A novel allele of the Arabidopsis phytochelatin synthase 1 gene conferring high sensitivity to arsenic and antimony

To identify genes involved in arsenite [As(III)] tolerance, we screened for As(III)-sensitive mutants using ethyl methanesulfonate (EMS)-mutagenized seeds of Arabidopsis thaliana. One mutant with high sensitivity to As(III) was isolated from the screening. It had a mutation in Glu⁵² of phytochelatin synthase 1 (AtPCS1), and introduction of the wild-type AtPCS1 gene rescued the phenotype, confirming that the mutant represented a new allele of atpcs1 (termed as cad1-7). The expression of AtPCS1 from cad1-7 and cad1-3, a second allele of atpcs1, complemented As-sensitive yeast, but not plants. Our study suggests that AtPCS1 activity is differentially regulated between plants and yeast.     

两种退耕还林模式对土壤微生物优势类群的影响

以两种退耕还林模式的桦木林地、苦竹林地和农耕地(对照)为对象,对各样地土壤微生物优势类群数量的季节性动态变化、垂直分布特征及其与土壤酶活性的相关性进行了研究。结果表明,土壤微生物的优势类群为微球菌属(Micrococcus)、芽孢杆菌属(Bacillus)、链霉菌属(Streptomyces)、游动放线菌属(Actinoplanes)、酵母菌(未定属)、木霉菌属(Trichoderma)这6类。两种退耕还林模式下土壤优势微生物类群数量存在差异,春、夏、秋三季均为苦竹林所占比例最大,冬季为桦木林最大。两种林地和农耕地土壤微生物优势类群数量季节性变化规律为微球菌和酵母菌在夏、冬季较大,春、秋季较少;芽孢杆菌在秋季最大,夏季最小;游动放线菌与木霉菌数量在四季中变化不大;链霉菌是夏季最高,冬季最少。各优势微生物类群数量的垂直分布特征表现为微球菌、游动放线菌、木霉菌随土壤层的加深其数量逐渐降低,芽孢杆菌、链霉菌数量则随土壤层的加深而逐渐增加,酵母菌随土壤层的加深其数量减少,但趋势不明显。两种林地对各优势微生物类群数量的根际效应为:链霉菌和木霉菌的R/S值大于1,酵母菌的R/S值小于1,微球菌和游动放线菌在桦木林的R/S值大于1而在苦竹林小于1。在优势微生物类群和土壤酶之间,微球菌与转化酶,芽孢杆菌与纤维素酶,木霉和游动放线菌与过氧化氢酶,酵母与脲酶呈显著正相关关系。     

Nitrogen use Rationalization and Boosting Wheat Productivity by Applying Packages ‎of Humic,Amino Acids,and Microorganisms

ABSTRACT

Integrated management of soil organic matter and nutritional status of crop plants is essential to sustain the production of organic farming systems. Thus, a 2–year field experiment was conducted to examine the effects of soil additions (192 kg N ha^–1, humic+192 kg N ha^–1, humic+144 kg N ha^–1 and humic+96 kg N ha^–1) and foliar applications (amino acids, Azotobacter+yeast, and amino acids plus Azotobacter+yeast) as various fertilizer resources on growth and yield of wheat. Results showed that humic+192 kg N ha^–1 × amino acids plus Azotobacter+yeast were the effective combination for producing the highest values of flag leaf area, total dry weight, tiller number m^–2, spike weight m^–2, and grain yield ha^–1. Under foliar application of amino acids plus Azotobacter+yeast, reducing N supply from recommended rate (192 kg N ha^–1) to 144 kg N ha^–1+ humic achieved higher values of all yield traits, with a saving of 25% of applied mineral nitrogen as well as enhancing nitrogen use efficiency.     

Suppression of rice blast,cabbage black leaf spot,and tomato bacterial wilt diseases by Meyerozyma guilliermondii TA-2 and the nature of protection

This study was conducted to evaluate the efficacy of yeast strain TA-2 for controlling rice blast, cabbage black leaf spot, and tomato bacterial wilt diseases. Microscopic and phylogenetic analyses based on rDNA-internal transcribed region (ITS) and rDNA-D1/D2 sequences indicated that yeast strain TA-2 is Meyerozyma guilliermondii. Pretreatment with TA-2 by soil drenching significantly reduced the severity of black leaf spot disease caused by Alternaria brassicicola and leaf blast disease caused by Magnaporthe oryzae. Symptom development of tomato bacterial wilt caused by Ralstonia solanacearum in both soil drench and needle inoculation tests was significantly reduced in TA-2-pretreated plants under soil drenching. Disease severity and R. solanacearum growth were significantly reduced in tomato plants pretreated with yeast culture, cell suspension, or culture filtrate of TA-2 under soil drenching. TA-2 does not produce antibiotics. The present study indicates that disease suppression is systemic, as the roots were treated with TA-2 and the pathogens were inoculated onto leaves or stems, thereby separating the two spatially. M. guilliermondii TA-2 could become a promising natural antimicrobial agent against rice blast, cabbage black leaf spot, and tomato bacterial wilt diseases and might be useful as an eco-friendly control measure, contributing to sustainable agriculture.     

Role of Hydrogen Peroxide Produced by Baker's Yeast on Dough Rheology

Baker's yeast, Saccharomyces cerevisiae, has a well-known effect on dough rheology during breadmaking. During a 3-hr fermentation, hydrogen peroxide (H₂O₂) produced by yeast (0.76%, fwb) increased from 1.09 to 2.32 μmol/g of flour. The spread test, a measure of a dough's rheological properties, showed that yeast had an effect on dough rheology similar to that of H₂O₂, an oxidant that makes flour-water dough more elastic. In additional experiments (spread test and H₂O₂ measurement), glucose oxidase, an enzyme that produces H₂O₂, gave results similar to those with yeast. The fact that catalase, an enzyme that destroys H₂O₂, reversed the rheological effect of added H₂O₂ but did not reverse the effect of either yeast or glucose oxidase suggested that either wheat flour contains an inhibitor to catalase or H₂O₂ was not the active component. A series of experiments, including use of defatted flour, remixing, and mixing dough under nitrogen, all indicated that catalase was inhibited by peroxides in the lipid fraction of flour. These results also suggested that H₂O₂ is responsible for the effects of yeast and glucose oxidase on dough.     

Changes in Phytate Content in Whole Meal Wheat Dough and Bread Fermented with Phytase‐Active Yeasts

The degradation of inositol hexakisphosphate (IP6) was evaluated in whole meal wheat dough fermented with baker's yeast without phytase activity, different strains of Saccharomyces cerevisiae (L1.12 or L6.06), or Pichia kudriavzevii with extracellular phytase activity to see if the degradation of IP6 in whole meal dough and the corresponding bread could be increased by fermentation with phytase‐active yeasts. The IP6 degradation was measured after the dough was mixed for 19 min, after the completion of fermentation, and in bread after baking. Around 60–70% of the initial value of IP6 in the flour (10.02 mg/g) was reduced in the dough already after mixing, and additionally 10–20% was reduced after fermentation. The highest degradation of IP6 was seen in dough fermented with the phytase‐active yeast strains S. cerevisiae L1.12 and P. kudriavzevii L3.04. Activity of wheat phytase in whole meal wheat dough seems to be the primary source of phytate degradation, and the degradation is considerably higher in this study with a mixing time of 19 min compared with earlier studies. The additional degradation of IP6 by phytase‐active yeasts was not related to their extracellular phytase activities, suggesting that phytases from the yeasts are inhibited differently. Therefore, the highest degradation of IP6 and expected highest mineral bioavailability in whole meal wheat bread can be achieved by use of a phytase‐active yeast strain with less inhibition. The strain S. cerevisiae L1.12 is suitable for this because it was the most effective yeast strain in reducing the amount of IP6 in dough during a short fermentation time.     

Limitations of colony morphology and antibiotic resistance in the identification of a Bradyrhizobium sp. (Lotus) strain in soil

Summary We investigated the reliability of antibiotic resistance and colony morphology of clones of a Bradyrhizobium sp. (Lotus) strain for strain identification in nodulation competitiveness experiments in soil. There was no difference in nodulation competitiveness between the wild type strain and each of five mutants resistant to streptomycin and spectinomycin at the time of their isolation from antibiotic-containing media. However, these mutants were significantly less competitive when tested 4 months later. The apparent instability of the newly isolated mutants and their subsequently decreased nodulation competitiveness show that mutants must be examined carefully after being allowed time to stabilize. Two clones of the Bradyrhizobium sp. (Lotus) strain that differed in colony morphology on yeast mannitol medium did not differ in antigenic properties, whole cell protein electrophoresis profiles, mean cell generation times in yeast mannitol medium, N₂-fixing ability, nodulation of Lotus pedunculatus in growth pouches, or in nodulation competitiveness. Both clones retained their colony morphology after numerous transfers on yeast mannitol agar over 3 years and after at least 6 months in soil. A limiting factor, which may restrict the use of colony morphology as a marker for strain identification in competition experiments, is the problem of detecting double-infected nodules when the small colony type comprises a relatively small portion of the total nodule population.     

Promotion of plant growth by an auxin-producing isolate of the yeast Williopsis saturnus endophytic in maize (Zea mays L.) roots

A plant-growth-promoting isolate of the yeast Williopsis saturnus endophytic in maize roots was found to be capable of producing indole-3-acetic acid (IAA) and indole-3-pyruvic acid (IPYA) in vitro in a chemically defined medium. It was selected from among 24 endophytic yeasts isolated from surface-disinfested maize roots and evaluated for their potential to produce IAA and to promote maize growth under gnotobiotic and glasshouse conditions. The addition of l-tryptophan (L-TRP), as a precursor for auxins, to the medium inoculated with W. saturnus enhanced the production of IAA and IPYA severalfold compared to an L-TRP-non-amended medium. The introduction of W. saturnus to maize seedlings by the pruned-root dip method significantly (P<0.05) enhanced the growth of maize plants grown under gnotobiotic and glasshouse conditions in a soil amended with or without L-TRP. This was evident from the increases in the dry weights and lengths of roots and shoots and also in the significant (P<0.05) increases in the levels of in planta IAA and IPYA compared with control plants grown in L-TRP-amended or non-amended soil. The plant growth promotion by W. saturnus was most pronounced in the presence of L-TRP as soil amendment compared to seedlings inoculated with W. saturnus and grown in soil not amended with L-TRP. In the glasshouse test, W. saturnus was recovered from inside the root at all samplings, up to 8 weeks after inoculation, indicating that the roots of healthy maize may be a habitat for the endophytic yeast. An endophytic isolate of Rhodotorula glutinis that was incapable of producing detectable levels of IAA or IPYA in vitro failed to increase the endogenous levels of IAA and IPYA and failed to promote plant growth compared to W. saturnus, although colonization of maize root tissues by R. glutinis was similar to that of W. saturnus. Both endophytic yeasts, W. saturnus and R. glutinis, were incapable of producing in vitro detectable levels of gibberellic acid, isopentenyl adenine, isopentenyl adenoside or zeatin in their culture filtrates. This study is the first published report to demonstrate the potential of an endophytic yeast to promote plant growth. This is also the first report of the production of auxins by yeasts endophytic in plant roots.     

Pizza Dough Differentiation by Principal Component Analysis of Alveographic,Microbiological, and Chemical Parameters

We contrast two types of pizza dough produced in Naples. One was prepared in accordance with pizza dough regulations drawn up by the Ente Nazionale Italiano di Unificazione (UNI). Different pizza doughs were studied using a set of parameters linked to mechanical, microbiological, and chemical properties. The pizza doughs were analyzed by alveographic tests following Approved Methods of AACC International. In particular, alveographic values of parameters ΔP_max, V_r, ΔP_max/V_r, E_r1, E_r2, E_rT, as well as the nondimensional ratios E_r1/E_rT and E_r2/E_rT were obtained. Microbial counts of bacteria and yeast were determined and the bacteria‐to‐yeast ratio was also calculated. Total titratable acidity (TTA) and pH were determined by standard methods. Principal component analysis (PCA) applied to some technological parameters (V_r, E_r1, E_r1/E_rT, bacteria‐to‐yeast ratios, pH, and TTA), preliminarily selected on the basis of the variance explained by the components, allowed us to discriminate typical pizza dough of Naples from the other types considered in the study, demonstrating that PCA could help recognize different types of pizza dough. Hence, this method could be used to reduce the empirical component of the pizza‐making process to produce pizza dough with standardized quality.     

Metabolite Analysis Allows Insight into the Differences in Functionality of 25 Saccharomyces cerevisiae Strains in Bread Dough Fermentation

During dough fermentation, yeast (Saccharomyces cerevisiae) changes the physical properties of the dough matrix. In this study, we investigate if different yeast strains have an impact on dough rheology and on the gas holding capacity of fermenting dough. Furthermore, we analyze whether observed differences are linked to the metabolite profiles of the yeast strains. More specifically, the impact of 25 yeast strains on dough spread, dough fermentation properties, and dough metabolite profile was analyzed. Our results demonstrate large differences in the fermentation ability and metabolite profile of the 25 strains. Analysis of metabolites in fermented dough confirmed that acetic acid and succinic acid are likely responsible for the lowering of dough pH during fermentation and that the onset of CO₂ release from dough is related to dough pH rather than to the volume of CO₂ within the dough. Our results further suggest that the spread test is an inadequate tool to quantify rheological differences observed for strains with different fermentation profiles.     

Regulation and function of AtNRAMP4 metal transporter protein

The NRAMP gene family encodes integral membrane proteins mediating the transport of a broad range of transition metals in bacteria, fungi, plants, and animals. We studied the regulation of AtNRAMP4 in Arabidopsis. In a previous study, we showed that AtNRAMP3 and AtNRAMP4 transport manganese (Mn), iron (Fe), and cadmium (Cd). In this study, we show that, in contrast to AtNRAMP3, AtNRAMP4 complements the growth phenotype of the zrt1zrt2 Zn uptake deficient yeast mutant. In a previous study, we have shown that, under Fe starvation, AtNRAMP4 mRNA levels are up-regulated in Arabidopsis. To analyze the regulation of AtNRAMP4 at the protein level, we generated specific antibodies against AtNRAMP4 protein. The antiserum was able to recognize a tagged version of AtNRAMP4 expressed in yeast. The antibody did not reveal any change in AtNRAMP4 protein level upon Fe starvation in Arabidopsis thaliana ecotype Columbia plants. In AtNRAMP4 overexpressing plants, high levels of AtNRAMP4 protein could be detected. AtNRAMP4 overexpressing plants display cadmium hypersensitivity in a medium containing 50 μm FeEDTA as Fe source. However, despite the constitutive accumulation of AtNRAMP4 protein, AtNRAMP4 over-expressing plants did not display Cd hypersensitivity under high Fe supply (100 μm FeHBED). AtNRAMP4 over-expressing lines displayed the same sensitivity to Zn as controls under all conditions tested. Our results suggest a translational level for the regulation of AtNRAMP4. Over-expression of AtNRAMP4 in Arabidopsis thaliana confers a slight hypersensitivity to Cd but not to Zn.     

Inhibition of growth of Bradyrhizobium japonicum bacteroid by spermidine and spermine in yeast extract

Abstract

Bacteroids are defined as the symbiotic forms of Rhizobium or Bradyrhizobium cells in the root nodules of their legume host. The differentiation to bacteroids involves various physiological changes and may be associated with some genetical changes. Single-colony isolates from a nodule formed by a Bradyrhizobium strain often differed in their effectiveness and intrinsic antibiotic resistance (Weaver and Wright 1987; Ozawa unpublished data). Detailed analysis of the changes in the gene structure requires the isolation of individual bacteroids from a nodule. However it has been reported that the viability of bacteroids in culture is very low and only a small fraction of a bacteroid population could produce colonies on yeast extract-mannitol (YEM) agar (Sutton et al. 1977). Bergersen (1974) concluded that the reversion of bacteroids to the vegetative, growing form is very rare.     

Effects of yeast on the growth and reproduction of the saprophagous millipedePolydesmus angustus (Diplopoda, Polydesmidae)

The millipedePolydesmus angustus was reared in the laboratory from hatching to maturity, reproduction and death. Two food types were used: dead leaves alone or dead leaves supplemented monthly with dry food yeast (Saccharomyces cerevisiae) at a rate not exceeding 5% of leaf dry weight. Growth, survival, adult live weight and fertility were compared between females reared on the two diets. Although the species was able to complete its life cycle on dead leaves alone, several parameters were strongly affected by the addition of yeast: growth was significantly faster, adult females became significantly larger and there was a 4.3-fold increase in fertility. Only survival was unaltered by the addition of yeast. The comparison between these laboratory results and field data on female fertility and live weight suggests that the natural diet of millipedes includes foods of higher quality than dead leaves. Possible sources of high-quality food in natural conditions are discussed.     

Assessing biological activity of agricultural biostimulants: Bioassays for plant growth regulators in three soil additives

Abstract

Laboratory bioassays were used to investigate plant growth‐regulating effects of three different experimental soil additives, designated EXP95, W91, and Z96. A yeast growth test was used as a general assay of bioactivity, responses to soil additives were compared to those of known plant growth regulators [indoleacetic acid (IAA), gibberellic acid (GA₃), and kinetin]. A corn coleoptile elongation test was used to assay for auxin‐like activity and a dwarf pea bioassay was used for gibberellin‐like activity. The three soil additives were tested at five solution concentrations ranging from 1 to 10,000 ppm (by volume). All three soil additives stimulated yeast growth, depending on the concentration of the test solutions. However, all three soil additives inhibited plant growth in the two plant bioassays. Although this study clearly demonstrated that the three soil additives had significant biological activity at very low concentrations, there was little evidence for auxin‐like or gibberellin‐like activity.     

Effect of Formulation and Processing Treatments on Viscosity and Solubility of Extractable Barley β‐Glucan in Bread Dough Evaluated Under In Vitro Conditions

β‐Glucan shows great potential for incorporation into bread due to its cholesterol lowering and blood glucose regulating effects, which are related to its viscosity. The effects of β‐glucan concentration, gluten addition, premixing, yeast addition, fermentation time, and inactivation of the flour enzymes on the viscosity of extractable β‐glucan following incorporation into a white bread dough were studied under physiological conditions, as well as, β‐glucan solubility in fermented and unfermented dough. β‐Glucan was extracted using an in vitro protocol designed to approximate human digestion and hot water extraction. The viscosity of extractable β‐glucan was not affected by gluten addition, the presence of yeast, or premixing. Fermentation produced lower (P ≤ 0.05) extract viscosity for the doughs with added β‐glucan, while inactivating the flour enzymes and increasing β‐glucan concentration in the absence of fermentation increased (P ≤ 0.05) viscosity. The physiological solubility of the β‐glucan concentrate (18.1%) and the β‐glucan in the unfermented dough (20.5%) were similar (P > 0.05), while fermentation substantially decreased (P ≤ 0.05) solubility to 8.7%, indicating that the reduction in viscosity due to fermentation may be highly dependent on solubility in addition to β‐glucan degradation. The results emphasize the importance of analyzing β‐glucan fortified foods under physiological conditions to identify the conditions in the dough system that decrease β‐glucan viscosity so that products with maximum functionality can be developed.     

DIRECT MEASUREMENTS OF THE DISTRIBUTION OF OXYGEN IN SOIL AGGREGATES AND IN COLUMNS OF FINE SOIL CRUMBS

The micro electrode of Evans and Naylor was modified to permit measurements of oxygen concentration in soil and was used to determine the distribution of oxygen in (a) agar which contained yeast and glucose and served as a model of a respiring water-saturated soil; (b) saturated spherical soil aggregates; (c) partly saturated columns of fine soil crumbs. Measurements in (a) and (b) but not in (c) agreed closely with predictions by diffusion theory. The disagreement appeared to result from water being unevenly distributed in the columns.     

Chemical and bacterial changes in a forest soil percolated with some amino acids and leaf litter extract

Chemical and bacterial changes in the forest soil percolated with glycine and glutamate solution and leaf litter extract were studied.

When the forest soil was percolated with 25 mM glycine or glutamate, it took about 20 days for the number of bacteria to reach the maximum in number and the amino acids to be completely ellhausted, the rate being much slower than that in the cultivated soil. Supplementation of 0.01% yeast extract to the percolate much rnhanced the growth of bacteria and degradation of glycine. In this percolation the metabolic pattern of glycine was comparable to that in the cultivated soil except for the absence of an appreciable amount of nitrate formation.

Growth of bacteria in the forest soil was also rapid when percolated with leaf litter extract and only 3 days were sufficient for reaching the maximum number. Neither ammonia nor nitrate was detected throughout the percolation period and only change observed in the solution was a slight rise in pH.

Characteristics of bacteria enriched in the forest soil percolated will) glycine with and without yeast extract, and leaf litter extract were studied, No one group was dominant in the soil before percolation. Bacteria enriched by glycine were almost occupied with Gram-negative nonmotile rods with rather complex nutritional requirement regarded as Achromobacter, which were characteristically unable to utilize glycine as a sole nitrogen source. When the soil was percolated with glycine supplemented with yeast extract, enriched bacteria were composed of many kinds. The soil percolated with leaf litter extract was occupied with bacteria with simple nutritional requirement, which were regarded as Pseudamonas.     

Probiotics and Prebiotics

Probiotics, bacteria from the genera Bifidobacterium and Lactobacillus, and yeast, Saccharomyces, as well as prebiotics belonging to the group of dietary fiber (inulin with low degree of polymerization, fructose‐derived oligosaccharides, and resistant starch) are natural factors useful in prophylaxis and therapy of several common diseases including some types of cancer. They are available commercially and can be introduced to produce so‐called functional food. Probiotics and prebiotics can be utilized either separately or jointly (as synbiotics or eubiotics). Mechanisms of both biotics are discussed. The role of cereals in probiosis is considered. Possibilities for extension of the uses of the original Chinese probiotic, chaw tofu, are also considered.     

Rhodotorulic acid enhances root colonization of tomato plants by arbuscular mycorrhizal (AM) fungi due to its stimulatory effect on the pre-symbiotic stages of the AM fungi

Exudates of Rhodotorula mucilaginosa, a yeast commonly found in the rhizosphere, increased hyphal length of the arbuscular mycorrhizal (AM) fungi Gigaspora rosea and Gigaspora margarita. Rhodotorulic acid (RA), a siderophore compound obtained from R. mucilaginosa exudates, increased hyphal length and branching. Thus, the increase in the number of entry points and the higher AM root colonization of tomato plants in the presence of RA can at least partially be explained by the positive effect of RA on the pre-symbiotic stages of the AM fungi.