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A member of the maize isopentenyl transferase gene family, Zea mays isopentenyl transferase 2 (ZmIPT2), encodes a cytokinin biosynthetic enzyme expressed during kernel development

Cytokinin biosynthesis in maize

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Abstract

Cytokinins (CKs) are plant hormones that regulate a large number of processes associated with plant growth and development such as induction of stomata opening, delayed senescence, suppression of auxin-induced apical dominance, signaling of nitrogen availability, differentiation of plastids and control of sink strength. In maize, CKs are thought to play an important role in establishing seed size and increasing seed set under normal and unfavorable environmental conditions therefore influencing yield. In recent years, the discovery of isopentenyl transferase (IPT) genes in plants has shed light on the CK biosynthesis pathway in plants. In an effort to increase our understanding of the role played by CKs in maize development and sink-strength, we identified several putative IPT genes using a bioinformatics approach. We focused our attention on one gene in particular, ZmIPT2, because of its strong expression in developing kernels. The expression of the gene and its product overlays the change in CK levels in developing kernels suggesting a major role in CK biosynthesis for kernel development. We demonstrate that at 8–10 days after pollination (DAP) the endosperm and especially the basal transfer cell layer (BETL) is a major site of ZmIPT2 expression, and that this expression persists in the BETL and the developing embryo into later kernel development stages. We show that ectopic expression of ZmIPT2 in calli and in planta created phenotypes consistent with CK overproduction. We also show that ZmIPT2 preferentially uses ADP and ATP over AMP as the substrates for dimethylallyl diphosphate (DMAPP) IPT activity. The expression pattern of ZmIPT2 in the BETL, endosperm and embryo during kernel development will be discussed with an emphasis on the suggested role of CKs in determining sink-strength and grain production in crop plants.

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Abbreviations

BETL:

Basal endosperm transfer layer

cis-Z:

cis-zeatin

IPT:

Isopentenyl transferase

iPAR:

Isopentenyl adenosine

iPMP:

Isopentenyl adenosine mono-phosphate

iPTP:

Isopentenyl adenosine tri-phosphate

PPCBE:

Pedicel/placental chalazal/basal endosperm

t-Z:

trans-zeatin

ZmIPT2:

Zea mays isopentenyl transferase 2

ZR:

Zeatin riboside

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Acknowledgments

The authors are indebted to Tom Davis for his help with ZmIPT2 recombinant protein purification and to Nic Bate and Shoba Sivasankar for critical reading of the manuscript.

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Author notes

  1. Sabrina Humbert

    Present address: Department of Molecular and Cellular Biology, Science Complex, University of Guelph, Guelph, ON, Canada, N1G 2W1

Authors and Affiliations

  1. Discovery Group/Agronomic Traits, Pioneer Hi-Bred International, Inc., A DuPont Business, 7300 NW 62nd Avenue, P.O. Box 552, Johnston, IA, 50131-0522, USA

    Norbert Brugière, Sabrina Humbert, Jennifer Bohn & Jeffrey E. Habben

  2. Analytical and Genomics Technologies, DuPont Experimental Station, Route 141, Henry, Clay Bldg #353, Wilmington, DE, 19880-0353, USA

    Nancy Rizzo

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  1. Norbert Brugière
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  2. Sabrina Humbert
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  3. Nancy Rizzo
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  4. Jennifer Bohn
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  5. Jeffrey E. Habben
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Corresponding author

Correspondence to Norbert Brugière.

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Supplemental Table 1S

ZmIPT Genbank accession numbers and corresponding MAGI and AZM contigs (XLS 22 kb)

Supplemental Fig. 1S

Southern blot using B73 and Mo17 genomic DNA and ZmIPT2 32P-dCTP as a probe. Only one band could be detected after hybridization when HindIII (H), EcoRI (E) or EcoRV (V) restriction enzymes were used suggesting that the gene is present at a single copy per haploid genome (PPT 925 kb)

Supplemental Fig. 2S

Chromosomic location of ZmIPT2 determined by PCR using oat-maize addition lines. ZmIPT2 was only found in the DNA of the oat-maize addition line containing maize chromosome 2 (OMAL 2). Position was further determined as chromosome 2 bin 4 using BAC clones and a proprietary genetic physical map (PPT 63 kb)

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Brugière, N., Humbert, S., Rizzo, N. et al. A member of the maize isopentenyl transferase gene family, Zea mays isopentenyl transferase 2 (ZmIPT2), encodes a cytokinin biosynthetic enzyme expressed during kernel development. Plant Mol Biol 67, 215–229 (2008). https://doi.org/10.1007/s11103-008-9312-x

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  • DOI: https://doi.org/10.1007/s11103-008-9312-x

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