Pectin Methylesterase Activity Research Articles

Rootstock effects on bitter pit incidence in 'Honeycrisp' apples are associated with changes in fruit's cell wall chemical properties.

Bitter pit (BP) is a physiological disorder of apples that often appears during or after cold storage. Despite being defined as a calcium deficiency disorder, BP is a complex process that is not only affected by the total Ca2+ content in the fruit but also by the proper cellular Ca2+ homeostasis and partitioning. Early investigations have also suggested that rootstocks could affect BP development and severity. In the present study, rootstock effects on BP development were assessed on ‘Honeycrisp’ trees that were grafted on 14 different rootstocks (B.10, G.11, G.202, G.214, G.30, G.41, G.935, G.969, M.26 EMLA, M.9, V.1, V.5, V.6, and V.7). We evaluated BP incidence at harvest, and three months after cold storage for four, and three growing seasons, respectively. BP incidence was significantly reduced in ‘Honeycrisp’ trees on B.10 compared to other rootstocks, whereas trees on V.6 showed the highest percentage of BP at harvest and after cold storage. ‘Honeycrisp’ apples were collected from three different rootstocks (B.10, G.41, and V.6) two months after cold storage and evaluated for mineral nutrient composition, Ca2+ homeostasis, and cell wall properties, e.g., pectin content, pectin de-esterification rate and pectin methylesterase (PME) activity. Water-soluble and insoluble pectin content was markedly higher in fruits from B.10 than in G.41 and V.6. We also observed increased PME enzyme activity and a greater degree of water-insoluble pectin de-esterification in ‘Honeycrisp’ apples from V.6 compared to those from B.10. A significantly higher Ca2+ was found in the fruits from B.10 than G.41 and V.6. Higher Ca2+ and lower Mg2+ levels were also observed in the cell wall and water-insoluble pectin fractions of the fruits from B.10 compared to G.41 and V.6. However, the ratio of cell wall-bound Ca2+ to total Ca2+ was lower in B.10 compared to G.41 and V.6. Together, our results indicate that the tolerance of B.10 to BP could be attributed to a reduced PME activity and lower pectin de-esterification level, which in turn reduced the amount of Ca2+ cross-linked with pectin, and probably increased the apoplastic free calcium concentrations that is essential for maintaining cell membrane integrity and reducing BP development.

Guar gum and chitosan-based composite edible coating extends the shelf life and preserves the bioactive compounds in stored Kinnow fruits

The present investigation aimed to elucidate the effects of chitosan (CH) and guar gum (GG) based composite edible coating on physicochemical quality and storability of Kinnow fruit under ambient conditions. Composite coating materials were prepared by using tamarind (TAM) seed starch and jackfruit (JACK) seed starch with CH and GG separately. The results indicated that GG + TAM coated fruits maintained significantly (P ≤ 0.01) lower physiological losses in weight (PLW) (3.12 %), decay incidence (0.83 %), and respiration rate (13.57 ml CO2 kg−1 h−1) compared to control. The reduced activity of pectin methylesterase (PME) (0.97 μmol min−1 g−1 FW) and lipoxygenase (LOX) (1.97 μmol min−1 g−1 FW) were noted in GG + TAM coated fruits. Also, the GG + TAM coated fruits retained higher fruit firmness (6.77 N), titratable acidity (TA) (0.94 %), ascorbic acid (AA) (27.83 mg 100 g−1), total phenols (213.05 mg GAE 100 g−1 FW), antioxidants activity (23.57 μmol TE g−1) along with higher sensory score over the control. Based on findings, it can be concluded that GG + TAM coating could be gainfully utilized for prolonging the shelf life of Kinnow fruits up to 25 days without losing desirable quality traits at ambient storage.

The calcium-mediated homogalacturonan pectin complexation in cell walls contributes the firmness increase in loquat fruit during postharvest storage

IntroductionPostharvest textural changes in fruit are mainly divided into softening and lignification. Loquat fruit could have severe lignification with increased firmness during postharvest storage. Pectin is mainly associated with the postharvest softening of fruit, but some studies also found that pectin could be involved in strengthening the mechanical properties of the plant. ObjectivesThis study focused on characterizing the dynamics of pectin and its complexation in the cell wall of lignified loquat fruit during postharvest storage, and how these changes could influence fruit firmness. MethodsThe homogalacturonan (HG) pectin in the cell wall of loquat fruit was identified using monoclonal antibodies. An oligogalacturonide (OG) probe was used to label the egg-box structure formed by Ca2+ cross-linking with low-methylesterified HG. An exogenous injection was used to verify the role of egg-box structures in the firmness increase in loquat fruit. ResultsThe JIM5 antibody revealed that low-methylesterified HG accumulated in the tricellular junctions and middle lamella of loquat fruit that had severe lignification symptoms. The pectin methylesterase (PME) activity increased during the early stages of storage at 0 °C, and the calcium-pectate content and flesh firmness constantly increased during storage. The OG probe demonstrated the accumulation of egg-box structures at the cellular level. The exogenous injection of PME and Ca2+ into the loquat flesh led to an increase in firmness with more low-methylesterified HG and egg-box structure signals. ConclusionPME-mediated demethylesterification generated large amounts of low-methylesterified HG in the cell wall. This low-methylesterified HG further cross-linked with Ca2+ to form egg-box structures. The pectin-involved complexations then contributed to the increased firmness in loquat fruit. Overall, besides being involved in fruit softening, pectin could also be involved in strengthening the mechanical properties of postharvest fruit. This study provides new ideas for obtaining a better texture of postharvest loquat fruits based on pectin regulation.

Physiological and transcriptomic analysis reveals the postharvest ripening differences between “Nantianhuang” and “Brazilian” banana fruit

Banana, as a typical climacteric fruit, undergoes a ripening process which impacts fruit quality. The Nantianhuang (NTH) banana fruit is a new cultivar resistant to Fusarium wilt, whose physiological characteristics and regulation mechanisms of fruit ripening are not clear. Ripening physiology and the related genes of the NTH fruit were compared with the Brazilian banana (BX) and investigated in this study. When compared with BX, NTH showed a higher hue value, firmness, and starch content. However, NTH exhibited lower total soluble solids, ethylene production, respiration rate, malondialdehyde content, and activities of pectin methyl esterase, polygalacturonase and 1-aminocyclopropane-1-carboxylate oxidase, indicating that the softening of NTH occurred later than that of BX. Moreover, transcriptome and gene expression analyses showed that NTH fruit exhibited lower expression levels of some ripening-related genes than BX. These results help understand the ripening characteristics of NTH and also form the basis for implementing measures to improve the ripening quality of NTH, eventually promoting the development of the banana industry.

Ammonium improved cell wall and cell membrane P reutilization and external P uptake in a putrescine and ethylene dependent pathway

Ammonium promotes rice P uptake and reutilization better than nitrate, under P starvation conditions; however, the underlying mechanism remains unclear. In this study, ammonium treatment significantly increased putrescine and ethylene content in rice roots under P deficient conditions, by increasing the protein content of ornithine decarboxylase and 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase compared with nitrate treatment. Ammonium treatment increased rice root cell wall P release by increasing pectin content and pectin methyl esterase (PME) activity, increased rice shoot cell membrane P release by decreasing phosphorus-containing lipid components, and maintained internal P homeostasis by increasing OsPT2/6/8 expression compared with nitrate treatment. Ammonium also improved external P uptake by regulating root morphology and increased rice grain yield by increasing the panicle number compared with nitrate treatment. The application of putrescine and ethylene synthesis precursor ACC further improved the above process. Our results demonstrate for the first time that ammonium increases rice P acquisition, reutilization, and homeostasis, and rice grain yield, in a putrescine- and ethylene-dependent manner, better than nitrate, under P starvation conditions.

Microcystin-LR and cyanobacterial extracts alter the distribution of cell wall matrix components in rice root cells

Cyanobacterial toxins (known as cyanotoxins) disrupt the plant cytoskeleton (i.e. microtubules and F-actin), which is implicated in the regulation of cell wall architecture. Therefore, cyanotoxins are also expected to affect cell wall structure and composition. However, the effects of cyanobacterial toxicity on plant cell wall have not been yet thoroughly studied. Accordingly, the alterations of cell wall matrix after treatments with pure microcystin-LR (MC-LR), or cell extracts of one MC-producing and one non-MC-producing Microcystis strain were studied in differentiated Oryza sativa (rice) root cells. Semi-thin transverse sections of variously treated LR-White-embedded roots underwent immunostaining for various cell wall epitopes, including homogalacturonans (HGs), arabinogalactan-proteins (AGPs), and hemicelluloses. Homogalacturonan and arabinan distribution patterns were altered in the affected roots, while a pectin methylesterase (PME) activity assay revealed that PMEs were also affected. Elevated intracellular Ca2+ levels, along with increased callose and mixed linkage glucans (MLGs) deposition, were also observed after treatment. Xyloglucans appeared unaffected and lignification was not observed. The exact mechanism of cyanobacterial toxicity against the cell wall is to be further investigated.

Postharvest Exogenous Melatonin Treatment of Table Grape Berry Enhances Quality and Maintains Bioactive Compounds during Refrigerated Storage

Table grape berries are classified as a perishable crop that deteriorates quickly after harvest. The application of melatonin after harvest was found to be effective for retarding senescence and slowing ripening. In the current study, we tested the influence of two melatonin concentrations (50 and 100 µmol) as a postharvest application on quality, bioactive compounds, and enzyme activities of grape berries cv “Crimson” stored at 0 ± 1 °C and 90% relative humidity (RH) for 35 days. Our results indicated that melatonin application extends the shelf-life of berries by reducing weight loss and maintaining total soluble solids (TSS), titratable acidity (TA), berry adherence strength, and firmness. Melatonin treatment also reduced pectin methyl esterase (PME) and polygalactouranase (PG) enzyme activities compared to the control. Moreover, O2•− and H2O2 rates in berries were reduced by high melatonin concentration. Moreover, peroxidase (POD) and catalase (CAT) enzyme activities were increased by melatonin application. Our findings suggested using melatonin postharvest to increase shelf life and maintain quality attributes during refrigerated storage, which could be advantageous on a large scale.

Tragacanth gum coating suppresses the disassembly of cell wall polysaccharides and delays softening of harvested mango (Mangifera indica L.) fruit

Mango is a climacteric fruit and is prone to high perishability. The rapid softening and ripening (due to degradation and disassembly of cell wall polysaccharides) are the major limitations in extending the storability of the harvested mango fruits. Various types of gum-based edible coatings have been reported for the shelf life extension of mango fruits. Tragacanth gum (TCG) also has appropriate coating properties. Its use as an edible coating has been reported on certain fruits. However, the effect of TCG coating in the regulation of harvested mango fruits ripening and softening has not been reported yet. So, the objective of this work was to investigate the effect of TCG (control, 0.5 %, 1 % and 1.5 %) coating on postharvest softening and ripening of harvested mango fruits. TCG coating affected the ripening and softening of mango in a dose-dependent manner. Results exhibited that mango fruits coated with 1.5 % TCG showed substantially lower disease incidence and weight loss. The 1.5 % TCG-coated mangoes showed substantially lower ethylene biosynthesis and respiration rate peaks as well as superoxide anion and hydrogen peroxide contents compared with the control. In the same way, 1.5 % TCG-coated mango fruits had markedly higher total chlorophyll content and lower L*, b* and a* along with substantially lower total carotenoids in peel tissues. Mango fruits coated with 1.5 % TCG exhibited markedly lower water-soluble pectin and higher chelate-soluble pectin, Na2CO3-soluble pectin, protopectin, cellulose and hemicellulose in flesh tissues compared with control. The activity of polygalacturonase (PG), cellulase (CX), pectin methylesterase (PME), β-galactosidase (β-Gal) and β-glucosidase (β-Glu) were significantly lower in flesh of 1.5 % TCG treated fruits along with substantially higher firmness in contrast with control. In addition, 1.5 % TCG coating treatment showed significantly higher activity of antioxidative enzymes and delayed the increase in soluble solids content (SSC) and ripening index (RI) along with considerably higher titratable acidity (TA) compared with the untreated control. So, pre-storage TCG based edible coating (1.5 %) could be applied to delay ripening and softening in mango fruit industry under postharvest ambient conditions.

Mechanical stress elicits kiwifruit ripening changes in gene expression and metabolic status

Mechanical stress of kiwifruit is of major importance since it is directly related to fruit quality and postharvest losses. However, the role of mechanical stress in kiwifruit postharvest metabolism remains poorly explored. The present work characterizes the impact of mechanical stress, applied as compression forces (147.1 N), in ‘Hayward’ kiwifruit at harvest as well as following short (14 days) or long (3 months) cold (0 ℃) storage. Using several experimental approaches, we documented that the applied compression-imposed stress conditions and induced the fruit ripening process at room temperature (20 ℃). Fluorescent antibody labeling showed that arabinogalactan proteins and xyloglucans were altered due to compression, while gel diffusion assay indicated that pectin methylesterase activity was reduced by the long cold exposure. Intracellular Fluo-3 AM-dependent Ca2+ signals were detected in proximal to the main compressed fruit areas just after mechanical treatment. Several primary metabolites changed (i.e. oxoproline, gamma-aminobutyric acid and malonic acid) in cold-stored pericarp and placenta tissues after 8 h of compression treatment. Transcriptomic data also disclosed numerous genes, which are mainly involved in sugar metabolism, TCA cycle, cell wall, ethylene and hormones signaling, that were affected by mechanical stress in pericarp and especially in placenta. The weighted correlation and network analysis based on the integration of metabolome and transcriptome datasets identified candidate modules involved in compression stress. We also detected potential TFs, including bZIP53, WRKY40 and GATA17, that are involved in compression stress while meta-data analysis enabled identification of several TFs that regulated by both compression and wounding in kiwifruit. This work provides a first characterization of the compression-affected changes in fruit tissues that offers insights into molecular basis of mechanical stress in kiwifruit.

Biochemical characterization of Pectin Methylesterase Inhibitor 3 from Arabidopsis thaliana

The de-methylesterification of the pectic polysaccharide homogalacturonan (HG) by pectin methylesterases (PMEs) is a critical step in the control of plant cell expansion and morphogenesis. Plants have large gene families encoding PMEs but also PME inhibitors (PMEIs) with differ in their biochemical properties. The Arabidopsis thaliana PECTIN METHYLESTERASE INHIBITOR 3 (PMEI3) gene is frequently used as a tool to manipulate pectin methylesterase activity in studies assessing its role in the control of morphogenesis. One limitation of these studies is that the exact biochemical activity of this protein has not yet been determined. In this manuscript we produced the protein in Pichia pastoris and characterized its activity in vitro. Like other PMEIs, PMEI3 inhibits PME activity at acidic pH in a variety of cell wall extracts and in purified PME preparations, but does not affect the much stronger PME activity at neutral pH. The protein is remarkable heat stable and shows higher activity against PME3 than against PME2, illustrating how different members of the large PMEI family can differ in their specificities towards PME targets. Finally, growing Arabidopsis thaliana seedlings in the presence of purified PMEI3 caused a dose-dependent inhibition of root growth associated with the overall inhibition of HG de-methylesterification of the root surface. This suggests an essential in vivo role for PME activity at acidic pH in HG de-methylesterification and growth control. These results show that purified recombinant PMEI3 is a powerful tool to study the connection between pectin de-methylesterification and cell expansion.

Effect of Chitosan/Thyme Oil Coating and UV-C on the Softening and Ripening of Postharvest Blueberry Fruits.

This study investigated the possible mechanism of softening and senescence of blueberry after harvest using chitosan/thyme oil coating combined with UV-C (short wave ultraviolet irradiation) treatment. On the 56th day of storage, the CBP, cellulose, and hemicellulose contents in the chitosan/thyme oil coating +UV-C-treated group were 1.41, 1.65, and 1.20 times higher than those in the control group. Compared with the control group, the activities of polygalacturonase (PG), pectin methylesterase (PME), β-glucosidase (β-Gal), and cellulose (Cx) were significantly reduced (p < 0.05) after chitosan/thyme oil coating +UV-C, and their maximum values decreased by 5.41 μg/h g, 5.40 U/g, 12.41 U/g, and 3.85 μg/h g, respectively. Moreover, chitosan/thyme oil coating combined with UV-C treatment inhibited the gene expression of PG, PME, Cx, and β-Gal and then regulated the decrease in PG, PME, Cx, and β-Gal activities, inhibited the degradation of cell wall polysaccharides, and delayed the softening and senescence of postharvest blueberries. The results showed that chitosan/thyme oil coating, UV-C, and chitosan/thyme oil coating + UV-C could significantly inhibit postharvest softening of blueberry; chitosan/thyme oil coating +UV-C had the best effect.

24-epibrassinolide enhances mango resistance to Colletotrichum gloeosporioides via activating multiple defense response

Brassinosteroids (BRs) are natural steroidal phytohormones that regulate many essential processes of plant growth, development and responses to stress. However, the role of BRs in the defense response against Colletotrichum gloeosporioides in mango fruit remains largely unknown. This study investigated the resistance against C. gloeosporioides and the possible mechanisms of exogenous 24-epibrassinolide (EBR) in mango fruit. Results showed 1 μmol L−1 EBR effectively reduced the disease incidence and disease severity on mango fruit after inoculating with C. gloeosporioides. However, EBR had no direct antifungal activity against C. gloeosporioides in vitro test. EBR significantly improved the activities of phenylalanine ammonia-lyase (PAL), peroxidase (POD), chitinase (CHT), cinnamate-4-hydroxylase (C4H), β-1,3-glucanase (GLU) and 4-hydroxycinnamate CoA ligase (4CL), as well as the accumulation of flavonoids, lignin and total phenolics in mango fruit. EBR treatment also increased hydrogen peroxide (H2O2) content in mango fruit at early storage stage after inoculation, while increased catalase (CAT), ascorbate peroxidase (APX) and superoxide dismutase (SOD) activities with low reactive oxygen species (ROS) level in later storage stage. Besides, EBR treatment markedly reduced the changes in pectin methylesterase (PME) and polygalacturonase (PG) activities, which slowed down the hydrolysis of pectin, thus maintaining cell wall integrity. The principal component analysis indicated that EBR treatment was strongly associated with the increase of defense enzyme activities and metabolite contents and the inhibition of pectin hydrolysis. These results indicated that exogenous EBR could be a promising strategy to increase resistance against C. gloeosporioides in mango fruit by improving the defense responses against pathogens.

Effect of High Hydrostatic Pressure and Pulsed Electric Fields Processes on Microbial Safety and Quality of Black/Red Raspberry Juice.

Black and red raspberries are fruits with a high phenolic and vitamin C content but are highly susceptible to deterioration. The effect of high hydrostatic pressure (HHP 400–600 MPa/CUT-10 min) and pulsed electric fields (PEF, frequency 100–500 Hz, pulse number 100, electric field strength from 11.3 to 23.3 kV/cm, and specific energy from 19.7 to 168.4 kJ/L) processes on black/red raspberry juice was studied. The effect on the inactivation of microorganisms and pectin methylesterase (PME) activity, physicochemical parameters (pH, acidity, total soluble solids (°Brix), and water activity (aw)), vitamin C and phenolic compounds content were also determined. Results reveal that all HHP-treatments produced the highest (p < 0.05) log-reduction of molds (log 1.85 to 3.72), and yeast (log 3.19), in comparison with PEF-treatments. Increments in pH, acidity, and TSS values attributed to compounds’ decompartmentalization were found. PME activity was partially inactivated by HHP-treatment at 600 MPa/10 min (22% of inactivation) and PEF-treatment at 200 Hz/168.4 kJ/L (19% of inactivation). Increment in vitamin C and TPC was also observed. The highest increment in TPC (79% of increment) and vitamin C (77% of increment) was observed with PEF at 200 Hz/168.4 kJ/L. The putative effect of HHP and PEF on microbial safety, enzyme inactivation, and phytochemical retention is also discussed in detail. In conclusion, HHP and PEF improve phytochemical compounds’ content, microbial safety, and quality of black/red raspberry juice.

Comparative effects of high hydrostatic pressure, pasteurization and nisin processing treatments on the quality of pickled radish

The quality of pickled vegetables is essentially safeguarded through various processing techniques. In this study the effects of high hydrostatic pressure processing (HPP), pasteurization (Pt), nisin, HPP + nisin and Pt + nisin on fermented radishes (Raphanus sativus L.) were compared. The effects of the different treatments on firmness were not immediately apparent, however, the HPP-treated pickles maintained their hardness best during storage. Compared with Pt, the non-thermal treatments were found to have moderate effects on the colors, both immediately after treatment and during storage. Both HPP and Pt reduced microbes, while HPP + nisin completely inactivated bacteria. The activities of polyphenol oxidase (PPO), peroxidase (POD) and pectin methylesterase (PME) were most inactivated via Pt, followed the action of HPP and then by that of the nisin. The sensory evaluation suggested that non-thermal pickled radish might be better accepted by consumers. Moreover, Pt had the strongest ability to reduce oxalic acid and sulfur-containing compounds, while the effects of HPP were relatively moderate, and HPP not only effectively reduced sulfur-containing compounds, but also helped to retain the favorable factors in the pickles. The findings of this study, thus, indicate that HPP could be a promising alternative to traditional pasteurization in the processing and storage of fermented vegetables.

Effect of ultra violet (UV-C) and cold storage on orange juice quality.

The effect of the UV-C treatment on the physico-chemical characteristics, pectin methylesterase activity (PME) as well as microbial quality of orange juice, compared to fresh juice, was studied. The juice samples were UV-C (254 nm) irradiated for different exposure times (15, 30, 45 and 60 min) and stored at 4 ± 1 °C for 30 days. UV-C treatment didn't significantly (p ≤ 0.05) affect pH values, titratable acidity, TSS (%), ascorbic acid content and PME activity in both fresh and stored samples. Increasing the exposure time from 5 to 60 min. showed no significant effect (p ≤ 0.05) on L* and a* values for both the fresh and the stored samples. On the contrary, negative relationship was observed between UV-C exposure time and b* values. Total bacterial counts were significantly (p ≤ 0.05) reduced from 2.69 to 0.93 log10 CFU/mL when the exposure time was increased from 0 to 60 min. The UV-C treatment showed similar trend on yeast and mold counts but to a lesser extend due to their resistance to UV. The sensory characteristics, i.e. odour, colour, taste, consistency and overall acceptability didn't change (p ≤ 0.05) as a result of UV-C treatment at any tested exposure times.

Laccaria bicolor pectin methylesterases are involved in ectomycorrhiza development with Populus tremula × Populus tremuloides.

The development of ectomycorrhizal (ECM) symbioses between soil fungi and tree roots requires modification of root cell walls. The pectin-mediated adhesion between adjacent root cells loosens to accommodate fungal hyphae in the Hartig net, facilitating nutrient exchange between partners. We investigated the role of fungal pectin modifying enzymes in Laccaria bicolor for ECM formation with Populus tremula × Populus tremuloides. We combine transcriptomics of cell-wall-related enzymes in both partners during ECM formation, immunolocalisation of pectin (Homogalacturonan, HG) epitopes in different methylesterification states, pectin methylesterase (PME) activity assays and functional analyses of transgenic L. bicolor to uncover pectin modification mechanisms and the requirement of fungal pectin methylesterases (LbPMEs) for ECM formation. Immunolocalisation identified remodelling of pectin towards de-esterified HG during ECM formation, which was accompanied by increased LbPME1 expression and PME activity. Overexpression or RNAi of the ECM-induced LbPME1 in transgenic L. bicolor lines led to reduced ECM formation. Hartig Nets formed with LbPME1 RNAi lines were shallower, whereas those formed with LbPME1 overexpressors were deeper. This suggests that LbPME1 plays a role in ECM formation potentially through HG de-esterification, which initiates loosening of adjacent root cells to facilitate Hartig net formation.

Genome-wide identification of the pectin methylesterase inhibitor genes in Brassica napus and expression analysis of selected members.

Pectin methylesterase inhibitors (PMEIs) modulate the status of pectin methylesterification by inhibiting the activity of pectin methylesterase (PME). Recent advances indicate PMEIs play an important role in regulating plant cell wall properties and defense responses. In this study, a genome-wide analysis of PMEI gene family in Brassica napus (B. napus) was conducted and the expression patterns of PMEI genes in response to Sclerotinia sclerotiorum (S. sclerotiorum) was investigated. A total of 190 PMEI proteins were identified from the genome of B. napus. Chromosomal location, gene structure and properties of the PMEI family were analyzed, and these features were compared with Arabidopsis thaliana (A. thaliana). A total of 123 syntenic ortholog pairs were detected from BnPMEI family by synteny analysis. Results showed the expansion of BnPMEI genes was likely predominately from whole-genome duplication (WGD) or segmental duplications. Multiple cis-elements related to plant growth and development, environmental stress responses, hormone responses were detected in the promoters of BnPMEI genes, implying they were regulated by both internal and external factors. Furthermore, expression analysis of transcriptome data combined with quantitative RT-PCR (qRT-PCR) validation identified several candidates that were strongly responsive to S. sclerotiorum infection. These BnPMEI genes are candidates for manipulation to breed novel and improved genotypes that are more resistant to sclerotinia stem rot (SSR). Extensive interactions were detected among 30 BnPMEI proteins, forming complex protein-protein interaction networks. Besides, 48 BnPMEIs showed interactions with other proteins including a range of cell wall structure-related enzymes. This study provides new insights into the evolution and function of PMEIs in B. napus and lays a foundation for breeding novel genotypes for crop improvement.

Starch-based composite coatings modulate cell wall modification and softening in Zaosu pears

Pear fruit is susceptible to softening and pulp disintegration leading to a short postharvest storage life. One possible method to alleviate softening is the use of edible coatings. Starch-based coatings and films were blended with cumin essential oil to investigate the film characteristics and the effect of the coating treatments on enzyme activities of Zaosu pears stored at 25 °C for 28 d. The essential oil incorporation enhanced the color, barrier properties and mechanical stability of the starch films. The coating treatment delayed fruit pulp degradation and retarded the activities of pectin methylesterase (PME), polygalacturonase (PG), cellulase (Cx), β-galactosidase (β-Gal), pectinesterase (PE), and polygalacturonase (PMG), during ambient storage for 28 d. The malondialdehyde (MDA) and membrane permeability (MP) activities of the coated fruits were lower compared to the control fruits. The composite coating improved the antioxidant enzymes; peroxidases (POD), phenylalanine ammonia lyase (PAL), superoxide dismutase (SOD), and catalase (CAT) and induced the fruit resistance against softening. The energy content; adenosine triphosphate (ATP) and energy charge (EC) and the activities of succinic dehydrogenase (SDH) and cytochrome c oxidase (CCO) enzymes were significantly higher in the coated fruits. It is viewed that the use of modified sweet potato starch with 0.2–0.4 % (v/v) cumin essential oil could be a useful tool in postharvest storage of fresh produce.

Relationship of the methanol production, pectin and pectinase activity during apple wine fermentation and aging

Understanding pectin structure and pectinase activity was important to control methanol content in apple wine. Therefore, this study compared inoculated fermentation (I), spontaneous fermentation (S) and inoculated fermentation combined with CaCl2 treatment (I & CaCl2) to explore their differences in methanol production, pectin peak molecular weight (Mp), and the activities of pectin methyl esterase (PME), pectin lyase (PL) and polygalacturonase (PG). The results showed that the activities of PME, PL and PG were intensively inhibited during fermentation; however, they still retained 3.41–5.84% (PME), 9.46–17.71% (PL) and 9.17–10.31% (PG) of the initial activities after aging for 30 days. Therefore, the methanol content was increased in all three aged wines even aging at 4 °C. CaCl2 promoted the PME and PL activities, and thus accelerated the methanol production. Because the pectin with Mp 3.07 kDa was retained by CaCl2, the highest pectin content was found in wine I & CaCl2 (160.69 mg/L), which was 95.47 mg/L higher than that in wine I, and 107.03 mg/L higher than that in wine S. In group S, the long lag period allowed pectin to withstand the pectinases inherent in apple juice for a long time, which was conducive to the cleavage of pectin to Mp lower than 3 kDa continuously, its further degradation led to the lowest pectin content (53.65 mg/L) in wine. Hence, inhibiting the pectinases activities, or shortening the aging period would play an important role in decreasing the methanol content in apple wine.

Auxin facilitates cell wall phosphorus reutilization in a nitric oxide-ethylene dependent manner in phosphorus deficient rice (Oryza sativa L.)

Auxin is involved in stress responses of plants, such as phosphorus (P) deficiency in rice. Studies on whether auxin participates in cell-wall inorganic phosphorous (Pi) reutilization in Pi-starved rice are scarce. This study explored the mechanisms underlying auxin-facilitated cell-wall Pi-reutilization in rice roots. Pi deficiency rapidly induced auxin accumulation in roots; exogenous auxin [α-naphthaleneacetic acid (NAA), a permeable analog of auxin] elevated soluble Pi content in roots and shoots by increasing pectin content by enhancing activity of pectin methylesterase, and upregulating the transcript level of PHOSPHORUS-TRANSPORTER-2, such that more Pi was translocated to the shoot. Irrespective of the Pi status, exogenous auxin induced nitric oxide (NO) and ethylene production, while exogenous sodium nitroprusside (an NO donor) and 1-aminocyclopropane-1-carboxylic acid (a precursor of ethylene) had no effect on auxin content, suggesting that auxin may act upstream of NO and ethylene. The beneficial effect of NAA in increasing soluble Pi content in roots and shoots disappeared when 2-(4-carboxyphenyl)− 4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (a scavenger of NO) or aminoethoxyvinylglycine (an inhibitor of ethylene) were applied, suggesting that auxin facilitates cell-wall Pi-reutilization in a NO-ethylene-dependent manner in Pi-deficient rice. Our study results suggest auxin application as an effective agronomic practice for improving plant Pi nutrition in P-deficient conditions.