Pectin Methylesterase Inhibitor Research Articles

Embryo sac cellularization defects lead to supernumerary egg cells and twin embryos in Arabidopsis thaliana

Embryo sac cellularization defects lead to supernumerary egg cells and twin embryos in Arabidopsis thaliana

Strigolactones alleviate AlCl3 stress by vacuolar compartmentalization and cell wall blocking in apple.

Poor management and excess fertilization of apple (Malus domestica Borkh.) orchards are causing increasingly serious soil acidification, resulting in Al toxicity and direct poisoning of roots. Strigolactones (SLs) are reported to be involved in plant responses to abiotic stress, but their role and mechanism under AlCl3 stress remain unknown. Here, we found that applying 1 μm GR24 (an SL analoge) significantly alleviated AlCl3 stress of M26 apple rootstock, mainly by blocking the movement of Al through cell wall and by vacuolar compartmentalization of Al. RNA-seq analysis identified the core transcription factor gene MdWRKY53, and overexpressing MdWRKY53 enhanced AlCl3 tolerance in transgenic apple plants through the same mechanism as GR24. Subsequently, we identified MdPMEI45 (encoding pectin methylesterase inhibitor) and MdALS3 (encoding an Al transporter) as downstream target genes of MdWRKY53 using chromatin immunoprecipitation followed by sequencing (ChIP-seq). GR24 enhanced the interaction between MdWRKY53 and the transcription factor MdTCP15, further increasing the binding of MdWRKY53 to the MdPMEI45 promoter and inducing MdPMEI45 expression to prevent Al from crossing cell wall. MdWRKY53 also bound to the promoter of MdALS3 and enhanced its transcription to compartmentalize Al in vacuoles under AlCl3 stress. We therefore identified two modules involved in alleviating AlCl3 stress in woody plant apple: the SL-WRKY+TCP-PMEI module required for excluding external Al by blocking the entry of Al3+ into cells and the SL-WRKY-ALS module allowing internal detoxification of Al through vacuolar compartmentalization. These findings lay a foundation for the practical application of SLs in agriculture.

Pectin methylesterase inhibitors GhPMEI53 and AtPMEI19 improve seed germination by modulating cell wall plasticity in cotton and Arabidopsis

The germination process of seeds is influenced by the interplay between two opposing factors: pectin methylesterase (PME) and pectin methylesterase inhibitor (PMEI), which collectively regulate patterns of pectin methylesterification. Despite the recognized importance of pectin methylesterification in seed germination, the specific mechanisms that govern this process remain unclear. In this study, we demonstrated that the overexpression of GhPMEI53 is associated with a decrease in PME activity and an increase in pectin methylesterification. This leads to the softening of the cell wall in seeds, which positively regulates cotton seed germination. AtPMEI19, the homologue in Arabidopsis thaliana, plays a similar role in seed germination to GhPMEI53, indicating a conserved function and mechanism of PMEI in seed germination regulation. Further studies revealed that GhPMEI53 and AtPMEI19 directly contribute to promoting radicle protrusion and seed germination by inducing cell wall softening and reducing mechanical strength. Additionally, the pathways of ABA and GA in the transgenic materials underwent significant changes, suggesting that GhPMEI53/AtPMEI19-mediated pectin methylesterification serves as a regulatory signal for the related phytohormones involved in seed germination. In summary, GhPMEI53 and its homologs alter the mechanical properties of cell walls, influencing the mechanical resistance of the endosperm or testa. Moreover, they impact cellular phytohormone pathways (e.g., ABA, GA) to regulate seed germination. These findings enhance our understanding of pectin methylesterification in cellular morphological dynamics and signaling transduction, and contribute to a more comprehensive understanding of the PME/PMEI super-gene family in plants.

Inactivation mechanisms on pectin methylesterase by high pressure processing combined with its recombinant inhibitor

High pressure processing (HPP) juice often experiences cloud loss during storage, caused by the activity of pectin methylesterase (PME). The combination of HPP with natural pectin methylesterase inhibitor (PMEI) could improve juice stability. However, extracting natural PMEI is challenging. Gene recombination technology offers a solution by efficiently expressing recombinant PMEI from Escherichia coli and Pichia pastoris. Experimental and molecular dynamics simulation were conducted to investigate changes in activity, structure, and interaction of PME and recombinant PMEI during HPP. The results showed PME retained high residual activity, while PMEI demonstrated superior pressure resistance. Under HPP, PMEI's structure remained stable, while the N-terminus of PME's α-helix became unstable. Additionally, the helix at the junction with the PME/PMEI complex changed, thereby affecting its binding. Furthermore, PMEI competed with pectin for active sites on PME, elucidating. The potential mechanism of PME inactivation through the synergistic effects of HPP and PMEI.

Functional dissection of the PME and PMEI super families in the protection of Populus pectin during Fusarium wilt

Integration of the pectin homogalacturonan (HG) integrity and remodeling into defense immunity facilitates building blocks of stress resilience within the host and pathogen interactions in the forests. Recent advances in apoplastic pectin methylesterases (PMEs) and coordinated PME inhibitors (PMEIs) deployed immune signaling, highlighting their critical roles in regulating biotic stress adaptation. Nevertheless, the PME and PMEI multiple isoenzyme families involved in defense priming in the model tree species are largely unknown. The in silico demonstration of the molecular profile identified 70 PME- (50 type-I proPME and 20 type-II PME) and 49 PMEI-encoding genes in the Populus genome. Evaluation of the transcript abundance verified 20 PtproPMEs and 18 PtPMEIs that showed predominant expression in the root and genetic redundancy for type-II PtPMEs. Dynamic changes in the transcriptome and experimental evaluation revealed a minuscule number of type-I proPMEs and PMEIs that showed marked promotions during a time-course Fusarium solani (Fs) infection. Using GC/LC-mass-based non-targeted metabolomics to inspect structure-related carbohydrates and PME activities suggested a correlation between proPME-PMEI co-expression and deformed pectin HG biosynthesis. Further functional examination of the enzyme inhibitory affinities of recombinant PtPMEI1 and 39 in vitro and apoplastic patterns suggested a clustering of the type-I proPME in the regulation of the innate immunity by maintaining homeostasis of the pectin HG de-methylesterification (DME) in the cell wall. The coherent omics-wide survey of the fungi pathogen-induced PME-PMEIs and disturbed carbohydrate accumulation provide theoretical cues for in-depth mining of the biological significance of the underlying immune signaling networks within an apoplastic niche in trees.

Pectin-Degrading Enzymes during Soursop (Annona muricata L.) Fruit Ripening: A Bioinformatics Approach

The rapid ripening of soursop (Annona muricata L.) fruits is owing to its high respiration rate. Several enzymes affect the fruit cell wall in this process, resulting in the depolymerization of pectin primarily in the homogalacturonan. The main group of enzymes affecting the pectin content of soursop fruits include polygalacturonase (PG), pectate lyase (PL), pectin methylesterase (PME), and PME inhibitors (PMEis). In this study, pectin-degrading enzymes were analyzed using bioinformatic tools to uncover the gaps in our knowledge of this fruit. In this context, 67 genes encoding PG, 33 PL, 58 PME, and 39 PMEi isoenzymes were found. These genes were categorized into several families based on the results of phylogenetic analysis. Regarding the analysis of gene expression, a total of 25 were identified as differentially expressed genes (DEGs) in PG, while 3, 21, and 15 were found for PL, PME, and PMEis, respectively. Likewise, functional enrichment analysis demonstrated that the DEGs are involved in the modification of the cell wall, specifically in the degradation of pectin. On the other hand, gene co-expression networks revealed that the genes PG32 and PG35 affect the expression of PGs, as well as PL19 of the PL family, PME20, PME32, and PME35 of the PME family, and PMEi04 of the PMEi family. This suggests that they have a significant impact on the softening of soursop fruits.

A novel PMEI3 in common bean suggests a key role of pectin methylesterification in Pseudomonas resistance.

The mechanisms that drives the common bean susceptibility against Pseudomonas syringae (Pph) has not been clarified yet. For this, 15-day-old common bean plants, variety riñón, were infected with Pph to analyze the transcriptomic changes at 2 and 9h post-infection. RNA-seq analysis showed an upregulation of genes involved in defense/signaling at 2h, most of them being downregulated at 9h, suggesting that Pph would inhibit the transcriptomic reprogramming of the plant. This trend was also observed in the modulation of 101 cell wall (CW)-related genes. CW composition changes at early stages of Pph infection were associated with homogalacturonan methylation and the formation of egg boxes. Among the CW genes modulated, a pectin methylesterase inhibitor 3 (PvPMEI3) gene, closely related to AtPMEI3, was detected. PvPMEI3 protein was located in the apoplast and its PME inhibitory activity was demonstrated. PvPMEI3 seems to be a good candidate to play a key role in Pph infection, which was supported by the analysis of Arabidopsis pmei3 mutant, that showed susceptibility to Pph, in contrast to resistant Col-0 plants. All these results point to a key role of the pectin methylesterification degree affecting host resistance to Pph during the first steps of the attack.

Uncovering the interactions between PME and PMEI at the gene and protein levels: Implications for the design of specific PMEI.

Pectin methylesterase inhibitor (PMEI) can specifically bind and inhibit the activity of pectin methylesterase (PME), which has been widely used in fruit and vegetable juice processing. However, the limited three-dimensional structure, unclear action mechanism, low thermal stability and biological activity of PMEI severely limited its application. In this work, molecular recognition and conformational changes of PME and PMEI were analyzed by various molecular simulation methods. Then suggestions were proposed for improving thermal stability and affinity maturation of PMEI through semi-rational design. Phylogenetic trees of PME and PMEI were established using the Maximum likelihood (ML) method. The results show that PME and PMEI have good sequence and structure conservation in various plants, and the simulated data can be widely adopted. In this work, MD simulations were performed using AMBER20 package and ff14SB force field. Protein interaction analysis indicates that H-bonds, van der Waals forces, and the salt bridge formed of K224 with ID116 are the main driving forces for mutual molecular recognition of PME and PMEI. According to the analyses of free energy landscape (FEL), conformational cluster, and motion, the association with PMEI greatly disrupts PME's dispersed functional motion mode and biological function. By monitoring the changes of residue contact number and binding free energy, IG35M/ IG35R: IT93F and IT113W/ IT113W: ID116W mutations contribute to thermal stability and affinity maturation of the PME-PMEI complex system, respectively. This work reveals the interaction between PME and PMEI at the gene and protein levels and provides options for modifying specific PMEI.

Genome-wide association identifies key loci controlling blackberry postharvest quality.

Blackberry (Rubus subgenus Rubus) is a soft-fruited specialty crop that often suffers economic losses due to degradation in the shipping process. During transportation, fresh-market blackberries commonly leak, decay, deform, or become discolored through a disorder known as red drupelet reversion (RDR). Over the past 50 years, breeding programs have achieved better fruit firmness and postharvest quality through traditional selection methods, but the underlying genetic variation is poorly understood. We conducted a genome-wide association of fruit firmness and RDRmeasured in 300 tetraploid fresh-market blackberry genotypes from 2019-2021 with 65,995 SNPs concentrated in genic regions of the R. argutus reference genome. Fruit firmness and RDR had entry-mean broad sense heritabilities of 68% and 34%, respectively. Three variants on homologs of polygalacturonase (PG), pectin methylesterase (PME), and glucan endo-1,3-β-glucosidase explained 27% of variance in fruit firmness and were located on chromosomes Ra06, Ra01, and Ra02, respectively. Another PG homolog variant on chromosome Ra02 explained 8% of variance in RDR, but it was in strong linkage disequilibrium with 212 other RDR-associated SNPs across a 23 Mb region. A large cluster of six PME and PME inhibitor homologs was located near the fruit firmness quantitative trait locus (QTL) identified on Ra01. RDR and fruit firmness shared a significant negative correlation (r = -0.28) and overlapping QTL regions on Ra02 in this study. Our work demonstrates the complex nature of postharvest quality traits in blackberry, which are likely controlled by many small-effect QTLs. This study is the first large-scale effort to map the genetic control of quantitative traits in blackberry and provides a strong framework for future GWAS. Phenotypic and genotypic datasets may be used to train genomic selection models that target the improvement of postharvest quality.

Genome-wide identification and functional characterization of pectin methylesterase inhibitors associated with male sterility in wheat

Pectin methylesterase inhibitors (PMEIs) can specifically bind to pectin methylesterases (PMEs) and regulate their enzymatic activities to control the status of pectin methyl esterification, which plays crucial roles in stabilizing the cell wall properties during plant growth and development. Although the evolution and biological functions of the PMEI gene family in various plants have been well characterized, the role of PMEI genes in wheat was poorly understood. In this study, a total of 237 PMEI genes (TaPMEIs) were identified in wheat (Triticum aestivum L.), which were dispersed irregularly over 22 chromosomes and tissue-specific in their expression. They were divided into nine groups based on similar motif number and composition. Using quantitative real-time PCR analysis, we discovered that TaPMEI23 as the most abundant-expressed PMEI in anther and significantly upregulated expression in fertile anthers. Moreover, subcellular localization analysis indicated that TaPMEI23 was localized on the plasma membrane or cell wall. Effective silencing of the TaPMEI23 significantly influences the anther dehiscence, pollen activity, seed setting rate, and male fertility, suggesting that PMEIs are necessary for pollen development and male sterility. Our work provides novel insights into the functions of PMEIs in wheat.

Identification and characterization of pectin remodeling gene families in Withania somnifera and their interaction during biotic stress

The Pectin methylesterase (PME), Pectin methylesterase inhibitor (PMEI), and Subtilase (SBT) are pectin remodeling enzymes that play an important role in plant growth and development as well as response to various biotic and abiotic stress conditions. Withania somnifera is an important medicinal plant that gained attention for its phytochemical properties, but relatively little in-depth genomic exploration has been available, including PME, PMEI, and SBT genes. In the present study, 54 PMEs, 19 PMEIs, and 51 SBTs genes were identified from the W. somnifera transcriptome. A detailed phylogeny was performed, followed by a conserved sequence signature analysis, which revealed the major phylogenetic division with Solanum lycopersicum. Insect infestation can cause a significant increase in the functions of these genes, which were identified using PME activity, methanol content, and emission, often by several orders of magnitude. WsPME28, WsPME32, and WsPME50 are putative genes that are significantly upregulated up to six fold during insect infestation, according to gene expression profile analysis. To study specific relative interactions for a PME-SBT pair, molecular docking analysis was performed and found that the serine residue of WsSBT37 interacts with the processing motif (RKLL) of WsPME28 and releases mature PME. As per our best knowledge, this is the first investigation into the identification and characterization of the PME, PMEI, and SBT gene families in W. somnifera. This study will lay the groundwork for future studies targeting these genes, thus giving a better understanding of the stress tolerance mechanism in W. somnifera, and will serve as a reference for determining the most specific SBT-PME pairs in development and stress conditions.

Genome and Transcriptome Identification of a Rice Germplasm with High Cadmium Uptake and Translocation.

The safe production of food on Cd-polluted land is an urgent problem to be solved in South China. Phytoremediation or cultivation of rice varieties with low Cd are the main strategies to solve this problem. Therefore, it is very important to clarify the regulatory mechanism of Cd accumulation in rice. Here, we identified a rice variety with an unknown genetic background, YSD, with high Cd accumulation in its roots and shoots. The Cd content in the grains and stalks were 4.1 and 2.8 times that of a commonly used japonica rice variety, ZH11, respectively. The Cd accumulation in the shoots and roots of YSD at the seedling stage was higher than that of ZH11, depending on sampling time, and the long-distance transport of Cd in the xylem sap was high. Subcellular component analysis showed that the shoots, the cell wall, organelles, and soluble fractions of YSD, showed higher Cd accumulation than ZH11, while in the roots, only the cell wall pectin showed higher Cd accumulation. Genome-wide resequencing revealed mutations in 22 genes involved in cell wall modification, synthesis, and metabolic pathways. Transcriptome analysis in Cd-treated plants showed that the expression of pectin methylesterase genes was up-regulated and the expression of pectin methylesterase inhibitor genes was down-regulated in YSD roots, but there were no significant changes in the genes related to Cd uptake, translocation, or vacuole sequestration. The yield and tiller number per plant did not differ significantly between YSD and ZH11, but the dry weight and plant height of YSD were significantly higher than that of ZH11. YSD provides an excellent germplasm for the exploration of Cd accumulation genes, and the cell wall modification genes with sequence- and expression-level variations provide potential targets for phytoremediation.

PECTIN METHYLESTERASE INHIBITOR18 functions in stomatal dynamics and stomatal dimension.

Pectin methylesterification in guard cell (GC) walls plays an important role in stomatal development and stomatal response to external stimuli, and pectin methylesterase inhibitors (PMEIs) modulate pectin methylesterification by inhibition of pectin methylesterase (PME). However, the function of PMEIs has not been reported in stomata. Here, we report the role of Arabidopsis (Arabidopsis thaliana) PECTIN METHYLESTERASE INHIBITOR18 in stomatal dynamic responses to environmental changes. PMEI18 mutation increased pectin demethylesterification and reduced pectin degradation, resulting in increased stomatal pore size, impaired stomatal dynamics, and hypersensitivity to drought stresses. In contrast, overexpression of PMEI18 reduced pectin demethylesterification and increased pectin degradation, causing more rapid stomatal dynamics. PMEI18 interacted with PME31 in plants, and in vitro enzymatic assays demonstrated that PMEI18 directly inhibits the PME activity of PME31 on pectins. Genetic interaction analyses suggested that PMEI18 modulates stomatal dynamics mainly through inhibition of PME31 on pectin methylesterification in cell walls. Our results provide insight into the molecular mechanism of the PMEI18-PME31 module in stomatal dynamics and highlight the role of PMEI18 and PME31 in stomatal dynamics through modulation of pectin methylesterification and distribution in GC walls.

Grapevine red blotch virus alters grape skin cell-wall composition impacting phenolic extractability during winemaking.

Grapevine red blotch virus (GRBV) is the causal agent of grapevine red blotch disease and is known to delay grape ripening. However, grape cell-wall modifications during GRBV infection are largely unknown, even though the cell wall plays a large role in pathogenicity, viral interactions with host plants, and phenolic extractability during winemaking. Understanding the impact of GRBV infection on cell-wall metabolism is important for the development of potential mitigations strategies. In this study, high-throughput transcriptome sequencing was conducted on Vitis vinifera L. 'Merlot' grapes during ripening. The cell-wall composition, phenolic content, and phenolic extractability at two different commercial harvest points were also determined. Log fold changes indicated a strong induction in diseased grapes at harvest of several transcripts involved in cell-wall solubilization and degradation. However, these observations did not translate to changes in cell-wall composition at either harvest point in diseased grapes, potentially suggesting post-transcriptional regulation. Moderate induction of pectin methylesterase inhibitor transcripts and transcripts associated with pathogenesis-related proteins coincided with increases in pectin and soluble proteins in cell walls of diseased grapes at harvest. Both pectin and pathogenesis-related proteins are known to retain phenolic compounds during winemaking. Our study corroborates this finding when the percentage extractability of flavonols in wines was significantly lower when made from GRBV-infected fruit. These results suggest GRBV alters the grape cell walls, consequently decreasing phenolic extraction during winemaking. © 2023 Society of Chemical Industry.

A Novel Strategy to Improve Cloud Stability of Orange-Based Juice: Combination of Natural Pectin Methylesterase Inhibitor and High-Pressure Processing.

This study investigated the prospect of producing cloud-stable orange-based juice by combining high-pressure processing (HPP) with a natural kiwifruit pectin methylesterase inhibitor (PMEI) during chilled storage. Kiwifruit is rich in a PMEI, which greatly improves the cloud loss caused by the pectin methylesterase (PME) demethylation of pectin. The results show that the cloud loss of orange juice occurred after 3 days, while the orange-kiwifruit mixed juice and kiwifruit puree were cloud stable during 28 days' storage. Although, the kiwifruit puree contained larger particles compared to the orange juice, its higher viscosity and solid-like behavior were dominant, improving the cloud stability of the juice systems. In addition, the particle size distribution and rheological properties were highly related to PME activity, PMEI activity, and pectin characterization. The kiwifruit PMEI showed higher resistance to HPP and storage time than PME. More water-solubilized pectin fractions with a high molecular mass were found in the kiwifruit puree, leading to its high viscosity and large particle size, but a more chelator-solubilized pectin fraction with a low esterification degree was observed in the orange juice, resulting in its cloud loss. In general, the outcome of this work provides a novel strategy to improve the cloud stability of orange-based juices using natural PMEIs and nonthermal processing technologies.

Ovule cell wall composition is a maternal determinant of grain size in barley.

In cereal species, seed and grain size is influenced by growth of the ovule integuments (seed coat), the spikelet hull (lemma and palea) and the filial endosperm. It has remained unclear whether a highly conserved ovule tissue, the nucellus, has any impact on grain size. Immunolabelling revealed that the barley nucellus comprises two distinct cell types that differ in terms of cell wall homogalacturonan (HG) accumulation. Transcriptional profiling of the nucellus identified two pectin methylesterase genes, OVULE PECTIN MODIFIER 1 (OPM1) and OPM2, which are expressed in the ovule but absent from the seed. Ovules from an opm1 opm2 mutant, and plants expressing an ovule-specific pectin methylesterase inhibitor (PMEI), exhibit reduced HG accumulation. This results in changes to ovule cell size and shape, and ovules that are longer than wild-type controls. At grain maturity, this is manifested as significantly longer grain. These findings indicate that cell wall composition during ovule development acts to limit ovule and seed growth. The investigation of ovule PME and PMEI activity reveals an unexpected role of maternal tissues in controlling grain growth prior to fertilisation, one that has been lacking from models exploring improvements in grain size.

CRISPR/Cas9 Mutant Rice Ospmei12 Involved in Growth, Cell Wall Development, and Response to Phytohormone and Heavy Metal Stress.

Pectin is one of the constituents of the cell wall, distributed in the primary cell wall and middle lamella, affecting the rheological properties and the cell wall stickiness. Pectin methylesterase (PME) and pectin methylesterase inhibitor (PMEI) are the most important factors for modifying methyl esterification. In this study, 45 PMEI genes from rice (Oryza sativa L.) were screened by bioinformatics tools, and their structure, motifs, cis-acting elements in the promoter region, chromosomal distribution, gene duplication, and phylogenetic relationship were analyzed. Furthermore, CRISPR/Cas9 was used to edit the OsPMEI12 (LOC_Os03G01020) and two mutant pmei12 lines were obtained to explore the functions of OsPMEI in plant growth and development, and under cadmium (Cd) stress. Compared to wild type (WT) Nipponbare, the second inverted internodes of the mutant plants shortened significantly, resulting in the reduction in plant height at mature stage. The seed setting rate, and fresh and dry weights of the mutants were also decreased in mutant plants. In addition, the pectin methylation of pmei12 lines is decreased as expected, and the pectin content of the cell wall increased at both seedling and maturity stages; however, the cellulose and hemicellulose increased only at seedling stage. Interestingly, the growth of the pmei12 lines was better than the WT in both normal conditions and under two phytohormone (GA3 and NAA) treatments at seedling stage. Under Cd stress, the fresh and dry weights were increased in pmei12 lines. These results indicated that OsPMEI12 was involved in the regulation of methyl esterification during growth, affected cell wall composition and agronomic traits, and might play an important role in responses to phytohormones and stress.

Genome-Wide Identification and Expression Analysis of the PME and PMEI Gene Families in Diospyros kaki: A Bioinformatics Study

Pectins are major components of cell walls in plants. Pectin methylesterases (PMEs) and pectin methylesterase inhibitors (PMEIs) play crucial roles in pectin synthesis and metabolism. Overall, 28 putative DkPMEs and 29 putative DkPMEIs were identified from the D. kaki genome. According to phylogenetic analysis, DkPME/DkPMEI proteins can be classified into four and five clades, respectively. Motif and gene structure analysis showed that DkPME/DkPMEI are highly conserved in the same clades, which indicates that the function of these DkPME/DkPMEI were similar. Besides, DkPME/DkPMEI genes were distributed unevenly on their corresponding chromosomes. Synteny analysis showed that PME or PMEI gene usually matched with more than one DkPME/DkPMEI in D. oleifera, D. lotus, and A. thaliana, implying that the function of these genes in D. kaki may be diverse. Expression analysis showed that DkPME/DkPMEI from the same clade exhibited diverse expression patterns, indicating that these genes might have diverse functions. Functional protein–protein interaction network analysis showed that DkPMEI21 and DkPMEI15 were core nodes and were, respectively, positive and negative regulators for carbohydrate metabolism, stress responses, and sugar signaling. This study provides a theoretical basis for the functional characteristics, evolutionary relationship, and role of these gene families in developing persimmon fruit.

Escaping drought: The pectin methylesterase inhibitor gene Slpmei27 can significantly change drought resistance in tomato

Drought stress will lead to a decrease in tomato yield and poor flavour, yield and quality, resulting in economic losses in agricultural production. Mining the key genes regulating tomato drought resistance is of great significance to improve the drought resistance of tomato plants. The cell wall can directly participate in the plant drought stress response as one of the main components of the cell wall, and the regulation of pectin content in plant drought resistance is still unclear. Here, the candidate gene Solyc08g006690 (Slpmei27) was obtained by fine mapping based on genome sequencing technology (BSA-seq) of late-maturing stress-resistant tomato mutants found in the field. Slpmei27 is expressed in the cell wall. The transient silencing of Slpmei27 by VIGS significantly improved the drought resistance of tomato. Meanwhile, Slpmei27 silencing could significantly change the cell wall structure of plants, change the stomatal pass rate, reduce the water loss rate of plants, improve the scavenging ability of reactive oxygen species, change the redox balance in plants, and thus improve the drought resistance of tomato. The promoter region of this gene contains a large number of hormone-response and stress-response binding sites. The promoter region of the Slpmei27 gene in the mutant could lower the expression of downstream genes. Through this study, the mechanism by which Slpmei27 improves tomato drought resistance was revealed, and the relationship between pectin methyl ester metabolism and plant drought resistance was established, providing a theoretical basis for the production of high-quality tomato materials with high drought resistance.

Transcriptome characteristics during cell wall formation of endosperm cellularization and embryo differentiation in Arabidopsis.

Embryonic and endosperm development are important biological events during Arabidopsis seed development, and are controlled by dynamic changes in a range of gene expression. Nevertheless, the regulatory mechanisms of endosperm cellularization and embryo differentiation remain unclear. Here, we characterized the early embryo and endosperm development of the naa15 mutant that had abnormal embryo differentiation and incomplete endosperm cellularization compared to WT of Arabidopsis, and comparatively investigated the changes of gene expressions in WT seeds at 3, 4, and 5 days after pollination (3W, 4W, and 5W) and the white homozygous aborted naa15 seeds at 5, 6, and 7 DAP (5M, 6M, and 7M) from naa15-1/+ siliques using RNA sequencing and qPCR assays. The transcriptome analyses showed that there were 2040 and 3630 differentially expressed genes (DEGs) in 4W (at endosperm cellularization initiation stage and heart embryo stage) vs 3W (at syncytium stage and globular embryo stage), and 5W (at end of endosperm cellularization stage and torpedo embryo stage) vs 4W, respectively. The KEGG and GO analyses showed that lipid metabolic processes and transmembrane transport related to cell wall biogenesis, cell division and differentiation, the plant hormone signaling pathway, photosynthesis, and transcription regulator activity were evidently enriched in WT and naa15. The heatmap and qPCR analyses showed that auxin response genes (ARFs), auxin transport genes (PINs) cytokinin synthesis genes (LOGs), cytokinin dehydrogenase genes (CKXs), cytokinin receptor, transcription factors (MYB, bHLH, MADS-box, and ERF) were significantly downregulated in naa15 compared to WT. A series of cell wall genes annotated to xyloglucan endotransglycosylase/hydrolase, pectin methyl esterase, and pectin methyl esterase inhibitor were also identified in these DEGs. Moreover, using an immunofluorescent assay, the features of cell walls displayed that cellulose fluorescence signals in the embryo and endosperm of naa15 were significantly decreased, and the signals of low- and high- methyl esterification of pectin were also obviously decreased in the endosperm of naa15. In summary, we identified a large number of DEGs and investigated the features of cell walls during endosperm cellularization and embryonic differentiation, which provided important information on transcription and gene expression to reveal their regulatory mechanisms.