Limits...
A roadmap for zinc trafficking in the developing barley grain based on laser capture microdissection and gene expression profiling.

Tauris B, Borg S, Gregersen PL, Holm PB - J. Exp. Bot. (2009)

Bottom Line: The amplified RNA was subsequently hybridized to Affymetrix 22K Barley GeneChips.Due to the short average length of the amplified transcripts and the positioning of numerous probe sets at locations more than 400 base pairs (bp) from the poly(A)-tail, a normalization approach was used where the probe positions were taken into account.On the basis of the expression levels of a number of metal homeostasis genes, a working model is proposed for the translocation of zinc from the phloem to the storage sites in the developing grain.

View Article: PubMed Central - PubMed

Affiliation: University of Aarhus, Facultry of Agricultural Sciences, Department of Genetics and Biotechnology, Forsøgsvej 1, 4200 Slagelse, Denmark.

ABSTRACT
Nutrients destined for the developing cereal grain encounter several restricting barriers on their path towards their final storage sites in the grain. In order to identify transporters and chelating agents that may be involved in transport and deposition of zinc in the barley grain, expression profiles have been generated of four different tissue types: the transfer cells, the aleurone layer, the endosperm, and the embryo. Cells from these tissues were isolated with the 'laser capture microdissection' technology and the extracted RNA was subjected to three rounds of T7-based amplification. The amplified RNA was subsequently hybridized to Affymetrix 22K Barley GeneChips. Due to the short average length of the amplified transcripts and the positioning of numerous probe sets at locations more than 400 base pairs (bp) from the poly(A)-tail, a normalization approach was used where the probe positions were taken into account. On the basis of the expression levels of a number of metal homeostasis genes, a working model is proposed for the translocation of zinc from the phloem to the storage sites in the developing grain.

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Related in: MedlinePlus

Laser capture of transfer cells (A) and aleurone cells (B). The cells of interest are marked with a line (A, B); the laser cuts along the line and catapults the isolated tissue into the lid of a tube (A′, B′); the captured tissue can be visualized in the cap (A′′, B′′). (C) Schematic drawing showing a cross-section of a cereal grain and the localization of the isolated tissues. (This figure is available in colour at JXB online.)
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fig1: Laser capture of transfer cells (A) and aleurone cells (B). The cells of interest are marked with a line (A, B); the laser cuts along the line and catapults the isolated tissue into the lid of a tube (A′, B′); the captured tissue can be visualized in the cap (A′′, B′′). (C) Schematic drawing showing a cross-section of a cereal grain and the localization of the isolated tissues. (This figure is available in colour at JXB online.)

Mentions: The purpose of the present study was to decide whether it is possible to establish a road map for zinc transport and deposition from the phloem into the barley grain based on laser capture microdissection and expression profiling. Barley (Hordeum vulgare) is a diploid cereal crop that, besides its importance as animal feed, human food, and as raw material for beer production, also serves as a model for the small-grained cereals. A substantial amount of genetic information is available for this crop, including more than 470 000 public ESTs. Seeds from the barley cultivar Golden Promise were harvested 20 dap. Cells from the unloading region (T_1 and T_2), the aleurone layer (A_1 and A_2), the endosperm (E_1 and E_2), and the embryo (Em_1 and Em_2) were isolated with the LCM technology (Fig. 1) and amplified RNA from each sample was hybridized to Affymetrix 22K Barley GeneChips.


A roadmap for zinc trafficking in the developing barley grain based on laser capture microdissection and gene expression profiling.

Tauris B, Borg S, Gregersen PL, Holm PB - J. Exp. Bot. (2009)

Laser capture of transfer cells (A) and aleurone cells (B). The cells of interest are marked with a line (A, B); the laser cuts along the line and catapults the isolated tissue into the lid of a tube (A′, B′); the captured tissue can be visualized in the cap (A′′, B′′). (C) Schematic drawing showing a cross-section of a cereal grain and the localization of the isolated tissues. (This figure is available in colour at JXB online.)
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2657541&req=5

fig1: Laser capture of transfer cells (A) and aleurone cells (B). The cells of interest are marked with a line (A, B); the laser cuts along the line and catapults the isolated tissue into the lid of a tube (A′, B′); the captured tissue can be visualized in the cap (A′′, B′′). (C) Schematic drawing showing a cross-section of a cereal grain and the localization of the isolated tissues. (This figure is available in colour at JXB online.)
Mentions: The purpose of the present study was to decide whether it is possible to establish a road map for zinc transport and deposition from the phloem into the barley grain based on laser capture microdissection and expression profiling. Barley (Hordeum vulgare) is a diploid cereal crop that, besides its importance as animal feed, human food, and as raw material for beer production, also serves as a model for the small-grained cereals. A substantial amount of genetic information is available for this crop, including more than 470 000 public ESTs. Seeds from the barley cultivar Golden Promise were harvested 20 dap. Cells from the unloading region (T_1 and T_2), the aleurone layer (A_1 and A_2), the endosperm (E_1 and E_2), and the embryo (Em_1 and Em_2) were isolated with the LCM technology (Fig. 1) and amplified RNA from each sample was hybridized to Affymetrix 22K Barley GeneChips.

Bottom Line: The amplified RNA was subsequently hybridized to Affymetrix 22K Barley GeneChips.Due to the short average length of the amplified transcripts and the positioning of numerous probe sets at locations more than 400 base pairs (bp) from the poly(A)-tail, a normalization approach was used where the probe positions were taken into account.On the basis of the expression levels of a number of metal homeostasis genes, a working model is proposed for the translocation of zinc from the phloem to the storage sites in the developing grain.

View Article: PubMed Central - PubMed

Affiliation: University of Aarhus, Facultry of Agricultural Sciences, Department of Genetics and Biotechnology, Forsøgsvej 1, 4200 Slagelse, Denmark.

ABSTRACT
Nutrients destined for the developing cereal grain encounter several restricting barriers on their path towards their final storage sites in the grain. In order to identify transporters and chelating agents that may be involved in transport and deposition of zinc in the barley grain, expression profiles have been generated of four different tissue types: the transfer cells, the aleurone layer, the endosperm, and the embryo. Cells from these tissues were isolated with the 'laser capture microdissection' technology and the extracted RNA was subjected to three rounds of T7-based amplification. The amplified RNA was subsequently hybridized to Affymetrix 22K Barley GeneChips. Due to the short average length of the amplified transcripts and the positioning of numerous probe sets at locations more than 400 base pairs (bp) from the poly(A)-tail, a normalization approach was used where the probe positions were taken into account. On the basis of the expression levels of a number of metal homeostasis genes, a working model is proposed for the translocation of zinc from the phloem to the storage sites in the developing grain.

Show MeSH
Related in: MedlinePlus