Pectin Methylesterase Gene Research Articles

A pollen expressed PME gene at Tcb1 locus confers maize unilateral cross-incompatibility.

A pollen expressed PME gene at Tcb1 locus confers maize unilateral cross-incompatibility.

Unraveling correlations between calcium deficiency and spongy tissue in mango fruit flesh

Spongy tissue, as one of internal flesh breakdown in mango fruits, is correlated with calcium (Ca) deficiency, and adversely affects fruit quality and yield worldwide. However, the underlying mechanism causing spongy tissue remains unknown. Here, we explored the effects of Ca deficiency on spongy tissue in a susceptible cultivar, ‘Keitt’, at the physiological and molecular levels. Nutrient analysis showed that flesh with spongy tissue had lower Ca content and higher N/Ca, K/Ca and Mg/Ca rations than healthy fruit flesh. Ca accumulation in the cell wall, especially in chelator-soluble pectin, was elevated due to increased pectin content, lower levels of pectin methylesterification, and increased pectin methylesterase (PME) activity. During spongy tissue development, levels of Ca oxalate (CaOx) and oxalic acid both increased, whereas soluble Ca content decreased in the flesh. Transcriptomic analysis of healthy flesh and flesh with spongy tissue revealed 2054 up-regulated and 996 down-regulated genes. Gene ontology term analysis showed that the up-regulated genes were mainly enriched in functions related to the transmembrane transporter activity, calcium ion binding, and the cell wall. Importantly, up-regulated genes in spongy tissue included five glucuronate 4-epimerase (GAE) genes and two galactosyltransferase (GALT) genes, which involved in pectin synthesis; four PME genes which control pectin demethylesterification; and eight autoinhibited calcium-transporting ATPase (ACA) genes and two vacuole H+/Ca2+ exchanger (CAX) genes which contribute to Ca influx in the vacuole. Our findings illustrate a mechanism by which Ca deficiency leads to spongy tissue, which will contribute to efforts to control this physiological disorder.

Systematic analysis of the pectin methylesterase gene family in Nicotiana tabacum and reveal their multiple roles in plant development and abiotic stresses.

The pectin methylesterases (PMEs) play multiple roles in regulating plant development and responses to various stresses. In our study, a total of 121 PME genes were identified in the tobacco genome, which were clustered into two groups based on phylogenetic analysis together with Arabidopsis members. The investigations of gene structure and conserved motif indicated that exon/intron and motif organizations were relatively conserved in each group. Additionally, several stress-related elements were identified in the promoter region of these genes. The survey of duplication events revealed that segmental duplications were critical to the expansion of the PME gene family in tobacco. The expression profiles analysis revealed that these genes were expressed in various tissues and could be induced by diverse abiotic stresses. Notably, NtPME029 and NtPME043, were identified as homologues with AtPME3 and AtPME31, respectively. Furthermore, NtPME029 was highly expressed in roots and the over-expression of the NtPME029 gene could promote the development of roots. While NtPME043 could be induced by salt and ABA treatments, and the over-expression of the NtPME043 gene could significantly enhance the salt-stress tolerance in tobacco. Overall, these findings may shed light on the biological and functional characterization of NtPME genes in tobacco.

Three types of genes underlying the Gametophyte factor1 locus cause unilateral cross incompatibility in maize

Unilateral cross incompatibility (UCI) occurs between popcorn and dent corn, and represents a critical step towards speciation. It has been reported that ZmGa1P, encoding a pectin methylesterase (PME), is a male determinant of the Ga1 locus. However, the female determinant and the genetic relationship between male and female determinants at this locus are unclear. Here, we report three different types, a total of seven linked genes underlying the Ga1 locus, which control UCI phenotype by independently affecting pollen tube growth in both antagonistic and synergistic manners. These include five pollen-expressed PME genes (ZmGa1Ps-m), a silk-expressed PME gene (ZmPME3), and another silk-expressed gene (ZmPRP3), encoding a pathogenesis-related (PR) proteins. ZmGa1Ps-m confer pollen compatibility. Presence of ZmPME3 causes silk to reject incompatible pollen. ZmPRP3 promotes incompatibility pollen tube growth and thereby breaks the blocking effect of ZmPME3. In addition, evolutionary genomics analyses suggest that the divergence of the Ga1 locus existed before maize domestication and continued during breeding improvement. The knowledge gained here deepen our understanding of the complex regulation of cross incompatibility.

Genome-wide identification of PME gene family and expression of candidate genes associated with aluminum tolerance in tea plant (Camellia sinensis)

BackgroundThe major aluminum (Al) detoxication mechanism of tea plant (Camellia sinensis), as an Al hyperaccumulator plant, is the fixation of almost 70% of Al in the cell walls. Pectin is the primary constituent of cell walls, a degree of methylation of pectin polysaccharides regulated by the pectin methylesterase (PME) genes can greatly affect the Al binding capacity. The knowledge on PME gene family in tea plant is still poor.ResultsWe identified 66 (CsPME1-CsPME66) PME genes from C. sinensis genome. We studied their protein characterization, conserved motifs, gene structure, systematic evolution and gene expression under Al treatments, to establish a basis for in-depth research on the function of PMEs in tea plant. Gene structures analysis revealed that the majority of PME genes had 2–4 exons. Phylogenetic results pointed out that the PME genes from the same species displayed comparatively high sequence consistency and genetic similarity. Selective pressure investigation suggested that the Ka/Ks value for homologous genes of PME family was less than one. The expression of CsPMEs under three Al concentration treatments was tissue specific, eight PME genes in leaves and 15 in roots displayed a trend similar to of the Al contents and PME activities under Al concentration treatments, indicating that the degree of pectin de-esterification regulated by PME was crucial for Al tolerance of tea plant.ConclusionsSixty-six CsPME genes were identified for the first time in tea plant. The genome-wide identification, classification, evolutionary and transcription analyses of the PME gene family provided a new direction for further research on the function of PME gene in Al tolerance of tea plant.

Transcriptome-Guided Identification of Pectin Methyl-Esterase-Related Enzymes and Novel Molecular Processes Effectuating the Hard-to-Cook Defect in Common Bean (Phaseolus vulgaris L.).

The hard-to-cook defect in common beans is dictated by the ability to achieve cell separation during cooking. Hydrolysis of pectin methyl-esters by the pectin methyl-esterase (PME) enzyme influences cell separation. However, the contributions of the PME enzyme and the cell wall to the hard-to-cook defect have not been studied using molecular tools. We compared relevant molecular processes in fast- and slow-cooking bean varieties to understand the mechanisms underpinning the hard-to-cook defect. A PME spectrophotometric assay showed minor differences in enzyme activity between varieties. Meanwhile, a PME HMMER search in the P. vulgaris genome unveiled 113 genes encoding PMEs and PME inhibitors (PMEIs). Through RNA sequencing, we compared the gene expression of the PME-related genes in both varieties during seed development. A PME (Phvul010g080300) and PMEI gene (Phvul005g007600) showed the highest expression in the fast- and slow-cooking beans, respectively. We further identified 2132 differentially expressed genes (DEGs). Genes encoding cell-wall-related enzymes, mainly glycosylphosphatidylinositol mannosyltransferase, xyloglucan O-acetyltransferase, pectinesterase, and callose synthase, ranked among the top DEGs, indicating novel relations to the hard-to-cook defect. Gene ontology mapping revealed hydrolase activity and protein phosphorylation as functional categories with the most abundant upregulated DEGs in the slow-cooking bean. Additionally, the cell periphery contained 8% of the DEGs upregulated in the slow-cooking bean. This study provides new insights into the role of pectin methyl-esterase-related genes and novel cell wall processes in the occurrence of the hard-to-cook defect.

Pectin methylesterase gene AtPMEPCRA contributes to physiological adaptation to simulated and spaceflight microgravity in Arabidopsis

SummaryPectin is biosynthesized in a highly methylated form and is partially de-methylated by pectin methylesterase (PME) activity. Plant PMEs play a critical role in cell wall remodeling in many physiological processes. Here, we studied Arabidopsis seedlings, which had been exposed to simulated or actual microgravity. Simulated microgravity inhibited total PME activity in Arabidopsis seedlings. We identified that AtPMEPCRA expression played a major role in the microgravity-induced inhibition of PME activity. atpmepcra mutants did not exhibit the enlarged leaf area of Arabidopsis seedlings observed under spaceflight microgravity. The downregulation of AtPMEPCRA expression in response to microgravity was due, in part, to changes in methylation patterns. The sexual offspring of the plants grown during spaceflight retained the methylation changes at AtPMEPCRA locus for one generation and thus contribute to the physiological adaptation to microgravity among F1 offspring seed generation. We conclude that AtPMEPCRA contributes to the spaceflight-induced transgenerational responses in Arabidopsis.

Remediation of Pear Iron Deficiency Chlorosis by Nanocellulose-iron Chelation and the Underlying Mechanism

Nanocrystal cellulose possesses a strong capability to chelate Fe due to its adsorptive properties. Iron deficiency chlorosis (IDC) is a mineral disorder that remarkably weakens pear photosynthesis, causing declines in plant yields and quality. Conventional methods for controlling IDC generally lack efficiency and overuse chemicals. Foliar application of nanocellulose (NC)-Fe chelate (NCFe) provides a new approach to remediate IDC in pear (Pyrus betulifolia). In this study, NC was prepared by acidic hydrolysis using 64 wt% H2SO4 at 45 °C for 45 minutes. NCFe was formulated based on the net charge density of NC and ferrous sulfate (FeSO4) solution. The nanoparticle properties were characterized by transmission electron microscopy (TEM), dynamic light scattering, and conductometry. Pyrus betulifolia seedlings were pre-etiolated in an improved Hoagland’s nutrient solution and treated with bicarbonate. Changes in chlorophyll content, active Fe content, and photosynthesis rate in NCFe-treated leaves were determined by SPAD values, spectrophotometry, and photosynthetic apparatus, respectively. Ferritin genes (PbFER) and pectin methylesterase genes (PbPME) were extracted from leaf tissue, and gene expression profiles were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). The results showed that NCFe particles maintained a whisker-like morphology; the Z-average hydrodynamic diameter and zeta potential of NCFe measured by dynamic light scattering were 107.4 ± 3.0 nm and −9.7 ± 0.4 mV, respectively. When NCFe was prepared at a mixing ratio of 1:3000, the total chlorophyll content, active Fe content, and net photosynthetic rate of plant leaves were significantly enhanced by 23.8%, 65.9%, and 40.4% after 72 hours of treatment, respectively, compared with FeSO4 spraying. Importantly, NCFe treatment also significantly downregulated the expression of PbPME and upregulated the expression of PbFER, which are key genes regulating the active Fe content.

Characterization of pectin methylesterase gene family and its possible role in juice sac granulation in navel orange (Citrus sinensis Osbeck)

BackgroundCitrus is one of the most important fresh fruit crops worldwide. Juice sac granulation is a physiological disorder, which leads to a reduction in soluble solid concentration, total sugar, and titratable acidity of citrus fruits. Pectin methylesterase (PME) catalyzes the de-methylesterification of homogalacturonans and plays crucial roles in cell wall modification during plant development and fruit ripening. Although PME family has been well investigated in various model plants, little is known regarding the evolutionary property and biological function of PME family genes in citrus.ResultsIn this study, 53 non-redundant PME genes were identified from Citrus sinensis genome, and these PME genes were divided into four clades based on the phylogenetic relationship. Subsequently, bioinformatics analyses of gene structure, conserved domain, chromosome localization, gene duplication, and collinearity were performed on CsPME genes, providing important clues for further research on the functions of CsPME genes. The expression profiles of CsPME genes in response to juice sac granulation and low-temperature stress revealed that CsPME genes were involved in the low temperature-induced juice sac granulation in navel orange fruits. Subcellular localization analysis suggested that CsPME genes were localized on the apoplast, endoplasmic reticulum, plasma membrane, and vacuole membrane. Moreover, yeast one-hybrid screening and dual luciferase activity assay revealed that the transcription factor CsRVE1 directly bound to the promoter of CsPME3 and activated its activity.ConclusionIn summary, this study conducts a comprehensive analysis of the PME gene family in citrus, and provides a novel insight into the biological functions and regulation patterns of CsPME genes during juice sac granulation of citrus.

The callus formation capacity of strawberry leaf explant is modulated by DNA methylation.

Shoot regeneration from leaf tissue requires the de-differentiation of cells from a highly differentiated state into an actively dividing state, but it remains unclear how this physiological transition occurs and is regulated, especially at the epigenetic level. Here, we characterized the DNA methylome represented by 5-methylcytosine (5mC) in leaf and callus tissue derived from leaf explants of woodland strawberry, Fragaria vesca. We detected an overall increase in DNA methylation and distinct 5mC enrichment patterns in the CG, CHG, and CHH sequence contexts in genes and transposable elements. Our analyses revealed an intricate relationship between DNA methylation and gene expression level in leaves or leaf-derived callus. However, when considering the genes involved in callus formation and shoot regeneration, e.g. FvePLT3/7, FveWIND3, FveWIND4, FveLOG4 and FveIAA14, their dynamic transcription levels were associated with differentially methylated regions located in the promoters or gene bodies, indicating a regulatory role of DNA methylation in the transcriptional regulation of pluripotency acquisition in strawberry. Furthermore, application of the DNA methyltransferase inhibitor 5′-azacytidine (5′-Aza) hampered both callus formation and shoot regeneration from the leaf explants. We further showed that 5′-Aza downregulated the expression of genes involved in cell wall integrity, such as expansin, pectin lyase, and pectin methylesterase genes, suggesting an essential role of cell wall metabolism during callus formation. This study reveals the contribution of DNA methylation to callus formation capacity and will provide a basis for developing a strategy to improve shoot regeneration for basic and applied research applications.

Integrative genomics analysis of the ever-shrinking pectin methylesterase (PME) gene family in foxtail millet (Setaria italica).

Pectin methylesterase (PME) plays a vital role in the growth and development of plants. Their genes can be classified into two types, with Type-1 having an extra domain, PMEI. PME genes in foxtail millet (Setaria italica L.) have not been identified, and their sequence features and evolution have not been explored. Here, we identified 41 foxtail millet PME genes. Decoding the pro-region, containing the PMEI domain, revealed its more active nature than the DNA encoding PME domain, easier to be lost to produce Type-2 PME genes. We inferred that the active nature of the pro-region could be related to its harbouring more repetitive DNA sequences. Further, we revealed that though whole-genome duplication and tandem duplication contributed to producing new copies of PME genes, phylogenetic analysis provided clear evidence of ever-shrinking gene family size in foxtail millet and the other grasses in the past 100 million years. Phylogenetic analysis also supports the existence of two gene groups, Group I and Group II, with genes in Group II being more conservative. Our research contributes to understanding how DNA sequence structure affects the functional innovation and evolution of PME genes.

The effect of 24-epibrassinolide on gene expression related to cell walls under boron deficiency and toxicity in the leaves of Arabidopsis thaliana

The changing composition of plant cell walls allows for the continuation of the existing structure under normal conditions and also the protection of physical integrity under altering environmental conditions. In this study, the possible effects of the 24-Epibrassinolide (EBL) hormone under boron (B) deficiency and toxicity conditions on the expression of cell wall-related genes [cellulose synthase (CESA), expansin (EXP), xyloglucan endotransglucosylase/hydrolase (XTH) and pectin methylesterase (PME)] were investigated in the rosette leaves of Arabidopsis thaliana. For this purpose, 0 or 3000 ?M of boric acid (BA) and/or 1 ?M of EBL were applied to the plants which were grown in a hydroponic medium for five and ten weeks. While B-toxicity elevated the mRNA levels of the CESA4 and CESA8 genes in the leaves of the five-week-old plants, B-stress (B-deficiency and -toxicity) caused an increase in the expression of the CESA4, CESA6, and CESA8 genes in the ten-week-old plants. The transcript levels of the EXPA5 gene increased under B-stress in the ten-week-old plants whereas the expression of the EXPA8 gene decreased when compared to the control at two developmental stages. Co-treatment of EBL and B-stress strongly elevated the transcript level of the EXPA5 gene in the ten-week-old plants and the EXPA8 gene at both developmental stages. The EXPA14 and XTH23 genes exhibited distinct expression profiles under B-deficiency and -toxicity in both the five- and ten-week-old plants. The transcript level of the XTH21 gene was upregulated in the leaves of the plants exposed to B-stress. The mRNA level of the PME2 and PME41 genes was generally upregulated in response to B-stress in both the five- and ten-week-old plants. 24-Epibrassinolide alone and in combination with B-stress led to a remarkable increase in the expression of the XTH and PME genes compared to the control. These results demonstrate that cell wall genes generally show a similar pattern of expression at both developmental stages and the EBL hormone induces changes in the expression levels of cell wall-related genes under B-stress.

Potassium enhances cadmium resistance ability of Panax notoginseng by brassinolide signaling pathway-regulated cell wall pectin metabolism

The mechanism of how potassium (K) attenuates cadmium (Cd)-induced demethylation and anabolism of cell wall (CW) pectin through the brassinolide (BR) signaling pathway was verified in Panax notoginseng (Burk.). The P. notoginseng pectin methylesterase gene (PnPME1) was cloned and functionally verified in tobacco. Pectin and BR metabolism, Cd content and the pectin methylation degree (PMD) were detected in response to K, 2,4-epibrassinolide (EBL), and brassinazole treatments of P. notoginseng and tobacco under Cd stress. Activity of the main root pectin methylesterase enzyme (PME) was promoted by 22.29% under the EBL treatment, and Cd content increased by 29.03% under Cd stress. Potassium reduced PME activity and Cd content in main root pectin by 61.03% and 50.73%, respectively, under the EBL and Cd co-treatment. Potassium inhibited the promoting effects of Cd stress on the expression of PnPME1 by 57.04%. Potassium also inhibited expression of BR synthesis genes PnDET2, PnROT3, PnCYP90A1, and PnBR6OX1 by 65.61%, 52.02%, 47.36%, and 55.16%, respectively, and reduced the accumulation of Cd. The PnPME1 was located in the CW. The activity of transgenic tobacco root PME was higher than that of the wild-type, while the PMD was significantly lower. The regulatory effects of K and EBL on tobacco root pectin metabolism were consistent with those in P. notoginseng. In conclusion, K downregulated the expression of BR synthesis genes in P. notoginseng roots under Cd stress and reduced the production of BRs, which inhibited PnPME1 expression. The reduction in PME activity increased the PMD, which reduced the accumulation of Cd.

Homogalacturonan and xylogalacturonan region specificity of self-cloning vector-expressed pectin methylesterases (AoPME1–3) in Aspergillus oryzae

Aspergillus oryzae is a safe microorganism that is commonly used in food production. We constructed a self-cloning vector capable of high expression in A. oryzae. Using the vector, three putative pectin methylesterase (PME) genes belonging to Carbohydrate Esterase family 8 derived from A. oryzae were expressed, and several characteristics of the gene products were examined. The effects of temperature and pH on the three enzymes (AoPME1, 2, and 3) were similar, with optimal reaction temperatures of 50 − 60 °C and optimal reaction pH range of 5 − 6. The specific activities of AoPME1, 2, and 3 for apple pectin were significantly different (34, 7,601, and 2 U/mg, respectively). When the substrate specificity was examined, AoPME1 showed high activity towards pectin derived from soybean and pea. Although AoPME2 showed little activity towards these pectins, it showed very high activity towards apple- and citrus-derived pectins. AoPME3 showed low specific activity towards all substrates tested. Sugar composition analysis revealed that apple- and citrus-derived pectins were rich in homogalacturonan, while soybean- and pea-derived pectins were rich in xylogalacturonan. When pea pectin was treated with endo-polygalacturonase or endo-xylogalacturonase in the presence of each PME, specific synergistic actions were observed (endo-polygalacturonase with AoPME1 or AoPME2 and endo-xylogalacturonase with AoPME1 or AoPME3). Thus, AoPME1 and AoPME3 hydrolyzed the methoxy group in xylogalacturonan. This is the first report of this activity in microbial enzymes. Our findings on the substrate specificity of PMEs should lead to the determination of the distribution of methoxy groups in pectin and the development of new applications in the field of food manufacturing.

Rice Putative Pectin Methyltransferase Gene OsPMT10 Is Required for Maintaining the Cell Wall Properties of Pistil Transmitting Tissues via Pectin Modification.

Precise directional control of pollen tube growth via mechanical guidance by pistil tissue is critical for the successful fertilization of flowering plants and requires active cell-to-cell communication and maintenance of softness in the transmitting tissue. However, the regulation of transmitting tissue softness as controlled by cell wall properties, especially pectin, has not been reported. Here we report that regulation of pectin methylesterification supports pollen elongation through pistil transmitting tissues in Oryza sativa. The rice pectin methylesterase gene OsPMT10 was strongly expressed in reproductive tissues, especially the pistil. The ospmt10 mutant did not have a significant effect on vegetative growth, but the fertility rate was reduced by approximately half. In the ospmt10 mutant, pollen tube elongation was observed in the transmitting tissue of the style, but approximately half of the pollen tubes did not extend all the way to the ovule. Tissue cross-sections of the upper ovary were prepared, and immunohistochemical staining using LM19 and LM20 showed that methylesterified pectin distribution was decreased in ospmt10 compared with the wild type. The decreased expression of methylesterified pectins in ospmt10 may have resulted in loss of fluidity in the apoplast space of the transmitting tissue, rendering it difficult for the pollen tube to elongate in the transmitting tissue and thereby preventing it from reaching the ovule.

Higher Cd-accumulating oilseed rape has stronger Cd tolerance due to stronger Cd fixation in pectin and hemicellulose and higher Cd chelation

Oilseed rape (Brassica napus) has potential as a hyperaccumulator in the phytoremediation of cadmium (Cd)-contaminated soils. Oilseed rape varieties with higher Cd accumulation ability and Cd tolerance are ideal candidates for the hyperaccumulation of excess Cd. To explore the physiological and molecular mechanisms underlying Cd tolerance and high Cd accumulation in oilseed rape leaves, we examined two genotypes, “BN067” (Cd-sensitive with lower Cd accumulation in leaves) and “BN06” (Cd-tolerant with higher Cd accumulation in leaves). We characterized the physiological morphology, structure, subcellular distribution of Cd, cell wall components, cell chelates, and the transcriptional levels of the related genes. Greater Cd accumulation was observed in the cell walls and vacuoles of Cd-tolerant leaves, reducing Cd toxicity to the lamellar structure of the chloroplast thylakoid and leaf stomata. Higher expression of PMEs genes and lower expression of pectin methylesterase inhibitors (PMEI) genes improved pectin methylesterase (PME) activity in leaves of Cd-tolerant genotype. Stronger demethylation of pectin along with higher pectin and hemicellulose levels induced by lower pectinase and hemicellulose activities in the leaves of the Cd-tolerant genotype, resulting in higher Cd retention in the cell walls. Under Cd toxicity, higher Cd sequestration within the vacuoles of Cd-tolerant leaves was closely related to greater accumulation of Cd chelates with stronger biosynthesis in protoplasts. The results highlight the importance of using hyperaccumulation by plants to remediate our environment, and also provide a theoretical basis for the development of Cd-tolerant varieties.

Search for Candidate Genes Causing the Excessive Ca Accumulation in Roots of Tipburn-Damaged Lisianthus (Eustoma grandiflorum) Cultivars

Occurrence of tipburn is a severe problem in the production of lisianthus cultivars. Previous studies have shown excessive Ca accumulation in the roots of tipburn-damaged cultivars, where the distribution of Ca to the tips of the top leaves is inhibited. However, few studies have investigated the association between Ca accumulation and gene expression in horticultural crops. To provide a list of candidate target genes that might be causing the excessive Ca accumulation in roots, we focused Ca2+ transporter and pectin methylesterase (PME) genes and RNA-seq of upper leaves and roots in tipburn-occurrence cultivar (“Voyage peach”: VP) and non-occurrence cultivar (“Umi honoka”: UH) was conducted. In both the upper leaves and roots of VP, genes encoding the glutamate receptors (GLRs), cation/Ca2+ exchangers 4 (CCX4), Na+/Ca2+ exchanger-like protein (NCL), and PMEs were upregulated, and a gene encoding the cyclic nucleotide-gated ion channel 9 (CNGC9) was downregulated. In contrast, genes encoding the vacuolar cation/proton exchanger 5 (CAX5), calcium-transporting ATPase 1 and 12 (ACA1 and ACA12) showed differential expression in each organ. Among them, only CAX5 was upregulated and ACA12 was downregulated in the roots of VP. Based on these results, we suggested that CAX5 and ACA12 are the candidate genes causing the excessive Ca accumulation in the roots of tipburn-occurrence lisianthus cultivars. Future studies should investigate the temporal changes in gene expression using quantitative PCR and conduct functional analysis of candidate genes in tipburn-damaged lisianthus cultivars.

Characterization of an acidic pectin methylesterase from Paenibacillus xylanexedens and its application in fruit processing

A pectinase-producing bacterial isolate, identified as Paenibacillus xylanexedens SZ 29, was screened by using the soil dilution plate with citrus pectin and congo red. A pectin methylesterase gene (Pxpme) was cloned and expressed in Escherichia coli. The gene coded for a protein with 334 amino acids and a calculated molecular mass of 36.76 kDa. PxPME showed the highest identity of 32.4% with the characterized carbohydrate esterase family 8 pectin methylesterase from Daucus carota. The recombined PxPME showed a specific activity with 39.38 U/mg against citrus pectin with >65% methylesterification. The optimal pH and temperature for PxPME activity were 5.0 and 45 °C. Its Km and Vmax value were determined to be 1.43 mg/mL and 71.5 μmol/mg·min, respectively. Moreover, PxPME could increase the firmness of pineapple cubes by 114% when combined with CaCl2. The acidic and mesophilic properties make PxPME a potential candidate for application in the fruit processing.

Genome-Wide Analysis of the Pectin Methylesterase Gene Family in Potato

Pectin methylesterase (PME) is a type of protein encoded by a multi-gene family and participates in seed germination, fruit ripening and stress response. To further study the function of the PME gene family in potato, this paper screened and identified potato PME gene family by bioinformatics methods, and analysed the structure, physicochemical properties, conserved domains and evolutionary relationships of the gene sequences. The results showed that the identified 54 potato PME genes can be divided into Group I (only PME domain) and Group II (both PME domain and PMEI domain). The phylogenetic tree constructed by the maximum likelihood method indicated that potato PME genes consisted of five subfamilies. The protein sequences of potato PME genes were compared with clustalx2.1 software. Most of these sequences were identified to have five conserved structural regions including Gx Yx E (Region I), QAVAxR (Region II), QDTL (Region III), GxxDFIFG (Region IV) and YLGRxWx (Region V). Conserved motifs of potato PME amino acid sequences were analysed by MEME online software. Motif1 contains three of the five conserved structural regions and motif2 and motif8 each contain one, indicating that these sequences are more conservative. The structure of 54 potato PME gene sequences was analysed by the GSDS website. It was found that there were only two parts, exon or coding sequence (CDS) between untranslated regions (UTRs), without intron structures in these sequences, indicating that the structure is relatively simple. It will provide a reference for the study on the function of potato PME gene.

Characterization of the pectin methyl-esterase gene family and its function in controlling pollen tube growth in pear (Pyrus bretschneideri)

Pectin methyl-esterases (PMEs) play crucial roles in plant growth. In this study, we identified 81 PbrPMEs in pear. Whole-genome duplication and purifying selection drove the evolution of PbrPME gene family. The expression of 47 PbrPMEs was detected in pear pollen tube, which were assigned to 13 clusters by an expression tendency analysis. One of the 13 clusters presented opposite expression trends towards the changes of methyl-esterified pectins at the apical cell wall. PbrPMEs were localized in the cytoplasm and plasma membrane. Repression of PbrPME11, PbrPME44, and PbrPME59 resulted in the inhibition of pear pollen tube growth and abnormal deposition of methyl-esterified pectins at pollen tube tip. Pharmacological analysis confirmed that reduced PbrPME activities repressed the pollen tube growth. Overall, we have explored the evolutionary characteristics of PbrPME gene family and found the key PbrPME genes that control the growth of pollen tube, which deepened the understanding of pear fertility regulation.