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Real-time RT-PCR profiling of transcription factors including 34 MYBs and signaling components in white lupin reveals their P status dependent and organ-specific expression

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Abstract

Phosphorus (P) is often a limiting macronutrient because of its low availability in soils. White lupin (Lupinus albus L.) plants are well adapted to growth under low-P conditions. White lupin acclimation to low-P conditions includes changes in root architecture and enhanced expression of numerous genes encoding for secreted acid phosphatases and phosphate transporters. However, information about transcription factors and signaling proteins that coordinate the P-starvation responses is limited in white lupin. In this study, cDNAs and ESTs encoding for transcription factors and signaling proteins were isolated and their transcription profiles were clarified to facilitate the identification of key signal transduction genes necessary to improve P acquisition, allocation, and use. 34 cDNA fragments of MYB-coiled coil (MYB-CC) and R2R3-MYB, and 26 ESTs encoding for transcription factors and signaling proteins were isolated. Four MYB-CC cDNAs showed high similarity to the transcription factor Phosphate starvation response 1 in Arabidopsis, which has been implicated in regulation of many P-starvation response genes. In addition, deduced amino acid sequences of 29 R2R3-MYB cDNAs showed similarities to Arabidopsis R2R3-MYB proteins. Transcription of the 60 genes, as measured by real-time reverse transcription-PCR, in normal roots, cluster roots, leaves, and shoot tips under P sufficient and low-P conditions revealed that six (10%) and two (3.3%) sequences were either induced or suppressed, respectively, by low-P condition. In addition, 36 genes (60%) showed an organ specific expression.

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Abbreviations

3′RACE:

Rapid amplification of cDNA 3′-end

CC:

coiled coil

DAE:

days after emergence

EST:

expressed sequence tag

LaMATE:

Lupinus albus multidrug and toxin efflux protein

LaPT1:

Lupinus albus phosphate transporter 1

LaSAP:

Lupinus albus secreted acid phosphatase

P:

Phosphorus

PHR:

phosphate starvation response

RT-PCR:

reverse transcription-PCR

TF:

transcription factor

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Financial source

This research was supported by a Grants-In-Aid for Scientific Research (No. 17658032), the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Author notes

  1. Keqin Zhou

    Present address: Key Laboratory of Mollisols Agroecology, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, Harbin, 150081, China

Authors and Affiliations

  1. Research Faculty of Agriculture, Hokkaido University, N9W9, Kita-ku, Sapporo, 060-8589, Japan

    Masumi Yamagishi & Mitsuru Osaki

  2. Graduate School of Agriculture, Hokkaido University, N9W9, Kita-ku, Sapporo, 060-8589, Japan

    Keqin Zhou

  3. USDA-ARS, Plant Science Research Unit, 1991 Upper Buford Circle, St. Paul, Minnesota, 55108, USA

    Susan S. Miller & Carroll P. Vance

  4. Department of Agronomy and Plant Genetics, University of Minnesota, 1991 Upper Buford Circle, St. Paul, Minnesota, 55108, USA

    Carroll P. Vance

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  1. Masumi Yamagishi
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  2. Keqin Zhou
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  3. Mitsuru Osaki
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Corresponding author

Correspondence to Masumi Yamagishi.

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Responsible Editor: Hans Lambers.

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Supplementary Fig. 1

The primary lateral root of white lupin at 6 DAE. A bleached root was stained by methylene blue The primary lateral root including rootlet primordia (very young proteoid root) was used for cDNA library construction. (GIF 149 kb)

Supplementary Fig. 2

(XLS 71 kb)

Supplementary Fig. 3-1

(GIF 258 kb)

Supplementary Fig. 3-2

(GIF 255 kb)

Supplementary Fig. 3-3

(GIF 279 kb)

Supplementary Fig. 3-4

Quantification of transcript abundance of genes encoding for TFs and signaling proteins in normal roots (NR), cluster roots (CR), leaves (Leaf), and shoot tips including shoot apical meristems (ST) of white lupin plants grown under P sufficient (white bars) and low-P (dark gray bars) growth conditions. Vertical bars show standard errors of the biological replicates. Within each figure, different letters above the bars represent significant differences at the 5% level. LaUbiquitin was used to normalize the expression. (GIF 230 kb)

Supplementary Table 1

List of cDNAs and ESTs analyzed in this study and primer sequences (XLS 35 kb)

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Yamagishi, M., Zhou, K., Osaki, M. et al. Real-time RT-PCR profiling of transcription factors including 34 MYBs and signaling components in white lupin reveals their P status dependent and organ-specific expression. Plant Soil 342, 481–493 (2011). https://doi.org/10.1007/s11104-010-0711-9

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