Ethylene reduces gas exchange and growth of lettuce plants under hypobaric and normal atmospheric conditions

Elevated levels of ethylene occur in controlled environment agriculture and in spaceflight environments, leading to adverse plant growth and sterility. The objectives of this research were to characterize the influence of ethylene on carbon dioxide (CO₂) assimilation (C_A), dark period respiration (DPR) and growth of lettuce ( Lactuca sativa L. cv. Buttercrunch) under ambient and low total pressure conditions. Lettuce plants were grown under variable total gas pressures of 25 kPa (hypobaric) and 101 kPa (ambient) pressure. Endogenously produced ethylene accumulated and reduced C_A, DPR and plant growth of ambient and hypobaric plants. There was a negative linear correlation between increasing ethylene concentrations [from 0 to around 1000 nmol mol⁻¹ (ppb)] on C_A, DPR and growth of ambient and hypobaric plants. Declines in C_A and DPR occurred with both exogenous and endogenous ethylene treatments. C_A was more sensitive to increasing ethylene concentration than DPR. There was a direct, negative effect of increasing ethylene concentration reducing gas exchange as well as an indirect ethylene effect on leaf epinasty, which reduced light capture and C_A. While the C_A was comparable, there was a lower DPR in hypobaric than ambient pressure plants – independent of ethylene and under non-limiting CO₂ levels (100 Pa pCO₂, nearly three-fold that in normal air). This research shows that lettuce can be grown under hypobaria (≅25% of normal earth ambient total pressure); however, hypobaria caused no significant reduction of endogenous ethylene production.     

Ethylene reduces plant gas exchange and growth of lettuce grown from seed to harvest under hypobaric and ambient total pressure

Naturally occurring high levels of ethylene can be a problem in spaceflight and controlled environment agriculture (CEA) leading to sterility and irregular plant growth. There are engineering and safety advantages of growing plants under hypobaria (low pressure) for space habitation. The goals of this research were to successfully grow lettuce (Lactuca sativa cv. Buttercrunch) in a long-term study from seed to harvest under hypobaric conditions, and to investigate how endogenously produced ethylene affects gas exchange and plant growth from seed germination to harvest under hypobaric and ambient total pressure conditions. Lettuce was grown under two levels of total gas pressure [hypobaric or ambient (25 or 101 kPa)] in a long-term, 32-day study. Significant levels of endogenous ethylene occurred by day-15 causing reductions in photosynthesis, dark-period respiration, and a subsequent decrease in plant growth. Hypobaria did not mitigate the adverse ethylene effects on plant growth. Seed germination was not adversely affected by hypobaria, but was reduced by hypoxia (6 kPa pO₂). Under hypoxia, seed germination was higher under hypobaria than ambient total pressure. This research shows that lettuce can be grown from seed to harvest under hypobaria (≅25% of normal earth ambient total pressure).     

Separating the effects of hypobaria and hypoxia on lettuce: growth and gas exchange

The objectives of this research were to determine the influence of hypobaria (reduced atmospheric pressure) and reduced partial pressure of oxygen (pO2) [hypoxia] on carbon dioxide (CO2) assimilation (C(A)), dark-period respiration (DPR) and growth of lettuce (Lactuca sativa L. cv. Buttercrunch). Lettuce plants were grown under variable total gas pressures [25 and 101 kPa (ambient)] at 6, 12 or 21 kPa pO2)(approximately the partial pressure in air at normal pressure). Growth of lettuce was comparable between ambient and low total pressure but lower at 6 kPa pO2 (hypoxic) than at 12 or 21 kPa pO2. The specific leaf area of 6 kPa pO2 plants was lower, indicating thicker leaves associated with hypoxia. Roots were most sensitive to hypoxia, with a 50-70% growth reduction. Leaf chlorophyll levels were greater at low than at ambient pressure. Hypobaria and hypoxia did not affect plant water relations. While hypobaria did not adversely affect plant growth or C(A), hypoxia did. There was comparable C(A) and a lower DPR in low than in ambient total pressure plants under non-limiting CO2 levels (100 Pa pCO2, nearly three-fold that in normal air). The C(A)/DPR ratio was higher at low than at ambient total pressure, particularly at 6 kPa pO2- indicating a greater efficiency of C(A)/DPR in low-pressure plants. There was generally no significant interaction between hypoxia and hypobaria. We conclude that lettuce can be grown under subambient pressure ( congruent with25% of normal earth ambient total pressure) without adverse effects on plant growth or gas exchange. Furthermore, hypobaric plants were more resistant to hypoxic conditions that reduced gas exchange and plant growth.     

Germination and growth of lettuce (Lactuca sativa) at low atmospheric pressure

The response of lettuce ( Lactuca sativa L. cv. Waldmann's Green) to low atmospheric pressure was examined during the initial 5 days of germination and emergence, and also during subsequent growth to vegetative maturity at 30 days. Growth took place inside a 66-l-volume low pressure chamber maintained at 70 kPa, and plant response was compared to that of plants in a second, matching chamber that was at ambient pressure (approximately 101 kPa) as a control. In other experiments, to determine short-term effects of low pressure transients, plants were grown at ambient pressure until maturity and then subjected to alternating periods of 24 h of low and ambient atmospheric pressures. In all treatments the partial pressure of O₂ was maintained at 21 kPa (approximately the partial pressure in air at normal pressure), and the partial pressure of CO₂ was in the range 66.5–73.5 Pa (about twice that in normal air) in both chambers, with the addition of CO₂ during the light phase. With continuous exposure to low pressure, shoot and root growth was at least as rapid as at ambient pressure, with an overall trend towards slightly greater performance at the lower pressure. Dark respiration rates were greater at low pressure. Transient periods at low pressure decreased transpiration and increased dark respiration but only during the period of exposure to low pressure. We conclude that long-term or short-term exposure to subambient pressure (70 kPa) was without detectable detriment to vegetative growth and development.     

Seed production of Arabidopsis thaliana under hypobaric conditions.

In the present study, we examined carbohydrate contents of Arabidopsis thaliana seeds during seed development under hypobaric conditions in order to characterize the mechanism of low pressure-induced seed damage, and to determine critical pressures for seed development under low total and/or low oxygen partial pressures. We analyzed contents of starch, sucrose, glucose, and fructose in seeds at different developmental stages at 101 kPa total pressure with 21 kPa O2 partial pressure (control conditions), and at various low pressure conditions (23 kPa Ptot/21 kPa pO2, 101 kPa Ptot/2 kPa pO2, 53 kPa Ptot/2 kPa pO2, 23 kPa Ptot/2 kPa pO2, 12 kPa Ptot/10 kPa pO2, and 12 kPa Ptot/2 kPa pO2). Our results indicate that maintaining an adequate oxygen partial pressure inside the siliques is necessary for seed production under hypobaric conditions.     

Effects of elevated CO<Subscript>2</Subscript> and nitrogen on wheat growth and photosynthesis

The effects of nitrogen [75 and 150 kg (N) ha⁻¹] and elevated CO₂ on growth, photosynthetic rate, contents of soluble leaf proteins and activities of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and nitrate reductase (NR) were studied on wheat (Triticum aestivum L. cv. HD-2285) grown in open top chambers under either ambient (AC) or elevated (EC) CO₂ concentration (350 ± 50, 600 ± 50 μmol mol⁻¹) and analyzed at 40, 60 and 90 d after sowing. Plants grown under EC showed greater photosynthetic rate and were taller and attained greater leaf area along with higher total plant dry mass at all growth stages than those grown under AC. Total soluble and Rubisco protein contents decreased under EC but the activation of Rubisco was higher at EC with higher N supply. Nitrogen increased the NR activity whereas EC reduced it. Thus, EC causes increased growth and P_N ability per unit uptake of N in wheat plants, even if N is limiting.     

Effects of hypobaric growth conditions on morphogenic potential and antioxidative enzyme activities in Saussurea involucrata

Effects of reduced atmospheric pressure on morphogenic potential and antioxidative enzyme activities in regenerated tissues of Saussurea involucrata were evaluated. Leaf explants were cultured at atmospheric pressure 30, 60 or 101 kPa on Murashige and Skoog (MS) medium with several plant growth regulators (PGRs). Oxygen and carbon dioxide partial pressures were maintained at 21 and 0.038 kPa, respectively. At 60 kPa, 12 shoots per explant were recorded, which was 1.5 and 2.1-folds higher than at 101 and 30 kPa, respectively. A shooting frequency of 80 % was observed at 60 and 101 kPa. Rooted plantlets were obtained on MS medium with indoleacetic acid. At 30, 60 and 101 kPa, rooting of shoots was 49, 72 and 85.6 %, respectively. The rooted plantlets were successfully acclimatized to soil. Activities of all of antioxidative enzymes determined in present study were affected by hypobaric conditions.     

Bioluminescent enzyme assay for the indication of plant stress in enclosed life support systems

The application of bioluminescent sensors for monitoring key metabolites and enzymes that are indicators of stress in plants is demonstrated. The sensitivity of bioluminescent assay for NAD(P)H and NAD(P)(+) was about 0.5 and 1 nmol, respectively. The levels of NAD(P)H and NAD(P)(+) in radish (Raphanus sativus) root extracts from controls and from stress-induced conditions were compared. To induce environmental stress, the plants were grown in enclosed environmental chambers with low pressure (9 or 32 kPa), high humidity (>80%) and low oxygen partial pressure (down to 3.3-6.5 kPa). The concentrations of NAD(P)(+) and NAD(P)H in plants varied under stress conditions. Decreasing both total pressure from 101.5 to 32 or 9 kPa and partial pressure of oxygen increased the ratio of NAD(P)(+) /NAD(P)H from 0.2 to 4 or 6, respectively. The increase in this ratio suggests that plants are undergoing stress in these hypobaric environments. The developed bioluminescent assay for quantification of pyridine nucleotides in plant tissues is rapid, low-cost and easily performed.     

Impact of light-emitting diode irradiation on photosynthesis,phytochemical composition and mineral element content of lettuce cv. Grizzly

Light-emitting diodes (LEDs) are a promising technology with a potential to improve the irradiance efficiency, light quality, and the light spectrum for increasing plant yield and quality. In this experiment, we investigated the impacts of various LED light qualities, including 100% red, 100% blue, 70% red + 30% blue, and 100% white, on the growth and photosynthesis, phytochemical contents, and mineral element concentrations in lettuce (Lactuca sativa L. cv. ‘Grizzly’) in comparison to normal greenhouse conditions. Photon flux of 300 µmol m^?2 s^?1 was provided for 14 h by 120 LEDs set on a 60 cm × 60 cm sheet of aluminum platform in the growth chambers, where plants were grown for 60 d. Fresh mass per plant was significantly higher when grown under 100% blue and 70% red + 30% blue LEDs compared to the other environments including greenhouse conditions. Phytochemical concentrations and a nutritive value of lettuce were also significantly affected by the light treatments. Chlorophyll and carotenoid concentrations increased in the plants grown under 70% red + 30% blue LEDs compared to those grown in the greenhouse. Vitamin C content was 2.25-fold higher in the plants grown under 100% blue LEDs compared to those grown in the greenhouse. Higher photosynthesis and maximal quantum yield of PSII photochemistry were also observed in the plants treated with LED lights. The application of LED light led to the elevated concentrations of macro-and micronutrients in lettuce possibly because of the direct effect of LED light and lower stress conditions in the growth chambers compared to the greenhouse. Although the mechanism of the changes in lettuce grown under LED is not well understood, the results of this study demonstrated that LED light could be used to enhance the growth and nutritional value of lettuce in indoor plant production facilities.     

Effects of vapour pressure deficit on growth of temperate and tropical evergreen rainforest trees of Australia

Little is known about the effect of vapour pressure deficit (VPD) on the growth of trees. Rainforest trees of eastern Australia provide an opportunity to investigate responses to VPD in species that occur in high precipitation areas but have contrasting dry seasons—summer in the temperate south and winter in the tropical north. Growth responses to VPD were measured in eight species of Australian rainforest trees from different latitudes to investigate possible differences in their response to atmospheric drought. Previous work on these species found that the tropical species have large reductions in gas exchange with increasing VPD whereas the temperate species were mainly unresponsive to increasing VPD. Plants were grown in glasshouses for a year under either low VPD or ambient conditions of a temperate climate. All species had non-significant increases in growth rates (1–9%) of plants grown under low VPD compared with plants grown under ambient VPD. In addition, growing the species under low VPD had no effect on allocation of biomass (leaf area ratio, leaf weight ratio and root/shoot ratio). Therefore, the high sensitivity of gas exchange to increasing VPD found in the tropical rainforest trees did not have a significant, long-term effect on growth under high VPD.     

Physiological,elemental, and stable isotope responses of the organs of mungbean to reduced atmospheric pressure

The effects of hypobaric conditions on stable isotope and mineral element concentrations during the germination of mungbean [Vigna radiata (Linn.) Wilczek] were evaluated. Mungbean seeds were cultured in lower atmospheric pressure (60 kPa) and normal air pressure (101 kPa) conditions, respectively. Oxygen and carbon dioxide partial pressures were maintained at 21 and 0.04 kPa, respectively. At 60 kPa, the fresh weight (FW) and dry weight (DW) of plants significantly increased by 5.41 and 9.62%, respectively, compared to those at 101 kPa after culturing for 7 d. Twelve mineral elements were compared among three organs (leaf, stem, and root) from seedlings grown under hypobaric and normal atmospheric conditions. This showed that lower air pressure generally improved element accumulation in the plant. A significantly lower value of δ ¹³C was observed at 60 kPa compared to that at 101 kPa. In addition, a significant increase in δ ¹⁵N value was detected in three different organs of plants grown under 60 kPa. Our survey provides a foundation for future field and laboratory studies on the influence of air pressure on plants, particularly in terms of stable isotope and mineral elements.     

Heterologous expression of the mutated melon ethylene receptor gene Cm-ERS1/H70A produces stable sterility in transgenic lettuce (Lactuca sativa)

The mutated melon ethylene receptor gene Cm-ERS1/H70A was introduced into tobacco and induced stable sterility in transgenic lines. This gene contains a missense mutation that converts the His(70) residue to Ala in the melon ethylene receptor gene Cm-ERS1. To test the applicability of this inducible sterility system to other plants, lettuce (Lactuca sativa) was transformed with the gene using Agrobacterium, and putative transformants containing Cm-ERS1/H70A were obtained. Thirteen randomly selected putative transformants were grown in a growth room under constant conditions, and seven of the lines showed sterility or significantly reduced fertility. DNA gel blot analysis confirmed the integration of the Cm-ERS1/H70A gene into the genomes of the putative transformants, and RT-PCR and protein gel blot analysis confirmed the expression of Cm-ERS1/H70A mRNA and protein in all of the transformants. Five transformants showing sterility or reduced fertility when grown in a growth room under constant conditions were randomly selected to be grown in an open-air greenhouse under various environmental conditions. All five showed stable sterility under the various conditions. These results suggest that Cm-ERS1/H70A can induce sterility in heterologous transgenic plants.     

Oxygen control of ethylene biosynthesis during seed development in Arabidopsis thaliana (L.) Heynh

An unforeseen side-effect on plant growth in reduced oxygen is the loss of seed production at concentrations around 25% atmospheric (50 mmol mol-1 O2). In this study, the model plant Arabidopsis thaliana (L.) Heynh. cv. 'Columbia' was used to investigate the effect of low oxygen on ethylene biosynthesis during seed development. Plants were grown in a range of oxygen concentrations (210 [equal to ambient], 160, 100, 50 and 25 mmol mol-1) with 0.35 mmol mol-1 CO2 in N2. Ethylene in full-sized siliques was sampled using gas chromatography, and viable seed production was determined at maturity. Molecular analysis of ethylene biosynthesis was accomplished using cDNAs encoding 1-aminocyclopropane-1-carboxylic acid (ACC) synthase and ACC oxidase in ribonuclease protection assays and in situ hybridizations. No ethylene was detected in siliques from plants grown at 50 and 25 mmol mol-1 O2. At the same time, silique ACC oxidase mRNA increased three-fold comparing plants grown under the lowest oxygen with ambient controls, whereas ACC synthase mRNA was unaffected. As O2 decreased, tissue-specific patterning of ACC oxidase and ACC synthase gene expression shifted from the embryo to the silique wall. These data demonstrate how low O2 modulates the activity and expression of the ethylene biosynthetic pathway during seed development in Arabidopsis.     

Very high CO2 reduces photosynthesis, dark respiration and yield in wheat

Although terrestrial CO2 concentrations, [CO2] are not expected to reach 1000 micromoles mol-1 for many decades, CO2 levels in closed systems such as growth chambers and glasshouses, can easily exceed this concentration. CO2 levels in life support systems in space can exceed 10000 micromoles mol-1 (1%). Here we studied the effect of six CO2 concentrations, from ambient up to 10000 micromoles mol-1, on seed yield, growth and gas exchange of two wheat cultivars (USU-Apogee and Veery-l0). Elevating [CO2] from 350 to 1000 micromoles mol-1 increased seed yield (by 33%), vegetative biomass (by 25%) and number of heads m-2 (by 34%) of wheat plants. Elevation of [CO2] from 1000 to 10000 micromoles mol-1 decreased seed yield (by 37%), harvest index (by 14%), mass per seed (by 9%) and number of seeds per head (by 29%). This very high [CO2] had a negligible, non-significant effect on vegetative biomass, number of heads m-2 and seed mass per head. A sharp decrease in seed yield, harvest index and seeds per head occurred by elevating [CO2] from 1000 to 2600 micromoles mol-1. Further elevation of [CO2] from 2600 to 10000 micromoles mol-1 caused a further but smaller decrease. The effect of CO2 on both wheat cultivars was similar for all growth parameters. Similarly there were no differences in the response to high [CO2] between wheat grown hydroponically in growth chambers under fluorescent lights and those grown in soilless media in a glasshouse under sunlight and high pressure sodium lamps. There was no correlation between high [CO2] and ethylene production by flag leaves or by wheat heads. Therefore, the reduction in seed set in wheat plants is not mediated by ethylene. The photosynthetic rate of whole wheat plants was 8% lower and dark respiration of the wheat heads 25% lower when exposed to 2600 micromoles mol-1 CO2 compared to ambient [CO2]. It is concluded that the reduction in the seed set can be mainly explained by the reduction in the dark respiration in wheat heads, when most of the respiration is functional and is needed for seed development.     

The effect of calcium on the production of ethylene and abscisic acid by fungus Fusarium culmorum and by wheat seedlings infected with that pathogen

Wheat seedlings infected and non-infected with Fusarium culmorum were grown on mediums with different content of calcium (0, 2, 4, 8 mM). It was found that the higher the content of calcium in the medium, the greater the amounts of ethylene produced in both non-infected and infected wheat seedlings, whereas the level of ABA in their tissues was decreased. Taking into consideration the fact that ethylene inhibits, whereas ABA stimulates the growth and development of Fusarium culmorum, it may be assumed that the production of ethylene increased under the influence of calcium and the decreased level of ABA in wheat seedlings causes the reduction growth and development of pathogen and as a result it lowers the degree of infection of wheat seedlings by this fungus. Thus, on the base of the obtained results it may be concluded that the inhibiting influence of calcium on injurying the wheat seedlings by Fusarium culmorum may be caused by the influence of this cation on the hormone balance in the plant.     

Effects of red light and ethylene on growth of etiolated lettuce seedlings

Low concentrations of ethylene inhibit hypocotyl elongation of etiolated lettuce seedlings (Lactuca sativa cv. Grand Rapids), whereas red light does not inhibit it. The plumular hook tightens in response to either ethylene or red light. A combination of these two factors gives an additive response. Red light has no effect on ethylene production and red light will cause hook closure even under hypobaric pressure which removes endogenous ethylene. This suggests that ethylene and red light act independently in causing hook closure.     

Trace gases generated in closed plant cultivation systems and their effects on plant growth.

Interactions between plants and trace gases, especially ethylene, were investigated from two different viewpoints; ethylene is toxic for plant growth, whereas the ethylene release rate of plants can be utilized as a plant growth indicator. When lettuce plants and shiitake mushroom mycelium were cultivated in closed chambers, ethylene concentration increased with time. Ethylene was released both from lettuce plant and from shiitake mushroom mycelium. Dioctyl phthalate (DOP) and Dibutyl phthalate (DBP) were detected, and these concentrations reached 3.7 ngL-1 for DOP and 2.4 ngL-1 for DBP 4 days after closing. Organic solvents such as xylene and toluene and organic siloxane were detected with GCMS. Visible injury was observed in lettuce plants cultivated in the chambers and it seemed to result from trace contaminants such as DOP, DBP, organic solvents, dimethylsiloxane polymer, and ethylene. In order to obtain basic data of ethylene evolution from plants, ethylene concentration in a closed chamber in which the plants were cultivated under a controlled environment (25 degrees C air temperature, 60-70% relative humidity, 250-300 micromoles m-2 s-1 photosynthetic photon flux density (PPFD)) was measured. Lettuce (Lactuca sativa L. cv. Okayama) released ethylene more than Brassica rapa var. pervidis, Brassica campestris var. communis, and Brassica campestris var. narinosa. Ethylene release rate of intact lettuce plant was highly correlated with plant growth parameters such as dry weight, leaf area and photosynthetic rate. Ethylene release rates of intact lettuce plant were affected by cultivation conditions such as ambient CO2 concentration, light intensity and light/dark period. Increase in ambient ethylene level influenced lettuce growth even at the concentration of 0.1 microliter L-1. The level of ethylene inhibited leaf expansion and slightly accelerated chlorophyll degradation. It did not affect photosynthesis and transpiration, and also little affected dry matter accumulation. Thus, ethylene release characteristics were clarified and an effect of ethylene on lettuce growth was revealed. These findings are useful for determination of a threshold level of ethylene and a capacity of ethylene removal system in CELSS. On the other hand, a possibility of plant growth diagnosis by measuring ethylene concentrations was evaluated. As a result, it became clear that the measurement of ethylene concentration in CELSS is one of the useful non-destructive measurement methods for plant growth diagnosis. Further research is needed to investigate the applicability of the method to environmental stresses other than Ni and Co in nutrient solution.     

Warming chambers stimulate early season growth of an arctic sedge: results of a minirhizotron field study

We examined the effects of passive open-top warming chambers on Eriophorum vaginatum production near Toolik Lake, Alaska, USA. During the 2002 growing season, chamber warming was consistent with the magnitude and seasonality observed in recent decades throughout northwestern North America. Leaf-growth rates were higher in late May and early June; maximum growth rates in each leaf cohort occurred earlier and peak biomass was observed 20 days earlier within the chambers. Consequently, plants within the chambers maintained more live leaf biomass during the period of highest photosynthetically active radiation. Annual leaf production within the chambers (21±2 mg tiller) was not significantly different than under ambient conditions (17±2 mg tiller) (P=0.2256) despite higher early-season growth rates. Root growth began earlier; growth rates were higher in late May and early June, and maximum growth rates occurred earlier within the chambers. Therefore, plants within the chambers maintained greater root biomass during what earlier studies have identified as a period of relatively high nutrient availability. Annual root production within the chambers (191±42 g m^–2) was not significantly different than under ambient conditions (119±48 g m^–2) (P=0.1979), although there was a trend toward higher production within the chambers. The tendency toward higher root production within the chambers is consistent with previous laboratory experiments and with the predictions of biomass allocation theory.     

Growth and Mitochondrial Respiration of Mungbeans (Phaseolus aureus Roxb.) Germinated at Low Pressure

Mungbean (Phaseolus aureus Roxb.) seedlings were grown hypobarically to assess the effects of low pressure (21-24 kilopascals) on growth and mitochondrial respiration. Control seedlings grown at ambient pressure (101 kilopascals) were provided amounts of O₂ equivalent to those provided experimental seedlings at reduced pressure to factor out responses to O₂ concentration and to total pressure. Respiration was assayed using washed mitochondria, and was found to respond only to O₂ concentration. Regardless of total pressure, seedlings grown at 2 millimoles O₂ per liter had higher state 3 respiration rates and decreased percentages of alternative respiration compared to ambient (8.4 millimoles O₂ per liter) controls. In contrast, seedling growth responded to total pressure but not to O₂ concentration. Seedlings were significantly larger when grown under low pressure. While low O₂ (2 millimoles O₂ per liter) diminished growth at ambient pressure, growth at low pressure in the same oxygen concentration was enhanced. Respiratory development and growth of mungbean seedlings under low pressure is unimpaired whether oxygen or air is used as the chamber gas, and further, low pressure can improve growth under conditions of poor aeration.     

Low-pressure Storage of Hibiscus Cuttings. Effect on Stomatal Opening and Rooting

Cuttings of Hibiscus rosa-sinensis L. cv. Moesiana were storedat 15 or 760 mmHg (2 or 101.3 kPa) for 3, 5, 8 or 11 weeks.No ethylene production could be detected regardless of treatment.No chlorophyll degradation, proline accumulation or respiratoryincrease were seen during storage. Stomata were open under lowpressure and closed under normal pressure. The open stomatacaused water loss from cuttings after removal from storage becausethey were unable to respond normally to low humidity. Rootingability decreased as storage time increased, but satisfactoryresults were obtained up to a storage period of five weeks. Adventitious rooting, cuttings, ethylene production, Hibiscus rosa-sinensis L., hypobaric storage, low-pressure storage, senescence, stomatal resistance