Molecular Genetics and Genomics

, Volume 286, Issue 2, pp 119–133 | Cite as

Identification of genes necessary for wild-type levels of seed phytic acid in Arabidopsis thaliana using a reverse genetics approach

Original Paper


The majority of phosphorus (P) in seeds is found in phytic acid (InsP6) which accumulates as the mixed salt phytate. InsP6 is generally considered to be an anti-nutrient and the development of low phytic acid (lpa) seed crops is of significant interest. We have employed a reverse genetics approach to examine the impact of disrupting genes involved in inositol phosphate metabolism on Arabidopsis seed InsP6 levels. Our analysis revealed that knockout mutations in three genes (AtITPK1, AtITPK4, and AtMIK/At5g58730) reduced seed InsP6 in addition to knockouts of four previously reported genes (AtIPK1, AtIPK2β, AtMRP5, and At5g60760). Seeds of these lpa mutants also exhibited reduced germination under various stress conditions. The greatest reduction in InsP6 (>70%) was observed in atmrp5 seeds which were also among the least sensitive to the stresses examined. Expression analysis of the lpa genes revealed three distinct patterns in developing siliques consistent with their presumed roles. Disruption of each lpa gene resulted in changes in the expression in some of the other lpa genes indicating that transcription of lpa genes is modulated by other constituents of InsP6 metabolism. While all the lpa genes represent possible targets for genetic engineering of low phytate seed crops, mutations in AtMRP5, AtMIK, and At5g60760 may be most successful for conventional approaches such as mutation breeding.


Phytic acid Arabidopsis Inositol phosphate metabolism Kinases Reverse genetics 



Basic local alignment search tool protein–protein


Days after fertilization


Glyceraldehyde 3-phosphate dehydrogenase


High-performance ion chromatography




Inositol (poly)phosphate




Low phytic acid




Inorganic phosphate


Reverse transcription-polymerase chain reaction


Syngenta Arabidopsis insertion lines


Salk Institute


Transfer DNA



This 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 UC DAVIS DANR analytic lab for assisting with HPIC analysis and to V. Raboy and C. Andaya for critical reading of the manuscript and helpful suggestions for improvement. Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. USDA is an equal opportunity provider and employer.

Supplementary material

438_2011_631_MOESM1_ESM.pdf (191 kb)
Supplementary material 1 (PDF 190 kb)


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Copyright information

© Springer-Verlag (outside the USA) 2011

Authors and Affiliations

  1. 1.Crops Pathology and Genetics Research Unit, U.S. Department of Agriculture, Agricultural Research Service, Department of Plant Sciences – MS 1University of CaliforniaDavisUSA

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