Differential metabolism of pectic galactan in tomato and strawberry fruit: detection of the LM26 branched galactan epitope in ripe strawberry fruit

Posé, Sara; Marcus, Susan E.; Knox, John

doi:10.1111/ppl.12748

Cited by 13 publications

(11 citation statements)

References 49 publications

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“…This crude cell wall preparation was then used in the fractionation studies. The cell wall materials were sequentially extracted (10 mg in 1 mL) with water, CDTA, 4 M KOH, and with a cellulase treatment of the final insoluble residue to release polysaccharides associated with cellulose microfibrils as described in Posé et al (2018). Solubilized extracts at dilutions ranging from 250-fold to 31,250-fold were used to coat microtiter plates before ELISA procedures as described in Willats et al (1998) and Posé et al (2018).…”

Section: Immunofluorescence and Immunocytochemistry Proceduresmentioning

confidence: 99%

Characterisation of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato

et al. 2018

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Tomato (Solanum lycopersicum) is a globally important crop with an economic value in the tens of billions of dollars, and a significant supplier of essential vitamins, minerals, and phytochemicals in the human diet. Shelf life is a key quality trait related to alterations in cuticle properties and remodeling of the fruit cell walls. Studies with transgenic tomato plants undertaken over the last 20 years have indicated that a range of pectin-degrading enzymes are involved in cell wall remodeling. These studies usually involved silencing of only a single gene and it has proved difficult to compare the effects of silencing these genes across the different experimental systems. Here we report the generation of CRISPR-based mutants in the ripening-related genes encoding the pectin-degrading enzymes pectate lyase (PL), polygalacturonase 2a (PG2a), and b-galactanase (TBG4). Comparison of the physiochemical properties of the fruits from a range of PL, PG2a, and TBG4 CRISPR lines demonstrated that only mutations in PL resulted in firmer fruits, although mutations in PG2a and TBG4 influenced fruit color and weight. Pectin localization, distribution, and solubility in the pericarp cells of the CRISPR mutant fruits were investigated using the monoclonal antibody probes LM19 to deesterified homogalacturonan, INRA-RU1 to rhamnogalacturonan I, LM5 to b-1,4-galactan, and LM6 to arabinan epitopes, respectively. The data indicate that PL, PG2a, and TBG4 act on separate cell wall domains and the importance of cellulose microfibril-associated pectin is reflected in its increased occurrence in the different mutant lines.

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Section: Immunofluorescence and Immunocytochemistry Proceduresmentioning

confidence: 99%

Characterisation of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato

et al. 2018

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“…Among them, the most apparent change is the reduction in fruit firmness. Cell wall disassembly is considered to be the major determinant of fruit softening during the ripening process in fleshy fruits [ 50 , 51 ], which results from a decrease in cell turgor pressure as well as cell wall polysaccharide remodeling and metabolism [ 52 ].…”

Section: Discussionmentioning

confidence: 99%

Changes in Homogalacturonan Metabolism in Banana Peel during Fruit Development and Ripening

Ning

Chen

et al. 2021

IJMS

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Though numerous studies have focused on the cell wall disassembly of bananas during the ripening process, the modification of homogalacturonan (HG) during fruit development remains exclusive. To better understand the role of HGs in controlling banana fruit growth and ripening, RNA-Seq, qPCR, immunofluorescence labeling, and biochemical methods were employed to reveal their dynamic changes in banana peels during these processes. Most HG-modifying genes in banana peels showed a decline in expression during fruit development. Four polygalacturonase and three pectin acetylesterases showing higher expression levels at later developmental stages than earlier ones might be related to fruit expansion. Six out of the 10 top genes in the Core Enrichment Gene Set were HG degradation genes, and all were upregulated after softening, paralleled to the significant increase in HG degradation enzyme activities, decline in peel firmness, and the epitope levels of 2F4, CCRC-M38, JIM7, and LM18 antibodies. Most differentially expressed alpha-1,4-galacturonosyltransferases were upregulated by ethylene treatment, suggesting active HG biosynthesis during the fruit softening process. The epitope level of the CCRC-M38 antibody was positively correlated to the firmness of banana peel during fruit development and ripening. These results have provided new insights into the role of cell wall HGs in fruit development and ripening.

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“…The plant cell wall of flesh fruit tissue is a heterogeneous matrix of interacting and cross-linked polysaccharides, which provides structural integrity for the whole fruit. Being a Type I primary cell wall, it is a network of xyloglucan-cellulose microfibrils, embedded and closely linked with an amorphous matrix of pectin polysaccharides [1][2][3][4]. Although the xyloglucan-cellulose network is considered to be the basic load-bearing component of the plant cell wall, the enzyme-induced modifications of the pectin matrix are now gaining more recognition as the major cause of postharvest quality losses in apples [5,6].…”

Section: Introductionmentioning

confidence: 99%

An Atomic Force Microscopy Study on the Effect of β-Galactosidase, α-l-Rhamnosidase and α-l-Arabinofuranosidase on the Structure of Pectin Extracted from Apple Fruit Using Sodium Carbonate

Pieczywek

Cybulska

Zdunek

2020

IJMS

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The enzyme driven changes in plant cell wall structure during fruit ripening result in debranching, depolymerization and solubilization of pectin polysaccharides, which has an effect in terms of the postharvest quality losses in fruit. Atomic force microscopy (AFM) has revealed that diluted alkali soluble pectins (DASP) from fruit and vegetables have an interesting tendency to self-assemble into regular structures. However, the mechanism is not yet fully understood. The current study is aimed at investigating the role of neutral sugars, namely galactose, rhamnose and arabinose in the formation of the branched structure of DASP. β-galactosidase, α-l-rhamnosidase and α-l-arabinofuranosidase enzymes were used for the treatment of DASP extracted from Golden Delicious apple flesh (Malus domestica cv. Golden Delicious). The effects of the selective degradation of pectic polysaccharides after 15, 30, 60, 90 and 120 min of incubation were observed using AFM. The α-l-rhamnosidase enzyme activity on pectin extracted with Na2CO3 did not cause any visible or measurable degradation of the molecular structure. The moderate effects of β-galactosidase enzymatic treatment suggested the possible role of galactose in the branching of DASP molecules deposited on mica. Data obtained for α-l-arabinofuranosidase indicated the crucial role of arabinose in the formation and preservation of the highly branched structure of the DASP fraction.

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Differential metabolism of pectic galactan in tomato and strawberry fruit: detection of the LM26 branched galactan epitope in ripe strawberry fruit

Cited by 13 publications

References 49 publications

Characterisation of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato

Characterisation of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato

Changes in Homogalacturonan Metabolism in Banana Peel during Fruit Development and Ripening

An Atomic Force Microscopy Study on the Effect of β-Galactosidase, α-l-Rhamnosidase and α-l-Arabinofuranosidase on the Structure of Pectin Extracted from Apple Fruit Using Sodium Carbonate

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