Rapid Enzymatic Method for Pectin Methyl Esters Determination

Łękawska-Andrinopoulou, Lucyna; Vasiliou, Efstathios; Georgakopoulos, Dimitrios G.; Yialouris, Constantinos P; Georgiou, Constantınos A.

doi:10.1155/2013/854763

Cited by 8 publications

(3 citation statements)

References 26 publications

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“…To identify whether the increased DM in erf4 seeds was a result of decreased PME activity, total proteins were extracted from whole seeds of wild-type, erf4 , erf4-2 : 35S::ERF4 and 35S::ERF4 overexpressors, and PME activity was determined as previously described by Lekawska-Andrinopoulou et al (2013) with the average wild-type PME activity being normalized to 100% (=1). The results showed that PME activity was reduced by ~13% and ~17% in the erf4-1 and erf4-2 seeds, whereas it was elevated by 5~14% in 35S::ERF4 lines compared to wild-type (Figure 2E and F).…”

Section: Resultsmentioning

confidence: 99%

Arabidopsis ERF4 and MYB52 Transcription Factors Play Antagonistic Roles in Regulating Homogalacturonan De-methylesterification in Seed Coat Mucilage

Ding

Tang

Han

et al. 2020

Preprint

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One-sentence summary: Arabidopsis ERF4 and MYB52 transcription factors interact 18 and play antagonistic roles in regulating homogalacturonan de-methylesterification 19 related genes in the seed coat mucilage.ABSTRACT 24 The Arabidopsis (Arabidopsis thaliana) seed coat mucilage is a specialized cell wall with 25 pectin as its major component. Pectin is synthesized in the Golgi apparatus with 26 homogalacturonan fully methylesterified, but it must undergo de-methylesterification by 27 pectin methylesterase (PME) after being secreted into the cell wall. This reaction is 28 critical for pectin maturation, but the mechanisms of its transcriptional regulation remain 29 largely unknown. Here, we show that the Arabidopsis ERF4 transcription factor 30 positively regulates pectin de-methylesterification during seed development and directly 31 suppresses the expression of PME INHIBITOR13 (PMEI13), 14, 15 and 32 SUBTILISIN-LIKE SERINE PROTEASE 1.7 (SBT1.7). The erf4 mutant seeds showed 33 repartitioning of mucilage between soluble and adherent layers as a result of decreased 34 PME activity and increased degree of pectin methylesterification. ERF4 physically 35 associates with and antagonizes MYB52 in activating PMEI6, 14 and SBT1.7 and 36 MYB52 also antagonizes ERF4 activity in the regulation of downstream targets. Gene 37 expression studies revealed that ERF4 and MYB52 have opposite effects on pectin 38 de-methylesterification. Genetic analysis indicated that the erf4-2 myb52 double mutant 39 seeds show mucilage phenotype similar to wild-type. Taken together, this study 40 demonstrates that ERF4 and MYB52 antagonize each other's activity to maintain the 41 appropriate degree of pectin methylesterification, expanding our understanding of how 42 pectin de-methylesterification is fine-tuned by the ERF4-MYB52 transcriptional complex 43 in the seed mucilage. 44 130 6 stress (Liu et al., 2017). ERF4 is expressed ubiquitously in Arabidopsis (Winter et al., 131 2007), suggesting that it has other functions in addition to those mentioned above. For 132 example, its regulatory role in cell wall biology has not been reported yet. 133Here, we provide evidence that ERF4 positively regulates pectin de-methylesterification 134 in the seed mucilage. We find a repartitioning of seed mucilage polysaccharides in the 135 erf4 seeds under vigorous shaking conditions which can be attributed to an increase in 136 DM. ERF4 binds to the cis-regulatory elements and directly suppresses the expression of 137 PMEI13, 14, 15 and SBT1.7. Moreover, ERF4 and MYB52 were found to interact, and 138 play completely opposite roles in pectin de-methylesterification by antagonizing each 139 other's transcriptional activity, which is confirmed by gene expression and genetic 140 evidence. Overall, these findings provide a fine-tuned mechanism where ERF4 and 141 MYB52 antagonistically control mucilage modification related genes. 142 RESULTS 143Expression pattern of ERF4 is correlated with seed mucilage deposition 144 Previous studies revea...

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Section: Resultsmentioning

confidence: 99%

Arabidopsis ERF4 and MYB52 Transcription Factors Play Antagonistic Roles in Regulating Homogalacturonan De-methylesterification in Seed Coat Mucilage

Ding

Tang

Han

et al. 2020

Preprint

View full text Add to dashboard Cite

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“…Several PME genes have been identified and investigated (Tieman et al, 1992;Gaffe et al, 1997;Wen et al, 1999;Hongo et al, 2012). PMT activity has been described, and the enzyme properties characterized, in several plant species (Goubet et al, 1998;Goubet and Mohnen, 1999;Ishikawa et al, 2000;Łękawska-Andrinopoulou et al, 2013). However, functions of the PMT gene are not well understood, especially during the reproductive process.…”

Section: Introductionmentioning

confidence: 99%

Rice Putative Methyltransferase Gene OsPMT16 Is Required for Pistil Development Involving Pectin Modification

et al. 2020

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Pectin synthesis and modification are vital for plant development, although the underlying mechanisms are still not well understood. Furthermore, reports on the function of pectin in the pistil are limited. Herein, we report the functional characterization of the OsPMT16 gene, which encodes a putative pectin methyltransferase (PMT) in rice. The cell walls of rice leaves contain less pectin, and chemical analysis of pectin in the flower organ had not been previously performed. Therefore, in the present study, the amount of pectin in the reproductive tissues of rice was investigated. Of the reproductive tissues, the pistil was especially rich in pectin; thus, we focused on the pistil. OsPMT16 expression was confirmed in the pistil, and effects of pectin methylesterification regulation on the reproductive stage were investigated by studying the phenotype of the T-DNA insertion mutant. The ospmt16 mutant showed significantly reduced fertility. When the flowers were observed, tissue morphogenesis was abnormal in the pistil. Immunofluorescence staining by pectin-specific monoclonal antibodies of the pistil revealed that total pectin and esterified pectin were decreased among ospmt16 mutants. These results indicate that OsPMT16 contributes significantly to pistil development during reproductive growth.

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“…Among the above detection techniques, spectrophotometry presents several advantages, including its flexibility to be automated using flow injection analysis (FIA) [ 12 , 13 ], methodology that has been widely used to develop analytical procedures [ 14 , 15 ]. Spectrophotometry technique allows the use of cost-effective instrumentation, such as homemade photometers, assembled using light emitting diode (LED) and photodiode [ 16 , 17 ].…”

Section: Introductionmentioning

confidence: 99%

A Highly Sensitive Multicommuted Flow Analysis Procedure for Photometric Determination of Molybdenum in Plant Materials without a Solvent Extraction Step

Santos

Reis

2017

Journal of Analytical Methods in Chemistry

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A highly sensitive analytical procedure for photometric determination of molybdenum in plant materials was developed and validated. This procedure is based on the reaction of Mo(V) with thiocyanate ions (SCN−) in acidic medium to form a compound that can be monitored at 474 nm and was implemented employing a multicommuted flow analysis setup. Photometric detection was performed using an LED-based photometer coupled to a flow cell with a long optical path length (200 mm) to achieve high sensitivity, allowing Mo(V) determination at a level of μg L−1 without the use of an organic solvent extraction step. After optimization of operational conditions, samples of digested plant materials were analyzed employing the proposed procedure. The accuracy was assessed by comparing the obtained results with those of a reference method, with an agreement observed at 95% confidence level. In addition, a detection limit of 9.1 μg L−1, a linear response (r = 0.9969) over the concentration range of 50–500 μg L−1, generation of only 3.75 mL of waste per determination, and a sampling rate of 51 determinations per hour were achieved.

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Rapid Enzymatic Method for Pectin Methyl Esters Determination

Cited by 8 publications

References 26 publications

Arabidopsis ERF4 and MYB52 Transcription Factors Play Antagonistic Roles in Regulating Homogalacturonan De-methylesterification in Seed Coat Mucilage

Arabidopsis ERF4 and MYB52 Transcription Factors Play Antagonistic Roles in Regulating Homogalacturonan De-methylesterification in Seed Coat Mucilage

Rice Putative Methyltransferase Gene OsPMT16 Is Required for Pistil Development Involving Pectin Modification

A Highly Sensitive Multicommuted Flow Analysis Procedure for Photometric Determination of Molybdenum in Plant Materials without a Solvent Extraction Step

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