Jocelyn R. Eason

Programmed cell death during flower senescence: isolation and characterization of cysteine proteinases from Sandersonia aurantiaca

Cysteine protease inhibitors delayed the senescence of Sandersonia aurantiaca Hook. flowers. Tepal fading and wilting occurred later in the 2,2´ -dipyridyl-treated flowers, and these flowers had a greater soluble protein content and less active endoproteases compared with control flowers that were held in water. Biochemical analysis revealed the presence of several protease-active bands in the soluble protein fraction of Sandersonia tepals. Activity of the polypeptides increased as flower senescence progressed. Western analysis with an antibody raised against the castor bean cysteine proteinase identified homologous proteins in Sandersonia flowers (ca 46, 41 and 31kDa). Three cDNAs encoding cysteine proteinases were isolated from Sandersonia tepals (PRT5, PRT15 and PRT22). Expression of all three increased in tepals as senescence progressed. mRNAs for PRT5 were detected only in senescing flower tissue, whereas PRT15 and PRT22 were expressed in leaf, stem and root tissue. PRT5 has significant homology to C-terminus KDEL proteins, which have a role in the degradation of plant cell contents during programmed cell death. PRT15 is most similar to cysteine proteinases with a long C-terminal extension, whereas PRT22 is homologous to stress-induced cysteine proteinases.

Amino acid metabolism in senescing Sandersonia aurantiaca flowers: cloning and characterization of asparagine synthetase and glutamine synthetase cDNAs

We have studied nitrogen metabolism during senescence of the ethylene-insensitive flower Sandersonia aurantiaca Hook. We report here on the isolation and characterization of two cDNAs from senescing sandersonia tepals. SAND1 encodes asparagine synthetase (AS; EC 6.3.5.4) and SAND3 encodes glutamine synthetase (GS; EC 6.3.1.2). The accumulation of SAND3 mRNAs was consistently high throughout flower development and senescence and was not associated with senescence per se. SAND1 mRNAs started to accumulate in mature flowers just prior to visual signs of senescence. Sucrose feeding of individual flowers delayed tepal senescence and altered amino acid metabolism. The levels of Asn, Gln and Glu were higher and Asp levels were lower in the sucrose-fed flowers. The senescence-associated peak in Gln was delayed in sucrose-fed flowers compared to controls that were held in water. Sucrose feeding had no effect on the initiation of SAND1 transcription at the onset of flower senescence; however, the later decline in SAND1 mRNA abundance was delayed in sucrose-fed flowers. Elimination of GS activity and the subsequent reduction in Gln levels caused by phosphinothricin treat-ment was associated with a delay of SAND1 mRNA accumulation in senescing flowers.

Galactosidases in opening, senescing and water-stressed Sandersonia aurantiaca flowers

Three glycosyl hydrolase family 35 β-galactosidase-encoding cDNAs, SaGAL1 (full-length), SaGAL2 and SaGA3L (both partial), have been isolated from Sandersonia aurantiaca (Hook.) SaGAL1 protein was functionally expressed in E. coli and β-galactosidase identity confirmed by activity assay. All three clones are primarily expressed in tepal tissues of senescing sandersonia flowers. In order to identify relationships between tepal texture and galactose metabolism, cut sandersonia flowers were treated with sucrose, periods of dryness or PEG and parameters associated with galactose metabolism and firmness were monitored. Sucrose supplementation, known to increase tepal firmness, delayed expression of SaGAL1 and SaGAL3 in opening (stage 5) flowers, whereas the response to periods of dryness followed by rehydration depended on the maturity of the flower. These treatments also tended to hasten the onset of processes associated with programmed cell death, monitored by PRT5 (a senescence-associated protease) expression. Galactosidase activity and cell wall galactose content were also affected but in an inconsistent manner. PEG supplied to opening flowers for 1 d followed by water, induced a long period of wilt, and intensive PRT5 expression. However, β-galactosidase gene expression and activity was delayed in these flowers, and cell-wall galactose content changed apparently independently of galactosidase activity. We have not been able to demonstrate a causal connection between the change in petal texture and concurrent induction of galactose mobilisation in sandersonia during normal development and senescence.

Australian consumer awareness of health benefits associated with vegetable consumption

AIM The present study investigated the perceived health benefits of specific vegetable consumption to guide the use of nutrition and health claims on vegetable marketing collateral. METHODS Free elicitation and consumer ranking data were collected through an online survey of 1000 adults from across Australia and analysed for the perceived importance of vegetables in the daily diet, number of serves consumed per day, knowledge about health-related benefits of specific vegetables and perceived health benefits of vegetable consumption. RESULTS The importance of vegetables in the diet and daily vegetable consumption was higher in people from an English-speaking background, females, people aged 45 years and over and people living in non-metropolitan areas. Digestion was selected as the major health benefit from consumption of specific vegetables. However, understanding of the health benefits of specific vegetable consumption was relatively low among consumers. Half of the respondents were not sure of the health benefits associated with specific vegetables, except for carrots and spinach. Some respondents volunteered nutrient content or other information. CONCLUSIONS There was no clear indication that consumers understand the specific health benefits conferred by consumption of vegetables. Nutrient and health benefit labelling therefore has the capacity to enhance knowledge of vegetable consumers. It is recommended that health benefit labelling be tailored to promote greater consumption of vegetables in those demographic groups where vegetable consumption was lower. The present study assists the Australian vegetable industry in helping consumers make more informed consumption choices.