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Expression of 1-Aminocyclopropane-1-Carboxylate Oxidase during Leaf Ontogeny in White Clover1

Donald A. Hunter2, Sang Dong Yoo2, Stephen M. Butcher, and Michael T. McManus*

Institute of Molecular BioSciences, Massey University, Private Bag 11222, Palmerston North, New Zealand

We examined the expression of three distinct 1-aminocyclopropane-1-carboxylic acid oxidase genes during leaf ontogeny in white clover (Trifolium repens). Significant production of ethylene occurs at the apex, in newly initiated leaves, and in senescent leaf tissue. We used a combination of reverse transcriptase-polymerase chain reaction and 3'-rapid amplification of cDNA ends to identify three distinct DNA sequences designated TRACO1, TRACO2, and TRACO3, each with homology to 1-aminocyclopropane-1-carboxylic acid oxidase. Southern analysis confirmed that these sequences represent three distinct genes. Northern analysis revealed that TRACO1 is expressed specifically in the apex and TRACO2 is expressed in the apex and in developing and mature green leaves, with maximum expression in developing leaf tissue. The third gene, TRACO3, is expressed in senescent leaf tissue. Antibodies were raised to each gene product expressed in Escherichia coli, and western analysis showed that the TRACO1 antibody recognizes a protein of approximately 205 kD (as determined by gradient sodium dodecyl sulfate-polyacylamide gel electrophoresis) that is expressed preferentially in apical tissue. The TRACO2 antibody recognizes a protein of approximately 36.4 kD (as determined by gradient sodium dodecyl sulfate-polyacylamide gel electrophoresis) that is expressed in the apex and in developing and mature green leaves, with maximum expression in mature green tissue. No protein recognition by the TRACO3 antibody could be detected in senescent tissue or at any other stage of leaf development.

1   This work was funded by the New Zealand Foundation for Research, Science and Technology (contract no. C10 635), by the New Zealand Agricultural and Pastoral Research Institute (AgResearch) with provision of a Ph.D. study award to D.H., and by the New Zealand Crop and Food Research Institute with provision of a Ph.D. study award to S.M.B.
2   These authors contributed equally to this work.
*   Corresponding author; e-mail m.t.mcmanus{at}massey.ac.nz; fax 64-6-350-5694.

Plant Physiol. (1999) 120: 131-142
Copyright Clearance Center:   0032-0889/99/120//12
© 1999 American Society of Plant Physiologists




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M. T. McManus
Further Examination of Abscission Zone Cells as Ethylene Target Cells in Higher Plants
Ann. Bot., January 1, 2008; 101(2): 285 - 292.
[Abstract] [Full Text] [PDF]


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