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Expression analysis of a sucrose carrier in the germinating seedling of Ricinus communis

Expression analysis of a sucrose carrier in the germinating seedling of Ricinus communis
Expression analysis of a sucrose carrier in the germinating seedling of Ricinus communis
This study describes the expression of a sucrose carrier at various developmental stages in Ricinus communis. A partial-length cDNA clone, RcSUT1, was isolated by RT-PCR from Ricinus seedling RNA. This is almost identical to a sucrose carrier cDNA, Rscr1, which has previously been isolated by library screening. However, we have observed a very different expression pattern in the seedling to that previously reported. Northern analysis, with RcSUT1 as a probe, revealed high expression of a 2 kb transcript in the cotyledons of the germinating seedling; transcript levels were similar in cotyledons harvested 3–6 days after germination. A much lower level of this transcript was detected in the root, hypocotyl and endosperm RNA of the seedling and very low levels were also present in the sink and source leaves of the mature plant. This pattern of expression was also reflected at the protein level with an antipeptide antibody raised to part of the RcSUT1 deduced amino acid sequence.

Tissue print hybridisation analysis of the hypocotyl revealed that the sucrose carrier transcripts were localised to the phloem cells of the vascular bundles. A more detailed analysis of sucrose carrier gene expression in the cotyledons of the germinating seedling was carried out by in situ hybridisation; the strongest signals were observed from the lower epidermal layer and the phloem, consistent with an active loading role for these cells. An ultrastructural study of the cells in the lower epidermis showed that they have wall ingrowths which are characteristic of transfer cells. The results are discussed in relation to the physiological role of the sucrose carrier in the Ricinus seedling and to the pathways of sucrose movement from endosperm to the sieve elements in the cotyledons
expression analysis, in situ hybridisation, phloem, Ricinus, sucrose carrier
0167-4412
425-435
Bick, J.A.
953907d5-dc2f-468f-80db-801edb7a75fc
Neelam, A
76df415b-27d8-42b2-b180-7c387139fb97
Smith, Esther
eaec4be8-0ef3-4429-8c45-0ded94511e29
Nelson, S.J.
b8566936-ff4c-4268-9128-cb3efac03e9a
Hall, J.L.
24cd62e9-4050-4514-9446-5a03949007f4
Williams, LE
79ee1856-3732-492b-8ac5-239749c85d9e
Bick, J.A.
953907d5-dc2f-468f-80db-801edb7a75fc
Neelam, A
76df415b-27d8-42b2-b180-7c387139fb97
Smith, Esther
eaec4be8-0ef3-4429-8c45-0ded94511e29
Nelson, S.J.
b8566936-ff4c-4268-9128-cb3efac03e9a
Hall, J.L.
24cd62e9-4050-4514-9446-5a03949007f4
Williams, LE
79ee1856-3732-492b-8ac5-239749c85d9e

Bick, J.A., Neelam, A, Smith, Esther, Nelson, S.J., Hall, J.L. and Williams, LE (1998) Expression analysis of a sucrose carrier in the germinating seedling of Ricinus communis. Plant Molecular Biology, 38 (3), 425-435. (doi:10.1023/A:1006040306581).

Record type: Article

Abstract

This study describes the expression of a sucrose carrier at various developmental stages in Ricinus communis. A partial-length cDNA clone, RcSUT1, was isolated by RT-PCR from Ricinus seedling RNA. This is almost identical to a sucrose carrier cDNA, Rscr1, which has previously been isolated by library screening. However, we have observed a very different expression pattern in the seedling to that previously reported. Northern analysis, with RcSUT1 as a probe, revealed high expression of a 2 kb transcript in the cotyledons of the germinating seedling; transcript levels were similar in cotyledons harvested 3–6 days after germination. A much lower level of this transcript was detected in the root, hypocotyl and endosperm RNA of the seedling and very low levels were also present in the sink and source leaves of the mature plant. This pattern of expression was also reflected at the protein level with an antipeptide antibody raised to part of the RcSUT1 deduced amino acid sequence.

Tissue print hybridisation analysis of the hypocotyl revealed that the sucrose carrier transcripts were localised to the phloem cells of the vascular bundles. A more detailed analysis of sucrose carrier gene expression in the cotyledons of the germinating seedling was carried out by in situ hybridisation; the strongest signals were observed from the lower epidermal layer and the phloem, consistent with an active loading role for these cells. An ultrastructural study of the cells in the lower epidermis showed that they have wall ingrowths which are characteristic of transfer cells. The results are discussed in relation to the physiological role of the sucrose carrier in the Ricinus seedling and to the pathways of sucrose movement from endosperm to the sieve elements in the cotyledons

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More information

Published date: 1 October 1998
Keywords: expression analysis, in situ hybridisation, phloem, Ricinus, sucrose carrier

Identifiers

Local EPrints ID: 483153
URI: http://eprints.soton.ac.uk/id/eprint/483153
ISSN: 0167-4412
PURE UUID: d26a9957-b20b-4371-8ede-9d7dfd7b3e07

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Date deposited: 25 Oct 2023 17:01
Last modified: 17 Mar 2024 05:17

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Contributors

Author: J.A. Bick
Author: A Neelam
Author: Esther Smith
Author: S.J. Nelson
Author: J.L. Hall
Author: LE Williams

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