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  • Author or Editor: Toyoki Kozai x
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Growth and net photosynthetic rate of potato (Solanum tuberosum L.) `Benimaru' plantlet in vitro were studied under a conventional photomixotrophic condition [with 20 g sucrose/liter in the medium and under 70 μmol·m-2·s-1 photosynthetic photon flux (PPF)] with minimal ventilation (MV) and under photoautotrophic conditions (without sugar in the medium and under 190 μmol·m-2·s-l PPF) with enhanced natural ventilation using an air diffusive filter (DV) or with forced ventilation (FV). Fresh weight of the plantlets cultured in the FV and DV treatments was 2.4 times that of the plantlets cultured in the MV treatment. Net photosynthetic rate and dry weight per plantlet were the highest in FV followed by DV. For photoautotrophic micropropagation, FV was superior to DV.

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A photoautotrophic or sugar-free medium micropropagation system (PAM) using five large culture vessels (volume = 120 L each) with a forced ventilation unit for supplying CO2-enriched air was developed and applied to commercial production of calla lily (Zantedeschia elliottiana) and china fir (Cunninghamia lanceolata) plantlets. The culture period of calla lily plantlets in the PAM was reduced by 50%, compared with that in a conventional, photomixotrophic micropropagation system (PMM) using small vessels each containing a sugar-containing medium. Percent survival ex vitro of calla lily plantlets from the PAM was 95%, while that from the PMM was 60%. The production cost of calla lily in the PAM was reduced by about 40%, compared with that in the PMM, and the initial investment per plantlet for the PAM was ≈10% lower than that for the PMM. The sales price of ex vitro acclimatized calla lily plantlet was increased by 25% due to its higher quality, compared with plantlets produced in the PMM.

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Broccoli (Brassica oleracea L. Botrytis Group `Ryokurei') plantlets, cultured photoautotrophically (without sugar in the medium) in vitro for 3 weeks at 23C and 160 μmol·m–2·s–1 photosynthetic photon flux (PPF), were stored for 6 weeks at 5, 10, or 15C under 0 (darkness) or 2 μmol·m–2·s–1 PPF (continuous lighting) supplied by fluorescent lamps (white light). Dry weight of the plantlets stored for 6 weeks at 5 or 10C in light was not significantly different from that of the plantlets before storage. Dry weight of the plantlets decreased as temperature increased and was maintained at higher levels in light than in darkness. Chlorophyll concentrations of the plantlets were higher at the lower temperatures. Chlorophyll fluorescence kinetics indicated higher activities of chlorophyll of the plantlets stored in light than in darkness. Lighting at as low as 2 μmol·m–2·s–1 PPF was important to preserve photosynthetic and regrowth abilities and dry weight of the plantlets during low-temperature storage.

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Commercial transplant production in Japan has been increasing rapidly since 1985. Transplant production began with plug seedlings for bedding plants, followed by carnation and Chrysanthemum plug transplants vegetatively-propagated using cuttings. Next, production more recently includes plug seedlings of lettuce and cabbage, and micropropagated tubers of potato plants and grafted transplants of tomato, eggplant, cucumber, and watermelon plants. The reasons for the rapid increase in commercial production of transplants will be reviewed. The current “cutting edge” practices include hardening before shipping or planting. The pros and cons of current transplant production systems in Japan will be discussed. Recent research advances in production of micropropagated, grafted and seedling transplants are reviewed with special reference to environmental control for hardening or acclimatization. Research on robotic or automated systems for micropropagation, grafting, and transplanting currently developed in Japan are described.

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A storage method of transplants in vitro was developed using light compensation points in conjunction with low temperatures. Broccoli (cv. Ryokurei) plantlets, aseptically germinated and cultured for three weeks in vitro, were used as model transplants. Culture conditions were: 23C air temperature, 160 μmol m-2s-1 PPF, and 3.6 air exchanges per hour of the vessel. Prior to storage, light compensation points were determined at 3, 5, 10, and 15C for the plantlets cultured with or without 20 g liter-1 sugar in the medium. Plantlets were stored for six weeks at 5, 10, and 15C under either 0 or 2 μmol m-2s-1 continuous PPF. The light compensation points varied with air temperature and with medium sugar level. Plantlet dry weight during storage was best maintained by keeping CO2 exchange rate of the plantlets close to zero throughout the storage period. High transplant qualities were successfully preserved at light compensation points: 2 μmol m-2s-1 PPF at 5-10C without sugar, and at 5C with sugar in the medium. This method may be applicable for storage of other crop transplants, plug seedlings and cuttings as well.

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We report the results of serial studies aimed at clarifying several factors affecting organogenesis in rhizome culture of temperate Cymbidium species and their hybrids. The growth patterns and regeneration ability of rhizomes derived from asymbiotic seed or shoot tip culture vary according to media composition, kinds and concentrations of plant growth regulators, culture conditions, and species and varieties. N6-benzyladenine was the best cytokinin for inducing shoot formation, for switching rhizome tissues into protocorm-like bodies, and for directly forming multiple shoots from branched rhizomes. Activated charcoal appeared to be necessary for producing healthy plantlets and for stimulating shoot growth at levels of 0.1% to 0.3% but concomitantly decreased rhizome growth. Sucrose at 5% was the most effective concentration for shoot induction from rhizomes. The above results support the conclusion that organogenic pathways between tropical, subtropical, and temperate Cymbidium species may be controlled by the genetic backgrounds of the species or cultivars.

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Providing continuous light (24-h photoperiod) at a relatively low photosynthetic photon flux (PPF) is one possible way to reduce both initial and operational costs for lighting and cooling during transplant production with an artificial light. However, physiological disorders (i.e., chlorosis and necrosis) are often observed in several species under continuous light with a constant temperature. The objective of this study was to find an effective air-temperature regime under the continuous light to avoid such physiological disorders, and simultaneously enhance floral development, using tomato [Lycopersicon esculentum Mill.] as a model. The seedlings with fully expanded cotyledons were grown for 15 d at a PPF of 150 μmol·m–2·s–1, a relative humidity of 70%, and a CO2 concentration of about 380 μmol·mol–1 (atmospheric standard). Leaf chlorosis was observed when the air temperature was constant regardless of average air temperature (16, 22,or 28 °C). Neither leaf chlorosis nor necrosis was observed when the air temperatures were alternated [periods of high (28 °C) and low (16 °C) air temperatures of 16/8, 12/12, and 8/16 h·d–1]. Faster floral development was observed in the seedlings grown at lower average air temperatures. These results indicated that physiological disorders of tomato seedlings grown under continuous light could be avoided, and at the same time floral development could be enhanced, by lowering the average air temperature through modification of the periods of high and low air temperatures.

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Potato (Solanum tuberosum L. cv. Benimaru) plantlets were cultured under four lighting cycles (photoperiod/dark period: 16 h/8 h, 4 h/2 h, 1 h/0.5 h, and 0.25 h/0.125 h) photoautotrophically (without sugar in the medium), and photomixotrophically (with sugar in the medium) in vitro for 28 days. Simulations of time courses of CO2 concentration in the vessel (Ci) and dry weight accumulation of the plantlets cultured photoautotrophically were conducted using a previously developed model (Niu and Kozai, 1997). While underestimation and overestimation of time courses of Ci in some treatments were observed, the simulated results of Ci and dry weight accumulation of the plantlets generally agreed with the measured ones. The difference of net photosynthetic rate response to Ci throughout the culture period was examined between the plantlets cultured photoautotrophically and photomixotrophically. Quantitative relationship between daily net photosynthetic rate (daily net production) and vessel ventilation rate per plantlet was simulated under various CO2 levels outside the vessel for given sizes of potato plantlets cultured photoautotrophically in vitro to aid appropriate CO2 enrichment and vessel design in commercial micropropagation.

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