Amino acid transporter expression and localisation studies in pea (Pisum sativum)
Mechthild Tegeder A D , Qiumin Tan A , Aleel K. Grennan A B and John W. Patrick CA School of Biological Sciences, Centre for Integrated Biotechnology, Centre for Reproductive Biology, Washington State University, Pullman, WA 99164, USA.
B Present address: Department of Plant Biology, University of Illinois, Urbana, IL 61801, USA.
C School of Environmental and Life Sciences, The University of Newcastle, NSW 2308, Australia.
D Corresponding author. Email: tegeder@wsu.edu
Functional Plant Biology 34(11) 1019-1028 https://doi.org/10.1071/FP07107
Submitted: 26 April 2007 Accepted: 7 September 2007 Published: 1 November 2007
Abstract
Expression of the amino acid permeases PsAAP1 and PsAAP2 was analysed in developing pea (Pisum sativum L.) plants. Both transporters were expressed in seed coats and cotyledon epidermal transfer cells and storage parenchyma cells. AAP expression is developmentally regulated and coincides with the onset of storage protein synthesis. Nitrogen was shown to induce AAP expression and AAP transcript levels were upregulated during the photoperiod. Analysis of Arabidopsis thaliana AAP1 promoter activity in pea, using promoter-β-glucuronidase (promotor-GUS) studies, revealed targeting of GUS to seed coats and cotyledon epidermal transfer cells. Expression was found in the nutritious endosperm during the early stages of seed development, whereas GUS staining in embryos was detected from the heart stage onward. In addition, AAP1 expression was observed in the phloem throughout the plant. This finding equally applied to PsAAP1 expression as shown by in situ mRNA hybridisation, which also demonstrated that PsAAP1 expression was localised to companion cells. Overall, PsAAP1 expression patterns and cellular localisation point to a function of the transporter in phloem loading of amino acids for translocation to sinks and in seed loading for development and storage protein accumulation.
Additional keywords: grain legume, pea, amino acid transport, seed loading, transfer cells, phloem loading.
Acknowledgements
We are grateful for the support of Dr Thomas J. V. Higgins (CSIRO, Canberra, Australia) in producing the transgenic pea lines and Chuck Cody (Washington State University) with growing the plants. The valuable help of Dr Hélène Pélissier and Erin Beneski (Washington State University) with some tissue preparation is much appreciated. We thank the National Research Initiative Competitive Grants Program–USA Department of Agriculture (grant no. 2001-35318-10990 to M. Tegeder), the National Science Foundation (grants IBN 0135344 and IOB 0448506 to M. Tegeder) and the Australian Research Council (A19530955 to J. W. Patrick) for financial support.
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