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Genhua Niu, Toyoki Kozai and Chieri Kubota

A system was designed for measuring the CO2 exchange rates [net photosynthetic rate (Pn) and dark respiration rate] of in vitro plantlets in situ (in the vessel with natural ventilation). The system, excluding gas cylinders, was placed in a growth chamber so that the desired photosynthetic photon flux (PPF) and temperature could be maintained during the measurement. The CO2 concentration inside the culture vessel (Ci) was indirectly controlled by controlling the CO2 concentration outside the vessel (Co). The Pn of the plantlets was estimated based on the measured Ci and Co at steady state using a gas chromatograph according to the method described by Fujiwara et al. (1987). The performance of the system was demonstrated by measuring the in situ Pn of sweetpotato [Ipomoea batatas (L.) Lam., cv. Beniazuma] and tomato (Lycopersicon esculentum Mill., cv. Hana Queen) plantlets in vitro under a range of CO2 concentrations and PPF. The photosynthetic parameters of the Pn model (Niu and Kozai, 1997) for the plantlets were then estimated based on the measured Pn. The preliminary measurements demonstrated the potential application of the system.

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Tomomi Eguchi, Ricardo Hernández and Chieri Kubota

Intumescence injury is an abiotic-stress-induced physiological disorder associated with abnormal cell enlargement and cell division. The symptom includes blister- or callus-like growths on leaves, which occur on sensitive cultivars of tomato when they are grown under ultraviolet (UV)-deficit light environment, such as light-emitting diodes (LEDs). Previous studies suggest that intumescence can be reduced by increasing far-red (FR) or blue light. In the present study, effects of end-of-day FR (EOD-FR) light and high blue photon flux (PF) ratio during the photoperiod on intumescence injury were examined using ‘Beaufort’ interspecific tomato rootstock seedlings (Solanum lycopersicum × Solanum habrochaites), a cultivar highly susceptible to intumescence injury. Our study showed that EOD-FR light treatment moderately suppressed intumescence injury. Using EOD-FR light treatment, the percent number of leaves exhibiting intumescences was reduced from 62.0–70.7% to 39.4–43.1%. By combining high blue PF ratio (75%) during the photoperiod and EOD-FR light treatment, the percent number of leaves exhibiting intumescences was further suppressed to 5.0%. Furthermore, the combination of high blue PF ratio and EOD-FR light treatment inhibited undesirable stem elongation caused by EOD-FR light treatment. We found that high blue PF ratio during the photoperiod combined with a small dose of EOD-FR lighting (≈1 mmol·m−2·d−1 provided by 5.2 µmol·m−2·s−1 FR PF for 3.3 minutes) could inhibit the problematic intumescence injury of tomato plants grown under LEDs without negatively influencing growth or morphology.

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Sandra B. Wilson, Jeongwook Heo, Chieri Kubota and Toyoki Kozai

Sweetpotato [Ipomoea batatas (L.) Lam., `Beniazuma'] plantlets were grown photoautotrophically (without sugar) for 12 days in an improved forced ventilation system designed with air distribution pipes for uniform spatial distributions of carbon dioxide (CO2) concentration. Enriched CO2 conditions and photosynthetic photon flux (PPF) were provided at 1500 μmol·mol-1 and 150 μmol·m-2·s-1, respectively. The forced (F) ventilation treatments were designated high (FH), medium (FM), and low (FL), corresponding to ventilation rates of 23 mL·s-1 (1.40 inch3/s), 17 mL·s-1 (1.04 inch3/s), and 10 mL·s-1 (0.61 inch3/s), respectively, on day 12. The natural (N) ventilation treatment was extremely low (NE) at 0.4 mL·s-1 (0.02 inch3/s), relative to the forced ventilation treatments. Total soluble sugar (TSS) and starch content were determined on day 12. Total soluble sugars (sucrose, glucose, fructose) of FH plantlets were lowest in leaf tissue and highest in stem tissue as compared to other ventilation treatments. Starch concentration was higher in leaf tissue of FH or FM plantlets as compared to that of FL or NE plantlets. Plantlets subjected to FH or FM treatments exhibited significantly higher net photosynthetic rates (NPR) than those of the other treatments; and on day 12, NPR was almost five times higher in the FH or FM treatment than the FL or NE treatments. Carbohydrate concentration of plantlets was also influenced by the position of the plantlets in the vessel. Within the forced ventilation vessels, leaf TSS of FH and FM plantlets was similar regardless of whether plantlets were located near the inlet or outlet of CO2 enriched air. However, under FH or FM conditions, leaf starch concentration was higher in plantlets located closest to the CO2 inlet as compared to the outlet.

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Ryo Matsuda, Chieri Kubota, M. Lucrecia Alvarez and Guy A. Cardineau

Using greenhouse tomato (Solanum lycopersicum) as a model system to produce pharmaceutical proteins, electrical conductivity (EC) of hydroponic nutrient solution was examined as a possible factor that affects the protein concentration in fruit. Transgenic tomato plants, expressing F1-V protein, a plant-made candidate subunit vaccine against plague (Yersinia pestis), were grown hydroponically at high (5.4 dS·m−1) or conventional EC [2.7 dS·m−1 (control)] with a high-wire system in a temperature-controlled greenhouse. There was no significant difference in plant growth and development including final shoot dry weight (DW), leaf area, stem elongation rate, or leaf development rate between high EC and control. Net photosynthetic rate, transpiration rate, and stomatal conductance (g S) of leaves were also not significantly different between EC treatments. For both EC treatments, immature green fruit accumulated DW at a similar rate, but dynamics observed in fruit total soluble protein (TSP) and F1-V during the fruit growth were different between the two ECs. Fruit TSP concentration per unit DW decreased while TSP content per whole fruit increased as fruit grew, regardless of EC. However, TSPs were significantly lower in high EC than in control. Fruit F1-V concentration per unit DW and F1-V content per whole fruit were also lower in high EC than in control. Our results found that increasing EC of nutrient solution decreased TSP including the vaccine protein in fruit, suggesting that adjusting nutrient solution EC at an appropriate level is necessary to avoid salinity stress in this transgenic tomato.

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Chieri Kubota, Cynthia A. Thomson, Min Wu and Jamal Javanmardi

Plants produce various phytochemicals that are of nutritional and medicinal value to humans. Phytochemicals having antioxidant capacity are drawing increased interest from consumers. Population studies among Americans have consistently demonstrated inadequate consumption of fruit and vegetables. Improving intake of fruit and vegetables has been a major public health effort for many years with minimal success. Given this, it seems opportunistic to consider other approaches to enhance the nutritional quality of the American diet. One plausible approach is the development of fresh produce containing a greater concentration of phytochemicals known to improve health, thus while consuming fewer servings of produce, Americans would still have significant exposure to health-promoting food constituents. Controlled environments provide a unique opportunity to modify the concentrations of selected phytochemicals in fruit and vegetables, yet practical information is limited regarding methods effective in optimizing antioxidant capacity. Our research at the University of Arizona Controlled Environment Agriculture Program has shown that application of moderate salt stress to tomato plants can enhance lycopene and potentially other antioxidant concentrations in fruit. The increase in lycopene in response to salt stress in the tomato fruit was shown to be cultivar specific, varying from 34% to 85%. Although the specific biological mechanisms involved in increasing fruit lycopene deposition has not been clearly elucidated, evidence suggests that increasing antioxidant concentrations is a primary physiological response of the plant to the salt stress. Another experiment showed that low temperature during postharvest increased antioxidant capacity of tomato fruit while it maintained the lycopene concentration. More detailed study in this area is needed including accumulation of antioxidant phytochemicals as affected by environmental conditions during the cultivation and the postharvest.

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Ryo Matsuda, Chieri Kubota, M. Lucrecia Alvarez and Guy A. Cardineau

Changes in the amounts of F1-V, an antigen fusion protein and a candidate subunit vaccine against plague, and total soluble protein (TSP) in green fruit of transgenic tomato plants were investigated to identify the optimum harvest timing to maximize the F1-V yields. Two T 2 progenies of the transgenic plant, ‘22.11.21’ and ‘22.11.5’, were grown. The F1-V concentration rapidly decreased at the beginning of the green stage and decreased to less than 5% of the initial concentration at the late green stage in ‘22.11.21’. The F1-V concentration also decreased as fruit size increased in ‘22.11.5’, but the pattern of the decrease was linear and different from that in ‘22.11.21’. The concentration of TSP also decreased with fruit growing in both plants. When calculated on a whole fruit basis, the F1-V content linearly decreased with increasing fruit size in ‘22.11.21’. In ‘22.11.5’, the F1-V content per fruit also tended to decrease from the middle to late green stage. Based on these observations, collecting small green fruits without pruning was proposed as a harvest practice that may maximize the F1-V yields. Thus, the optimum protocols for harvesting and pruning for plant-made pharmaceutical production may be substantially different from those currently used in commercial hydroponic greenhouses for fresh market tomato.

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Myles Lewis, Chieri Kubota, Russell Tronstad and Young-Jun Son

Grafting of fruiting vegetables is a relatively new advent in the United States with promise as a technology to improve both yields and the environment. However, investing in a commercial-sized grafting enterprise requires substantial capital investment and is a risky endeavor. A tool to help evaluate grafting costs for different production technologies and sizes of operation is a useful decision aid for individuals investing in new or modifying existing operations to produce grafted plants. Using a combination of engineering and financial equations, a scenario-based analysis was completed to obtain approximate capital and variable costs per plant for both new and existing production facilities. For exemplary purposes, four scenarios consisting of two different crops (tomato and watermelon) at two production sizes with different technology levels [low-volume manual grafting (one million plants per year) and high-volume fully automated grafting (100 million plants per year)] are presented to compare costs. For simplification purpose, consistent weekly production was assumed in the cost simulation. Total capital costs were $115,127 and $118,974 for low-volume production for grafted tomato and watermelon plants, respectively. They were $21.6 million and $16.7 million under high-volume production for tomato and watermelon, respectively. Among the four scenarios evaluated, variable costs per plant (costs of plants produced) were lowest for watermelons with high-volume production ($0.089 per plant), suggesting that production costs of grafted plants could decrease by scaling up production and introducing automation. Sensitivity analyses for high-volume production of tomato showed that the electricity rate, grafting clip price, and grafting robot speed were factors with the greatest influence on costs of plants. Scenario-based cost analysis was shown to be an effective tool for developing strategies to reduce the price of grafted plants.

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Chieri Kubota, Nihal C. Rajapakse and Roy E. Young

Broccoli (Brassica oleracea L. Botrytis Group `Green Duke') and Hosta tokudama F. Maekawa `Newberry Gold' plantlets, which were ready for transplanting after photoautotrophic (sugar-free) culture, were stored 4 to 6 weeks at 5C under various light qualities and photosynthetic photon fluxes (PPF). Illumination during storage maintained quality, photosynthetic ability, and regrowth potential of plantlets stored at low temperature. PPF affected quality of broccoli and Hosta plantlets. Broccoli plantlets responded to storage light quality, while Hosta did not. White light maintained the quality of broccoli plantlets better during 6 weeks of storage than did red or blue light. Red and blue light caused an increase in internode length and reduction in chlorophyll concentrations compared to white light. Photosynthetic and regrowth potentials of plantlets were not affected by spectral quality during storage. Considering changes in dry weight, stem length, and leaf yellowing, the quality of broccoli plantlets was best maintained under white light at 2 μmol·m–2·s–1 PPF. PPF and light quality were shown to be important factors in the preservation of transplant quality and suppression of growth of the plantlets during low-temperature storage.

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Chieri Kubota, Nihal C. Rajapakse and Roy E. Young

`Green Duke' broccoli plantlets, which were ready for transplanting after 2 weeks of photoautotrophic (sugar free) culture under the conditions of 1100 μmol·mol–l CO2 (outside the vessel), 22 + 4C air temperature, and 140 μmol·m–2·s–1 photosynthetic photon flux (PPF), were stored for 6 weeks at 5C in darkness or in white, red, or blue light at 2 μmol·m–2·s–l PPF (light compensation point at 5C). Photoperiod was set at 24 hour/day during storage. Spectral quality significantly affected plantlet quality: stem length was longer and chlorophyll concentration of leaves was lower in red or in blue light than in white light or in darkness after 6 weeks in storage. Regardless of the spectral quality, light in storage maintained plantlet dry weight at a level comparable to that before storage; dry weight was reduced significantly in dark-stored plantlets. Spectral quality did not significantly affect the photosynthetic and regrowth potential of plantlets. All plantlets stored in light, regardless of light spectra, grew preferably and had similar dry weight and stem length after 9 weeks of transplanting to the greenhouse under natural light.

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Katsumi Ohyama, Koji Manabe, Yoshitaka Omura, Toyoki Kozai and Chieri Kubota

To evaluate the potential use of a 24-hour photoperiod for transplant production in a closed system, tomato (Lycopersicon esculentum Mill.) plug transplants were grown for 17 days either under a 24-hour photoperiod with a photosynthetic photon flux (PPF) of 200 μmol·m-2·s-1 or under a 16-hour photoperiod with a PPF of 300 μmol·m-2·s-1, resulting in the same daily integrated PPF (17.3 mol·m-2). Air temperatures were alternated between 28 °C during the first 16 hours and 16 °C for the subsequent 8 hours of each day. Fresh weight, dry weight and leaf area were 41%, 25%, and 64% greater, respectively, under the 24-hour photoperiod than under the 16-hour photoperiod. Physiological disorders (e.g., chlorosis and/or necrosis) were not observed under the 24-hour photoperiod, probably due to the alternating air temperature. Floral development of plants originating from both treatments did not differ significantly. Electric energy use efficiency of the closed system was 9% greater under the 24-hour photoperiod than under the 16-hour photoperiod. These results suggest that using a 24-hour photoperiod with relatively low PPF can reduce both initial and operational costs for transplant production in a closed system due to the reduction in the number of lamps.