A hybridization strategy for certain coloration could be developed based on accurate histological information of parental material together with the knowledge of heritability of color and color intensity. A sample of 12 Anthurium species and hybrids were histologically examined for pigmentation in spathes using a new method employing vacuum infiltration of spathe tissue with polyethylene glycol (PEG) prior to cross-sectioning. PEG infiltration displaces intercellular air spaces between cells. This method greatly improved the clarity of the cross sections and consequently improved observations of spatial localization of anthocyanins and chloroplasts. This infiltration method accurately identified the spatial localization of pigments for future breeding reference, notably among Anthurium species.
Jennifer A. Ehrenberger* and Adelheid R. Kuehnle
Masooma Ali-Abmad and Harrison Hughes
Scanning electron microscopy was used to study stomatal function of grape (Vitis sp. `Valiant') plantlets grown in vitro, polyethylene glycoltreated (PEG) in vitro and greenhouse. Fully open stomata were observed in in vitro grown plants with large aperture (13.5μm) as compared to narrow stomatal opening and small aperture in PEG-treated (4.9pm) and greenhouse grown plants (3.2μm). Furthermore, stomates of persistent leaves initiated during in vitro culture remained fully open with large apertures (12.8μm) two weeks after transplanting in the greenhouse. In contrast, newly-formed leaves produced in the greenhouse from in vitro cultured plants showed narrow stomatal opening with small apertures (3.3μm). In vitro produced leaves exhibited rapid wilting followed by irreversible tissue damage and severe desiccation within three hours of transplantation into the greenhouse. However, PEG-treated plantlets showed a reduced stomatal opening with associated minimal stress when directly transferred into the greenhouse. Thus use of an osmotic agent, PEG, induced more normal stomata1 function as well as improved survival after transfer to the greenhouse.
Zhongchun Wang and Gary W. Stutte
Previous results showed that active sorbitol accumulation occurs under water stress. We tested the hypotheses that sorbitol accumulation is due to reduced sorbitol export from leaves or from increased synthesis of glucose to sorbitol. To test the hypotheses, 230 μl 14C-sucrose was introduced through the stems to detached `Jonathan' apple shoots which had either water stress or no stress. Following uptake of 14C-sucrose, 0% or 10% PEG was applied to shoots for 24 hours. The results showed that 73% of 14C-sucrose in non-stressed leaves was broken down within 1 hour and 44% was recovered in sorbitol. PEG initially stimulated the breakdown of 14C-sucrose to glucose and fructose, but further conversion to sorbitol was reduced. However, the percentage of 14C-sorbitol in mature leaves increased gradually in 10% PEG until it exceeded that of control at 24 hours. In contrast to mature leaves, young leaves and stems showed significantly less sorbitol under 10% PEG 24 hours after treatment. These results supported the hypothesis that sorbitol accumulation under water stress was due to the reduced sorbitol transport.
William J. Carpenter
Salvia (Salvia spendens, F. Sellow ex Roem & Schult.) seeds imbibed in distilled water at 6C for 6 days germinated earlier and with fewer days to 50% of total germination (T50) than non-imbibed seeds. Drying imbibed seeds for 1 to 5 days at 5C and 45% RH before sowing signficiantly reduced seed viability. Priming seeds in a hypertonic osmotic solution of aerated polyethylene glycol 8000 (PEG) at —0.8 MPa for 10 days at 15C improved germination of the three cultivars tested. In laboratory and plant growth chamber trials, seeds primed with PEG 8000 and nonprimed seeds had similar total germination at 20 and 25C, but primed seeds had significantly higher germination at 10, 15, and 30C. At 35C, PEG-primed seeds had 44% to 65% germination, while nonprimed seeds failed to germinate. Alternating 10 and 20C or 20 and 30C diurnally at 12-hr cycles did not increase total germination regardless of seed treatment. Seeds primed with PEG had lower T50 than nonprimed seeds at 10, 15, 20, 25, and 30C, with the largest difference at the most unfavorable temperatures for germination. Primed seeds stored at 5C for 1 to 16 weeks reduced total germination and the potential capacity for rapid germination.
Dariusz Swietlik, R. F. Korcak, and Miklos Faust
‘York Imperial’ apple seedlings (Malus domestica Borkh.) grown in nutrient solution cultures with decreased water potential to− 1.0 bar by polyethylene glycol (PEG) increased water consumption, photosynthesis rate (Pn), and stomatal conductance (Cs). High light preconditioning of the plants used in this experiment was probably the reason why− 1.0 bar water potential in the nutrient solution was not low enough to induce apple seedling responses typical of water-stressed plants. However, application of PEG stress (−1.0 bar), to K-sprayed (K2SO4, −0.5%) trees lowered seedling water consumption Pn, and Cs. Potassium sprays alone did not significantly affect water consumption, Pn or Cs. When the water potential of the nutrient solution of PEG stressed plants was further decreased to −2.5 bars, unsprayed trees started to wilt within 2 days while sprayed trees did not. It is proposed that earlier stomatal closure of K-sprayed trees when stressed, already at low level of water stress (−1.0 bar), prevented plant water depletion when stress level was increased. This in turn delayed commencement of plant wilting. Potassium sprays also increased root:shoot ratio and root K concentration in PEG-stressed plants. These responses of K-sprayed trees could also contribute to greater tolerance to higher levels of water stress.
Stephanie E. Burnett, Svoboda V. Pennisi, Paul A. Thomas, and Marc W. van Iersel
Polyethylene glycol 8000 (PEG-8000) was applied to a soilless growing medium at the concentrations of 0, 15, 20, 30, 42, or 50 g·L-1 to impose controlled drought. Salvia (Salvia splendens F. Sellow. ex Roem & Shult.) seeds were planted in the growing medium to determine if controlled drought affects morphology and anatomy of salvia. Polyethylene glycol decreased emergence percentage and delayed emergence up to 5 days. Stem elongation of salvia treated with the five lowest concentrations was reduced up to 35% (21 days after seeding), and salvia were a maximum of 53% shorter and the canopy was 20% more narrow compared to nontreated seedlings 70 days after seeding. These morphological changes were attributed to PEG-8000 mediated reduction in leaf water potential (Ψw). The growing medium Ψw ranged from -0.29 to -0.85 MPa in PEG-8000 treated plants, and plant height was positively correlated with Ψw 21 days after seeding. Stem diameter of PEG-treated seedlings was reduced up to 0.4 mm mainly due to reductions in vascular cross-sectional area. Xylem cross-sectional area decreased more than stem and phloem cross-sectional area. Polyethylene glycol 8000 reduced vessel element number, but not diameter.
Dariusz Swietlik, R. F. Korcak, and Miklos Faust
Low- and high-K pretreated ‘York Imperial’ apple seedlings (Malus domestica Borkh.) were grown in nutrient solution cultures. Addition of polyethylene glycol (PEG) to the nutrient solution to reduce water potential to −1.0 bar reduced water consumption, fresh weight, specific leaf weight (SLW), and leaf water potential and increased the amount of water consumed per unit of fresh weight gain. High-K pretreatment increased water consumption of unstressed seedlings but decreased water consumption of PEG-stressed plants. Daily sprays with 0.5% KCl applied in early afternoon had no effect on water consumption rate in apple seedlings. However, sprays probably induced wider stomatal opening, since K-sprayed trees had lower leaf water potential when measured at noon than unsprayed trees. This effect was not observed when water potential was measured in the morning (0800 hr). High-K plants had higher leaf water potential than low-K plants in the morning. Potassium pretreatment and PEG stress as well as K-sprays had numerous effects on plant mineral composition. The K-pretreatment or K-sprays did not alleviate the detrimental effects of PEG-induced water stress despite the effects of K-pretreatment and K-sprays on mineral composition and leaf water potential.
Charles F. Forney
1 Storage Physiologist. Contribution no. 2142. I thank K.B. McRae (statistician) for advice on experiment planning, statistical analysis, and presentation of results; Peg Dixon for technical assistance; and Bruce
Masooma Ali-Ahmad and Harrison Hughes
Scanning electron microscopic (SEM) studies and gravimetric analysis of in vitro cultured leaf surfaces showed reduced epicuticular wax (EW) structurally and quantitatively as compared to greenhouse plants. However, leaves of in vitro plantlets subjected to polyethylene glycol-treatment (PEG) showed an increase in quantitative and structural EW which was similar to that of greenhouse plants. Furthermore, leaves initiated during in vitro culture and which persisted, when transferred to the greenhouse, showed an increase in structural wax as well as in amount, 30 days after transplanting in the greenhouse. Similarly, leaves newly-formed in the greenhouse from in vitro cultured plants developed more dense crystalline structure and greater levels of wax than those leaves observed immediately after removal from culture. A correlation between density of structural EW and amount of EW were observed in in vitro cultured, PEG-treated in vitro cultured and greenhouse grown leaves.
Leigh E. Towill
Papaya shoot tips, obtained either from seedlings or from in vitro plants, survived liquid nitrogen (-196°C) exposure using a vitrification procedure. Vitrification is a technically simple method but requires large concentrations of cryoprotectants. These were added in two steps, first slow addition of dimethylsulfoxide (DMSO) and PEG-8000, and subsequent fast addition of ethylene glycol (PG). The final concentration before cooling was 40% EG, 7.8% DMSO, and 10% PEG-8000. Both rapid cooling and rapid warming rates were required. Differential scanning calorimetry (DSC) was used to determine that the external solution vitrified upon cooling. It could not be demonstrated by DSC that cells within the shoot-tip vitrified, but since both DMSO and EG rapidly permeate plant cells, vitrification within the cells seems a likely explanation for retention of viability.