1Pheophytinase (PPH) activity and gene expression of chlorophyll (Chl)-degrading 2 enzymes relating to UV-B treatment in postharvest broccoli (Brassica oleracea L. Italica 3 Group) florets were determined. PPH is involved in the dephytylation of Mg-free Chl a, 4 pheophytin (Phy) a. However, chlorophyllase (Chlase, EC.3.1.1.14) also uses Phy a as a 5 substrate to produce pheophorbide (Pheide) a by dephytylation. For an accurate 6 determination of PPH activity, the PPH protein fraction was separated from Chlase protein by 7 ammonium sulfate precipitation. The protein precipitated by 45-60% saturated ammonium 8 sulfate was included a little bit Chlase activity and was suitable for PPH determination.

Eight Japanese bunching onion (Allium fistulosum L.)-shallot (Allium cepa L. Aggregatum group) monosomic addition lines (MALs, FF + 1A-FF + 8A) were used to reveal the effects of single alien chromosomes of A. cepa on the production of amino acids and S-alk(en)yl-L-cysteine sulfoxides (ACSOs) in leaf tissues of A. fistulosum. Amino acid and ACSO contents in MAL leaf blades were determined once every 3 months from August 2005 to May 2006. The amino acid found in the greatest amount in all of the MALs throughout the year was cysteine, except for FF + 7A accumulated glycine as the greatest amount of amino acid (Nov. 2005). All of the MALs contained three ACSOs in varying amounts and proportions, and all accumulated S-1-propenyl CSO as a major ACSO but hardly produced S-2-propenyl CSO throughout the year. FF + 3A showed greatly increased proportions of Smethyl CSO in total ACSO, suggesting that anonymous genes controlling S-methyl CSO production are located on chromosome 3A of shallot. High accumulation of total ACSOs in the monosomic additions FF + 3A, FF + 4A, FF + 5A, and FF + 8A was observed during different growth periods. Using PCR-based marker analysis, sulfate transporter, adenosine 5'-phosphosulfate reductase (APSR), serine acetyltransferase, O-acetylserine thiol-lyase, glutamylcysteine synthase, glutathione synthase, and γ-glutamyl transpeptidase candidate genes related to sulfur assimilation and ACSO biosynthesis were allocated to chromosomes 7A, 2A, 7A, 7A, 2A, 7A, and 4A, respectively. This result showed little association between the chromosomal locations of these genes and ACSO accumulation, suggesting that anonymous genes controlling ACSO accumulation were dispersed on the 2A, 3A, 4A, 5A, 7A, and 8A chromosomes of shallot. APSR gene expression was inhibited by 5A chromosome additions, suggesting that one of the regulatory genes was located on a 5A chromosome and inhibited APSR expression.

Eight kinds of japanese bunching onion [Allium fistulosum L. (genomes FF)]-shallot [Allium cepa L. Aggregatum group (genomes AA)] monosomic addition line [MAL (FF+1A-FF+8A)] were used to study the effects of single alien chromosomes from A. cepa on the production of carbohydrates in the leaf tissues of A. fistulosum. Carbohydrate contents in green leaf blades of these MALs were measured during alternate months from May 2005 to Mar. 2006. The determination of soluble sugar content from leaf blades of each MAL and A. fistulosum revealed that nonreducing sugars (sucrose and fructan) accumulated in winter leaf blades. Reducing sugar (fructose) in the leaf blades of each MAL was lower than A. fistulosum in almost every time period. In the leaf blades of FF+4A, high fructan accumulation was observed from Nov. 2005 to Mar. 2006. A series of determinations on the pectin content showed that amounts of NaOH-soluble pectin and HCl-soluble pectin remained at low levels at all time periods. High pectin accumulations in FF+7A and FF+8A occurred in September and slightly decreased in November. A decrease in hexametaphosphoric acid-soluble pectin content was associated with the maturity of MALs from autumn to winter, whereas the water-soluble pectin content increased. The pectin methylesterase and polygalactronase genes of shallot were assigned to chromosome 7A and 4A, respectively. These results demonstrate that important genes related to pectin metabolism in shallot are located on chromosomes 4A, 7A, and 8A of shallot.