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Genetic analysis of two OsLpa1-like genes in Arabidopsis reveals that only one is required for wild-type seed phytic acid levels

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Abstract

Phytic acid (inositol-1,2,3,4,5,6-hexakisphosphate or InsP6) is the primary storage form of phosphorus in plant seeds. The rice OsLpa1 encodes a novel protein required for wild-type levels of seed InsP6 and was identified from a low phytic acid (lpa) mutant exhibiting a 45–50% reduction in seed InsP6. OsLpa1 is highly conserved in plants and Arabidopsis contains two OsLpa1-like genes, At3g45090 and At5g60760. Analysis of homozygous T-DNA insertion mutants of At5g60760 revealed significantly reduced levels of seed InsP6 while no changes were observed in seeds of At3g45090 mutants. A double knockout mutant of At5g60760 and At3g45090 was created and its seed InsP6 content was similar to that of the At5g60760 mutant indicating that At3g45090 does not provide functional redundancy. OsLpa1 was confirmed to be the ortholog of At5g60760 by complementation of a knockout mutant with a cDNA clone corresponding to the largest of three alternative transcripts of OsLpa1. The spatial and temporal expression of At5g60760 during seed development is consistent with its involvement in seed InsP6 biosynthesis.

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Abbreviations

At :

Arabidopsis thaliana

BLASTp:

Basic local alignment search tool protein–protein

cDNA:

Complementary deoxyribonucleic acid

DAF:

Days after fertilization

GADPH:

Glyceraldehyde 3-phosphate dehydrogenase

GUS:

β-Glucuronidase

HPIC:

High-performance ion chromatography

InsP:

Inositol (poly)phosphate

lpa :

Low phytic acid

Os :

Oryza sativa

P:

Phosphorus

Pi:

Inorganic phosphate

RT-PCR:

Reverse transcription-polymerase chain reaction

SAIL:

Syngenta Arabidopsis Insertion Lines

SALK:

Salk Institute

T-DNA:

Transfer DNA

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Acknowledgments

This study was supported by USDA Agricultural Research Service CRIS Project 5306-21000-016/017-00D (T.H.T.) and National Research Initiative Competitive Grant 2005-35301-15708 from the USDA Cooperative State Research, Education, and Extension Service (T.H.T.). We are thankful to the UC Davis DANR Analytical Lab for assistance with HPIC analysis and to Victor Raboy, Cynthia Andaya, and Erin Easlon for critical reading of the manuscript and helpful suggestions for improvement.

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  1. Crops Pathology and Genetics Research Unit, US Department of Agriculture, Agricultural Research Service, Department of Plant Sciences, Mail Stop 1, University of California, Davis, CA, 95616, USA

    Sang-Ic Kim & Thomas H. Tai

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  1. Sang-Ic Kim
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  2. Thomas H. Tai
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Correspondence to Thomas H. Tai.

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Kim, SI., Tai, T.H. Genetic analysis of two OsLpa1-like genes in Arabidopsis reveals that only one is required for wild-type seed phytic acid levels. Planta 232, 1241–1250 (2010). https://doi.org/10.1007/s00425-010-1243-5

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