Anthocyanins and flavonols were analyzed by HPLC using inbread lines and varieties of Pansy (Viola × wittrockiana). Each of the six main anthocyanins and flavonols were detected by HPLC. Four anthocyanins were identified by co-chromatography to be Dp3RG, Cy3RG, Dp3pCRG5G (nasunin) and Cy3pCRG5G. One of the flavonols was identified by co-chromatography to be Qu3RG (rutin). Yellow and white lines did not contain anthocyanin. Nasunin, rutin and five flavonols were detected in bluish color. There were differences in the amounts of major flavonols between lines. This result indicated the possibility of anthocyanin-flavonol copigmentation in variations of bluish color. Cyanidin-3-ramnosylglucoside, rutin and five flavonols were detected in reddish flowers. Major flavonols were No. 3, No. 4, and rutin in red lines. Another anthocyanin (Cy3pCRG5G) was detected in reddish-purple line. Differences in anthocyanin influenced the color variation of reddish colors. These findings indicated the importance of HPLC analysis in pansy breeding.
To develop a DNA fingerprinting technique suitable for edible tissues of vegetables, the reliable DNA extraction methods followed by fluorescent AFLP were proposed for each material. DNA was extracted from edible tissues of 33 vegetables using the miniprep DNA isolation method. In all materials, the DNA concentration and purity (A260/280 ratios of more than 1.8, A230/260 ratios of less than 2.95) estimated by UV spectrophotometry were thought to be sufficient for the next PCR step, but the browned DNA solution extracted from leaf lettuce or edible burdock inhibited PCR. The subsequent PCR was successful after purification of the crude DNA solutions, including 2-Mercaptethanol at a final concentration of 10mM, using Nucleon PhytoPure Kit (Amersham Biosciences Corp.). A total of 65 to 249 peaks were obtained by fluorescent AFLP analysis using six primer combinations in 33 vegetables. The technique developed in this study seems to have potential for cultivar identification in various vegetables. Furthermore, a similar AFLP analysis using bulked DNA samples from 16 plants each in three cultivars and 13 strains of Japanese bunching onion (Allium fistulosum L.) was conducted to examine the possibility of cultivar identification in outcrossing species. The DNA fingerprint obtained in this study allowed these cultivars and strains to be distinguished from each other.
To more easily evaluate longevity and ethylene sensitivity in spray-type carnation flowers, the following evaluation methods were investigated. Though the vase life was negatively correlated with room temperature, differences in vase life among varieties were confirmed, irrespective of room temperature. In some varieties of spray-type carnation with several flowers on one stem, there was a difference in vase life due to the flower setting position, which was caused by a difference in flowering stage at the flower setting position. If the flowering stages were the same, the variation in vase life decreased among flowers cut to about 3 cm at the peduncle and evaluation of the original longevity of the variety was possible to control. The petal soak treatment in ethephon solution was effective for evaluation of exogenous ethylene sensitivity because it could test the mass easily and simultaneously. Furthermore, the longevity and exogenous ethylene sensitivity in the main 47 varieties of spray-type carnation was clarified using new evaluation methods.
A tetraploid of Meiwa kumquat (Fortunella crassifolia Swingle) was selected from nucellar seedlings using root morphology, and its morphological characteristics were investigated. One seedling with thick short rootlets was obtained from 457 seedlings. Flow cytometry and chromosome observation demonstrated that the seedling was tetraploid (2n=4X=36). This tetraploid was presumed by Randam Amplified Polymorphic DNA (RAPD) analysis to have derived from nucellar embryos. The tetraploid showed similar morphology to other Citrus tetraploids such as thick, round leaves and larger stomata, flowers, and pollen grains compared with the original diploid. The tetraploid fruit was almost equal in weight and size to that of the diploid, but had a thicker peel and smaller number of developed seeds than the diploid fruit. In the future, the tetraploid obtained in the present study will be important material as a new cultivar and a parent of triploid breeding.
We investigated the relationship between petal brightness under ultraviolet light (365 nm) and flavonoid concentrations using 75 pale color lines of Eustoma grandiflorum. Flavonoid concentrations of the dark color petals were above 10 mg/g f. w., while those of the bright color petals were below 3 mg/g f. w. Several lines showed an image of bright petals with dark edges. A purple picotee petal and a white petal derived from ‘Candy Marin’ also showed such images. The edge tissue contained a 30- times higher concentration of flavonoids than the other tissue in both petals, suggesting that the same mechanism affects the flavonoid distribution. We can roughly estimate flavonoid concentration in pale color petals of Eustoma using this method. This method could be applied to other plants including Dianthus and Petunia. Because information on the flavonoid concentration in many flowers can be obtained immediately and non-destructively, this method could contribute to breeding selection and studies on the regulation of flavonoid-anthocyanin biosynthesis.
Inter- and intra-specific variation of capsaicinoid concentration in five cultivated Capsicum species were investigated. Capsaicinoid concentrations in 145 accessions of C. annuum, C. chinense, C. frutescens, C. baccatum, C. pubescens collected widely around the world were quantified using HPLC. Three components of capsaicinoid, capsaicin (CAP), dihydrocapsaicin (DC) and nordihydrocapsaicin (NDC) were detected by HPLC. C. annuum accessions showed comparatively low concentrations, half of which were for vegetable use with less than 1000 μg/gDW of capsaicinoid. C. chinense and C. frutescens had a range of variation wider than C. annuum, from non-detectable to a remarkably high concentration, Therefore, accessions of these two species are useful breeding materials for varieties with various degrees of pungency. C. baccatum included many low concentration accessions available for breeding vegetable-use varieties. Capsaicinoid composition differed between C. chinense and C. frutescens (CAP>DC) and C. pubescens (CAP<DC). C. annuum showed a broad variation of composition covering the ranges of the other four species and showing a geographical difference. Accessions of CAP<DC were found with high frequency only in Japan. The significant negative correlations among capsaicinoid concentration and fruit size characteristics were recognized in C. annuum and C. baccatum. However, these are not genetic correlations but phenotypic correlation due to negative correlation between the ratio of placenta to fruit and fruit size.
The direction of flower cluster extension was investigated in micropropagated strawberry plants with no stolons. If micropropagated plants were planted at an angle of 20 to 25°, primary flower clusters developed in an inclined direction. However, primary flower clusters developed in the opposite direction if inclined pots were rotated by 180°, 40 days after the first inclination treatment. These results show that the direction of extension of primary flower clusters is determined geotropically during the period between flower bud differentiation and anthesis. Furthermore, it is likely that micropropagated plants will extend flower clusters to the aisle side if they are planted at an angle of 20 to 25° toward the aisle side.
Shoot tops (10 to 25 cm from apices) of one- and two-year-old ‘Fuji’ and ‘Akibae’ apple trees grafted onto M. 9 Nagano and Pajam 2 rootstocks were sprayed one to five times with 6-benzyladenine (BA, 300 ppm) to induce branch development in the nursery. It was observed that repeated spraying with BA increased the average number of lateral shoots per tree in both varieties of trees. Trees sprayed three to five times with BA produced significantly more branches than those receiving a single spray or untreated controls. The most effective treatments, five applications of BA to the new shoot tops of two-year-old ‘Fuji’ apple trees, produced an average of 14.3 lateral shoots (1 cm and longer) per tree, while single application produced 7.2 laterals per tree. Trunk diameter and tree height were not affected by the treatments.
The experiments were conducted to investigate the effects of plug cell shape on growth of seedlings and development of root systems in cabbage after transplanting. A plug cell with a convex inner wall, a cell with a slit wall, and a normal cell were used for examination. There was no remarkable difference in seedling growth due to cell shape, though the growth of seedlings in convex cells was slightly suppressed. There was no influence of cell shape on cabbage head weight. Convex cells and slit cells were found to suppress root circling. The roots of seedlings grown in convex cells extended deeply after transplanting and uprooting resistance of the plants increased. However, the roots of seedlings grown in slit and normal cells did not extend deeply after transplanting.
Experiments were conducted to determine the effect of solid supporting materials and culture media on the growth of regenerated plantlets from root segments of Aralia elata Seemann in in vitro culture. The plantlets, subcultured on MS medium, were removed from a culture tube and transplanted aseptically into four kinds of solid supporting materials. They were cultured on these materials for 2 months at 25°C in a 16 h light (3,000 lx) condition. A mixture of equivalent volumes of vermiculite and perlite or sphagnum was the most effective material for raising high-quality plants, out of the four kinds of solid supporting materials. Hydroponics with 3% sucrose supported better growth of the plantlets than MS medium with 3% sucrose. The plantlets directly transplanted to vermiculite in a planting cell tray also easily grew into young plants after 2 months in a closed culture container. These results indicate that the use of solid supporting materials and culture media are effective for promoting the growth of plantlets from in vitro culture.
To research the cyclic use of the drainage solution in a closed system of rockwool cultivation of cut roses, we examined the effects of diluting the drainage solution with water, both with and without additions of NH4-N, Fe and Mn nutrient components following the slow sand and active charcoal filtration of the drainage solution. The yields of the cut flower stems were almost the same as those in the conventional cultivation, at 35 per plant, when the drainage solution was recirculated, and whether or not we added nutrient components. The cut flower stem quality was almost the same as those in the conventional cultivation, in each drainage cycle. Under the conventional cultivation system, the total nitrogen quantity applied per 1,000 m2 cultural facilities for 1 year was 290.3 kg, and the quantity for disposal was 135.9 kg. Under the closed system, using the added nutrient components, the total nitrogen quantity was 151.8 kg, and with no added nutrients components, the total for disposal was only 110.5 kg. In addition, under the closed systems, there was no nitrogen and phosphoric acid to dispose of in each drainage cycle. PO4-P, Mn, Fe and SO4-S concentrations in the drainage solution were lowered, after the drainage solution was slowly filtered. Na and Cl concentrations in the solution rose remarkably when the filtered drainage solution was circulated. However, no symptoms of mineral excess or deficiency were observed in the rose plants. In a closed system of rock wool rose cultivation, it was shown that there was no neccessity to supplement the nutrient components, when the drainage solution was diluted by 2.5 times after slow filtration process.
The effect of night break on flowering time of Lilium × formolongi was examined. The flowering time of ‘Sakigake-raizan’, ‘White Lancer’ and ‘Inakei’ was accelerated to June by night break treatment for 4 hours for cultivars planted in February., When night break was started during the period from February to April, the flowering time of ‘White Lancer’ was late June. More than four weeks of night break are required to harvest flowers in June.
To harvest Lilium × formolongi cv. Hayachine flowers in April and May, effects of night break and heating treatment were examined. The flowering time of ‘Hayatine’lily was accelerated to the middle of April by night-break and heating treatment, but to late May by night-break treatment alone. When liles were planted in September and October, flowering time was earlier and the resulting quality of cut flowers was better, compared to those planted at other times. There was little difference in the quality of cut flowers regardless of the starting time of the night break treatment.
To clarify the relationship between flower color and growing season in pink flower genotypes of spray chrysanthemum (Dendranthema grandiflorum), the flower color in each cropping season was evaluated by the use of color different meter and HPLC. Although there were significant differences in the value of L*a*b* among the cropping season in yellow and white flower genotypes, there was no flower color change during the cropping season recognized by visible observation. However, pink flower genotypes except ‘Chatoo’ showed low L* and b* values and remarkably high a* and c* values in the autumn, then reversed the tendency of each value was observed in the summer season. In ‘Chatoo’, the difference of each value among cropping season was low although it showed the same tendency as other pink flower genotypes. The two main anthocyanins, cyanidin 3-O-(6”-O-monomalonyl-β-glucopyranoside) and cyanidin 3-O-(3”, 6”-O-dimalonyl-β-glucopyranoside), were detected in pink flower genotypes. There was no change in the component ratio of anthocyanins observed among the cropping season, suggesting that flower color in pink flower genotypes of spray type chrysanthemums is determined by changes in the two main flower pigments.
Effects of rootstocks on tree mortality rate and canopy volume expansion in young Zanthoxylum piperitum (L.) DC. f. inerme Makino (Budousanshou) Japanese pepper tree were compared among Z. schinifolium Sieb. et Zucc. (Inuzanshou), Z. alatum Roxb. var. planispinum Rehd. et Wils. (Fuyuzanshou) and Fagara ailanthoides Engl. (Karasuzanshou) rootstocks for high viability after grafting and an early canopy establishment. The tree mortality rate was significantly higher on the Inuzanshou rootstock than on Fuyuzanshou or Karasuzanshou rootstock. All trees on the Inuzanshou rootstock died within 3 years after planting. The volume of tree canopy 5 years after planting was significantly larger on the Karasuzanshou rootstock than on the Fuyuzanshou rootstock. The canopy volume on the Karasuzanshou rootstock was twice as large as that of Fuyuzanshou rootstock. Overgrowth of rootstock was observed on the Karasuzanshou rootstock during the 5- year experiment, but there was no imbalance in the growth rate between rootstock and scion diameter. Numbers of flower clusters produced on 5-year old trees were significantly larger on the Fuyuzanshou than on the Karasuzanshou rootstock.
The difference in oxalic acid content between tart and sweet cultivars of carambola (Averrhoa carambola L.) was examined, specifically the difference of total oxalic acid content in leaves, and the proportion of insoluble oxalic acid of total oxalic acid content in fruits. We thought that taste acidity could be reduced by decreasing the proportion of the soluble oxalic acid content in the total oxalic acid content of carambola fruits. The relationships among oxalic acid, potassium, calcium and magnesium contents in leaves were examined in hybrid seedlings of tart and sweet cultivars of carambola. Total oxalic acid content closely correlated with insoluble oxalic acid content. The correlation coefficient between insoluble oxalic acid and calcium content was highly significant compared with potassium and magnesium. These results suggested that the total oxalic acid content in leaves was strongly influenced by the calcium content, and it appears to be decreased by the regulation of calcium content.
The rhizomes of Polygonatum macranthum (Maxim.) Koidz. undergo a period of endodormancy after the formation of new rhizomes in summer. The decrease in the dormancy level with time and the chilling requirements for breaking dormancy are examined in this paper. The rhizome dormancy was deepest in early July or earlier, and after this the level of dormancy decreased gradually with time. After late November the decrease was more progressive, as a result of exposure to low outdoor temperatures. The rhizome dormancy was almost broken by late February to late March. The optimum temperature range, for sprouting of rhizomes almost released from dormancy, was 25 to 30°C. Chilling was effective in breaking the dormancy of rhizomes when the rhizomes were dug in late August or later. The rhizomes dug in late September required a 120-day period of chilling at 5°C to break dormancy completely, whereas a 90-day period of chilling at 5°C was enough to completely break the dormancy of those dug in late October. When connected rhizomes with many buds were planted, half the number of buds did not expand. However, all buds expanded when divided rhizomes with only one bud were planted.
Effects of a high soil temperature using plastic mulch and cultivation of the resistant cultivar, ‘Peking’ on the population of soybean cyst nematode (Heterodera glycines) were investigated in the field and in an incubator with vegetable soybean (Edamame). Soybean cyst nematode eggs were inoculated into seedlings of the susceptible cultivar, ‘Fuki’, grown in incubators with soil temperatures of 25, 29 and 33°C. Soybean cyst nematode did not produce cysts containing eggs at a soil temperature of 33°C, although it did produce eggs at 25 and 29°C. In field experiments, susceptible ‘Fuki’ and the resistant soybean cultivar, ‘Peking’, were cultivated for about 3 months with or without plastic mulch. When the accumulated duration of soil temperature above 33°C was 200 hr or longer, the egg reproduction ratio was significantly suppressed and edamame yield was high the following year. After ‘Peking’ cultivation, the egg number in the soil was significantly suppressed compared to that after ‘Fuki’, and edamame yield the following year was also high.
Properties of nine kinds of anthocyanidins and 24 kinds of anthocyanins were compared by thin-layer chromatography (TLC) data and UV-VIS spectrophotometer data with high performance liquid chromatography (HPLC) data including photo diode array (PDA) detector data. The measuring time is about one hour with HPLC analysis though it takes over twelve hours using TLC combined with UV-VIS measurement. Moreover, it is possible to analyze even by a small amount of material using the HPLC analysis, and the efficiency is also high. In addition, absorption spectrum measurement is possible using PDA at the same time. In this study, spectrum data obtained with a PDA detector, which is expected to increase the efficiency of the identification of anthocyanin by HPLC analysis, was presented.
The antioxidative activities of normal and injured eggplant ‘Mizu-nasu’ fruit were evaluated using Trolox, which is a 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging reagent. In normal ‘Mizu-nasu’ fruit, there was a positive correlation (r=0.93, n=9) between anthocyanin concentration and the radical scavenging activity of the peel of ‘Mizu-nasu’ fruit. The amount of chlorogenic acid in the pulp of ‘Mizu-nasu’ fruit was 1.4 μmol/g · f.w. About half of the radical scavenging activity of the pulp was attributed to the chlorogenic acid in the pulp. The peel contributed 15.9% of the activity of its whole fruit, and the pulp contributed 84.1%. The injured part (6.69-8.05μmol of Trolox equivalent/g) had stronger activity than the normal part (3.46-4.73μmol of Trolox equivalent/g) of the fruit. These results suggest that injured fruit could be worth using as a polyphenol-rich-food.