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      Characterization and analysis of CCR and CAD gene families at the whole-genome level for lignin synthesis of stone cells in pear ( Pyrus bretschneideri) fruit

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          ABSTRACT

          The content of stone cells has significant effects on the flavour and quality of pear fruit. Previous research suggested that lignin deposition is closely related to stone cell formation. In the lignin biosynthetic pathway, cinnamoyl-CoA reductase (CCR) and cinnamyl alcohol dehydrogenase (CAD), dehydrogenase/reductase family members, catalyse the last two steps in monolignol synthesis. However, there is little knowledge of the characteristics of the CCR and CAD families in pear and their involvement in lignin synthesis of stone cells. In this study, 31 CCRs and 26 CADs were identified in the pear genome. Phylogenetic trees for CCRs and CADs were constructed; key amino acid residues were analysed, and three-dimensional structures were predicted. Using quantitative real-time polymerase chain reaction (qRT-PCR), PbCAD2, PbCCR1, -2 and - 3 were identified as participating in lignin synthesis of stone cells in pear fruit. Subcellular localization analysis showed that the expressed proteins (PbCAD2, PbCCR1, -2 and -3) are found in the cytoplasm or at the cell membrane. These results reveal the evolutionary features of the CCR and CAD families in pear as well as the genes responsible for regulation of lignin synthesis and stone cell development in pear fruit.

          Abstract

          Summary: The characteristics of CCR and CAD families were systematically analyzed, and candidate members related to lignin synthesis and stone cell development were screened out in this study.

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          Most cited references54

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          The genome of the pear ( Pyrus bretschneideri Rehd.)

          The draft genome of the pear ( Pyrus bretschneideri ) using a combination of BAC-by-BAC and next-generation sequencing is reported. A 512.0-Mb sequence corresponding to 97.1% of the estimated genome size of this highly heterozygous species is assembled with 194× coverage. High-density genetic maps comprising 2005 SNP markers anchored 75.5% of the sequence to all 17 chromosomes. The pear genome encodes 42,812 protein-coding genes, and of these, ∼28.5% encode multiple isoforms. Repetitive sequences of 271.9 Mb in length, accounting for 53.1% of the pear genome, are identified. Simulation of eudicots to the ancestor of Rosaceae has reconstructed nine ancestral chromosomes. Pear and apple diverged from each other ∼5.4–21.5 million years ago, and a recent whole-genome duplication (WGD) event must have occurred 30–45 MYA prior to their divergence, but following divergence from strawberry. When compared with the apple genome sequence, size differences between the apple and pear genomes are confirmed mainly due to the presence of repetitive sequences predominantly contributed by transposable elements (TEs), while genic regions are similar in both species. Genes critical for self-incompatibility, lignified stone cells (a unique feature of pear fruit), sorbitol metabolism, and volatile compounds of fruit have also been identified. Multiple candidate SFB genes appear as tandem repeats in the S -locus region of pear; while lignin synthesis-related gene family expansion and highly expressed gene families of HCT , C3′H , and CCOMT contribute to high accumulation of both G-lignin and S-lignin. Moreover, alpha-linolenic acid metabolism is a key pathway for aroma in pear fruit.
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            Genome-wide characterization of the lignification toolbox in Arabidopsis.

            Lignin, one of the most abundant terrestrial biopolymers, is indispensable for plant structure and defense. With the availability of the full genome sequence, large collections of insertion mutants, and functional genomics tools, Arabidopsis constitutes an excellent model system to profoundly unravel the monolignol biosynthetic pathway. In a genome-wide bioinformatics survey of the Arabidopsis genome, 34 candidate genes were annotated that encode genes homologous to the 10 presently known enzymes of the monolignol biosynthesis pathway, nine of which have not been described before. By combining evolutionary analysis of these 10 gene families with in silico promoter analysis and expression data (from a reverse transcription-polymerase chain reaction analysis on an extensive tissue panel, mining of expressed sequence tags from publicly available resources, and assembling expression data from literature), 12 genes could be pinpointed as the most likely candidates for a role in vascular lignification. Furthermore, a possible novel link was detected between the presence of the AC regulatory promoter element and the biosynthesis of G lignin during vascular development. Together, these data describe the full complement of monolignol biosynthesis genes in Arabidopsis, provide a unified nomenclature, and serve as a basis for further functional studies.
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              Transcriptional networks for lignin biosynthesis: more complex than we thought?

              Lignin is an aromatic heteropolymer and the second most abundant plant biopolymer after cellulose. It is deposited mostly in the secondary cell walls of vascular plants and is essential for water transport, mechanical support and for plant pathogen defense. Lignin biosynthesis is a highly energy-consuming and irreversible process that responds to many developmental and environmental cues, including light, sugar content, circadian clock, plant hormones and wounding. During the past decade, many transcription factors involved in lignin biosynthesis have been identified and characterized. In this review, we assess how these transcriptional activators and repressors modulate lignin biosynthesis, and discuss crosstalk between the lignin biosynthesis pathway and other physiological processes. Copyright © 2010 Elsevier Ltd. All rights reserved.
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                Author and article information

                Journal
                Biol Open
                Biol Open
                bio
                biolopen
                Biology Open
                The Company of Biologists Ltd
                2046-6390
                15 November 2017
                15 November 2017
                15 November 2017
                : 6
                : 11
                : 1602-1613
                Affiliations
                [1 ]School of Life Science, Anhui Agricultural University , No. 130, Changjiang West Road, Hefei 230036, China
                [2 ]Horticultural Institute, Anhui Academy of Agricultural Sciences , Hefei, Anhui 230031, China
                Author notes
                [*]

                These authors contributed equally to this work

                []Author for correspondence ( ypcaiah@ 123456163.com )
                Author information
                http://orcid.org/0000-0002-0842-6968
                Article
                BIO026997
                10.1242/bio.026997
                5703608
                29141952
                7f217cb3-9693-41ab-aed5-d4a8d0a99719
                © 2017. Published by The Company of Biologists Ltd

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

                History
                : 17 May 2017
                : 18 September 2017
                Funding
                Funded by: National Natural Science Foundation of China, http://dx.doi.org/10.13039/501100001809;
                Award ID: 31640068
                Funded by: Anhui Agriculture University, http://dx.doi.org/10.13039/501100006462;
                Award ID: 2017yjs-32
                Award ID: 201610364013
                Categories
                Research Article

                Life sciences
                pyrus bretschneideri,stone cells,lignin synthesis,ccr and cad gene families,expression analysis,subcellular localization

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