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Aqueousmethanol extracts of Cit rus junos, C. unshiu and C. sudachi fruit peel in hib ited the growth of the roots and hypocotyls of alfalfa (Medicago sativa L.), cress (Lepidiumsativum L.) and lettuce (Lactuca sativa L.) seedlings. Signifi cantreductions in the root and hypocotyl growth were observed as the extract concentration in creased in all bioassays. The in hibitory activity of C. junos ex tract on the growth of test plants was 3.3-to 17.9-fold and 3.6- to 20.6-fold greater than that of C. unshiu and C. sudachi extracts, respectively. Theconcent rationin C. junos was 3.5- and 4.9-fold greater than that in C. unshiu and C. sudachi, re spectively. Thus, there was a good correlation-between abscisic acid-b-D-glucopyranosylester (ABA-GE) concentrationsin C. junos, C. unshiu and C. sudachi fruit peel and theinhibito ryactivities of the irextracts. The concentration of ABA-GE in C. junos fruit peel was in creased with fruit maturityasgrowthinhibit oryactivity of C. junos fruit peel was reported to be in creased with fruit maturity, in dicating that the concentrations of ABA-GEin C. junos fruit peel was correlated withgrowthinhibit oryactivity of C. junos fruit peel in time course of fruit maturation. These findings suggest that ABA-GE may be involved in the growth inhibitory effect of C. junos, C. unshiu and C. sudachi fruit peel.

The allelopathic potential of an aqueous methanol extract of Citrus junos leaves was investigated under laboratory conditions. The extract inhibited the growth of roots and shoots of cress (Lepidium sativum L.), lettuce (Lactuca sativa L.) and alfalfa (Medicago sativa L.) seedlings. Significant reductions in the root and shoot growth were observed as the extract concentration increased. Since abscisic acid -β-D-glucopyranosyl ester (ABA-GE) was found to be a major inhibitor in fruit peel of C. junos, ABA-GE in C. junos leaves was determined and the concentration was 540 μg kg-1 fresh weight in the leaves harvested on September 26, 2002. These results suggest that C. junos leaves possess allelopathic activity and could work as a weed inhibiting agent, which should be investigated further in the field for practical application.

The allelopathic potential of Citrus junos fruit waste after juice extraction was investigated. Aqueous methanol extracts of peel, inside and seeds separated from the fruit waste inhibited the growth of the roots and shoots of alfalfa (Medicago sativa L.), cress (Lepidium sativum L.), crabgrass (Digitaria sanguinalis L.), lettuce (Lactuca sativa L.), timothy (Pheleum pratense L.), and ryegrass (Lolium multiflorum Lam.). The inhibitory activity of the peel extract was greatest and followed by that of the inside and seed extracts in all bioassays. Significant reductions in the root and shoot growth were observed as the extract concentration was increased. The concentrations of abscisic acid-β- d -glucopyranosyl ester (ABA-GE) in peel, inside and seeds separated from the C. junos fruit waste were determined, since ABA-GE was found to be one of the main growth inhibitors in C. junos fruit. The concentration was greatest in the peel, followed by the inside and seeds; there was a good correspondence between these concentrations and the inhibitory activities of the extracts. This suggests that ABA-GE may also be involved in the growth inhibitory effect of C. junos waste. These results suggested that C. junos waste may possess allelopathic potential, and the waste may be potentially useful for weed management.

Aqueous methanol extracts of Citrus junos, C. unshiu and C. hassaku fruit peel inhibited the growth of the roots and hypocotyls of lettuce (Lactuca sativa L.) seedlings. Significant reductions in the root and hypocotyl growth were observed as the extract concentration increased in all bioassays. The inhibitory activity of C. junos extract on the growth of lettuce roots and hypocotyls was about 13- and 24-fold greater than that of C. unshiu and C. hassaku extracts, respectively. The concentration of abscisic acid-β-D-glucopyranosyl ester (ABA-GE) in fruit peel of C. junos, C. unshiu and C. hassaku was determined since ABA-GE had been found to be a possible cause of the growth inhibitory effect of C. junos. The concentration was 132, 10.6 and 5.0 µg g−1 dry weight in C. junos, C. unshiu and C. hassaku fruit peel, respectively. Thus, there was a good correlation between ABA-GE concentrations in C. junos, C. unshiu and C. hassaku fruit peel and the inhibitory activities of their aqueous extracts, which suggests that ABA-GE may be involved in the growth inhibitory effect of C. junos, C. unshiu and C. hassaku fruit peel.

Exogenously applied ABA-beta-D-glucopyranosyl ester (ABA-GE) inhibited hypocotyl growth of Arabidopsis seedlings at concentrations greater than 0.3 micromol/L, and the concentration for 50% inhibition of hypocotyl growth was 1.8 micromol/L. ABA-beta-D-glucosidase activity in Arabidopsis seedlings was 17 nmol/mg protein/mim and increased by exogenously applied ABA-GE. The pH optimum of this enzyme in crude extract of Arabidopsis seedlings was 6.0 for the assay in the ABA-GE to ABA direction and its K(m) value for ABA-GE (pH 6.0) was 0.41 mmol/L. These results suggests that exogenously applied ABA-GE may be absorbed by roots and hydrolyzed by ABA-beta-D-glucosidase and librated free ABA may induce growth inhibition in Arabidopsis hypocotyls.

The inhibitory effect of Citrus junos peel on plant growth using lettuce (Lactuca sativa L.) as a bioassay material was investigated, since the powder of the peel had been found to inhibit growth of weeds. Basic, neutral and acidic fractions were separated from the aqueous fraction obtained from the methanol extract of C. junos peel. All fractions inhibited the growth of lettuce seedlings, but by far the greatest inhibition was observed with the neutral fraction. Thus, the latter was further purified and an allelopathically active substance was isolated. The structure of the substance was determined from high-resolution MS and 1H and 13C NMR spectral data as abscisic acid-beta-D-glucopyranosyl ester (ABA-GE). ABA-GE inhibited hypocotyl and root growth of lettuce seedlings at concentrations greater than 0.3 microM, and the concentrations for 50% inhibition of hypocotyl and root growth were 2.3 and 1.4 microM, respectively. The effectiveness of ABA-GE on inhibition of growth and the occurrence of ABA-GE in the peel itself suggested that ABA-GE may play an important role in the allelopathic potential of C. junos peel. The peel may be potentially useful for weed management in a field setting.