Optimization of potassium for proper growth and physiological response of Houttuynia cordata Thunb.

Houttuynia cordata Thunb. is an edible herb with a variety of pharmacological activities, but only limited information is available about its response towards potassium supplementation. Sterile plantlets were cultured in media with different potassium levels, and parameters related to growth, foliar potassium, water and chlorophyll contents, photosynthesis, transpiration, H₂O₂ contents and antioxidative enzyme activities were determined after a month. Results showed that 1.28 mM potassium was the optimum for H. cordata as highest values of dry weight, shoot height, root length and number were obtained at this concentration. The optimum potassium concentration resulted in the maximum net photosynthetic rate which could be associated with the highest chlorophyll content rather than limited stomatal conductance. The supply of surplus potassium resulted in higher content of foliar potassium, but negatively correlated with the biomass. Both potassium starvation (0 mM) and high potassium (>1.28 mM) could lead to water loss through high transpiration rate and low water absorption, respectively, and resulted in H₂O₂ accumulation and increased activities of catalase and peroxidase, which suggested induction of oxidative stress. Moreover, H. cordata showed the minimum of H₂O₂ content and the maximum of superoxide dismutase activity on 1.28 mM potassium, implying its role in inducing tolerance against oxidative stress.     

Role of 5-aminolevulinic acid on growth,photosynthetic parameters and antioxidant enzyme activity in NaCl-stressed <Emphasis Type="Italic">Isatis indigotica</Emphasis> Fort.

5-aminolevulinic acid (ALA) is a key precursor for the biosynthesis of porphyrins such as heme and chlorophyll. ALA alleviates salinity stress damage in germinating seeds and improves seedling growth. Exogenous application of ALA at low concentrations has been shown to enhance salt tolerance in a number of plants. In the present study, we studied the effect of exogenous application of ALA on enhancing salt stress tolerance in Isatis indigotica Fort. (Anhui population as S1, Shanxi population as S2). A foliar application of 0, 12.5, 16.7, 25.0, and 50.0 mg/L ALA was given to the leaves of I. indigotica plants treated with 100 mmol/L NaCl. The fresh weight of leaves and roots; chlorophyll relative content (SPAD value); photosynthetic parameters, such as net photosynthetic rate (Pn), stomatal conductance (Gs), intercellular carbon dioxide concentration (Ci) and water use efficiency of the treated plants were determined. The third leaf of each treated plant was used to determine the activities of antioxidant enzymes. Superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), glutamate synthase (GOGAT), nitrate reductase (NR) activities and the malondialdehyde (MDA) content increased in response to 100 mmol/L NaCl in both S1 and S2 plants. However, the fresh weight of leaf and root, chlorophyll relative content, Pn, Gs, Ci decreased in response to salt stress in both S1 and S2 plants. In all foliar application of ALA in S1 plants, the MDA content, and the activities of SOD and POD were the highest in response to 50.0 mg/L foliar application of ALA. GOGAT and NR activities were the highest in response to 16.7 mg/L foliar ALA. Chlorophyll content and Pn were the highest in S1 plants treated with by 25.0 mg/L ALA. In S2 plants, plant fresh weight, chlorophyll relative content, SOD, CAT, NR activities and Pn treated with 16.7 mg/L ALA were higher than that of the control (CK0). POD, MDA, GOGAT activities in S2 plants treated with 25.0 mg/L ALA were the highest among all treatments. Thus, our results showed that the optimal concentration of ALA (16.7 ~ 25.0 mmol/L) increases the activity of antioxidant enzymes, which in turn helps to abate the damage caused by salt stress in I. indigotica seedlings. Furthermore, ALA also results in an increase in chlorophyll content, Pn and the activities of GOGAT and NR.     

Inhibition of estrogen signaling through depletion of estrogen receptor alpha by ursolic acid and betulinic acid from Prunella vulgaris var. lilacina

Extracts of Prunella vulgaris have been shown to exert antiestrogenic effects. To identify the compounds responsible for these actions, we isolated the constituents of P. vulgaris and tested their individual antiestrogenic effects. Rosmarinic acid, caffeic acid, ursolic acid (UA), oleanolic acid, hyperoside, rutin and betulinic acid (BA) were isolated from the flower stalks of P. vulgaris var. lilacina Nakai (Labiatae). Among these constituents, UA and BA showed significant antiestrogenic effects, measured as a decrease in the mRNA level of GREB1, an estrogen-responsive protein; the effects of BA were stronger than those of UA. UA and BA were capable of suppressing estrogen response element (ERE)-dependent luciferase activity and expression of estrogen-responsive genes in response to exposure to estradiol, further supporting the suppressive role of these compounds in estrogen-induced signaling. However, neither UA nor BA was capable of suppressing estrogen signaling in cells ectopically overexpressing estrogen receptor α (ERα). Furthermore, both mRNA and protein levels of ERα were reduced by treatment with UA or BA, suggesting that UA and BA inhibit estrogen signaling by suppressing the expression of ERα. Interestingly, both compounds enhanced prostate-specific antigen promoter activity. Collectively, these findings demonstrate that UA and BA are responsible for the antiestrogenic effects of P. vulgaris and suggest their potential use as therapeutic agents against estrogen-dependent tumors.     

RESPONSE OF BUSH BEAN EXPOSED TO ACID MIST

Bush bean plants (Phaseolus vulgaris L. cv. Contender) were treated once a week for six weeks with simulated acid mist at five pH levels ranging from 5.5 to 2.0. Leaf injury developed on plants exposed to acid concentrations below pH 3 and many leaves developed a flecking symptom similar to that caused by ozone. An adaxial, interveinal bleached area resembling SO₂ injury also developed on some trifoliate leaves at the low pH treatments. Microscopic observation of injured trifoliates indicated that the palisade cells were plasmolyzed and that the chloroplasts lost structural integrity. Reductions in plant weight and chlorophyll content were detected across the pH gradient. Seed and pod growth were reduced at some intermediate acid depositions even though no visible foliar injury developed. Foliar losses of nitrogen, calcium, magnesium and phosphorous increased with decreases in acid mist pH, whereas foliar potassium concentrations were unaffected by acid mist treatment.     

The relationship between transpiration and chlorophyll synthesis in etiolated squash cotyledons

Etiolated cotyledons of squash (Cucurbita moschata Duch. var.melonaeformis Makino cv. Tokyo) treated with kinetin, KCl orfusicoccin in the dark increased both transpiration in the darkand chlorophyll synthesis after exposure to light. The minimumperiod of kinetin treatment to stimulate transpiration was similarto that to accelerate chlorophyll synthesis. On the other hand,treatment of cotyledons with vaseline or abscisic acid markedlyinhibited both transpiration and chlorophyll synthesis, suggestingthat cytokinin-induced stimulation of chlorophyll synthesiscould be mediated through the effect on the stomatal opening. (Received December 7, 1977; )     

Studies on plant growth-regulating substances

The dwarfing of seedlings of Phaseolus vulgaris cv. Masterpiece by treatment with solutions of 4-chlorobenzyltributylammonium bromide (B₄) as a foliar spray or by application to the roots has been demonstrated. Dwarfing resulted from a reduction of internodal and petiolar lengths but there was no evidence of delayed leaf or flower development. A reduction in transpiration rate was accompanied by some protection against wilting and an increased ability to recover from this condition. No changes in transpiration coefficient were observed. The primary leaves of treated seedlings retained chlorophyll longer than those of control seedlings, but in other leaves, chlorophyll and carotenoid pigment levels tended to fall following treatment with B₄. Evidence was obtained that B₄ treatment delayed root nodule formation, an effect which could be associated with the antibacterial properties of quaternary ammonium salts.     

Separate and combined effects of silicon and selenium on salt tolerance of wheat plants

Soil salinity is the leading global abiotic stress which limits agricultural production with an annual increment of 10%. Therefore; a pot experiment was conducted with the aim to alleviate the salinity effects on wheat seedlings through exogenous application of silicon (Si) and selenium (Se). Treatments included in the study were viz. (Ck) control (no NaCl nor Si and Se added), only salinity (50 mM NaCl), salinity + Si (50 mM NaCl with 40 mM Si), salinity + Se (50 mM NaCl with 40 mM Se) and salinity + Si + Se (50 mM NaCl + 40 mM Si + 40 mM Si). The salt stress impaired the growth (root and shoot dry weight, root: shoot ratio, seedlings biomass), water relations, photosynthetic attributes, transpiration rate and chlorophyll contents of wheat seedlings. Nonetheless, the foliar application of Si and Se alone and in combination improved the growth, water relations, photosynthetic attributes, transpiration rate and chlorophyll contents of wheat seedlings under stressed conditions. Moreover, an increase in antioxidant enzyme activity and accumulation of osmo-protectants (proline, soluble protein and soluble sugar) was noted under stressed conditions, which was more pronounced in wheat seedling which experienced combined application of Si and Se. To conclude that, foliar application of Si alone mitigated the adverse effect of salinity, while the combined application of Si and Se was proved to be even more effective in alleviating the toxic effects of salinity stress on wheat seedlings.     

Modulation in growth,photosynthetic efficiency,activity of antioxidants and mineral ions by foliar application of glycinebetaine on pea (Pisum sativum L.) under salt stress

A pot experiment was carried out to explore the role of glycinebetaine (GB) as foliar spray foliar on two pea (Pisum sativum L.) varieties (Pea 09 and Meteor Fsd) under saline and non-saline conditions. Thirty-two-day-old plants were subjected to two levels 0 and 150 mM of NaCl stress. Salt treatment was applied in full strength Hoagland’s nutrient solution. Three levels 0, 5 and 10 mM of GB were applied as foliar treatment on 34-day-old pea plants. After 2 weeks of foliar treatment with GB data for various growth and physiochemical attributes were recorded. Rooting-medium applied salt (150 mM NaCl) stress decreased growth, photosynthesis, chlorophyll, chlorophyll fluorescence and soluble protein contents, while increasing the activities of enzymatic (POD and CAT) and non-enzymatic (ascorbic acid and total phenolics) antioxidant enzymes. Foliar application of GB decreased root and shoot Na⁺ under saline conditions, while increasing shoot dry matter, root length, root fresh weight, stomatal conductance (g _s), contents of seed ascorbic acid, leaf phenolics, and root and shoot Ca²⁺ contents. Of three GB (0, 5, 10 mM) levels, 10 mM proved to be more effective in mitigating the adverse effects of salinity stress. Overall, variety Pea 09 showed better performance in comparison to those of var. Meteor Fsd under both normal and salinity stress conditions. GB-induced modulation of seed ascorbic acid, leaf phenolics, g _s, and root Ca²⁺ values might have contributed to the increased plant biomass, reduction of oxidative stress, increased osmotic adjustment and better photosynthetic performance of pea plants under salt stress.     

Physiological and Metabolic Effects of 5-Aminolevulinic Acid for Mitigating Salinity Stress in Creeping Bentgrass

The objectives of this study were to determine whether foliar application of a chlorophyll precursor, 5-aminolevulinic acid (ALA), could mitigate salinity stress damages in perennial grass species by regulating photosynthetic activities, ion content, antioxidant metabolism, or metabolite accumulation. A salinity-sensitive perennial grass species, creeping bentgrass (Agrostis stolonifera), was irrigated daily with 200 mM NaCl for 28 d, which were foliar sprayed with water or ALA (0.5 mg L⁻¹) weekly during the experiment in growth chamber. Foliar application of ALA was effective in mitigating physiological damage resulting from salinity stress, as manifested by increased turf quality, shoot growth rate, leaf relative water content, chlorophyll content, net photosynthetic rate, stomatal conductance and transpiration rate. Foliar application of ALA also alleviated membrane damages, as shown by lower membrane electrolyte leakage and lipid peroxidation, which was associated with increases in the activities of antioxidant enzymes. Leaf content of Na⁺ was reduced and the ratio of K⁺/Na⁺ was increased with ALA application under salinity stress. The positive effects of ALA for salinity tolerance were also associated with the accumulation of organic acids (α-ketoglutaric acid, succinic acid, and malic acid), amino acids (alanine, 5-oxoproline, aspartic acid, and γ -aminobutyric acid), and sugars (glucose, fructose, galactose, lyxose, allose, xylose, sucrose, and maltose). ALA-mitigation of physiological damages by salinity could be due to suppression of Na⁺ accumulation and enhanced physiological and metabolic activities related to photosynthesis, respiration, osmotic regulation, and antioxidant defense.     

Short-term UV-B radiation effects on morphology,physiological traits and accumulation of bioactive compounds in Prunella vulgaris L.

Prunella vulgaris plants (full-bloom stage) were subjected to short-term (15 days) UV-B radiation in a growth chamber. UV-B radiation was effective at enhancing morphological and biomass characteristics and decreasing chlorophyll contents of P. vulgaris. Treatment of P. vulgaris with artificial UV-B radiation significantly increased peroxidase (POD), superoxide dismutase (SOD) and glutathione (GSH) activities compared to the control treatment. UV-B radiation significantly increased the levels of hydrogen peroxide (H₂O₂), malondialdehyde (MDA) and proline in leaves of P. vulgaris compared to those of control plants. In addition, the contents of total flavonoids, rosmarinic acid, caffeic acid and hyperoside significantly increased under UV-B radiation. The total phenolic levels also increased under UV-B treatment. These results demonstrated that short-term UV-B radiation can enhance production of secondary metabolites in P. vulgaris, resulting in increased spica yield compared to that of control plants.     

Microwave assisted extraction of ursolic acid and oleanolic acid from Ocimum sanctum

Present study deals with the microwave assisted extraction (MAE) of ursolic acid (UA) and oleanolic acid (OA) from Ocimum sanctum leaves. UA and OA have been reported to possess significant medicinal properties. Various experimental parameters such as selection of solvent, solvent composition, irradiation time, microwave power, solid to solvent ratio, preleaching time and number of cycles were investigated to optimize the extraction process. Under optimum conditions of irradiation time (3 min), microwave power (272 W), solid to solvent ratio (1:30), preleaching time (10 min), maximum UA and OA has been extracted in one extraction cycle with ethanol: water (80:20) as a solvent. Maximum 86.76 and 89.64% of UA and OA was extracted under above mentioned optimized experimental conditions. MAE was also compared with the batch and ultrasound assisted extraction (UAE) method. As compared to batch and UAE, higher extraction yield of these important phytochemicals have been obtained through MAE in only 3 min.     

Potassium salts influence the fidelity of mRNA translation initiation in rabbit reticulocyte lysates: unique features of encephalomyocarditis virus RNA translation.

It is widely assumed that in vitro translation of mRNA is more efficient in the presence of potassium acetate rather than KCl, that the optimum concentration of potassium acetate is higher than for KCl, and that uncapped RNAs exhibit a lower optimum salt concentration than capped mRNAs. When these assumptions were examined using several different mRNA species in four batches of rabbit reticulocyte lysate, some notable exceptions were found. The translation of encephalomyocarditis virus (EMCV) RNA exhibited a salt optimum unusually high for an uncapped mRNA, and was very much more efficient and accurate with KCl rather than potassium acetate. It was also unique in being strongly activated by low concentrations (5-10 mM) KSCN in the presence of 90 mM potassium acetate. For the translation of other uncapped RNAs (poliovirus RNA, cowpea mosaic virus (CPMV) M RNA and bacteriophage MS2 RNA) amino acid incorporation at the optimum potassium acetate level was significantly greater than could be achieved using KCl. However, KCl was found to be restrictive and potassium acetate permissive for the synthesis of abnormal products thought to arise from initiation at incorrect sites, with the result that KCl gave a product pattern closer to that observed in vivo. In the particular case of the reticulocyte lysate system, accurate translation therefore requires the use of KCl rather than potassium acetate, but the choice of salt was found to be less critical in cell-free extracts from HeLa or L-cells.     

Salvia suspension cultures as production systems for oleanolic and ursolic acid

Oleanolic acid (OA) and ursolic acid (UA) are triterpenic acids with diverse biological activities that are of interest to the pharmaceutical industry. To investigate the scope for producing these compound using cell suspension cultures of Salvia species, calli from Salvia officinalis, S. virgata and S. fruticosa were induced using several plant growth regulator combinations. Eleven lines were selected for suspension induction from a pool of calli. Six suspension cultures were established successfully and cultivated in the respiration activity monitoring system (RAMOS^®) to obtain online data on their growth kinetics and to establish appropriate sampling schedules for the determination of their OA and UA production. Based on their observed growth behaviour, OA and UA contents, and aggregation properties, one suspension culture from each studied Salvia species was selected for further optimisation. The µ_max values for these suspension cultures ranged from 0.20 to 0.37 day^?1, their OA and UA contents were greater than 1.3 and 1.2 mg g^?1, respectively, and they afforded maximum volumetric yields of 21.0 mg l^?1 for OA and 32.8 mg l^?1 for UA. These results will be useful in the development of a refined Salvia suspension-based process for OA and UA production.     

适合于悬浮培养的枇杷愈伤组织诱导及状态调控

以枇杷不同品种和不同器官为外植体,通过对愈伤组织诱导率、诱导状态、熊果酸（UA）和齐墩果酸（OA）含量的综合考虑,筛选适合于悬浮培养的枇杷愈伤组织,并通过调整2,4-D、6-BA浓度,同时加入酸水解酪蛋白（CH）、谷氨酰胺（Gln）、KCl等物质对花丝愈伤组织状态进行调控。结果表明：枇杷‘早钟6号’幼胚诱导的愈伤组织最适用于枇杷悬浮培养,愈伤诱导率为83.70%,愈伤组织嫩黄、疏松、颗粒明显,UA、OA含量分别为6.90、2.34 mg/g dw。在MS中附加2,4-D 6.0 mg/L、6-BA 0.8 mg/L、CH 200 mg/L、KCl 500 mg/L,花丝诱导的坚硬愈伤出现细软、嫩黄的愈伤组织,但多次继代后水渍化,状态不佳。     

5-Aminolevulinic acid improves photosynthetic gas exchange capacity and ion uptake under salinity stress in oilseed rape (Brassica napus L.)

Salinity is one of the major constraints in oilseed rape (Brassica napus L.) production. One of the means to overcome this constraint is the use of plant growth regulators to induce plant tolerance. To study the plant response to salinity in combination with a growth regulator, 5-aminolevulinic acid (ALA), oilseed rape plants were grown hydroponically in greenhouse conditions under three levels of salinity (0, 100, and 200 mM NaCl) and foliar application of ALA (30 mg/l). Salinity depressed the growth of shoots and roots, and decreased leaf water potential and chlorophyll concentration. Addition of ALA partially improved the growth of shoots and roots, and increased the leaf chlorophyll concentrations of stressed plants. Foliar application of ALA also maintained leaf water potential of plants growing in 100 mM salinity at the same level as that of the control plants, and there was also an improvement in the water relations of ALA-treated plants growing in 200 mM. Net photosynthetic rate and gas exchange parameters were also reduced significantly with increasing salinity; these effects were partially reversed upon foliar application with ALA. Sodium accumulation increased with increasing NaCl concentration which induced a complex response in the macro-and micronutrients uptake and accumulation in both roots and leaves. Generally, analyses of macro- (N, P, K, S, Ca, and Mg) and micronutrients (Mn, Zn, Fe, and Cu) showed no increased accumulation of these ions in the leaves and roots (on dry weight basis) under increasing salinity except for zinc (Zn). Foliar application of ALA enhanced the concentrations of all nutrients other than Mn and Cu. These results suggest that under short-term salinity-induced stress (10 days), exogenous application of ALA helped the plants improve growth, photosynthetic gas exchange capacity, water potential, chlorophyll content, and mineral nutrition by manipulating the uptake of Na⁺.     

The Effect of Calcium Chloride on Growth,Photosynthesis, and Antioxidant Responses of Zoysia japonica under Drought Conditions

Few attempts have been made to study the alleviating effects of signal molecules on zoysiagrass ( Zoysia japonica ) under drought stress. Calcium chloride has been shown to ameliorate the adverse effects of drought stress on many plants. It is necessary to investigate how to enhance drought tolerance of zoysiagrass using calcium chloride. The study elucidated the effects of calcium chloride on zoysiagrass under drought conditions by investigating the following parameters: biomass, chlorophyll (Chl) content, net photosynthetic rate (Pn), chlorophyll fluorescence, antioxidant enzymes, proline content, and malondialdehyde (MDA) content. Experimental conditions consisted of an aqueous CaCl₂ solution at 5, 10, and 20 mM sprayed on zoysiagrass leaves for 3 d, following by an inducement of drought conditions by withholding water for 16 d. Under drought conditions, all CaCl₂ pretreatments were found to increase the above-ground fresh biomass, as well as below-ground fresh and dry biomass. The resulting Chl (a, b, a+b) contents of the 5 and 10 mM CaCl₂ pretreatment groups were higher than those of the control. In the later stages of drought conditions, the chlorophyll fluorescence parameter Fv/Fm was higher in leaves treated with 10 mM CaCl₂ than in the leaves of the other two treatment groups. Zoysiagrass pretreated with 10 mM CaCl₂ possessed both the maximum observed Pn and antioxidant enzyme activities. Meanwhile, lower MDA and proline contents were recorded in the plants pretreated with 5 and 10 mM CaCl₂ under drought conditions. As a whole, the drought tolerance of zoysiagrass was improved to some extent by the application of a moderate calcium concentration.     

Senescence of attached bean leaves accelerated by sprays of silicone oil antitranspirants

During an investigation into the use of oil emulsions in foliar sprays, it was found that silicone oil emulsions accelerated the senescence of the primary leaves of bean (Phaseolus vulgaris) plants. It was shown that accelerated senescence was not a result of the reduced transpiration rates found in silicone-sprayed leaves. Furthermore, the silicone oil emulsions did not induce leakiness in plant cell membranes. The senescence-enhancing effect seems to be connected with the ability of the silicone oil emulsions to penetrate into the leaf interior.     

Effectiveness of cyanobacteria and green algae in enhancing the photosynthetic performance and growth of willow (<Emphasis Type="Italic">Salix viminalis</Emphasis> L.) plants under limited synthetic fertilizers application

The physiological response of plants to triple foliar biofertilization with cyanobacteria and green algae under the conditions of limited use of chemical fertilizers was investigated. Triple foliar biofertilization with intact cells of Microcystis aeruginosa MKR 0105, Anabaena sp. PCC 7120, and Chlorella sp. significantly enhanced physiological performance and growth of plants fertilized with a synthetic fertilizer YaraMila Complex (1.0, 0.5, and 0.0 g per plant). This biofertilization increased the stability of cytomembranes, chlorophyll content, intensity of net photosynthesis, transpiration, stomatal conductance, and decreased intercellular CO2 concentration. Applied monocultures augmented the quantity of N, P, K in plants, the activity of enzymes, such as dehydrogenases, RNase, acid or alkaline phosphatase and nitrate reductase. They also improved the growth of willow plants. This study revealed that the applied nontoxic cyanobacteria and green algae monocultures have a very useful potential to increase production of willow, and needed doses of chemical fertilizers can be reduced.     

Rapid solubility determination of the triterpenes oleanolic acid and ursolic acid by UV-spectroscopy in different solvents

Salvia triloba (Greek sage) has been used for the treatment of various diseases and contains two bioactive triterpene acids of major interest: oleanolic acid (OA) and ursolic acid (UA). The determination of the solubility of OA and UA in different solvents is a prerequisite to select the optimal solvent. The main goal of this work was to develop a quick method of predicting the solubility of OA/UA in different solvents to get a first indication of which solvents could be considered suitable for extraction from any plant material containing at least one of these triterpenes. A novel and simple ultra-violet spectroscopy method was developed for this purpose.The best solubilities were determined in THF, dioxane and n-butanol as well as in blends of dioxane and n-butanol.