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Open Access Highly Access Research article

Phloem small RNAs, nutrient stress responses, and systemic mobility

Anja Buhtz1†, Janin Pieritz2†, Franziska Springer2 and Julia Kehr1*

Author Affiliations

1 Centro de Biotecnología y Genómica de Plantas (UPM-INIA), Campus de Montegancedo, M40 (km38), 28223 Pozuelo de Alarcón/Madrid, Spain

2 Max Planck Institute of Molecular Plant Physiology, Department Lothar Willmitzer, 14476 Potsdam, Germany

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BMC Plant Biology 2010, 10:64 doi:10.1186/1471-2229-10-64

Published: 13 April 2010

Additional files

Additional file 1:

Comparison of miRNA abundance in phloem sap vs. inflorescence stem. Comparison of sRNA microarray analysis of stem tissue (green) and phloem sap (blue) of Brassica napus. Only known miRNAs present on the commercial array, only one member per family are depicted. The upper graphs show the signal intensities on the array while the lower depict the log2 differences between phloem and inflorescence stem. Insets show RNA gel blot analyses of selected miRNAs from an independent experiment. Numbers indicate the number of sequences that were previously obtained by phloem sap sequencing [1], asterisks (*) indicate sequences from miRNA stars.

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Additional file 2:

Comparison of sRNA abundances in phloem, leaves and roots. sRNA microarray comparison of phloem (blue), leaf (green) and root (red) tissue of Brassica napus plants from biologically independent replications (n = 3). To allow inter-array comparison, signal intensities were normalized to the median signal of each sample. Only known miRNAs present on the commercial array and only one member per family are depicted.

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Additional file 3:

Transcript analysis of known nutrient stress-specific genes. Transcript analysis of known nutrient stress-specific genes in leaf and root tissue of hydroponically grown Brassica napus plants by semi-quantitative RT-PCR after 25, 30 and 35 cycles under -S, -Cu and -Fe compared to full nutrition (FN).

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Additional file 4:

Accumulation of three miR395 targets in WT and hen1-1 shoots grown under full nutrition. Levels of the targets SULTR2;1, APS1 and APS4 in shoots as detected by sqRT-PCR (35 cycles, UBC10, At5g53300 served as a control). FN: full nutrition.

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Additional file 5:

Sequences of the unknown phloem sap sRNAs represented on the microarrays. Phloem sap small RNA sequences of Brassica napus (Bn_PsRNAs) that were contained on the sRNA microarray (sequences were derived from high-throughput sequencing of B. napus phloem sap published in [1]).

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Additional file 6:

List of oligonucleotides used. Oligonucleotide sequences used for the detection of nutrient stress-specific miRNAs by RNA gel blots or by semi-quantitative RT-PCR, for the analysis of miR395 target genes, and for transcript detection of nutrient-responsive genes by semi-quantitative RT-PCR.

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