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      Apoplastic sugar may be lost from grape berries and retrieved in pedicels

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          Abstract

          In ripening grape ( Vitis sp.) berries, the combination of rapid sugar import, apoplastic phloem unloading, and water discharge via the xylem creates a potential risk for apoplastic sugar to be lost from the berries. We investigated the likelihood of such sugar loss and a possible sugar retrieval mechanism in the pedicels of different Vitis genotypes. Infusion of D-glucose-1- 13C or L-glucose-1- 13C to the stylar end of attached berries demonstrated that both sugars can be leached from the berries, but only the nontransport sugar L-glucose moved beyond the pedicels. No 13C enrichment was found in peduncles and leaves. Genes encoding 10 sugar transporters were expressed in the pedicels throughout grape ripening. Using an immunofluorescence technique, we localized the sucrose transporter SUC27 to pedicel xylem parenchyma cells. These results indicate that pedicels possess the molecular machinery for sugar retrieval from the apoplast. Plasmodesmata were observed between vascular parenchyma cells in pedicels, and movement of the symplastically mobile dye carboxyfluorescein demonstrated that the symplastic connection is physiologically functional. Taken together, the chemical, molecular, and anatomical evidence gathered here supports the idea that some apoplastic sugar can be leached from grape berries and is effectively retrieved in a two-step process in the pedicels. First, sugar transporters may actively retrieve leached sugar from the xylem. Second, retrieved sugar may move symplastically to the pedicel parenchyma for local use or storage, or to the phloem for recycling back to the berry.

          Abstract

          Grape berry pedicels may retrieve sugar that is lost via the xylem following apoplastic phloem unloading in the berries.

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          Most cited references73

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          An optimized grapevine RNA isolation procedure and statistical determination of reference genes for real-time RT-PCR during berry development

          Background Accuracy in quantitative real-time RT-PCR is dependent on high quality RNA, consistent cDNA synthesis, and validated stable reference genes for data normalization. Reference genes used for normalization impact the results generated from expression studies and, hence, should be evaluated prior to use across samples and treatments. Few statistically validated reference genes have been reported in grapevine. Moreover, success in isolating high quality RNA from grapevine tissues is typically limiting due to low pH, and high polyphenolic and polysaccharide contents. Results We describe optimization of an RNA isolation procedure that compensates for the low pH found in grape berries and improves the ability of the RNA to precipitate. This procedure was tested on pericarp and seed developmental series, as well as steady-state leaf, root, and flower tissues. Additionally, the expression stability of actin, AP47 (clathrin-associated protein), cyclophilin, EF1-α (elongation factor 1-α), GAPDH (glyceraldehyde 3-phosphate dehydrogenase), MDH (malate dehydrogenase), PP2A (protein phosphatase), SAND, TIP41, α-tubulin, β-tubulin, UBC (ubiquitin conjugating enzyme), UBQ-L40 (ubiquitin L40) and UBQ10 (polyubiquitin) were evaluated on Vitis vinifera cv. Cabernet Sauvignon pericarp using three different statistical approaches. Although several of the genes proved to be relatively stable, no single gene outperformed all other genes in each of the three evaluation methods tested. Furthermore, the effect of using one reference gene versus normalizing to the geometric mean of several genes is presented for the expression of an aquaporin and a sucrose transporter over a developmental series. Conclusion In order to quantify relative transcript abundances accurately using real-time RT-PCR, we recommend that combinations of several genes be used for normalization in grape berry development studies. Our data support GAPDH, actin, EF1-α and SAND as the most relevant reference genes for this purpose.
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            The phloem, a miracle of ingenuity

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              Membrane-transport systems for sucrose in relation to whole-plant carbon partitioning.

              Brian Ayre (2011)
              Sucrose is the principal product of photosynthesis used for the distribution of assimilated carbon in plants. Transport mechanisms and efficiency influence photosynthetic productivity by relieving product inhibition and contribute to plant vigor by controlling source/sink relationships and biomass partitioning. Sucrose is synthesized in the cytoplasm and may move cell to cell through plasmodesmata or may cross membranes to be compartmentalized or exported to the apoplasm for uptake into adjacent cells. As a relatively large polar compound, sucrose requires proteins to facilitate efficient membrane transport. Transport across the tonoplast by facilitated diffusion, antiport with protons, and symport with protons have been proposed; for transport across plasma membranes, symport with protons and a mechanism resembling facilitated diffusion are evident. Despite decades of research, only symport with protons is well established at the molecular level. This review aims to integrate recent and older studies on sucrose flux across membranes with principles of whole-plant carbon partitioning.
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                Author and article information

                Contributors
                Journal
                Plant Physiol
                Plant Physiol
                plphys
                Plant Physiology
                Oxford University Press
                0032-0889
                1532-2548
                September 2022
                01 June 2022
                01 June 2022
                : 190
                : 1
                : 592-604
                Affiliations
                Department of Horticulture, Irrigated Agriculture Research and Extension Center, Washington State University , Prosser, WA, USA
                Ste. Michelle Wine Estates , Prosser, WA, USA
                Department of Horticulture, Irrigated Agriculture Research and Extension Center, Washington State University , Prosser, WA, USA
                Department of Horticulture, Irrigated Agriculture Research and Extension Center, Washington State University , Prosser, WA, USA
                INRAE, University of Bordeaux, ISVV , Villenave d’Ornon, France
                Institute of Botany, Chinese Academy of Sciences , Beijing, China
                INRAE, University of Bordeaux, ISVV , Villenave d’Ornon, France
                Department of Horticulture, Irrigated Agriculture Research and Extension Center, Washington State University , Prosser, WA, USA
                Author notes
                Author for correspondence: mkeller@ 123456wsu.edu
                [†]

                Summerland Research and Development Centre, Agriculture & Agri-Food Canada, Summerland, BC V0H 1Z0, Canada.

                [‡]

                Belgrano 471, 5561 Tupungato, Mendoza, Argentina.

                [ # ]

                Senior author

                Author information
                https://orcid.org/0000-0001-7500-3191
                https://orcid.org/0000-0002-9319-2042
                https://orcid.org/0000-0002-8098-5801
                https://orcid.org/0000-0002-7625-8337
                https://orcid.org/0000-0001-9602-0441
                https://orcid.org/0000-0003-2144-2388
                Article
                kiac262
                10.1093/plphys/kiac262
                9434297
                35642904
                7d01f69e-e538-4b74-ae50-d12fd50da1c4
                © The Author(s) 2022. Published by Oxford University Press on behalf of American Society of Plant Biologists.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 07 April 2022
                : 07 May 2022
                Page count
                Pages: 13
                Funding
                Funded by: Washington State University’s College of Agricultural, Human, and Natural Resource Sciences;
                Funded by: Department of Horticulture, and Viticulture and Enology Program; the Chateau Ste;
                Funded by: Michelle Distinguished Professorship in Viticulture; the Institut National de la Recherche Agronomique and the University of Bordeaux;
                Categories
                Focus Issue on Evolution of Plant Structure and Function
                Research Articles
                Genes, Development and Evolution
                AcademicSubjects/SCI02286
                AcademicSubjects/SCI02287
                AcademicSubjects/SCI01270
                AcademicSubjects/SCI01280
                AcademicSubjects/SCI02288

                Plant science & Botany
                Plant science & Botany

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