Changes in populations of rhizosphere bacteria associated with take-all disease of wheat

Take-all, caused by Gaeumannomyces graminis var. tritici, is one of the most important fungal diseases of wheat worldwide. Knowing that microbe-based suppression of the disease occurs in monoculture wheat fields following severe outbreaks of take-all, we analyzed the changes in rhizosphere bacterial communities following infection by the take-all pathogen. Several bacterial populations were more abundant on diseased plants than on healthy plants, as indicated by higher counts on a Pseudomonas-selective medium and a higher fluorescence signal in terminal restriction fragment length polymorphism analyses of amplified 16S ribosomal DNA (rDNA). Amplified rDNA restriction analysis (ARDRA) of the most abundant cultured populations showed a shift in dominance from Pseudomonas to Chryseobacterium species in the rhizosphere of diseased plants. Fluorescence-tagged ARDRA of uncultured rhizosphere washes revealed an increase in ribotypes corresponding to several bacterial genera, including those subsequently identified by partial 16S sequencing as belonging to species of alpha-, beta-, and gamma-proteobacteria, sphingobacteria, and flavobacteria. The functional significance of some of these populations was investigated in vitro. Of those isolated, only a small subset of the most abundant Pseudomonas spp. and a phlD(+) Pseudomonas sp. showed any significant ability to inhibit G. graminis var. tritici directly. When cultured strains were mixed with the inhibitory phlD(+) Pseudomonas strain, the Chryseobacterium isolates showed the least capacity to inhibit this antagonist of the pathogen, indicating that increases in Chryseobacterium populations may facilitate the suppression of take-all by 2,4-diacetylphloroglucinol-producing phlD(+) pseudomonads.     

Stacking pairs of disease resistance genes in wheat populations using telocentric chromosomes

Resistance of wheat to diseases such as fusarium head blight (FHB) and leaf rust is more effective and durable when resistance genes are stacked. This study investigated whether pairs of disease resistance genes will become fixed at higher frequencies in subsequent generations when placed in the hemizygous condition using telocentric chromosomes. Three pairs of telocentric chromosomes were tested for their male and female transmission to predict the fixation rate of hemizygous chromosome arms using reciprocal testcrosses. Hemizygous arm transmission was about 50% through ovules and about 75% through pollen because of pollen certation. To test if a corresponding increase in disease resistance could be observed in populations utilizing telocentric chromosomes, three resistance gene pairs were analyzed separately in three populations. These pairs were Lr16/Lr34 and Lr22a/Lr52 for resistance to leaf rust and Fhb1/Qfhs.ifa-5A for FHB resistance. Each of these gene combinations was involved in a crossing and selection scheme that identified F₁ plants that were either dihybrid or double monotelodisomic (DMTD). For each resistance gene combination F₃ families were produced for phenotypic testing. The Lr16/Lr34 and Lr22a/Lr52 F₃ populations both showed a sharp increase in leaf rust resistance among families derived from DMTD F₁ plants compared to those from dihybrid F₁ plants. A smaller increased resistance was found in the FHB population. The increased frequency of resistance was attributed to pollen certation and zygotic selection against the ditelosomic and double ditelosomic conditions. We conclude that telocentric chromosomes are a viable breeding tool to fix gene stacks.     

Assignment of rainbow trout linkage groups to specific chromosomes

The rainbow trout genetic linkage groups have been assigned to specific chromosomes in the OSU (2N=60) strain using fluorescence in situ hybridization (FISH) with BAC probes containing genes mapped to each linkage group. There was a rough correlation between chromosome size and size of the genetic linkage map in centimorgans for the genetic maps based on recombination from the female parent. Chromosome size and structure have a major impact on the female:male recombination ratio, which is much higher (up to 10:1 near the centromeres) on the larger metacentric chromosomes compared to smaller acrocentric chromosomes. Eighty percent of the BAC clones containing duplicate genes mapped to a single chromosomal location, suggesting that diploidization resulted in substantial divergence of intergenic regions. The BAC clones that hybridized to both duplicate loci were usually located in the distal portion of the chromosome. Duplicate genes were almost always found at a similar location on the chromosome arm of two different chromosome pairs, suggesting that most of the chromosome rearrangements following tetraploidization were centric fusions and did not involve homeologous chromosomes. The set of BACs compiled for this research will be especially useful in construction of genome maps and identification of QTL for important traits in other salmonid fishes.     

DNA-binding proteins related to the dosage of specific yeast chromosomes

A comparison has been made of the $\text{in vitro}$ DNA-binding proteins of specific aneuploid and isogenic euploid cells of $\text{Saccharomyces cerevisiae}$ by DNA-cellulose chromatography. We have been able to detect changes in the level of a small fraction of the yeast DNA-binding proteins which can be related to the dosage of specific yeast chromosomes. At least four proteins show a dosage related to the cellular level of chromosome I and at least one protein shows a dosage related to the level of chromosome VI.     

Effects of roxithromycin on ammonia-oxidizing bacteria and nitrite-oxidizing bacteria in the rhizosphere of wheat

In a pot-cultural experiment, the impact of the antibiotic roxithromycin (ROX) addition was assessed on the diversities of microbial structure and function communities, especially involved in ammonia and nitrite oxidation in wheat rhizosphere soil with and without the addition of earthworms. The abundances of ammonia-oxidizing bacteria (AOB), nitrite-oxidizing bacteria (NOB), and total bacteria were surveyed by the quantitative PCR. The quantities of total bacteria, AOB, and NOB with earthworms were higher than those without earthworms because of the synergistic effect. ROX inhibited the growth of AOB in all treatments, although the quantities of AOB were in a light increase in medium and heavy polluted treatments compared with that in the light polluted treatments. Different from AOB, the quantities of NOB were lowest in light polluted treatments, but the quantities of NOB were rapidly increased in medium and heavy polluted treatments compared with that in the control. These results indicated that the application of ROX principally had a negative effect on nitrification performance by affecting the abundances and relative ratios of both AOB and NOB in soil communities, which affected the N cycle in an agricultural ecosystem. According to the metabolic diversities evaluated by the biologic assay, the tendency of metabolic diversities was quite contrary to the quantities of NOB in all treatments and showed the contrast growing relation of autotrophic and heterotrophic bacteria under ROX pollution pressure in agricultural ecosystems.     

The role of calcium in improving photosynthesis and related physiological and biochemical attributes of spring wheat subjected to simulated acid rain

The response of photosynthesis parameters, catalase, superoxide dismutase and peroxidase activity, malondialdehyde, proline, chlorophyll, yield and yield components to foliar application of calcium and simulated acid rain in wheat were investigated. Foliar treatment of calcium led to significant increases in the photosynthesis rate, transpiration rate, stomatal conductance, proline, chlorophyll, yield and yield components in plants subjected to acid rain. Antioxidant enzyme activity and lipid peroxidation in the wheat leaves decreased because of calcium foliar application. Calcium hindered degradation of the rubisco subunits under acid rain treatment compared with water-treated plants. Results suggest that acid rain induces the production of free radicals resulting in lipid peroxidation of the cell membrane so that significant increase in antioxidant enzyme activity was observed. In addition, photosynthetic parameters i.e. photosynthesis rate, transpiration rate and stomatal conductance were drastically suppressed by acid rain. The cellular damage caused by free radicals might be reduced or prevented by a protective metabolism including antioxidative enzymes and calcium. We report that foliar application of calcium before acid rain may ameliorate the adverse effects of acid rain in wheat plants.     

Reproductive allocation patterns in different density populations of spring wheat

The effects of increased intraspecific competition on size hierarchies （size inequality） and reproductive allocation were investigated in populations of the annual plant, spring wheat （Triticurn aestivurn）. A series of densities （100, 300, 1 000, 3 000 and 10 000 plants/m^2） along a gradient of competition intensity were designed in this experiment. The results showed that average shoot biomass decreased with increased density. Reproductive allocation was negatively correlated to Gini coefficient （R^2 = 0.927）, which suggested that reproductive allocation is inclined to decrease as size inequality increases. These results suggest that both vegetative and reproductive structures were significantly affected by intensive competition. However, results also indicated that there were different relationships between plant size and reproductive allocation pattern in different densities. In the lowest density population, lacking competition （100 plants/m^2）, individual reproductive allocation was size independent but, in high density populations （300, 1 000, 3 000 and 10 000 plants/m^2）, where competition occurred, individual reproductive allocation was size dependent： the small proportion of larger individuals were winners in competition and got higher reproductive allocation （lower marginal reproductive allocation; MRA）, and the larger proportion of smaller individuals were suppressed and got lower reproductive allocation （higher MRA）. In conclusion, our results support the prediction that elevated intraspecific competition would result in higher levels of size inequality and decreased reproductive allocation （with a negative relationship between them）. However, deeper analysis indicated that these frequency- and size-dependent reproductive strategies were not evolutionarily stable strategies.     

Differential reactions of wheat and pea genotypes to root inoculation with growth-affecting rhizosphere bacteria

Spring wheat (Triticum aestivum L.) and pea (Pisum spp.) genotypes were tested for reaction to root inoculation with rhizosphere bacteria affecting plant growth. Plant response was studied in greenhouse experiments after treatment of seedlings with bacteria suspended in nutrient broth. Significant genotype variation was found in both wheat and pea in terms of shoot dry weight and severity of bacteria-induced leaf symptoms. For most bacterial isolates tested, there was good correlation between ratings of leaf symptoms 7 to 14 days after inoculation and growth inhibition measured after four weeks. Interactions between isolates and plant genotypes were significant in both wheat and pea (P=0.0024 and 0.0001, respectively), but genotypes with sensitivity or tolerance to most isolates could be distinguished. In an outdoor pot experiment, two of the bacterial isolates caused delayed plant development and differential decreases in grain yield of wheat genotypes. The hypothesis that the reaction of wheat genotypes to the tested becteria was related to their influence on bacterial establishment in the rhizosphere could not be substantiated.     

The determination of chromosomes involved in controlling epicuticular wax,water statues and stomatal characteristics using selected wheat substitution lines under water-stress conditions

Some selected chromosomal substitution lines were used to determine the role of their relevant chromosomes in affecting variation of epicuticular wax, water statues and stomatal characteristics under water-deficit conditions. Recipient and donor parents of the substitution lines were Chinese spring (CS) and Timstein, respectively. Analyses of variance revealed highly significant variations among the candidate substitution lines for leaf relative water content (LRWC), excised leaf water lost and grain weight. However, no significant variation was found for epicuticular wax (ECW). In the case of stomatal characteristics, analyses of variance indicated significant variation for stomatal frequency only and no significant variation was found for other stomatal characteristics. Comparison between the substitution lines and their recipient parent (CS) revealed that none of the substitution lines was significantly different with the recipient parent (CS) for ECW, thus indicating none of the candidate chromosomes involved in controlling this character. However, the results indicated the effects of chromosomes 1A, 3D and 7D from donor parent (Tim) in controlling LRWC. Chromosomes 7D of Timstein also had a significant effect in enhancing LRWC when substituted into CS background. In addition, it was observed that none of the characters correlated with grain yield in water-stressed experiments.     

Survival of potentially pathogenic human-associated bacteria in the rhizosphere of hydroponically grown wheat

Plants may serve as reservoirs for human-associated bacteria (H-AB) in long-term space missions containing bioregenerative life support systems. The current study examined the abilities of five human-associated potential pathogens, Pseudomonas aeruginosa, Pseudomonas cepacia, Staphylococcus aureus, Streptococcus pyogenes and Escherichia coli, to colonize and grow in the rhizosphere of hydroponically grown wheat, a candidate crop for life support. All of these bacteria have been recovered from past NASA missions and present potential problems for future missions. The abilities of these organisms to adhere to the roots of axenic five-day-old wheat (Triticum aestivum L. cv. Yecora rojo) were evaluated by enumeration of the attached organisms after a one hour incubation of roots in a suspension (approximately 10(8) cfu ml-1) of the H-AB. Results showed that a greater percentage of P. aeruginosa cells adhered to the wheat roots than the other four H-AB. Similarly incubated seedlings were also grown under attempted axenic conditions for seven days to examine the potential of each organism to proliferate in the rhizosphere (root colonization capacity). P. cepacia and P. aerogiunosa showed considerable growth, E. coli and S. aureus showed no significant growth, and S. pyogenes died off in the wheat rhizosphere. Studies examining the effects of competition on the survival of these microorganisms indicated that P. aeruginosa was the only organism that survived in the rhizosphere of hydroponically grown wheat in the presence of different levels of microbial competition.     

Changes in physiological groups of microorganisms in soil following wildfire

Abstract: Physiological groups of soil microorganisms were investigated in a forest ( Pinus pinaster Sol.) to asses their response to wildfire-induced soil changes. Microbial fluctuations were recorded 1 month and 1 year after the fire, both in the field and during controlled soil incubations. In both the burned and the unburned soil, starch-mineralizing microbes predominated over cellulose-mineralizing microbes; there were a relatively high number of ammonium-producers, whereas nitrite and nitrate producers were scarce. In the short term, burning produced a decreasing to nearly undetectable number in cellulase-producers whilst amylase-producers, and especially, ammonifying microbes increased, and the nitrifying groups did not change. One year after the wildfire, the burning effect was slightly overcome by cellulolytic microorganisms and the amylolytic population was slightly decreased; the improvement of ammonifiers was reduced, ammonium oxidizers were positively affected and nitrite oxidizers continued to be unaffected by the fire. The trends of populations during soil incubation indicated that, in the long term, the effect of burning will probably be nil on ammonifiers, somewhat negative on cellulolytic and amylolytic microbes and slightly positive on nitrite- and nitrate-formers.     

Irrigation differentially impacts populations of indigenous antibiotic-producing pseudomonas spp. in the rhizosphere of wheat

This work determined the impact of irrigation on the seasonal dynamics of populations of Pseudomonas spp. producing the antibiotics phenazine-1-carboxylic acid (Phz(+)) and 2,4-diacetylphloroglucinol (Phl(+)) in the rhizosphere of wheat grown in the low-precipitation zone (150 to 300 mm annually) of the Columbia Plateau of the Inland Pacific Northwest. Population sizes and plant colonization frequencies of Phz(+) and Phl(+) Pseudomonas spp. were determined in winter and spring wheat collected during the growing seasons from 2008 to 2009 from selected commercial dryland and irrigated fields in central Washington State. Only Phz(+) bacteria were detected on dryland winter wheat, with populations ranging from 4.8 to 6.3 log CFU g(-1) of root and rhizosphere colonization frequencies of 67 to 100%. The ranges of population densities of Phl(+) and Phz(+) Pseudomonas spp. recovered from wheat grown under irrigation were similar, but 58 to 100% of root systems were colonized by Phl(+) bacteria whereas only 8 to 50% of plants harbored Phz(+) bacteria. In addition, Phz(+) Pseudomonas spp. were abundant in the rhizosphere of native plant species growing in nonirrigated areas adjacent to the sampled dryland wheat fields. This is the first report that documents the impact of irrigation on indigenous populations of two closely related groups of antibiotic-producing pseudomonads that coinhabit the rhizosphere of an economically important cereal crop. These results demonstrate how crop management practices can influence indigenous populations of antibiotic-producing pseudomonads with the capacity to suppress soilborne diseases of wheat.     

转Bt基因玉米根际微生物和细菌生理群多样性

大田栽培条件下,以转Bt玉米Mon810及其亲本非转基因玉米为研究对象,在玉米的不同生育期测定根际土壤微生物的数量变化,并对细菌群落结构及多样性进行了分析。结果表明,各生育期内Bt玉米与对照相比根际土壤真菌无显著差异,但细菌在抽丝期,放线菌在苗期二者有显著差异。同种细菌功能群的数量变化在Bt玉米和对照各生育期内趋势一致,不同的菌群表现不同。亚硝化细菌、好气固氮菌、氨化细菌和钾细菌的数量变化波动较大,其中Bt玉米和对照相比好气固氮菌的数量在生育期内有5个生育期有显著差异,而氨化细菌、钾细菌和亚硝酸菌仅分别在乳熟期、喇叭口期及拔节期和抽雄期差异显著。好气纤维分解菌和硝化细菌在苗期、拔节期和喇叭口期差异显著,无机磷分解菌在乳熟期和完熟期数量差异显著。整个生育期内3种群落特征参数均保持基本稳定,除乳熟期和完熟期外Bt玉米根际微生物群落特征参数均高于对照。     

Morphological and physiological changes in roots of various wheat genotypes as related to cytoskeleton disruption

A depolymerizing effect of anti-microtubule drug oryzalin on the roots of three winter wheat (Triticum aestivum L.) cultivars contrasting in their frost-resistance was studied. The influence of plant cold acclimation (3°C, 7 days) and ABA treatment (30 μM) on oryzalin action was evaluated. Plant growing in the presence of 10 μM oryzalin under optimum temperature of 23°C resulted in the root-length decrease by 19–24% and root-apex swelling. All cells, especially in the root cortex, changed their radial dimensions. The cells acquired a rounded or irregular shape and increased in size. This indicates the loss of correct cell growth polarity. Most pronounced changes in the root apex diameter and most severe linear growth suppression were observed in the cultivar of moderate frost-resistance. The roots of this cultivar contained the highest amounts of actin and tubulins, as was evident from the immunoblot analysis. The effect of oryzalin on root growth and apex swelling was correlated with the content of actin in the roots of different wheat cultivars. Cold acclimation and exogenous ABA reduced (or prevented) oryzalin action on roots in a cultivar-specific manner. The conclusion was made that the bulk of the cytoskeletal net determined the efficiency of the cytoskeletal control of plant growth and morphogenesis. During autumn and winter periods, this is important for a better adaptation to temperature fluctuations of moderately frost-resistant plants, which are characterized by a high ecological plasticity.     

土霉素暴露对小麦根际抗生素抗性细菌及土壤酶活性的影响

通过培养的方法研究了土霉素暴露和小麦根际抗性细菌的数量、种类、分布特征及土壤酶活性之间的剂量效应关系。结果表明,土霉素暴露下小麦根际单一抗生素抗性细菌数量和抗土霉素—链霉素双重抗性细菌数都明显增加,且与暴露剂量呈正效应关系;同时,土壤磷酸酶、脱氢酶活性下降,但与土霉素的剂量效应关系不明显。从土霉素暴露的土壤中分离到50株抗性细菌,经形态观察、RFLP分组和16S rDNA序列测定与分析,将它们聚集在Actinobacteria、Bacilli、Alphaproteobacteria、Gammaproteobacteria 和Sphingobacteria类群。其中放线菌最多（15株）,占抗性菌总数的30 %;其次是Bacillus属细菌（9株）和Pseudomonas属细菌（8株）,分别占18 %和16 %。同时,具有抗性的人类机会致病菌Pseudomonas、Sphingomonas和Stenotrophomonas属细菌在土霉素暴露的样品中均被分离到,分别占抗性菌株总数的16 %、8 %和4 %。值得注意的是,随着土霉素暴露剂量的增加,小麦根际优势促生菌Bacillus属细菌的抗性检出率逐步降低;但具有抗生素抗性的人类机会致病菌Pseudomonas、Sphingomonas和Stenotrophomonas属细菌的检出率却明显增加,提示可能会进一步增大其机会致病性。     

持续干旱对樱桃根际土壤细菌数量及结构多样性影响

以1年生吉塞拉实生容器苗为试材,采用绿色荧光蛋白基因标记技术,研究了干旱胁迫(连续干旱0、7、14、21、28 d和35 d)对樱桃根际促生细菌YT3的标记菌YT3-gfp数量的影响,同时结合平板计数法和末端限制性片段长度多态性分析(terminal restriction fragment length polymorphism,T-RFLP)技术,研究了干旱对樱桃土壤中的微生物数量及细菌群落结构多样性影响。结果表明:樱桃根际土壤中的YT3-gfp数量是非根际土壤中的8.75—28.77倍,随着持续干旱强度的增加,YT3-gfp的数量先增加后减小。干旱对根际土壤中YT3-gfp数量的影响大于对非根际土壤的影响,分别在持续干旱至第21天和28天时,YT3-gfp的数量达到最大值。随着持续干旱强度的增加,根际土壤中细菌和放线菌数量先增加后减小,而真菌的数量一直减少。此外,持续干旱至第21天或28天时,樱桃根际土壤具有最高的丰富度指数、多样性指数和最低的优势度指数。基于T-RFLP的主成分分析结果显示持续干旱14—35 d时,其细菌群落结构成为一个相对独立的群,群落结构趋于多样性;而持续干旱7 d和42 d构成另外两个相对独立的群,群落结构趋于简单。以上分析可知,干旱对土壤微生物影响显著,一定强度的干旱可提高细菌和放线菌数量,提高细菌群落结构多样性,适当干旱对维持根际土壤细菌群落结构多样性是有益的。     

Changes in rhizosphere bacterial populations during phytoextraction of heavy metal contaminated soil with poplar

Abstract

The objective of this paper was to study the response of rhizosphere ammonia‐oxidizing bacterial (AOB) populations during phytoextraction. Hybrid poplars were grown in compartmented root containers with an aged heavy metal (HM)‐contaminated soil for 13 weeks. Bulk and poplar rhizosphere soils were analyzed by denaturing gradient gel electrophoresis (DGGE) of amoA gene fragments. DGGE patterns revealed that amoA‐containing populations in the contaminated soils were markedly different from those in the uncontaminated soils. AmoA profiles appeared to be stable over time in the bulk soils. In contrast, contaminated rhizosphere soils revealed a clear shift of populations with removal of HMs. Rhizosphere AOB populations of the HM‐contaminated soils became similar to the populations of the uncontaminated soils during phytoextraction. The effect of phytoextraction was, however, not evident in the bulk samples, which still contained large amounts of HMs. This study suggests that rhizosphere AOB populations are able to recover after the relief of HM stress by phytoextraction practices.     

Plant genotype, micronutrient fertilization and take-all infection influence bacterial populations in the rhizosphere of wheat

The relationship between micronutrient efficiency of four wheat (Triticum aestivum L.) genotypes, tolerance to take-all disease (caused by Gaeumannomyces graminis (Sacc.) Arx and Olivier var. tritici Walker), and bacterial populations in the rhizosphere was tested in soil fertilized differentially with Zn and Mn. Plant growth was reduced by Mn or Zn deficiency and also by take-all. There was an inverse relationship between micronutrient efficiency of wheat genotypes when grown in deficient soils and the length of take-all lesions on roots (efficient genotypes had shorter lesions than inefficient ones). In comparison to the rhizosphere of control plants of genotypes Aroona and C8MM receiving sufficient Mn and Zn, the total numbers of bacterial cfu (colony forming units) were greater in the rhizosphere of Zn-efficient genotype Aroona under Zn deficiency and in Mn-efficient genotype C8MM under Mn deficiency. These effects were not observed in other genotypes. Take-all decreased the number of bacterial cfu in the rhizosphere of fully-fertilized plants but not of those subjected to either Mn or Zn deficiency. In contrast, the Zn deficiency treatment acted synergistically with take-all to increase the number of fluorescent pseudomonads in the rhizosphere. Although numbers of Mn-oxidising and Mn-reducing bacteria were generally low, take-all disease increased the number of Mn reducers in the rhizosphere of Mn-efficient genotypes Aroona and C8MM. Under Mn-deficiency conditions, the number of Mn reducers in the rhizosphere increased in Aroona but not in C8MM wheat. The results suggest that bacterial microflora may play a role in the expression of Mn and Zn efficiency and tolerance to take-all in some wheat genotypes.