Effects of Salinity on the Extensibility and Ca Availability in the Expanding Region of Growing Barley Leaves

The growth of barley (Hordeum vulgare L.) leaves is reduced by salinity. We used the Instron extensometric technique to measure the reversible and irreversible compliance of the expanding regions of growing barley leaves from plants exposed to 1, 40, 80 and 120 mM NaCl in nutrient solution. Two barley cultivars differing in salinity resistance (cv ‘Arivat’ and cv ‘Briggs’) were compared over 5d of leaf growth. During the period of most active leaf expansion, salinity reduced reversible compliance and increased compliance in the leaf segments, although responses to salinity were complex and changed over the course of leaf expansion. Salinity increased irreversible compliance more in the salt-sensitive cultivar Arivat than in the more salt-tolerant cultivar Briggs. Elemental analysis of the basal leaf segments used for extensometry revealed an accumulation of Na and a depletion of Ca in segments from salinized plants, resulting in very high Na: Ca ratios in salinized expanding tissue. The concentrations of K and Mg in basal leaf tissue were elevated by salinity. Our data do support the hypothesis that the inhibition of leaf expansion by salinity stress is mediated by a decline in irreversible extensibility. We suggest that reduced Ca availability in expanding leaf tissue may contribute to growth reduction in salt-stressed barley seedlings.     

Intracellular Localization of Jasmonate-Induced Proteins in Barley Leaves

The plant growth substance jasmonic acid and its methyl ester (JA-Me) induce a set of proteins (jasmonate-induced proteins, JIPs) when applied to leaf segments of barley (Hordeum vulgare L. cv. Salome). Most of these JIPs could be localized within different cell compartments by using a combination of biochemical and histochemical methods. Isolation and purification of various cell organelles of barley mesophyll cells, the separation of their proteins by one-dimensional polyacrylamide gel electrophoresis and the identification of the major abundant JIPs by Western blot analysis, as well as the immuno-gold labelling of JIPs in ultrathin sections were performed to localize JIPs intracellularly. JIP-23 was found to be in vacuoles, peroxisomes, and in the granular parts of the nucleus as well as within the cytoplasm; JIP-37 was detected in vacuoles and in the nucleoplasm; JIP-66 is a cytosolic protein. Some less abundant JIPs were also localized within different cell compartments: JIP-100 was found within the stromal fraction of chloroplasts; JIP-70 is present in the peroxisome and the nucleus; JIP-50 and JIP-6 accumulate in vacuoles. The location of JIP-66 and JIP-6 confirms their possible physiological role deduced from molecular analysis of their cDNA.     

Regulation of Magnesium Chelatase Activity during Excessive Accumulation of Porphyrins in Green Barley Leaves

Activity of magnesium chelatase was studied in green barley leaves treated with 5-aminolevulinic acid (ALA). After this treatment, leaves accumulated excessive amounts of porphyrinic precursors of chlorophyll : protoporphyrin IX (PP), magnesium-protoporphyrin IX (MgPP), its monomethyl ester (MgPPE), and protochlorophyllide. The enzyme activity was found to be inversely dependent on the amount of MgPP formed from exogenous ALA. A conclusion was drawn about the existence of a mechanism for the regulation of the enzyme activity in vivo via its inhibition by the reaction product.     

Protective Action of Abscisic Acid Against the Inhibition of Photosynthesis of Barley Leaves by Bisulphite

The inhibition of photosynthetic activity by bisulphite was studied in intact leaves of abscisic acid (ABA)-treated and non-treated (control) barley plants. ABA inhibited the photosynthetic process as evidenced by lower values of chlorophyll fluorescence kinetic parameters F_v/F_m (photosystem 2 activity) and R_fd (vitality index, related to the whole photosynthetic activity) compared with ABA-non-treated plants. After bisulphite treatment, the extent of inhibition was smaller in ABA-treated plants than in the control ones indicating a protective effect of ABA. The protective action sites of ABA were the Q_A reduction and the Calvin cycle. This revised version was published online in September 2006 with corrections to the Cover Date.     

The Effect of High Temperatures on the Photosynthetic Activity of Intact Barley Leaves at Low and High Irradiance

Light curves of CO₂ fixation by barley seedling leaves preliminarily heated at 30–43°C for 5 min were measured. The slope of the linear part of the light curve decreased after leaf heating at temperatures above 35°C; whereas, at a high light level, the photosynthesis rate decreased only at temperatures of 40°C and higher. The linear relationships between the photosynthetic CO₂-fixation rate and a photon flux density up to 1400 mol/(m² s) were found in leaves preheated at 42°C; this indicates the strong nonphotochemical dissipation of absorbed light quanta. The lowering of the oxygen concentration from 21 to 1% led to a CO₂ fixation maximum quantum yield and a photosynthesis-rate increase at the highest light intensity in leaves preheated at temperatures above 40°C as compared to the control leaves. Nevertheless, the linear relationship between the photosynthetic CO₂ fixation and the light intensity was found in leaves heated at 42°C at O₂ concentrations of both 21 and 1%. The latter fact suggests that the proton gradient of the thylakoid membrane, which causes an increase in the nonphotochemical dissipation of the quanta absorbed, could also be formed due to the cyclic electron transport over photosystem I.     

Interaction Between Root and Leaf Disease Development in Barley Cultivars after Inoculation with Different Isolates of Bipolaris sorokiniana

The relationship between root and leaf infection in 11 cultivars of barley ( Hordeum vulgare ) by different isolates of Bipolaris sorokiniana was investigated in young plants. Roots of 10-day-old seedlings, grown in filterpaper rolls, and the third leaf of 17-day-old seedlings were inoculated with the different isolates and a Disease Development Index (DDI) was calculated.
The rate of lesion development in leaves was higher than in roots, indicated by generally higher DDI after leaf inoculation than after root inoculation. Significant differences in resistance were found among the barley cultivars. Inoculation with different isolates of B. sorokiniana caused significant differences in DDI for both roots and leaves. In the leaves, but not in the roots, a significant cultivar–isolate interaction was found. No significant correlations, neither in isolate aggressiveness nor in cultivar reaction between root and leaf, were observed.     

Sensitivity of Resistance to Net Blotch in Barley

The aim of this study was to demonstrate various methods of analysing terminal net blotch, Pyrenophora teres Drechs. f. teres Smedeg., severity data from 15 spring barleys, Hordeum vulgare L., grown in Finnish official variety trials in five environments. The analyses have been developed and used principally by plant breeders for assessing crop yield, but lend themselves to use by plant pathologists. Pyrenophora teres is the major barley phytopathogen in Finland and improved resistance to it is sought. Joint regression analysis (JRA) and an additive main effects and multiplicative interaction (AMMI) model were used to investigate the data. Statistically significant genotype by environment (GE) interaction for resistance was indicated, and this included qualitative (crossover) interactions among genotypes over environments. A stable, non-sensitive, response to net blotch over environments, combined with a low mean score for terminal severity of the disease characterized the six-row barley 'Thule' which showed statistically significant crossover interaction only with 'Tyra'. 'Kustaa' exhibited the lowest mean terminal net blotch severity, but was relatively sensitive to net blotch. 'Arve' exhibited severe terminal net blotch in all environments, was relatively sensitive to environment and exhibited no crossover interaction with other genotypes. AMMI analysis appeared to represent a useful method for analysing these disease severity data, facilitating the selection of useful sources of resistance. Plots of AMMI-adjusted mean net blotch severities against first principal component axis (PCA) scores were informative for differentiating genotype response over environments, and are therefore potentially useful to plant pathologists and barley breeders seeking to gauge and subsequently improve the resistance status of barley to net blotch.     

A Simple Technique for Determining the Reaction of Barley Genotypes to Pyrenophora graminea

An in vitro technique was used to determine the reaction of 10 barley genotypes to Pyrenophora graminea, the seed‐borne pathogen causing barley leaf stripe disease. Determination was based on the percentage of inoculated seeds that produced fungal hyphae when cultured on potato dextrose agar. The technique allows low, intermediate and absolute levels of resistance to leaf stripe to be determined. Genotypes CI‐5791 and Banteng were resistant, Thibaut, Igri and PK (30‐531) were moderately resistant, Gollf was moderately susceptible, and WI2291, Arabi Abiad, Furat 1 and Arrivate were susceptible. The in vitro and in field assessments were significant (correlation coefficient r=0.96), results indicating that repeated measurements for infected seeds by this in vitro method were very similar to those of field assessments.     

Molecular Cloning of emip,a Member of the Major Intrinsic Protein (MIP) Gene Family,Preferentially Expressed in Epidermal Cells of Barley Leaves

Cloning and characterization of a barley gene named emip is reported that encodes a member of the major intrinsic protein family. A λ-unizap cDNA library synthesized from poly A⁺?mRNA of leaf epidermis was screened differentially with epidermis-versus mesophyll-derived probes. One of the clones epi 3-2 was sequenced and further analyzed. The open reading frame of the full length clone codes for a polypeptide of 288 amino acids with a molecular mass of 30,634 Da exhibiting a high degree of homology with members of the major intrinsic protein family. Hydropathy analysis predicts six potential membrane-spanning helices. mRNA levels were high in the growing zone of barley leaves and declined towards the tip of the fully expanded leaf blade. Expression was high in epidermal strips, lower in roots and very low in the leaf mesophyll. In order of decreasing response, wilting, salt shock and heat shock resulted in stimulated expression. mRNA levels remained low during slow salting up experiments. The expressional pattern suggests a role of EMIP in turgor regulation, particularly under stress.     

Two-Dimensional Electrophoretic Analysis of Salicylic Acid-Induced Changes in Polypeptide Pattern of Barley Leaves

The salicylic acid-induced changes in the polypeptide patterns of barley (Hordeum vulgare L.) leaves have been analysed using two-dimensional gel electrophoresis. An optimized 2-D PAGE protocol was used and gave reproducible 2-D gels from leaf crude protein extracts with a high number of detected polypeptides. When applied for 24 h SA affected the expression of a number of soluble proteins. Most of them appeared to be down-regulated. Although no abundant expression of specific proteins was observed, we detected three polypeptides that were present only in SA-treated leaves.     

Formation of the Pigment Apparatus in Etiolated Barley Leaves under the Influence of Levulinic Acid

The effects of levulinic acid (LA) on the synthesis of pigments and the membrane system of etioplasts were studied in etiolated leaves of barley (Hordeum vulgare L.). Growing in the solution of LA during a six-day period, which started one day after the soaking of seeds, resulted in a retardation of leaf growth, more than a twofold decrease in the level of carotenoids and protochlorophyllide (Pd) in the leaf tissue, and suppression of the synthesis of long-wave form of Pd₆₅₅; these effects depended on the LA concentration. In etioplasts isolated from the seedlings treated with 50 M LA and containing predominantly a short-wave form of Pd with a peak of fluorescence at 632 nm (–196°C), there was a membrane fraction whose location in the sucrose density gradient was identical to that of prolamellar bodies (PLB) in the control plants. The content of Pd and carotenoids in this fraction calculated on a protein basis was 2.46 and 1.3 times lower than in control seedlings, while the relative content of Pd oxidoreductase (POR) essentially did not change. Thus, the suppression of Pd synthesis did not affect translocation of POR from the cytoplasm to the membranes of etioplasts. In the PLB membranes, there was no transfer of energy from the molecules of lipophilic fluorescent probe pyrene (excitation at 337, 278, and 296 nm) to Pd; however, under pigment deficiency, the production therein of pyrene excimer form at the expense of energy transfer from protein tryptophanyls (excitation at 278 and 296 nm) became more efficient, which indicated changes in protein–lipid interactions. The obtained results suggest that the short-wave form of Pd₆₃₂ accumulating in etioplasts under the suppressed synthesis of tetrapyrroles is not a free pigment.     

Effect of Elevated Temperatures on the Activity of Alternative Pathways of Photosynthetic Electron Transport in Intact Barley and Maize Leaves

The light-induced rise in chlorophyll fluorescence and the subsequent decay of fluorescence in darkness were measured in barley and maize leaves exposed to heat treatment. The redox conversions of the photosystem I primary donor P700, induced by far-red light, were also monitored from the absorbance changes at 830 nm. After heating of leaves at temperatures above 40°C, the ratio of variable and maximum fluorescence decreased for leaves of both plant species, indicating the inhibition of photosystem II (PSII) activity. A twofold reduction of this ratio in barley and maize leaves was observed after heating at 45.3 and 48.1°C, respectively, which suggests the higher functional resistance of PSII in maize. The amplitude of the slow phase in the dark relaxation of variable fluorescence did not change after the treatment of barley and maize leaves at temperatures up to 48°C. In leaves treated at 42 and 46°C, the slow phase of dark relaxation deviated from an exponential curve. The relaxation kinetics included a temporary increase in fluorescence to a peak about 1 s after turning off the actinic light. Unlike the slow component, the fast and intermediate phases in the dark relaxation of variable fluorescence disappeared fully or partly after the treatment of leaves at 46°C. The photooxidation of P700 in heat-treated leaves was saturated at much higher irradiances of far-red light than in untreated leaves. At the same time, the dark reduction of P700⁺ was substantially accelerated after heat treatment. The data provide evidence that the heating of leaves stimulated the alternative pathways of electron transport, i.e., cyclic transport around photosystem I and/or the donation of electrons to the plastoquinone pool from the reduced compounds located in the chloroplast stroma. The rate of alternative electron transport after the heat treatment was higher in maize leaves than in barley leaves. It is supposed that the stimulation of alternative electron transport, associated with proton pumping into the thylakoid, represents a protective mechanism that prevents the photoinhibition of PSII in leaves upon a strong suppression of linear electron transport in chloroplasts exposed to heat treatment.     

山西森林蔬菜资源及其开发利用研究

山西森林资源丰富,植物组成多样。通过野外调查和数据分析,山西森林蔬菜有426种,分为6类,隶属于92个科中的260个属,其中叶菜类多达215种,茎菜类最少为14种,6类森林蔬菜的数量由多到少依次为叶菜类>果菜类>花菜类>根菜类>菌藻类>茎菜类。在此基础上,评价了山西森林蔬菜的种类、特色、营养价值及药用价值等,并对其开发利用提出了建议。     

The Effect of Diuron and 5-Aminolevulinic Acid on the Maintenance of Stable Chlorophyll Content in Greening Barley Leaves

Chlorophyll (Chl) accumulation and delayed luminescence of PSII were compared in greening barley leaves pretreated and untreated with diuron (DCMU) in the etiolated state, and reactions of two photosystems were studied in the plastids isolated from the pretreated and untreated leaves. The effect of treatment in light of post-etiolated leaves after 40-h illumination with 5-aminolevulinic acid (ALA), on the content of Chl and its precursor, protochlorophyllide (PChld) was also studied. The pretreatment of etiolated leaves with DCMU did not affect the rate of greening and the stable level of Chl content in barley. ALA, when introduced to leaves after the termination of Chl accumulation, increased PChld, but not Chl level. We suppose that the primary cause of greening cessation in etiolated leaves is the inhibition and cessation of the synthesis of apoproteins of pigment–protein complexes. The exhaustion of binding sites for newly synthesized Chl molecules leads to their retention in the so-called retroinhibitory pool of Chl, thus resulting in the inhibition of ALA synthesis by a negative feedback mechanism.     

The Pool of Chlorophyllous Pigments in Barley Seedlings of Different Ages under Heat Shock and Water Deficit

The effects of a high temperature (3 h, 40°C) and water deficit (45 h on 3% PEG 6000) on the pool of chlorophyllous pigments in the leaves of 4-, 7-, and 11-day-old barley (Hordeum vulgare L.) seedlings were studied. Heating resulted in a decrease in the total content of chlorophylls (Chl) (a + b) in 4-day-old plants but not in the older leaves. Water deficit induced an increase in the pigment content in young seedlings but reduced it in the leaves of 11-day-old plants. In young seedlings, hyperthermia and dehydration affected similarly Chl (a + b) degradation, leading to a marked inhibition of the chlorophyllase (Chlase) activity hydrolyzing Chl to chlorophyllides and phytol. In old leaves, an activation of this enzyme was observed. The stress factors under study affected different stages of pigment biosynthesis. High temperature inhibited the activity of dark and light stages of Chl(a + b) biosynthesis. Dehydration did not change markedly the resynthesis of protochlorophyllide, while the enzymes of the light stage of Chl biosynthesis were activated in young but inhibited in old barley leaves. The results thus obtained allowed us to conclude that heat treatment and dehydration specifically affected the Chl biosynthesis. At the same time, the Chlase response was nonspecific.     

The Ability of Barley Powdery Mildew to Grow in vitro

A technique was developed for the in vitro culture of Blumeria graminis f. sp. hordei , an obligate biotrophic pathogen of barley. Optimal growth occurred at pH 5.6 on a medium containing 39 gl^–1 potato dextrose agar, 40 gl^–1 shredded fresh barley leaves, 20 gl^–1 sucrose, 13 mgl^–1 kanamycin and 80 mgl^–1 benzimidazole. At 20°C (90% relative humidity), conidia germinated 48 h after inoculation, producing an average colony diameter of 1 cm after 10 days. However, numerous colonies were present on the medium after 15 days. Light microscopy showed that there was a positive relationship between the amount of leaf in the medium and fungus growth. The fungus retained its virulence during 60 days of storage in vitro , and was able to infect barley. This is a useful and novel technique that could be beneficial in barley pathology breeding programs.     

大麦花药培养中海藻酸钠作用的初步探讨

本试验以普通大麦（Hordeum vulgare L.）品种单核晚期的花药为材料,研究了他们在液体培养过程中海藻酸钠的作用。结果表明：在供试的4个海藻酸钠浓度（1、2、3、6%）中,以浓度为2-3%的效果最佳。当他们代替20-30%的Ficoll时,无论是用BAC3培养基,还是用MS培养基,培养基中加NAA,还是家2,4-D,都能诱导出愈伤组织,但以每升培养基加2mg NAA和1 mg 6-BA的效果最佳,在含这种激素的培养基上,一些基因型的出愈率相当高,供试基因型都在100%以上,而且有些花粉粒能直接发育成能正常发芽出苗的胚状体。Barley anthers at the late uninucleate stage were cultured in order to study the use of sodium alginate in liquidmedium.The results showed that, the effect of 2-3% (W/V) sodium alginate was best in 4 concentration studied (1%,2%,3%and 6%). When the 2-3% sodium alginate was substituted for 20-30% ficoll, the calli could be induced from anthers of all genotypes whether on BAC3 medium or MS medium and whether adding NAA or 2,4-Din medium.2mg/L NAA+1mg/L 6-BA added to medium could give better results and on the medium with these hormones the callus rate of all genotypes were above 100%. Some pollens could grow into embryoids directly.