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Protocols in ecological and environmental plant physiology

 

Article << Previous     |     Next >>   Contents Vol 39(12)

Two Arabidopsis thaliana dihydrodipicolinate synthases, DHDPS1 and DHDPS2, are unequally redundant

Susan Jones-Held A, Luciana Pimenta Ambrozevicius A C, Michael Campbell B, Bradley Drumheller B, Emily Harrington B and Thomas Leustek A D

A Department of Plant Biology and Pathology, Rutgers University, New Brunswick, NJ 08901-8520, USA.
B School of Science, Penn State Erie, The Behrend College, P-1 Prischak Building, 4205 College Drive, Erie, PA 16563-0203, USA.
C Present address: Brazilian Ministry of Agriculture, Livestock and Food Supply, Viçosa 57700, Brazil.
D Corresponding author. Email: leustek@aesop.rutgers.edu

Functional Plant Biology 39(12) 1058-1067 http://dx.doi.org/10.1071/FP12169
Submitted: 7 June 2012  Accepted: 14 August 2012   Published: 1 October 2012


 
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Abstract

In Arabidopsis thalinana (L.) Heynh., DHDPS1 and DHDPS2 encode orthologous dihydrodipicolinate synthases (DHDPS), the first enzyme of the lysine (Lys) biosynthesis pathway. A TDNA insertion mutant of dhdps2 was previously reported to be viable and to accumulate free threonine (Thr). Analysis of additional TDNA insertion lines showed that dhdps1 and dhdps2 mutants are both viable and that whereas dhdps2 mutants accumulate Thr, dhdps1 plants do not. Thr-accumulation was complemented by heterologous expression of Escherichia coli DapA, indicating that the phenotype is due to reduced DHDPS activity in dhdps2. DHDPS1 contributes ~30% towards the total DHDPS activity in leaves of young plants and DHDPS2 contributes 70%; therefore, the threshold of activity resulting in Thr accumulation lies within this narrow range. dhdps1dhdps2 double mutants could not be isolated, even after exogenous feeding with Lys. Segregation analysis indicated that gametes lacking functional DHDPS genes are defective, as are embryos. Plants carrying only a single DHDPS2 gene do not accumulate Thr, but they show a gametophytic defect that is partially rescued by Lys application. Despite the accumulation of Thr, dhdps2 seedlings are no more sensitive than wild-type plants to growth inhibition by Lys or the Lys precursor diaminopimelate. They also are not rescued by methionine at growth-inhibitory Lys concentrations. Exogenous application of Lys and methionine to dhdps2 mutants did not reduce the accumulation of Thr.

Additional keywords: DHDPS, diaminopimelate, lysine.


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