Experimental and mathematical simulation of plant growth promoting rhizobacteria and plant interaction under cadmium stress

Bacterial inoculants of the commercially available plant growth promoting rhizobacteria (PGPR) Arthrobacter mysorens 7, Flavobacterium sp. L30, and Klebsiella mobilis CIAM 880 were selected to obtain ecologically safe barley crop production on cadmium (Cd) polluted soils. All the PGPR immobilized 24–68% soluble cadmium from soil suspension. A. mysorens 7 and K. mobilis CIAM 880 were highly resistant to Cd and grew in up to 1 and 3 mmol CdCl₂ on DAS medium respectively. All PGPR were able to fix nitrogen (276–1014 nmol mg^–1 bacterial DW) and to produce indole acetic acid (IAA) (126–330 nmol mg^–1 bacterial DW) or ethylene (4.6–13.5 nmol bacterial DW). All the PGPR actively colonized barley root system and rhizosphere and significantly stimulated root elongation of barley seedlings (up to 25%), growing on soil containing 5 or 15 mg Cd kg^–1 of soil. Created in the simulation mathematical model confirms our hypothesis that PGPR beneficial effect on barley growing under Cd-stress is a complex process. One of mechanisms underlying this effect might be increase of bacterial migration from rhizoplane to rhizosphere, where PGPR bind soluble free Cd ions in biologically unavailable complex forms. Among the studied PGPR K. mobilis CIAM 880 was the most effective inoculant. Inoculation with K. mobilis CIAM 880 of barley plants growing on Cd contaminated soil (5 mg Cd kg^–1 of soil) under field conditions increased by 120% grain yield and 2-fold decreased Cd content in barley grain. The results suggest that the using K. mobilis CIAM 880 is an effective way to increase the plant yield on poor and polluted areas.     

Sugar beet and barley yields in relation to inoculation with N2-fixing and phosphate solubilizing bacteria

Recently, there has been a resurgence of interest in bioorganic fertilizers as part of sustainable agricultural practices to alleviate drawbacks of intensive farming practices. N₂-fixing and P-solubilizing bacteria are important in plant nutrition increasing N and P uptake by the plants, and playing a significant role as plant growth-promoting rhizobacteria in the biofertilization of crops. A study was conducted in order to investigate the effects of two N₂-fixing (OSU-140 and OSU-142) and a strain of P-solubilizing bacteria (M-13) in single, dual and three strains combinations on sugar beet and barley yields under field conditions in 2001 and 2002. The treatments included: (1) Control (no inoculation and fertilizer), (2) Bacillus OSU-140, (3) Bacillus OSU-142, (4) Bacillus M-13, (5) OSU-140 + OSU-142, (6) OSU-140 + M-13, (7) OSU-142 + M-13, (8) OSU-140 + OSU-142 + M-13, (9) N, (10) NP. N and NP plots were fertilized with 120 kg N ha^–1 and 120 kg N ha^–1 + 90 kg P ha^- for sugar beet and 80 kg N ha^–1 and 80 kg N ha^–1 + 60 kg P ha^–1 for barley. The experiments were conducted in a randomized block design with five replicates. All inoculations and fertilizer applications significantly increased leaf, root and sugar yield of sugar beet and grain and biomass yields of barley over the control. Single inoculations with N₂-fixing bacteria increased sugar beet root and barley yields by 5.6–11.0% depending on the species while P-solubilizing bacteria alone gave yield increases by 5.5–7.5% compared to control. Dual inoculation and mixture of three bacteria gave increases by 7.7–12.7% over control as compared with 20.7–25.9% yield increases by NP application. Mixture of all three strains, dual inoculation of N₂-fixing OSU-142 and P-solubilizing M-13, and/or dual inoculation N₂-fixing bacteria significantly increased root and sugar yields of sugar beet, compared with single inoculations with OSU-140 or M-13. Dual inoculation of N₂-fixing Bacillus OSU-140 and OSU-142, and/or mixed inoculations with three bacteria significantly increased grain yield of barley compared with single inoculations of OSU-142 and M-13. In contrast with other combinations, dual inoculation of N₂-fixing OSU-140 and P-solubilizing M-13 did not always significantly increase leaf, root and sugar yield of sugar beet, grain and biomass yield of barley compared to single applications both with N₂-fixing bacteria. The beneficial effects of the bacteria on plant growth varied significantly depending on environmental conditions, bacterial strains, and plant and soil conditions.     

Isolation of Components of the Peptide Antibiotic Virginiamycin and Breeding of Their Producer,Streptomyces virginiae

A method for chromatographic separation and quantitative determination of individual components of the antibiotic virginiamycin, produced by microbiological synthesis (Streptomyces virginiaestrain 147), is described. The components, M_1–2and S_1–5, were isolated from fermentation broth and identified by HPTLC and HPLC (the results obtained using the two methods correlate well with each other). The conditions of culturing of the producer and the composition of nutritive media were optimized. Using UV irradiation as a mutagenic factor, the producer was selected for increased level of synthesis of the antibiotic; this was achieved by inducing mutations that impart resistance to virginiamycin and meta-fluorophenylalanine, an analog of phenylalanine.     

Phospholipid metabolism and lysosomal enzyme secretion by leukocytes Effects of dibutyryl cyclic adenosine 3′ : 5′-monophosphate and ATP

The effects of dibutyryl cyclic adenosine 3′ : 5′-monophosphate and ATP on isotope incorporation into phospholipids and the release of β-glucuronidase into the extracellular medium were studied in polymorphonuclear leukocytes from guinea pig peritoneal exudates. Exogenous dibutyryl cyclic adenosine 3′ : 5′-monophosphate (0.1–1.0 mM) reduced β-glucoronidase release induced by cytochalasin B in the absence of inert particles. It selectively inhibited ³²P_i incorporation into phosphatidic acid and the phosphoinositides and the incorporation of myo-[2-³H]inositol into the phosphoinositides. Added ATP (0.1–1.0 mM), but not other nucleotides, was found to potentiate β-glucuronidase release provoked by cytochasin B, but it impaired the labeling of the phosphoinositides by myo-[2-³H]inositol. The mechanism of the inhibition of the isotope incorporation into these acidic phospholipids by the two nucleotides has not been defined. Dibutyryl cyclic adenosine 3′ : 5′-monophosphate at 2–4 mM concentration was not found to appreciably alter the incorporation of [γ-³²P]ATP into phosphatidic acid, phosphatidylinositol, diphosphoinositide, and triphosphoinositide.     

Estimation of residual N effect of faba bean and pea on two succeeding cereals using15N methodology

A field experiment was conducted using¹⁵N methodology to study the effect of cultivation of faba bean (Vicia faba L.), pea (Pisum sativum L.) and barley (Hordeum vulgare L.) on the N status of soil and their residual N effect on two succeeding cereals (sorghum (Sorghum vulgare) followed by barley). Faba bean, pea and barley took up 29.6, 34.5 and 53.0 kg N ha^–1 from the soil, but returned to soil through roots only 11.3, 10.8 and 5.7 kg N ha^–1, respectively. Hence, removal of faba bean, pea and barley straw resulted in a N-balance of about –18, –24, and –47 kg ha^–1 respectively. A soil nitrogen conserving effect was observed following the cultivation of faba bean and pea compared to barley which was of the order of 23 and 18 kg N ha^–1, respectively. Cultivation of legumes resulted in a significantly higher A_N value of the soil compared to barley. However, the A_N of the soil following fallow was significantly higher than following legumes, implying that the cultivation of the legumes had depleted the soil less than barley but had not added to the soil N compared to the fallow. The beneficial effect of legume cropping also was reflected in the N yield and dry matter production of the succeeding crops. Cultivation of legumes led to a greater exploitation of soil N by the succeeding crops. Hence, appreciable yield increases observed in the succeeding crops following legumes compared to cereal were due to a N-conserving effect, carry-over of N from the legume residue and to greater uptake of soil N by the succeeding crops when previously cropped to legumes.     

Analysis of Qualitative Contribution of Assimilatory and Non-Assimilatory De-Excitation Processes to Adaptation of Photosynthetic Apparatus of Barley Plants to High Irradiance

The adaptation of barley (Hordeum vulgare L. cv. Akcent) plants to low (LI, 50 µmol m^–2 s^–1) and high (HI, 1000 µmol m^–2 s^–1) growth irradiances was studied using the simultaneous measurements of the photosynthetic oxygen evolution and chlorophyll a (Chl a) fluorescence at room temperature. If measured under ambient CO₂ concentration, neither increase of the oxygen evolution rate (P) nor enhancement of non-radiative dissipation of the absorbed excitation energy within photosystem 2 (PS2) (determined as non-photochemical quenching of Chl a fluorescence, NPQ) were observed for HI plants compared with LI plants. Nevertheless, the HI plants exhibited a significantly higher proportion of Q_A in oxidised state (estimated from photochemical quenching of Chl a fluorescence, q_P), by 49–102 % at irradiances above 200 µmol m^–2 s^–1 and an about 1.5 fold increase of irradiance-saturated PS2 electron transport rate (ETR) as compared to LI plants. At high CO₂ concentration the degree of P stimulation was approximately three times higher for HI than for LI plants, and the irradiance-saturated P values at irradiances of 2 440 and 2 900 µmol m^–2 s^–1 were by 130 and 150 % higher for HI plants than for LI plants. We suggest that non-assimilatory electron transport dominates in the adaptation of the photosynthetic apparatus of barley grown at high irradiances under ambient CO₂ rather than an increased NPQ or an enhancement of irradiance-saturated photosynthesis.     

Responses of alfalfa and Barley to foliar application of N and P on a coal mine spoil

Summary To ensure adequate growth of plants on the highly impoverished and erodable surface mined lands, the application of N and P fertilizers by suitable methods is essential. In the present study, five growth chamber experiments were conducted to evaluate the relative efficacy of foliar and spoil application of N and P using alfalfa (Medicago sativa L. var. Erand) and barley (Hordeum vulgare L. var. Manker) as test crops on a freshly exposed coal mine spoil collected from western North Dakota. In general, barley responded to both N and P, but alfalfa mainly to P. Growth responses of barley to foliar or spoil-applied N+P were substantial and similar in magnitude. However, the yields were much higher when the plants received 3–4 sprays of 1.5–2.2% urea, with P supplied through the spoil. Increasing the number of 2.2% urea sprays from 1 to 3 increased the growth response from 40 to 243%. In another study, increasing the concentration of foliar-applied urea from 0 through 1% resulted in further increases in the dry weights of barley at all the levels of spoil-applied (0, 25, 75, 225 g/g) N.Foliar sprays of 0.5–1.0% NaH₂PO₄ increased the dry weights of alfalfa and barley by an average of 366% and 86%, respectively. However, the yield response of alfalfa to spoil-applied P (100 g/g) was as high as 782% compared to only 117% for barley. Alfalfa responded significantly to increasing concentrations of H₃PO₄ (0–0.3%) in foliar sprays only in the absence of spoil-applied P. With increasing rates of spoil-applied P, alfalfa yields increased steadily, but additional supply of P sprays caused leaf burning which intensified as the P concentration in sprays increased.The results of chemical analyses indicated that foliar applications were more effective than soil applications in increasing the concentration of N or P in the plants. Moreover, urea sprays increased the uptake of K, Zn, and Fe in barley, whereas spraying alfalfa with P compounds caused increases in its K and Fe content and decreases in those of Zn and Na. The results of these experiments indicated that the nutritional requirements of plants grown on coal mine spoils can be met through foliar fertilization as effectively as, or better than, through conventional soil fertilization methods.Presented at the Annual Meeting, American Society of Agronomy, Chicago, Illinois, Dec. 3–8, 1978.     

Studies on Bacillus licheniformis endo-\-1,3-1,4-d-glucanase: characterization and kinetic analysis

Summary A new purification procedure for endo-\-1,3-1,4-d-glucanase from Bacillus licheniformis is described. The secreted enzyme was purified both from B. licheniformis and from recombinant Escherichia coli harbouring the cloned gene by ion exchange chromatography on a CM-Sepharose matrix at pH 5.6. The mature enzyme was resistant to proteolysis by trypsin and chymotrypsin but it was slowly digested by protease V8. It showed a continuous trimming where no large-limit polypeptides were noticeable thus supporting a monodomain structure. Former appearing peptides have been assigned theoretically according to the protein sequence and predictive methods of accessible areas. Kinetic parameters for the hydrolysis of barley \-glucan and lichenan by measuring the net release of reducing sugars at the optimum pH (7.02) and temperature (55° C) are k _cat=3500 ±800 s^–1 (turnover number) and K _m=1.45±0.21 mg/ml for barley \-glucan and k _cat=3000±750 s^–1 and K _m=1.98±0.40 mg/ml for lichenan. Correspondence to: E. Querol     

The fate of15N-labelled organic nitrogen in sheep manure applied to soils of different texture under field conditions

The fate of nitrogen from¹⁵N-labelled sheep manure and ammonium sulfate in small lysimeters and plots in the field was studied during two growth seasons. In April 1991,¹⁵N-labelled sheep faeces (87 kg N ha^–1) plus unlabelled (NH₄)₂SO₄ (90 kg N ha^–1), and (¹⁵NH₄)₂SO₄ (90 kg N ha^–1) were each applied to three soils; soil 1 (100% soil + 0% quartz sand), soil 2 (50% soil + 50% quartz sand) and soil 3 (25% soil + 75% quartz sand). The lysimeters were cropped with spring barley (Hordeum vulgare L.) and undersown ryegrass (Lolium perenne L.). The barley crop recovered 16–17% of the labelled manure N and 56% of the labelled (NH₄)₂SO₄-N. After 18 months 30% of the labelled manure N and 65% of the labelled (NH₄)₂SO₄-N were accumulated in barley, the succeeding ryegrass crop and in leachate collected below 45 cm of soil, irrespective of the soil-sand mixture. Calculating the barley uptake of manure N by difference of N uptake between manured and unmanured soils, indicated that 4%, 10% and 14% of the applied manure N was recovered in barley grown on soil-sand mixtures with 16%, 8% and 4% clay, respectively. The results indicated that the mineralization of labelled manure N was similar in the three soil-sand mixtures, but that the manure caused a higher immobilization of unlabelled ammonium-N in the soil with the highest clay content. Some of the immobilized N apparently was remineralized during the autumn and the subsequent growth season. After 18 months, 11–19% of the labelled manure N was found in the subsoil (10–45 cm) of the lysimeters, most of this labelled N probably transported to depth as organic forms by leaching or through the activities of soil fauna. In unplanted soils 67–74% of the labelled manure N was recovered in organic form in the 0–10 cm soil layer after 4 months, declining to 55–64% after 18 months. The lowest recovery of labelled N in top-soil was found in the soil-sand mixture with the lowest clay content. The mass balance of¹⁵N showed that the total recovery of labelled N was close to 100%. Thus, no significant gaseous losses of labelled N occurred during the experiment.     

Foliar and root absorption of Na+ and Cl− in maize and barley: Implications for salt tolerance screening and the use of saline sprinkler irrigation

Above-canopy sprinkler irrigation with saline water favours the absorption of salts by wetted leaves and this can cause a yield reduction additional to that which occurs in salt-affected soils. Outdoor pot experiments with both sprinkler and drip irrigation systems were conducted to determine foliar ion accumulation and performance of maize and barley plants exposed to four treatments: nonsaline control (C), salt applied only to the soil (S), salt applied only to the foliage (F) and salt applied to both the soil and to the foliage (F+S). The EC of the saline solution employed for maize in 1993 was 4.2 dS m^–1 (30 mM NaCl and 2.8 mM CaCl₂) and for barley in 1994, 9.6 dS m^–1 (47 mM NaCl and 23.5 mM CaCl₂). The soil surface of all pots was covered so that in the F treatment the soil was not salinized by the saline sprinkling and drip irrigation supplied nutrients in either fresh (treatments C and F) or saline water (treatments S and F+S).Saline sprinkling increased leaf sap Na⁺ concentrations much more than did soil salinity, especially in maize, even though the saline sprinkling was given only two or three times per week for 30 min, whereas the roots of plants grown in saline soil were continuously exposed to salinity. By contrast, leaf sap Cl^– concentrations were increased similarly by saline sprinkling and soil salinity in maize, and more by saline sprinkling than saline soil in barley. It is concluded that barley leaves, and to a greater extent maize leaves, lack the ability to selectively exclude Na⁺ when sprinkler irrigated with saline water. Moreover, maize leaves selectively absorbed Na⁺ over Cl^– whereas barley leaves showed no selectivity. When foliar and root absorption processes were operating together (F+S treatment) maize and barley leaves accumulated 11–14% less Na⁺ and Cl^– than the sum of individual absorption processes (treatment F plus treatment S) indicating a slight interaction between the absorption processes. Vegetative biomass at maturity and cumulative plant water use were significantly reduced by saline sprinkling. In maize, reductions in biomass and plant water use relative to the control were of similar magnitude for plants exposed only to saline sprinkling, or only to soil salinity; whereas in barley, saline sprinkling was more detrimental than was soil salinity. We suggest that crops that are salt tolerant because they possess root systems which efficiently restrict Na⁺ and Cl^– transport to the shoot, may not exhibit the same tolerance in sprinkler systems which wet the foliage with saline water. ei]T J Flowers     

A constitutive cystatin-encoding gene from barley (Icy) responds differentially to abiotic stimuli

A barley cDNA clone encoding a cysteine proteinase inhibitor was characterized. The deduced amino acid sequence of this barley cystatin (Hv-CPI) contains the motif QXVXG conserved among members of the cystatin superfamily. The gene (Icy), located on chromosome 2, was expressed in embryos, developing endosperms, leaves and roots as assessed by northern blot analysis. Western blot analysis detected a slightly retarded band in leaves that was not present in roots or seeds. In these two organs a more precise location of Hv-CPI was done by immuno-histochemical analysis, with polyclonal antibodies raised against the recombinant CPI protein expressed in Escherichia coli. This protein efficiently inhibited papain (K _i 2.0×10^–8 M) and ficin (K _i 2.2×10^–8 M) and, to a lesser extent, chymopapain (K _i 1.6×10^–7 M) and was inactive against bromelain. The Icy mRNA expression in vegetative tissues increased in response to anaerobiosis, dark and cold shock (6 °C).these authors contributed equally to this work     

Gibberellin-like substances in root exudate of Vitis vinifera

Summary The levels of gibberellin (GA)-like activity in the root exudate of two seedless varieties of Vitis vinifera were examined by the barley endosperm assay, and compared with levels determined for other parts of the plant. That activity was due to GA-like substances was confirmed with dwarf-5 corn.When acidic, ethyl acetate soluble GA-like substances from sap and leaf extracts were chromatogrammed on thin layers of silica gel in chloroform/ethyl acetate/formic acid (50:50:1), activity moved to the same Rf as GA₃ and GA₁ (Rf 0.05–0.25). However, substances inhibitory to the barley endosperm assay were detected in both sap and leaf extracts. In the above solvent system the inhibitor(s) co-chromatogrammed with a GA₁/GA₇ mixture, and with abscisin II. The GA-like activity co-chromatogrammed with GA₃ on paper developed in isopropanol/ammonia/water (10:1:1).Calculations on the rate of gibberellin movement from the roots seemed to be compatible with levels of activity in the leaves, although these levels could also be a reflection of the general gibberellin level in the plant.The relevance of the findings is discussed.     

The substrate specificity of phosphoinositide phospholipase C in rat heart sarcolemma

In rat cardiac sarcolemmal membranes a phosphoinositide-specific phospholipase C (PLC) was found to be present. The enzyme hydrolysed exogenous [³H-]phosphatidylinositol 4,5-biphosphate ([³H-]PtdIns(4,5)P ₂) in an optimized assay mixture containing 15 leg SL protein, 100 mM NaCl, 1 mM free Ca²⁺,14 mM Na-cholate and 20 AM [³H-]PtdIns (4,5)P ₂ (400–500 dpm/gm-l) in 30 mM HEPES-Tris buffer (pH 7.0). The average specific activity was 9.14±0.55 nmol-mg^–1·2.5 min^–1. The addition of Mg²⁺ to the assay mixture did not change PLC activity but increased the relative amounts of dephosphorylated inositol products. In the absence of Na⁺ and at a low Ca²⁺ concentration (0.3 M), Mg²⁺ also enhanced the intraSL levels of PtdIns4P and PtdIns, and, moreover, inhibited PLC activity (IC₅₀0.07 mM). PtdIns4P seemd to be a good substrate for the rat SL PLC (23.07 ± 1.57 nmol·mg^–1·2.5 min^–1) whereas PtdIns was hydrolysed at a very low rate (0.36 ± 0.08 nmol·mg^–1·2.5 min^–1). Unlike PtdIns(4,5)P ₂, PLC-dependent PtdIns4P and PtdIns hydrolysis was not inhibited by Ca²⁺ concentrations over 1 mM. The possibility of distinct isozymes being responsible for the different hydrolytic activities is discussed. (Mol Cell Biochem116: 27–31, 1992).Abbreviations DAG sn-1,2-diacylglycerol - EGTA ethyleneglycol-O,O-bis(aminoethyl)-N,N,N,N,-tetraacetic acid - Ins(1,4,5)P ₃ inositol 1,4,5-trisphosphate - InsP inositol monophosphate (unidentified isomer) - InsP ₂ inositol bisphosphate (unidentified isomer) - InsP ₃ inositol trisphosphate (unidentified isomer) - InsP _x any inositol phosphate - PLC phospholipase C - PtdIns phosphatidylinositol - PtdIns(4,5)P ₂ phosphatidylinositol 4,5-bisphosphate - PtdIns4P phosphatidylinositol 4-monophosphate - SL sarcolemma     

RAPD-Based Analysis of Introgression of Barley Genetic Material into the Genome of Alloplasmic Wheat Lines (Hordeum geniculatumAll./Triticum aestivum L.)

Genomes of three alloplasmic wheat lines obtained on the basis of barley–wheat hybrid Hordeum geniculatumAll. (2n = 28) ×Triticum aestivumL. (2n = 42)(Pyrotrix 28) were examined using random amplified polymorphic DNA (RAPD) analysis. Line L-29 was obtained after first backcross of the initial hybrid with the wheat variety Pyrotrix 28 and ten subsequent self-pollinating generations. This line was represented by euploid plants with typical to the common wheat chromosome number (2n = 42), as well as by aneuploids, which contained an additional telocentric chromosome in the main karyotype (2n = 42 + t). Lines L-26 and L-27 were obtained by two backcrosses of one BC₁ plant with the wheat variety Novosibirskaya 67 and one subsequent self-polination of one BC₃ plant. Chromosome number in all these plants corresponded to 2n = 40 + 4t. RAPD analysis was carried out using seven primers, which were previously proved to be effective for identification of the barley genome fragments within hybrid genomes of alloplasmic lines. The presence of barley genome fragments in line L-29 was revealed by use of five primers, while in lines L-26 and L-27 these fragments were detected by use of one primer. The significant difference in the number of barley RAPD fragments in the genomes of alloplasmic lines obtained at different backcrossing stages suggests more intense displacement of barley genome during backcrossing compared to self-pollination in BC₁ plants.     

Characteristics of a photorespiratory mutant of barley (Hordeum vulgare L.) deficient in phosphogly collate phosphatase

A barley mutant RPr84/90 has been isolated by selecting for plants which grow poorly in natural air, but normally in air enriched to 0.8% CO₂. After 5 minutes of photosynthesis in air containing¹⁴CO₂ this mutant incorporated 26% of the¹⁴C carbon into phosphoglycollate, a compound not normally labelled in wild type (cv. Maris Mink) leaves.The activity of phosphoglycollate phosphatase (EC 3.1.1.18) was 1.2 nkat mg^–1 protein at 30°C in RPr 84/90 compared to 19.2 nkat mg^–1 protein in the wild-type leaves. Phosphoglycollate phosphatase activity was not detected after protein separation by electrophoresis of leaf extracts from the mutant on polyacrylamide gels; on linear 5% acrylamide gels three bands with enzyme activity were separated from extracts of wild type plants. Gradient gel electrophoresis followed by activity staining showed two bands in Maris Mink tracks of MW 86,000 and 96,000, but no bands in 84/90. This is the first report of isozymes of phosphoglycollate phosphatase in barley which were absent in the mutant extracts. Our results confirm an earlier report of isozymes of this phosphatase in Phaseolus vulgaris [18].The photosynthetic rate of RPr 84/90 in 1% O₂, 350 l CO₂ l^–1 was 9–12 mg CO₂ dm^–2 h^–1 at 20°C, whereas the wild-type rate was 27–29 mg CO₂ dm^–2 h^–1 at 20°C. In 21% O₂, 350 l CO₂ l^–1 the rate was 2–3 mg CO₂ dm^–2 h^–1 in the mutant and 20 mg CO₂ dm^–2 h^–1 in the wild type.Genetic analysis has shown that the mutation segregates as a single recessive nuclear gene.     

Specific Features of the Nuclear Genome Recombination in Backcross Progenies of Barley–Wheat Hybrids Hordeum vulgare L. (2n = 14) × Triticum aestivum L. (2n = 42)

The backcross progenies of the barley–wheat hybrids Hordeum vulgare L. (2n = 14) × Triticum aestivum L. (2n= 42) and two alloplasmic lines derived from them were studied using microsatellite markers of barley and wheat. The F₁ hybrids and first backcross plants BC₁ contained the genetic material of both cultivated barley and the cultivars of common wheat involved in developing of these hybrid genotypes. The genomes of BC₃, BC₄, and alloplasmic lines contained no microsatellite markers of the cultivated barley, whereas chromosomes of each homeologous group of common wheat were identified. In chromosomes of backcross progenies BC₃, BC₄, and alloplasmic lines yielded by backcrosses of hybrids and various common wheat cultivars, microsatellite markers of the parental wheat cultivars were shown to undergo recombination.     

Amino acid conjugates as metabolites of the plant growth regulator dihydrojasmonic acid in barley (Hordeum vulgare)

The biotransformation of [2-¹⁴C](±)9, 10-dihydrojasmonic acid (DJA) was studied in excised shoots of 6-day-old barley seedlings after 72 h. From the ethyl acetate extract, some minor metabolites were isolated and purified by DEAE-Sephadex A-25 chromatography, thin-layer chromatography (TLC), C₁₈-cartridges, and high-performance liquid chromatography (HPLC). The structural identification of these metabolites was performed by gas chromatography-mass spectrometry (GC-MS), circular dichroism (CD), and amino acid analysis, and the following amino acid conjugates were found:N-[(–)9,10-dihydrojasmonoyl]valine,N-[(–)9,10-dihydrojasmonoyl]isoleucine,N-[9,10-dihydrojasmonoyl]leucine,N-[11-hydroxy-9,10-dihydrojasmonoyl]valine,N-[11-hydroxy-9,10-dihydrojasmonoyl]isoleucine,N-[12-hydroxy-9,10-dihydrojasmonoyl]isoleucine; and the cucurbic acid-related compoundsN-{[3-hydroxy-2(4-hydroxypentyl)-cyclopent-1-yl]-acetyl}isoleucine andN-{[3-hydroxy-2(5-hydroxypentyl)-cyclopent-1-yl]-acetyl}isoleucine. The results suggest conjugation with isoleucine and valine, as well as preferential hydroxylation at position C-11 or hydrogenation at position C-6, as being important steps in the metabolism of (±)DJA in barley shoots.     

Barley Chromosome Identification Using Genomic in Situ Hybridization in the Genome of Backcrossed Progeny of Barley–Wheat Amphiploids [Hordeum geniculatum all. (2n = 28) × Triticum aestivum L. (2n = 42)] (2n = 70)

Genomic in situ hybridization (GISH) has been used to study characteristics of the formation of alloplasmic lines detected among self-pollinated backcrossed progeny (BC₁F₅–BC₁F₈) of barley–wheat amphiploids [Hordeum geniculatum All. (2n = 28) × Triticum aestivum L. (2n = 42)] (2n = 70). The chromosome material of the wild barley H. geniculatum has been shown to contribute to these lines. For example, fifth-generation plants (BC₁F₅) had genotypes (2n= 42w + 2g), (2n = 42w + 1g + 1tg), and (2n = 41w + 1g), where w is common wheat chromosomes, g is barley (H. geniculatum) chromosomes, and tg is the telocentric chromosome of wild barley. Beginning from theBC₁F₆ generation, alloplasmic telocentric addition lines (2n= 42 + 2tg) and (2n = 42 + 1tg) appear. This lines has been found cytogenetically unstable. The progeny of each of these cytological types include not only the (2n= 42 + 2tg) and (2n = 42 + 1tg) addition plants, but also plants with the monosomic (2n = 41 + 1tg) and the disomic (2n = 40 + 2tg) substitutions, as well as the (2n = 41 + 2tg) plants, which lack one wheat chromosome and have two telocentric barley chromosomes. It has been demonstrated that the selection for well-filled grains favors the segregation of telocentric addition lines (2n = 42 + 2tg) and (2n = 42 + 1tg).     

Brief pre- and post-irrigation sprinkling with fresh water reduces foliar salt uptake in maize and barley sprinkler irrigated with saline water

Brief pre- and post-irrigation sprinkling treatments using freshwater were tested to determine if these practices could reduce the uptake of salts through leaves when saline water is used to sprinkler irrigate crops. Maize and barley were sprinkler irrigated 2 to 3 times per week for 30 min with saline water (4.2 dS m^–1, 30 mmol L^–1 NaCl and 2.8 mmoles L^–1 CaCl₂ for maize and 9.6 dS m^–1, 47 mmoles L^–1 NaCl and 23.5 mmoles L^–1 CaCl₂ for barley) in separate experiments with plants grown in pots outdoors. The soil surface of all pots was covered to prevent salinization of the soil by the sprinkling water. One half of the sprinkled plants was grown in nonsaline soil to study the effects of pre-wetting and post-washing when ion uptake was primarily through leaves. The other half of the sprinkled plants was grown in soil salinized by drip irrigation, in order to evaluate the effects of pre-wetting and post-washing when Na⁺ and Cl^- uptake was through both leaves and roots.Post-washing with freshwater (5 min) reduced the leaf sap concentrations of Cl^- in saline-sprinkled plants from 56 to 43 mmol L^–1 in maize and from 358 to 225 mmol L^–1 in barley (averages for plants grown in nonsaline and saline soil). Na⁺ concentrations in leaf sap were reduced from 93 to 65 mmoles L^–1 (maize) and from 177 to 97 mmoles L^–1 (barley) by the post-washing. Pre-wetting had a small effect on ion uptake through leaves, the only significant reduction in seasonal means being in leaf Na⁺ concentrations for plants grown in nonsaline soil. Pre-wetting and post-washing, when combined, reduced leaf Cl^- concentrations to levels similar to those of nonsprinkled plants grown in saline soil; however, Na⁺ concentrations in leaves remained 3.5 times (maize) and 1.5 times (barley) higher than those of nonsprinkled plants. When pre-wetting and post-washing were not applied, sprinkled barley plants grown in saline soil had grain yields which were 58% lower than nonsprinkled plants grown in saline soil, but the reduction in grain yield was only 17% when the freshwater treatments were given. We conclude that a brief period of post-washing with freshwater is essential when saline water is employed in sprinkler irrigation. By comparison, the benefits from pre-wetting were small in these experiments. ei]T J Flowers     

Study of mitochondrial genomes of allopllasmic recombinant wheat lines constructed on the basis of barley-wheat hybrids

Using RFLP analysis with three probes homologous to specific regions of mitochondrial DNA genes and PCR analysis of the mitochondrial recombining-repeat-sequence 18S/5S region of cereals, five alloplasmic wheat lines of different origin and fertility expression were studied. These lines are self-pollinated progeny of BC₁-BC₄ generations of barley–wheat hybrids Hordeum geniculatum All. (2n = 28) × Triticum aestivum L. (2n = 42). It was found that recombinant alloplasmic lines characterized by partial fertility contain either maternal (barley) DNA fragments or maternal and paternal (wheat) DNA fragments simultaneously (heteroplasmy). In lines with stable expression of self-fertility, fragments of only paternal mitochondrial DNA were detected. It is assumed that in alloplasmic lines, there is the interrelation between the presence of definite fragments of the mitochondrial genome belonging to either parental type and fertility expression.Translated from Genetika, Vol. 41, No. 3, 2005, pp. 349–355.Original Russian Text Copyright © 2005 by Trubacheeva, Salina, Pershina.