile of CAZymes by their function and family, derived from the filamentous fungus Trichoderma longibrachiatum LMBC 172. The cultivation was performed through submerged fermentation with three distinct carbon sources: sugarcane bagasse, tamarind seeds, and a control simulating hemicellulose containing 0.5% beechwood xylan plus 0.5% oat spelt xylan. The secretome analysis revealed 206 distinct CAZymes. Each carbon source showed particularities and differences. Of these, 89 proteins were produced simultaneously with all the carbon sources; specifically, 41 proteins using only the hemicellulose simulation, 29 proteins when sugarcane bagasse was used as a carbon source, and only 3 when tamarind seeds were used. However, in this last condition, there was a high intensity of xyloglucanase GH74 production, thus reaffirming the richness of xyloglucan in the constitution of these seeds. When evaluating the proteins found in two conditions, 18 proteins were shown between the simulation of hemicellulose and sugarcane bagasse, 11 proteins between the simulation of hemicellulose and tamarind seeds, and 15 proteins between sugarcane bagasse and tamarind seeds. Among the proteins found, there are representatives of different families such as glycosyl hydrolases (GHs) that cleave cellulose, hemicellulose, pectin, or other components; carbohydrate esterases (CEs); polysaccharide lyases (PLs); carbohydrate-binding modules (CBMs); and auxiliary activity enzymes (AAs). These results demonstrate the importance of analyzing CAZymes secreted by microorganisms under different culture conditions. Full article
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<p>CAZymes from secretome analysis of <span class="html-italic">T. longibrachiatum</span> LMBC 172 in culture condition: hemicellulose simulation, or sugarcane bagasse, or tamarind seeds. (<b>A</b>) Total CAZymes found. (<b>B</b>) Venn plot correlating the CAZymes found in each culture condition.</p>
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14 pages, 3603 KiB  
Article
Xyloglucan–Cellulose Nanocrystals Mixtures: A Case Study of Nanocolloidal Hydrogels and Levers for Tuning Functional Properties
by Géraldine Rangel, Céline Moreau, Ana Villares, Christophe Chassenieux and Bernard Cathala
Gels 2024, 10(5), 334; https://doi.org/10.3390/gels10050334 - 15 May 2024
Viewed by 1067
Abstract
The development of fully biobased hydrogels obtained by simple routes and in the absence of toxic or environmentally harmful reagents is a major challenge in meeting new societal demands. In this work, we discuss the development of hydrogels made from cellulose nanocrystals (CNCs) [...] Read more.
The development of fully biobased hydrogels obtained by simple routes and in the absence of toxic or environmentally harmful reagents is a major challenge in meeting new societal demands. In this work, we discuss the development of hydrogels made from cellulose nanocrystals (CNCs) and xyloglucan (XG), two non-toxic, renewable, and biobased components. We present three strategies to fine-tune the functional properties. The first one consists in varying the XG/CNC ratio that leads to the modulation of the mechanical properties of hydrogels as well as a better comprehension of the gel mechanism formation. The second relies on tuning the XG chains’ interaction by enzymatic modification to achieve thermoresponsive systems. Finally, the third one is based on the increase in the hydrogel solid content by osmotic concentration. The high-solid-content gels were found to have very high mechanical properties and self-healing properties that can be used for molding materials. Overall, these approaches are a case study of potential modifications and properties offered by biobased nanocolloidal hydrogels. Full article
(This article belongs to the Special Issue Cellulose-Based Gels: Synthesis, Properties, and Applications)
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<p>Schematic view of the different levers that were investigated in this paper to tune XG/CNC nanocolloidal hydrogel properties. XG (green pearl necklaces) and CNC (blue rods) are not scaled. The first one corresponds to the variation in the XC/CNC weight ratio, the second one involves the enzymatic modification of the XG structure to change interaction, and finally, the increase in concentration that leads to higher crowding corresponds to the third lever.</p>
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<p>Principle of QCM−D (<b>2a</b>) and <span class="html-italic">ΔD</span> vs. <span class="html-italic">Δf</span> scheme (<b>2b</b>). Graph (<b>2e</b>) presents the <span class="html-italic">ΔD</span> vs. <span class="html-italic">Δf</span> plots for different injections of XG/CNC ratio corresponding to XG adsorption at low (<b>2c</b>) and high (<b>2d</b>) concentration. As the injection concentration increases (from 0.3 μg/mL up to 15 μg/mL), the slopes increase, indicating a more dissipative structure that can be attributed to the formation of loops and tails.</p>
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<p>Phase diagram of the XG/CNC system. The gelation concentration (XG + CNC) is plotted against de XG/CNC ratio. The continuous line is merely a guide for the eyes to visualize the gel and liquid states.</p>
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<p>(<b>a</b>) Storage modulus as a function of pulsation for neat XG and mixtures at various XG/CNC weight ratios as indicated in the legend of the figure. The total concentration is 20 g L<sup>−1</sup>. (<b>b</b>) Values of <span class="html-italic">G</span>′ (blue trace) and <span class="html-italic">G</span>″ (red trace) obtained for the same hydrogels at 1 Hz as a function of the XG/CNC ratio; lines are a guide for the reader.</p>
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<p>Enzymatic modification of XG by β-galactosidase and temperature dependence of the storage (<span class="html-italic">G</span>′) and loss (<span class="html-italic">G</span>″) moduli for three different XG/CNC ratios.</p>
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<p>CNC + XG concentration for hydrogels with various XG/CNC ratios obtained after osmotic dehydration starting from a concentration of 20 g L<sup>−1</sup>.</p>
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<p>(<b>a</b>) Frequency sweeps obtained at 20 °C for storage (blue circle) and loss (red diamond) moduli of hydrogels at various XG/CNC ratios for hydrogels obtained by dehydration; the concentrations were 165, 96, and 86 g L<sup>−1</sup>, respectively. (<b>b</b>) Strain sweeps obtained at 20 °C for storage (blue circle) and loss (red diamond) moduli for the same hydrogels. The blue traces correspond to an increase in the strain (dark blue <span class="html-italic">G</span>′/dark red <span class="html-italic">G</span>″), and the light blue (<span class="html-italic">G</span>′)/light red (<span class="html-italic">G</span>″) ones to a decrease. (<b>c</b>) Macroscopic pictures of composite hydrogels with XG/CNC = 0.25 and C = 165 g L<sup>−1</sup> before and after being left overnight at 4 °C in a patterned mold.</p>
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15 pages, 2523 KiB  
Article
Spectroscopic Investigation of Tomato Seed Germination Stimulated by Trichoderma spp.
by Igor Vukelić, Danka Radić, Ilinka Pećinar, Steva Lević, Daniela Djikanović, Ksenija Radotić and Dejana Panković
Biology 2024, 13(5), 340; https://doi.org/10.3390/biology13050340 - 13 May 2024
Cited by 1 | Viewed by 1868
Abstract
Seed germination is a complex process that can be negatively affected by numerous stresses. Trichoderma spp. are known as effective biocontrol agents as well as plant growth and germination stimulators. However, understanding of the early interactions between seeds and Trichoderma spp. remains limited. [...] Read more.
Seed germination is a complex process that can be negatively affected by numerous stresses. Trichoderma spp. are known as effective biocontrol agents as well as plant growth and germination stimulators. However, understanding of the early interactions between seeds and Trichoderma spp. remains limited. In the present paper, Fourier-transform infrared spectroscopy (FTIR) and Raman spectroscopy were used to reveal the nature of tomato seed germination as stimulated by Trichoderma. A rapid response of tomato seeds to Trichoderma spp. was observed within 48 h on Murashige and Skoog medium (MS) substrate, preceding any physical contact. Raman analysis indicated that both Trichoderma species stimulated phenolic compound synthesis by triggering plant-specific responses in seed radicles. The impact of T. harzianum and T. brevicompactum on two tomato cultivars resulted in alterations to the middle lamella pectin, cellulose, and xyloglucan in the primary cell wall. The Raman spectra indicated increased xylan content in NA with T9 treatment as well as increased hemicelluloses in GZ with T4 treatment. Moreover, T4 treatment resulted in elevated conjugated aldehydes in lignin in GZ, whereas the trend was reversed in NA. Additionally, FTIR analysis revealed significant changes in total protein levels in Trichoderma spp.-treated tomato seed radicles, with simultaneous decreases in pectin and/or xyloglucan. Our results indicate that two complementary spectroscopic methods, FTIR and Raman spectroscopy, can give valuable information on rapid changes in the plant cell wall structure of tomato radicles during germination stimulated by Trichoderma spp. Full article
(This article belongs to the Special Issue Beneficial Microorganisms for Plants)
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<p>Tomato seed (GZ) cultivated in the presence of <span class="html-italic">T. harzianum</span> on MS medium.</p>
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<p>Plant radicles imaged with a scanning electron microscope. GZ tomato radicle epidermis surface in control condition at 200× magnification (<b>A</b>) and at 2000× magnification (<b>C</b>); GZ tomato radicle epidermis surface after contact with <span class="html-italic">T. brevicompactum</span> at 200× magnification (<b>B</b>) and at 2000× magnification (<b>D</b>). Similar results were obtained for another tomato variety Narvik (not presented).</p>
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<p>Percentage of germinated tomato seeds, after 48 h on MS media under different treatments: control (C), <span class="html-italic">T. harzianum</span> (T4), <span class="html-italic">T. virens</span> (T7), <span class="html-italic">T. brevicompactum</span> (T9), <span class="html-italic">T. longibrachiatum</span> (T11), and <span class="html-italic">T. citrinoviride</span>s (T13). (<b>A</b>) Percentage of germinated GZ seeds; (<b>B</b>) Percentage of germinated NA seeds. One-way ANOVA and post hoc Tukey’s test were used for statistical analysis of data. Values marked with the same letter are not statistically significantly different according to Tukey’s test (<span class="html-italic">p</span> &lt; 0.05). The error lines represent ± standard deviation of the mean. Different letters (a, b and c) indicate statistically significant differences according to Tukey’s test (<span class="html-italic">p</span> &lt; 0.05). For each treatment, 28 Petri dishes of 30 seeds each were analyzed (<span class="html-italic">n</span> = 28).</p>
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<p>Averages of normalized Raman spectra of two tomato cultivar radicles GZ and NA in control conditions or in co-culture with either <span class="html-italic">T. harzianum</span> or <span class="html-italic">T. brevicompactum</span>. (<b>A</b>) Spectra of the Gruzanski zlatni cultivar radicles (GZ—control; GZT4—<span class="html-italic">T. harzianum</span>; GZT9—<span class="html-italic">T. brevicompactum</span>); (<b>B</b>) Spectra of the Narvik cultivar radicles (NA—control; NAT4—<span class="html-italic">T. harzianum</span>; NAT9—<span class="html-italic">T. brevicompactum</span>).</p>
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<p>(<b>A</b>) Score plot of the first principal component (PC1) versus the second principal component (PC2) of the tomato radicle from two cultivars (GZ and NA) treated with <span class="html-italic">T. harzianum</span> (T4) and <span class="html-italic">T. brevicompactum</span> (T9). (<b>B</b>) Loading plot corresponding to PC1 and PC2 responsible for the differences between the cultivars. N—closed cycle, NAT4—closed square, NAT9—closed triangle, GZ—open cycle, GZT4—open square, and GZT9—open triangle.</p>
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<p>FTIR spectra of homogenized Gruzanski zlatni (<b>A</b>) and Narvik (<b>B</b>) radicles under different treatments: control (GZ, NA), seeds treated with <span class="html-italic">T. harzianum</span> (GZT4, NAT4), seeds treated with <span class="html-italic">T. brevicompactum</span> (GZT9, NAT9).</p>
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