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RESEARCH ARTICLE
Contents Vol 32(10)

Senescence-associated down-regulation of 1-aminocyclopropane-1-carboxylate (ACC) oxidase delays harvest-induced senescence in broccoli

Nigel E. Gapper A B D H , Simon A. Coupe A E , Marian J. McKenzie A , Richard W. Scott B F , Mary C. Christey C , Ross E. Lill A , Michael T. McManus B and Paula E. Jameson B G
+ Author Affiliations
- Author Affiliations

A New Zealand Institute for Crop & Food Research Limited, Food Industry Science Centre, Private Bag 11600, Palmerston North, New Zealand.

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

C New Zealand Institute for Crop & Food Research Limited, Private Bag 4704, Christchurch, New Zealand.

D Current address: Produce Quality and Safety Laboratory, Beltsville Agricultural Research Centre, USDA / ARS, 10300 Baltimore Avenue, Beltsville, MD 20705, USA.

E Current address: Marks and Spencer Pl., Waterside House, 35 North Wharf Rd, London W2 1NW, UK.

F Current address: AgResearch Ltd, Grasslands Research Centre, Private Bag 11008, Palmerston North, New Zealand.

G Current address: School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch, New Zealand.

H Corresponding author. Email: gappern@ba.ars.usda.gov

Functional Plant Biology 32(10) 891-901 https://doi.org/10.1071/FP05076
Submitted: 30 March 2005  Accepted: 8 June 2005   Published: 5 October 2005

Abstract

To gain an in-depth understanding of the role of ethylene in post harvest senescence, we used broccoli (Brassica oleracea var. italica) as our model species. The senescence-associated asparagine synthetase (AS) promoter from asparagus was used to drive the expression of an antisense 1-aminocyclopropane-1-carboxylate oxidase (ACO) cDNA from broccoli, BoACO2, to reduce ethylene production following harvest. Physiological analyses revealed that transgenic broccoli lines harbouring the antisense BoACO2 gene construct (designated as AS-asACO) displayed delayed senescence in both detached leaves and detached heads as measured by hue angle. Harvested floret tissue from these plants also showed a delayed loss of chlorophyll, lower protease activity and higher total protein content, and changes in transcript levels of senescence marker genes when compared with wild type and transgenic lines transformed with an empty T-DNA. Genes that were down-regulated included those coding for cysteine protease (BoCP5), metallothionein-like protein (BoMT1), hexokinase (BoHK1), invertase (BoINV1) and sucrose transporters (BoSUC1 and BoSUC2). Northern analysis for BoACO1 and BoACO2, ACO assays and western analysis, revealed reduced ACO transcript, enzyme activity and protein accumulation, as well as reduced ethylene production in the transgenic AS-asACO lines when compared with controls, confirming that a key enzyme regulating ethylene biosynthesis was reduced in these plants. This, together with the changes observed in gene expression, confirm a significant role for ethylene in regulating the events leading to senescence in broccoli following harvest.

Keywords: ACC oxidase, antisense, asparagine synthetase promoter, Brassica oleracea, broccoli, ethylene, postharvest, senescence.


Acknowledgments

We acknowledge Duncan Hedderley for statistical analyses, Anthony Corbett for photography, Ian King for technical assistance and the New Zealand Foundation for Research, Science and Technology (FRST) for funding this work.


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