Evolutionary and functional characterization of leucoanthocyanidin reductases from Camellia sinensis

Wang, Peiqiang; Zhang, Lingjie; Jiang, Xiaolan; Dai, Xinlong; Xu, Lijuan; Li, Tong; Xing, Dawei; Li, Yanzhi; Li, Mingzhuo; Gao, Liping; Xia, Tao

doi:10.1007/s00425-017-2771-z

Cited by 79 publications

(72 citation statements)

References 50 publications

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“…Unlike grapevine and M. truncatula, there is no LAR gene in the Arabidopsis genome (Lepiniec et al, 2006). Although leucocyanidin is very unstable and difficult to trap by analytical instruments, based on previous studies it can be inferred that leucocyanidin does not "leak out" of a coupled DFR/ANS in Arabidopsis (Liu et al, 2013;Ferraro et al, 2014;Wang et al, 2018). If leucocyanidin is available in Arabidopsis, the seeds should contain PAs with (1)-catechin as extension units, which is not observed.…”

Section: Table 2 Identification Of Rossmann Folds and Prediction Of mentioning

confidence: 99%

“…If leucocyanidin is available in Arabidopsis, the seeds should contain PAs with (1)-catechin as extension units, which is not observed. Moreover, overexpression of pea (Pisum sativum) LAR or three tea (Camellia sinensis) LARs in wild-type Arabidopsis does not result in synthesis of (1)-catechin, although LAR activity to generate (1)catechin was detected in vitro (Ferraro et al, 2014;Wang et al, 2018), and expression of cacao LAR in the Arabidopsis ans mutant results in significant accumulation of (1)-catechin (Liu et al, 2013), indicating that ANS has a high affinity for leucocyanidin or associates with DFR very tightly. Based on the above, the absence of Cys-C in Arabidopsis might be due to the lack of availability of leucocyanidin in vivo due to channeling between DFR and ANS.…”

Section: Table 2 Identification Of Rossmann Folds and Prediction Of mentioning

confidence: 99%

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VvLAR1 and VvLAR2 Are Bifunctional Enzymes for Proanthocyanidin Biosynthesis in Grapevine

Jun

Duan

et al. 2019

Plant Physiol.

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Proanthocyanidins (PAs) in grapevine (Vitis vinifera) are found mainly in berries, and their content and degree of polymerization are important for the mouth feel of red wine. However, the mechanism of PA polymerization in grapevine remains unclear. Previous studies in the model legume Medicago truncatula showed that 4b-(S-cysteinyl)-epicatechin (Cys-EC) is an epicatechintype extension unit for nonenzymatic PA polymerization, and that leucoanthocyanidin reductase (LAR) converts Cys-EC into epicatechin starter unit to control PA extension. Grapevine possesses two LAR genes, but their functions are not clear. Here, we show that both Cys-EC and 4b-(S-cysteinyl)-catechin (Cys-C) are present in grapevine. Recombinant VvLAR1 and VvLAR2 convert Cys-C and Cys-EC into (1)-catechin and (2)-epicatechin, respectively, in vitro. The kinetic parameters of VvLARs are similar, with both enzymes being more efficient with Cys-C than with Cys-EC, the 2,3-cis conformation of which results in steric hindrance in the active site. Both VvLARs also produce (1)-catechin from leucocyanidin, and an inactive VvLAR2 allele reported previously is the result of a single amino acid mutation in the N terminus critical for all NADPH-dependent activities of the enzyme. VvLAR1 or VvLAR2 complement the M. truncatula lar:ldox double mutant that also lacks the leucoanthocyanidin dioxygenase (LDOX) required for epicatechin starter unit formation, resulting in increased soluble PA levels, decreased insoluble PA levels, and reduced levels of Cys-C and Cys-EC when compared to the double mutant, and the appearance of catechin, epicatechin, and PA dimers characteristic of the ldox single mutant in young pods. These data advance our knowledge of PA building blocks and LAR function and provide targets for grapevine breeding to alter PA composition.

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Section: Table 2 Identification Of Rossmann Folds and Prediction Of mentioning

confidence: 99%

Section: Table 2 Identification Of Rossmann Folds and Prediction Of mentioning

confidence: 99%

VvLAR1 and VvLAR2 Are Bifunctional Enzymes for Proanthocyanidin Biosynthesis in Grapevine

Jun

Duan

et al. 2019

Plant Physiol.

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“…Labels that are highlighted red, yellow, and blue represent seeds with (regular darkening) RD, SD, and ND phenotypes, respectively (color figure online) Reverse: GAG AAG AGA CCT TGG ATT TGT GTG darkening and non-darkening beans (Beninger et al 2005;Chen et al 2015;Freixas Coutin et al 2017). Proanthocyanidins are abundant in the brown seed coats of many plants such as A. thaliana (Dixon et al 2005) and ANR plays a key role in the biosynthesis of PAs in that species (Wang et al 2018). PvANR1 was found to be strongly associated with seed coat PA accumulation in darkening cranberry beans (Freixas Coutin et al 2017).…”

Section: Discussionmentioning

confidence: 99%

A R2R3-MYB gene-based marker for the non-darkening seed coat trait in pinto and cranberry beans (Phaseolus vulgaris L.) derived from ‘Wit-rood boontje’

Erfatpour

Pauls

2020

Theor Appl Genet

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Key message The gene Phvul.010G130600 which codes for a MYB was shown to be tightly associated with seed coat darkening in Phaseolus vulgaris and a single nucleotide deletion in the allele in Wit-rood disrupts a transcription activation region that likely prevents its functioning in this non-darkening genotype. Abstract The beige and white background colors of the seed coats of conventional pinto and cranberry beans turn brown through a process known as postharvest darkening (PHD). Seed coat PHD is attributed to proanthocyanidin accumulation and its subsequent oxidation in the seed coat. The J gene is an uncharacterized classical genetic locus known to be responsible for PHD in common bean (P. vulgaris) and individuals that are homozygous for its recessive allele have a non-darkening (ND) seed coat phenotype. A previous study identified a major colorimetrically determined QTL for seed coat color on chromosome 10 that was associated with the ND trait. The objectives of this study were to identify a gene associated with seed coat postharvest darkening in common bean and understand its function in promoting seed coat darkening. Amplicon sequencing of 21 candidate genes underlying the QTL associated with the ND trait revealed a single nucleotide deletion (c.703delG) in the candidate gene Phvul.010G130600 in non-darkening recombinant inbred lines derived from crosses between ND 'Witrood boontje' and a regular darkening pinto genotype. In silico analysis indicated that Phvul.010G130600 encodes a protein with strong amino acid sequence identity (70%) with a R2R3-MYB-type transcription factor MtPAR, which has been shown to regulate proanthocyanidin biosynthesis in Medicago truncatula seed coat tissue. The deletion in the 'Wit-rood boontje' allele of Phvul.010G130600 likely causes a translational frame shift that disrupts the function of a transcriptional activation domain contained in the C-terminus of the R2R3-MYB. A gene-based dominant marker was developed for the dominant allele of Phvul.010G130600 which can be used for marker-assisted selection of ND beans.Communicated by Albrecht E. Melchinger. Electronic supplementary materialThe online version of this article (https ://doi.org/10.1007/s0012 2-020-03571 -7) contains supplementary material, which is available to authorized users.

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“…NtLAR and NtANR s in CsMYB5b transgenic tobacco [70]. So the transport of anthocyanidins through GhANR, GhLAR into fiber cell will be the important link for genetic engineering of colored fiber molecular improvement.…”

Section: Lars Promoted the Biosynthesis Of Catechin Monomers And Inhimentioning

confidence: 99%

Functional analysis of GhCHS, GhANR and GhLAR in colored fiber formation of Gossypium hirsutum L.

Jianfang

Shen

Yuan

et al. 2019

Preprint

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Background The formation of natural colored fibers mainly results from the accumulation of different anthocyanidins and their derivatives in the fibers of Gossypium hirsutum L. Chalcone synthase (CHS) is the first committed enzyme of flavonoid biosynthesis, and anthocyanidins are transported into fiber cell after biosynthesis mainly by Anthocyanidin reductase (ANR) and Leucoanthocyanidin reductase (LAR) to present diverse colors with distinct stability. The biochemical and molecular mechanism of pigment formation in natural colored cotton fiber is not clear. Results The three key genes of GhCHS , GhANR and GhLAR were predominantly expressed in the developing fibers of colored cotton. In the GhCHSi , GhANRi and GhLARi transgenic cottons, the expression levels of GhCHS , GhANR and GhLAR significantly decreased in the developing cotton fiber, negatively correlated with the content of anthocyanidins and the color depth of cotton fiber. In colored cotton Zongxu1 (ZX1) and the GhCHSi , GhANRi and GhLARi transgenic lines of ZX1, HZ and ZH, the anthocyanidin contents of the leaves, cotton kernels, the mixture of fiber and seedcoat were all changed and positively correlated with the fiber color. Conclusion The three genes of GhCHS , GhANR and GhLAR were predominantly expressed early in developing colored cotton fibers and identified to be a key genes of cotton fiber color formation. The expression levels of the three genes affected the anthocyanidin contents and fiber color depth. So the three genes played a crucial part in cotton fiber color formation and has important significant to improve natural colored cotton quality and create new colored cotton germplasm resources by genetic engineering.

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Evolutionary and functional characterization of leucoanthocyanidin reductases from Camellia sinensis

Cited by 79 publications

References 50 publications

VvLAR1 and VvLAR2 Are Bifunctional Enzymes for Proanthocyanidin Biosynthesis in Grapevine

VvLAR1 and VvLAR2 Are Bifunctional Enzymes for Proanthocyanidin Biosynthesis in Grapevine

A R2R3-MYB gene-based marker for the non-darkening seed coat trait in pinto and cranberry beans (Phaseolus vulgaris L.) derived from ‘Wit-rood boontje’

Functional analysis of GhCHS, GhANR and GhLAR in colored fiber formation of Gossypium hirsutum L.

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