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In this study, effects of leaf age (20 to 240 days), plant age (4, 8, and 14 months after deflasking), and various day/night temperature regimes (16 to 33 °C) on photosynthesis of Phalaenopsis amabilis L. Blume var. formosa Shimadzu (Phal. TS97) leaves were investigated. The diurnal net CO2 uptake in Phal. TS97 leaves was measured and integrated to obtain total net CO2 uptake, which represents photosynthetic efficiency in plants performing crassulacean acid metabolism (CAM). Under all conditions, Phal. TS97 leaves performed typical CAM photosynthesis and reached their highest net CO2 uptake rate, ≈6 μmol·m-2·s-1, after 3 to 4 hours in the dark under a 12-hour photoperiod. When grown under 30 °C day/25 °C night temperature, the total net CO2 uptake of leaf increased with maturation and was highest at 80 days old, 20 days after full expansion. The CAM photosynthetic capacity of mature leaves remained high after maturation and began to decline at a leaf age of 240 days. The trend was consistent with malate fixation but the highest nocturnal malate concentration was observed in 100-day-old leaves. Young leaves or leaves from small juvenile plants had higher daytime CO2 fixation compared to mature leaves or large plants, suggesting that Phal. TS97 leaves progressed from C3-CAM to CAM during the course of maturation. The second newly matured leaf from the top had the highest net CO2 fixation when the newest leaf was 8 cm in length. Although plant age did not influence total CO2 uptake in the leaf, photosynthetic efficiency of leaves in small younger plants was more sensitive to high light intensity, 340 μmol·m-2·s-1 photosynthetic photon flux. The day/night temperature of 32/28 and 29/25 °C resulted in the highest total net CAM CO2 fixation in vegetative Phal. TS97 plants than higher (33/29 °C) and lower temperatures (21/16 °C).
The dancing-lady orchid, Oncidesa Gower Ramsey, is an important cultivar for cut-flower production, but it has low pollen fertility in breeding programs. In this study, we compared the pollen germination in vitro, sporad type, and pollinia development of Oncsa. Gower Ramsey and a diploid species, Oncidium sphacelatum Lindl (one of its grandparents). In Oncsa. Gower Ramsey, the pollen germination in vitro was lower as compared with those in Onc. sphacelatum. In addition, the frequency of abnormal sporads in Oncsa. Gower Ramsey was higher than those in Onc. sphacelatum. In Oncsa. Gower Ramsey, the middle layer and the tapetum were disorganized before meiosis, and subsequently they degenerated at the early tetrad stage. In contrast, the middle layer and the tapetum of Onc. sphacelatum began to degenerate at the early tetrad stage and fully disappeared at the bicellular pollen stage. These results suggested that the abnormal meiosis caused by unbalanced genomes and the premature degeneration of the middle layer and the tapetum could probably result in the abnormal pollen development and the low fertility of Oncsa. Gower Ramsey.
Photosynthetic rate is reduced during midday in some crops; this phenomenon has been termed as midday depression (MD). Oncidium also suffers greatly from MD in the summer, resulting in reduced growth and poor flowering quality. Since high radiation usually accompanies high temperature midday in the summer, it is difficult to figure out the key factor that promotes MD. We investigated the photosynthetic activities of Oncidium Gower Ramsey in the following conditions: environment-controlled and nonenvironment-controlled. In a growth chamber that simulated field growth conditions, photosynthesis declined dramatically when the temperature was higher than 32 °C. Photosynthesis was also reduced when photosynthetically active radiation (PAR) exceeded the saturating point of Oncidium. Gower Ramsey, which is about 250 μmol·m-2·s-1. However, the reduction was slight when PAR was under 500 μmol·m-2·s-1. Daily photosynthetic patterns were changed when Oncidium Gower Ramsey was grown under different environments. By regression, we found that MD was not directly associated with PAR within the range of 0–400 μmol·m-2·s-1. By contrast, photosynthesis was significantly reduced when temperature was higher than 32 °C. This explains the observation of greater photosynthetic reduction and earlier occurrence of MD when Oncidium Gower Ramsey was grown in rain-shelter rather than in phytotron and growth chamber, since temperature in the rain-shelter was not controlled, while the others were controlled at 25 °C. When Oncidium Gower Ramsey was moved from 35 °C to 25 °C, the photosynthetic depression was relieved.
Phalaenopsis has become one of the most important potted plants around the world. Thus, we used a key commercial Phalaenopsis amabilis cultivar, TS97, as a model to determine the light requirements for maximal carbon fixation and photosystem II (PSII) efficiency in its leaves and to investigate the effects of low irradiance and daylength on photosynthesis and flower development. In mature ‘TS97’ leaves, the daily total CO2 uptake capacity and net acid fixation increased with increasing photosynthetic photon flux (PPF) and saturated at ≈200 μmol·m−2·s−1, whereas the fluorescence ratio values were significantly reduced to 0.68 to 0.75 above 325 μmol·m−2·s−1 PPF, indicating photoinhibition of PSII. Positive assimilation of the nocturnal CO2 uptake occurred at a very low PPF (less than 5 μmol·m−2·s−1), suggesting highly efficient use of light energy by ‘TS97’ plants. Leaves developed under 30 μmol·m−2·s−1 PPF exhibited lower light requirement of 125 μmol·m−2·s−1 PPF to reach maximal CO2 uptake, below which the daytime CO2 uptake declined dramatically. Under a 12-hour daylength, exposing the leaves to a low PPF for 4 hours at any time during the day did not affect the photosynthetic capacity in ‘TS97’ leaves, suggesting that 8 hours of optimal irradiance is required for high-level photosynthesis, whereas the 12-hour daylength resulted in a higher CO2 uptake rate and the daily total CO2 uptake than the 8-hour daylength. Moreover, the 12-hour daylength promoted earlier flower formation and higher flower count compared with the 6- to 8-hour daylengths. Longer daylengths neither accelerated flowering formation nor enhanced total flower count. In conclusion, 8 hours of saturating PPF at 200 μmol·m−2·s−1 and a 12-hour daylength are sufficient for maximizing photosynthesis and flower production in ‘TS97’ plants.
Changes in endogenous abscisic acid (ABA) concentrations were investigated in developing seeds and the pretreated seeds of Calanthe tricarinata, a hard-to-germinate terrestrial orchid. ABA concentration was as low as 2.16 to 2.26 ng·mg−1 fresh weight at the proembryo stage [60 to 90 days after pollination (DAP)] and then continuously increased to 11.6 ng·mg−1 fresh weight at 210 DAP. Seed maturation was accompanied by a dramatic decrease in water content and a prominent accumulation of protein and lipid bodies within the embryo proper. The optimum time for asymbiotic seed germination was obtained from immature seeds at 150 DAP. At this stage, the embryo proper reached its maximum size, and the seedcoat became dehydrated and gradually shrunk into a thin layer. By 180 DAP, seed germination declined sharply as seed approached maturity. Mature seeds pretreated with ultrasound (45 min), 1% NaOCl (45 to 60 min), or 1N NaOH (45 min) were effective in improving the germination percentage and lowering seed ABA concentrations. Our results suggest that high concentrations of endogenous ABA in orchid seeds may play a critical role in arresting embryo growth and in preventing seed germination.
This investigation documents the key anatomical features in embryo development of Cypripedium formosanum Hayata, in association with the ability of embryos to germinate in vitro, and examines the effects of culture media and seed pretreatments on seed germination. A better understanding of zygotic embryogenesis for the Cypripedium L. species would provide insights into subsequent germination events and aid in the in vitro propagation of these endangered species. In seeds collected at 60 days after pollination (DAP), soon after fertilization, no germination was recorded. The best overall germination was found at 90 DAP (≈70%), at which time early globular to globular embryos with a single-celled suspensors can be observed. After 135 DAP, the seeds germinated poorly. At this time the inner integument shrinks and forms a tight layer, which encloses the embryo, the so-called “carapace.” Using Nile red stain, a cuticular substance was detected in the carapace, which may play a role in the impermeability of the mature seed and may help the seeds survive in the stringent environment. At maturity (after 210 DAP), the embryo proper has an average size of eight cells along its length and six cells across the width. Lipids and proteins are the main storage products within the embryo. To improve seed germination, experiments were conducted to test the suitability of various media and pretreatments of seeds. When different media were used, except for the Harvais medium at 120 DAP, there was no significant difference in seed germination at three different developmental stages tested. Soaking mature seeds in 1% NaOCl or treating them with ultrasound may slightly increase the germination percentage. For seed germination, our results indicate that the timing of seed collection outweighs the composition of medium and the seed pretreatments.
Phalaenopsis flowers are prone to wilting under ethylene (C2H4) stress. 1-Methylcyclopropene (1-MCP) can protect Phalaenopsis flowers against ethylene injury. In this study, we determined the residual effect of 1-MCP and how it is affected by temperature. The efficacy of multiple applications of 1-MCP was also investigated. The residual effect of 1-MCP was determined by pretreating blooming Phalaenopsis amabilis plants with 0.8 μL·L−1 1-MCP for 8 hours on Day 0 followed by 2 μL·L−1 ethylene fumigation for 12 hours on designated days. Without 1-MCP pretreatment, flowers began to wilt within 2 days after exposure to ethylene. Duration of the residual protection of 1-MCP on P. amabilis was ≈6 to 8 days during summer in Taiwan. Lower temperatures after 1-MCP application prolonged protection times. The full protection times under day/night temperatures of 25/20, 20/15, and 15/13 °C were 4 to 8, 10 to 13, and 13 to 17 days, respectively. Furthermore, multiple applications of 1-MCP extended the duration of 1-MCP protection against ethylene. Three applications increased the residual protection of P. amabilis by 1-MCP to at least 24 days.