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The Triticum aestivum non-specific lipid transfer protein (TaLtp) gene family: comparative promoter activity of six TaLtp genes in transgenic rice

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Abstract

Plant non-specific lipid transfer proteins (nsLTPs) are encoded by a multigene family and support physiological functions, which remain unclear. We adapted an efficient ligation-mediated polymerase chain reaction (LM-PCR) procedure that enabled isolation of 22 novel Triticum aestivum nsLtp (TaLtp) genes encoding types 1 and 2 nsLTPs. A phylogenetic tree clustered the wheat nsLTPs into ten subfamilies comprising 1–7 members. We also studied the activity of four type 1 and two type 2 TaLtp gene promoters in transgenic rice using the β-Glucuronidase reporter gene. The activities of the six promoters displayed both overlapping and distinct features in rice. In vegetative organs, these promoters were active in leaves and root vascular tissues while no β-Glucuronidase (GUS) activity was detected in stems. In flowers, the GUS activity driven by the TaLtp7.2a, TaLtp9.1a, TaLtp9.2d, and TaLtp9.3e gene promoters was associated with vascular tissues in glumes and in the extremities of anther filaments whereas only the TaLtp9.4a gene promoter was active in anther epidermal cells. In developing grains, GUS activity and GUS immunolocalization data evidenced complex patterns of activity of the TaLtp7.1a, TaLtp9.2d, and TaLtp9.4a gene promoters in embryo scutellum and in the grain epicarp cell layer. In contrast, GUS activity driven by TaLtp7.2a, TaLtp9.1a, and TaLtp9.3e promoters was restricted to the vascular bundle of the embryo scutellum. This diversity of TaLtp gene promoter activity supports the hypothesis that the encoded TaLTPs possess distinct functions in planta.

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Abbreviations

dpa:

Day post-anthesis

EST:

Expressed sequence tag

GUS:

β-Glucuronidase

LM-PCR:

Ligation mediated PCR

MATAB:

Mixed alkyl trimethyl ammonium bromide

nsLTP:

Non-specific lipid transfer protein

nsLtp :

Non-specific lipid transfer protein gene

SAR:

Systemic acquired resistance

TaLtp :

Triticum aestivum non-specific lipid transfer protein gene

uidA :

β-Glucuronidase gene

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Acknowledgments

The authors wish to thank Caroline Hartman for the wheat genomic library, Julie Petit for the CaMV35S::uidA construct, Jacques Escoute and Geneviève Conejero for helpful advice on histological analysis. We would also like to thank Emmanuelle Bourgeois for help in adapting the LM-PCR protocol to wheat. Freddy Boutrot was the recipient of a fellowship from the French Ministère de l’Education Nationale, de l’Enseignement Supérieur et de la Recherche. The support of the Génopole LR for containment greenhouse infrastructures is also acknowledged.

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Authors and Affiliations

  1. INRA, UMR 1096 PIA, 2 place Viala, 34060, Montpellier Cedex 01, France

    Freddy Boutrot, Philippe Joudrier & Marie-Françoise Gautier

  2. Cirad Amis, UMR 1096 PIA, TA40/03, Avenue Agropolis, 34398, Montpellier Cedex 05, France

    Donaldo Meynard & Emmanuel Guiderdoni

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  1. Freddy Boutrot
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  2. Donaldo Meynard
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  5. Marie-Françoise Gautier
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Correspondence to Marie-Françoise Gautier.

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Boutrot, F., Meynard, D., Guiderdoni, E. et al. The Triticum aestivum non-specific lipid transfer protein (TaLtp) gene family: comparative promoter activity of six TaLtp genes in transgenic rice. Planta 225, 843–862 (2007). https://doi.org/10.1007/s00425-006-0397-7

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