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JXB Advance Access published online on May 27, 2009
Journal of Experimental Botany, doi:10.1093/jxb/erp169
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© 2009 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details)

RESEARCH PAPER

A cytosolic invertase is required for normal growth and cell development in the model legume, Lotus japonicus

Tracey Welham1, Jodie Pike2, Irmtraud Horst1 *, Emmanouil Flemetakis3, Panagiotis Katinakis3, Takakazu Kaneko4, Shusei Sato4, Satoshi Tabata4, Jillian Perry1, Martin Parniske5 and Trevor L. Wang1,{dagger}

1Metabolic Biology, John Innes Centre, Norwich Research Park, Colney, Norwich NR4 7UH, UK
2The Sainbury Laboratory, Norwich Research Park, Colney, Norwich NR4 7UH, UK
3Agricultural Biotechnology, Agricultural University of Athens, Iera Odos 75, 118 55 Athens, Greece
4Kazusa DNA Research Institute, 2-6-7 Kazusa-kamatari, Kisarazu, Chiba 292-0818, Japan
5Department Biology I, Genetics, University of Munich (LMU), Grosshaderner Str. 2–4, D-82152 Planegg, Germany

{dagger} To whom correspondence should be addressed: E-mail:trevor.wang{at}bbsrc.ac.uk

Neutral/alkaline invertases are a subgroup, confined to plants and cyanobacteria, of a diverse family of enzymes. A family of seven closely-related genes, LjINV1–LjINV7, is described here and their expression in the model legume, Lotus japonicus, is examined. LjINV1 previously identified as encoding a nodule-enhanced isoform is the predominant isoform present in all parts of the plant. Mutants for two isoforms, LjINV1 and LjINV2, were isolated using TILLING. A premature stop codon allele of LjINV2 had no effect on enzyme activity nor did it show a visible phenotype. For LjINV1, premature stop codon and missense mutations were obtained and the phenotype of the mutants examined. Recovery of homozygous mutants was problematic, but their phenotype showed a severe reduction in growth of the root and the shoot, a change in cellular development, and impaired flowering. The cellular organization of both roots and leaves was altered; leaves were smaller and thicker with extra layers of cells and roots showed an extended and broader zone of cell division. Moreover, anthers contained no pollen. Both heterozygotes and homozygous mutants showed decreased amounts of enzyme activity in nodules and shoot tips. Shoot tips also contained up to a 9-fold increased level of sucrose. However, mutants were capable of forming functional root nodules. LjINV1 is therefore crucial to whole plant development, but is clearly not essential for nodule formation or function.

Key words: Cellular development, legume, Lotus japonicus, neutral/alkaline invertase, mutants, plant development, sucrose metabolism, TILLING

* Present address: The Bingham Laboratory, NIAB, Huntingdon Road, Cambridge CB3 0LE, UK.

Received 22 December 2008; Revised 29 April 2009 Accepted 30 April 2009


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