Changes in Amino-acid Content of Tomato Xylem Sap Following Infection with Strains of Verticillium albo-atrum

Xylem exudates of four tomato cultivars showing resistance andsusceptibility to the vascular pathogen Verticillium albo-atrum,were analysed for amino acids and amides by column chromatography42 days after inculation with different isolates of the fungus.Analysis of the exudate from healthy, resistant Loran Bloodshowed a substantial reduction in amino compounds compared withthe healthy, susceptible Potentate and Bonny Best. ResistantMoscow, however, gave similar results to Potentate. Infectionin all cultivars except Moscow led to a general increase inthe concentration of amino acids and predominantly in asparticand glutamic acids, threonine, serine, proline, tyrosine, ornithine,lysine, histidine, and phenylalanine. A reduction in prolinecontent of up to 90 per cent occurred in the resistant plantsin both pathogenic and non-pathogenic combinations. The resultsare discussed in relation to previously published data on vascularcolonization and disease development with the same tomato cultiversand fungal strains.     

Cellulose decomposing ability of Trichoderma in relation to their saprophytic survival

Abstract

Forty one strains of Trichoderma were isolated from four provinces in the Philippines and tested for their cellulose adequacy indices (CAI). T. harzianum strain no. 94-016 with the highest CAI was further tested for its saprophytic colonization in pots relative to the amount of inoculum. The higher the amount of inoculum of T. harzianum strain no. 94-016, the higher percentage colonization of rice straw in the soil. Further tests were done to compare the decomposition of rice straw on the surface and buried in the soil and the effect of moisture on soil decomposition. Buried rice straw inoculated with T. harzianum strain no. 94-016 watered on a daily basis provided better decomposition.     

Common Bean Reaction to Fusarium oxysporum f. sp. Phaseoli, the Cause of Severe Vascular Wilt in Central Africa

During the September‐December season of 1990, severe symptoms of Fusarium wilt were for the first time observed on a popular climbing bean (Phaseolus vulgaris L.) cultivar. G 2333. introduced within the previous 5 years. Seventy‐three bean genotypes were screened for resistance lo the disease, using artificial inoculation. The effect of inoculation density on the reaction of four selected genotypes was also investigated. Of the 29 climbing bean genotypes evaluated, 19 were resistant, including 11 of the 15 pre‐release or released cultivars. Of the 44 bush bean cultivars evaluated, 28 were resistant, five were intermediate and 11 were susceptible. All susceptible cultivars showed vascular discoloration. In both susceptible and resistant genotypes, the fungus spread almost equally from the entry points in inoculated roots to the base of the plants, but colonization and vertical spread within the vascular system were markedly less in resistant than in susceptible cultivars. At 20 and 30 cm above soil level, the fungus was only recovered from susceptible cultivars. Increasing inoculum density from 10² to 10⁷ conidia/ml did not affect the resistance of cultivars RWR 950 and G 685 but. in the susceptible cultivars G 2333 and MLB‐48‐89 A. it resulted in early appearance, high incidence and severity of the disease.     

The dark side of the mycorrhiza

Plant association with arbuscular mycorrhizal (AM) fungi is usually regarded as mutualistic. However, this positive effect could disappear if the benefit of the fungal-plant association changes with colonization density. In order to test the conditionality of this interaction, we evaluated plant performance and tolerance to defoliation across five levels of commercial AM fungal inoculum concentrations. Additionally, we evaluated if plant performance and tolerance were similarly affected by a whole soil community collected under a native congener. Along the gradient of inoculation, plant performance exhibited a peak at intermediate inoculum concentration, indicating the presence of an optimum level of AM fungal concentration that maximized AM fungal benefit. Root colonization by fungal hyphae increased linearly across the experimental inoculation gradient. Paralleling root colonization, plant tolerance to defoliation decreased linearly along the inoculum gradient. Plant performance was similar under the whole soil and commercial treatments. Our results show a negative correlation between tolerance to defoliation and AM fungal inoculum concentration, indicating that AM fungi colonization could constrain the evolution of plant tolerance to herbivory.Key words: compensation, defences, ecological interactions, herbivory, multitrophic interactions, mycorrhizal fungi, toleranceArbuscular mycorrhizal (AM) fungi occur in all ecosystems of the world and associate with the roots of about 70% of all vascular plants.¹ This association is typically regarded as mutualistic, because there is a bidirectional transfer of nutrients between the host plant and its fungal partners. Carbon compounds are passed from the plant to the fungus and, in return, there is a transfer of mineral nutrients, principally nitrate and phosphate.² However, this association also entails costs. The amount of carbon allocated to AM fungi is estimated to range from 4% to 20% of a plant''s total carbon budget.² Throughout the literature, there are examples of the conditionality of this relationship exemplified by a continuum of the effects of AM fungal colonization on hosts from positive, through null to negative.^3–5 Moreover, it has been suggested that the benefit of a plant associating with fungal symbionts depends not only on the identity of AM fungi⁴ and plant genotypes⁶ but also on hyphal colonization density in roots.⁷ In a recent greenhouse study, we examined components of the conditionality of plant interactions with soil biota.⁸ We were interested in knowing how the performance and tolerance to defoliation of the annual plant Datura stramonium varied along a concentration gradient of commercial AM fungal inoculum containing four Glomus species (Mycorrhizal Applications, Grants Pass, OR USA).We found a curvilinear relationship between AM fungal inoculum concentration and plant performance, as predicted by previous models.⁷ The quadratic decelerating function between inoculum concentration and plant performance indicates an optimum level of AM fungal concentration (1/24^th total pot volume) that maximizes AM fungal benefit (Fig. 1A). This result suggests that, in D. stramonium, positive associations between AM fungi and plant fitness may not be proportional and, that at high colonization densities, mycorrhizae may have detrimental effects, perhaps by competing with plants for nutrients, or by interfering with other essential interactions.^4,5 We also found, from root examination, that hyphal colonization of roots increased linearly with AM fungi inoculum concentration. Moreover, we found that tolerance to herbivory decreased linearly with increasing AM fungal inoculum concentration (r² = −0.40; F_1,27 = 5.89; p = 0.0222; Fig. 1B), suggesting that, in our system, at high densities, mycorrhizae may become parasitic and may compete for resources (e.g., carbon) with the induced host plant response to leaf damage.Open in a separate windowFigure 1Effect of a gradient in AM fungi inoculum concentration on D. stramonium performance. (A) Non-linear relation between seed production and inoculum concentration. In general, plants achieved their maximal performance at an inoculum concentration of 1/24^th total pot volume. (B) Tolerance to defoliation decreased linearly with inoculum concentration. Tolerance was calculated as the difference in standardized seed production between related damaged and undamaged genetically related plants corresponding to six genetic full-sib families.In order to know whether the effects we found in the greenhouse using commercial inoculum could be expected in the field, we addressed whether or not D. stramonium performance and tolerance were similarly influenced by whole soil field communities; including AM fungi, pathogens, root herbivores, etc. Unfortunately, D. stramonium is not native to the area where this research was undertaken, so we collected soil immediately below plants of a native congener Datura wrightii, a perennial herb that grows at the Putah Creek Reserve (UC, Davis). Pots were inoculated at a 1/12^th total pot volume with this live soil and plants were grown concurrently with those in the previous experiment. We compared plant performance and tolerance under the live soil treatment and the last level of the commercial AMF inoculum gradient (both inoculated at a 1/12^th total pot volume). Results indicated no differences in foliar area (F_1,94 = 1.18; p = 0.2782), root mass (F_1,94 = 0.99; p = 0.3222), flowering day (χ² = 0.31; p = 0.5804) and fitness (χ² = 0.03; p = 0.8691). Moreover, root colonization levels were (F_1,94 = 0.75; p = 0.3877) in both 1/12^th volume vs. live soil, as well as in the 0 AMF and sterilized soil (F_1,94 = 2.56; p = 0.1130). Despite these similarities, plant tolerance did differ significantly between AMF and live soil treatments (F_1,94 = 5.49; p = 0.0411), tolerance being greater under the live soil treatment (0.3755 ± 0.0311 tolerance) relative to the 1/12^th AM fungal treatment (−0.5744 ± 0.2714 tolerance). This result suggests that the expression of plant tolerance may also depend on the identity of AMF colonizing roots or the number and identities of soil bacteria. We did not know which microbial species were in the soils we collected.We show that, when inoculated over a gradient of abundance, Glomus AM fungal colonization consistently decreased tolerance to herbivory. The presence of mycorrhizae could therefore decrease the adaptive value of traits increasing tolerance. We also show here that though live soil inoculum had similar effects in magnitude and direction to those of commercial AMF incoculum on growth and fitness, live soil biota collected under a congener of D. stramonium increased tolerance to herbivory at the same levels of root colonization. Overall, the results of this study indicate that the interaction between soil biotic components and the response of D. stramonium to leaf damage is highly conditional; and can depend on amounts of root colonization, as well as perhaps identities of AM fungi and bacteria. In both cases, soil biota affected the impact of damage to leaves aboveground. AM fungi may mediate the efficacy of tolerance as a defense, and this effect may be especially important in light of herbivore adaptation, when tolerance may be favored over resistance as a plant defense strategy.¹⁰     

Response of tomato genotypes to bacterial speck (Pseudomonas syringae pv. tomato)

Two genotypes of tomato A 100 and Ontario 7710 which were inoculated separately with four strains of Pseudomonas syringae pv. tomato differed significantly in disease severity (susceptibility) to bacterial speck. At both concentrations of inoculum of each strain used (10⁷ and 10⁸ cfu/ml) A 100 appeared to be highly susceptible whereas Ontario 7710 showed very low or no susceptibility. The significant differences in virulence between strains and in response of tomato plants in three replicate experiments were found. Generally, concentration of inoculum 10⁷ cfu/ml was too low to induce consistent level of disease severity. The obtained results indicate the importance of consistent and favorable conditions for disease development in screening of tomato resistance to bacterial speck.     

Fungal inoculum properties and its effect on growth and enzyme activity of Trametes versicolor in soil

The effect of fungal inoculum properties on colonization of nonsterile soil by three isolates of the white-rot fungus Trametes versicolor was investigated. Fungal inoculum properties were examined in separate experiments and were fungal inoculum composition, age of fungal inoculum, concentration of the inoculum and inoculation method. The fungal inoculum composition study compared pine versus poplar sawdust as the basic carrier with varying amounts of corn grit, corn meal and starch. The age of the fungal inoculum studied ranged from 3 to 21 days. The inoculum concentration gradually increased from 0 to 50% (v/v). The study assessing inoculation method compared mixing with layering techniques. The effect of moisture conditions of soil, sawdust and sand in combination with two inoculation methods (mixing versus point source inoculation) on colonization by T. versicolor was also determined. Colonization of soil was always assessed visually and enzymatically monitoring mycelial growth, biological potential (fluorescein diacetate assay) and laccase levels. Generally, the three different assessment methods correlated (P < 0.05) with each other. A fungal inoculum based on pine sawdust supported white-rot fungal growth in soil better than a poplar sawdust basis. Colonization of soil by T. versicolor was improved by increasing the corn content of the fungal inoculum. Younger (<7 days old) fungal inoculum resulted in better soil colonization than older (>10 days). A strong correlation (P < 0.001) was observed between the amount of fungal inoculum used in the soil augmentation and white-rot fungal colonization of soil. Inoculation of the fungal inoculum into soil by mixing was preferable over application in layers or point source inoculation. Moisture level did not influence biological potential measurements, but affected mycelial growth and laccase expression.     

Electrolyte Leakage, Lipoxygenase, and Lipid Peroxidation Induced in Tomato Leaf Tissue by Specific and Nonspecific Elicitors from Cladosporium fulvum

Glycoprotein nonspecific elicitor (NSE) and a specific elicitor preparation from intercellular fluids (SE) of tomato (Lycopersicon esculentum Mill. cv Bonny Best or Potentate) infected with race 2.4.5 of Cladosporium fulvum Cooke [syn. Fulvia fulva (Cooke) Ciferri] were injected into cv Sonatine (resistant to race 2.4.5) to compare electrolyte leakage, lipoxygenase activity, and lipid peroxidation induced in response to these elicitors. Increased electrolyte leakage was induced by NSE or SE; the leakage due to NSE but not to SE was inhibited by the nonsteroidal antiinflammatory drug (NSAID) piroxicam. Under normal photoperiod conditions, higher levels of lipoxygenase activity were detected 6 hours after injection with either elicitor. This activity peaked by 12 hours with both elicitors and declined to control levels by 24 hours when visible necrosis could be detected. Both NSE and SE-induced lipoxygenase was inhibited by piroxicam in vitro. Lipid peroxidation in elicitor-treated tissue was also assayed at 6, 12, and 24 hours after injection using the TBA test for malonaldehyde. Increased peroxidation was detected in response to NSE or SE at 12 hours with similar values obtained at 24 hours. With plants incubated in the dark, lipoxygenase, and lipid peroxidation were similarly induced in SE-injected tissue whereas necrosis induction by SE was light dependent.     

Rapid Tomato Seedling Assay for Virulent Isolates of Fusarium oxysporum F. sp. Radicis-lycopersici (FORL), the Tomato Crown and Root Rot Pathogen

A rapid tomato seedling assay was developed for determining the relative wilt capacity of isolates of Fusarium oxysporum f. sp. radicis-lyco-persici (FORL), a virulent strain of the tomato pathogen. The procedures for the assay require that 5-day-old cv. Bonny Best tomato seedlings be dipped in 30-day cell-free concentrated culture filtrates of FORL isolates, which were grown in Czapek-Dox medium with 2% Bacto-casamino acids (CDA). The seedlings in the culture filtrates were then incubated at 30 C under artificial light (1200 ftcandles) at 28% relative humidity in a wind stream of 100–150 m min. The relative pathogenicity of the isolates was determined by inoculating the roots of 18-day-old seedlings with cultures of FORL isolates. The pattern of cell-free filtrate wilt among the isolates was the same as that for the disease caused by cultures of the isolates. The seedlings treated with the filtrate from the most virulent isolate (Harrow HRS-182) wilted in 20 min. The filtrates from less virulent isolates took progressively longer. up to 90 min. to cause comparable wilt. Isolate HRS-082 was the first isolate also to induce disease in 10-day-old seedling assay. Both assays indicate three levels of wilt and disease capacities amongthe isolates examined. The utility of the assay in research and breeding for resistance is discussed.     

Observations consistent with autocrine stimulation of hybridoma cell growth and implications for large-scale antibody production

Existence of autocrine growth factors (aGFs) may influence the serum requirement for growth of hybridoma cells and thus significantly influence process economics. For the murine hybridoma cell line S3H5/2bA2, critical inoculum density (cID) and serum requirement for growth were inversely related for cultivation in both T flasks and spinner flasks. In spinner flasks, an inoculum density of 10⁶ cells/ml was necessary for the cells to grow in RPMI 1640 medium without serum supplement, and an inoculum density of 10³ cell/ml was necessary in RPMI 1640 medium with 10% serum. In T flasks, where the local cell density is higher than in spinner flasks, an inoculum density of 10⁶ cells/ml was necessary for the cells to grow in RPMI 1640 medium without serum supplement, and an inoculum density of 1 cell/ml was also necessary in RPMI 1640 medium with 10% serum. Further, immobilized cells at high local cell density could grow under conditions where cells in T flasks at corresponding overall cell density could not grow. The cells at high inoculum density were less sensitive to shear induced by mechanical agitation than the cells at low inoculum density. Taken together these observations support the existence of secreted aGF(s) by the hybridoma cell line used. Since the specific MAb production rate was independent of cultivation method and inoculum density, the existence of autocrine growth factors would suggest that the use of immobilized cells should improve the economics of MAb production.     

Influence of Solid Surface, Adhesive Ability, and Inoculum Size on Bacterial Colonization in Microcosm Studies

Microcosm studies were performed to evaluate the effect of solid surfaces, bacterial adhesive ability, and inoculum size on colonization success and persistence of Pseudomonas fluorescens and Xanthomonas maltophilia, each with a Tn5 insertion that conferred resistance to kanamycin and streptomycin. Two types of microcosms were used: (i) a simple system that was colonized by Aeromonas hydrophila and a coryneform and (ii) a complex system produced from lake water enrichment cultures. Simple microcosms contained 100 ml of peptone- and yeast extract-supplemented artificial lake water or 60 ml of peptone- and yeast extract-supplemented artificial lake water with 70 g of 3-mm glass beads. Complex microcosms contained 100 ml of lake water with no nutrient additions or 100 ml of lake water with 70 g of glass beads. The microcosms were incubated for 35 days at 20°C. In lake water enrichment microcosms, the presence of beads increased the abilities of P. fluorescens or X. maltophilia to colonize, but their numbers decreased with time in microcosms both with and without beads. The adhesiveness of the bacteria, measured in an in vitro assay, did not relate to colonization success. In simple microcosms, the inoculum size (10, 10², or 10³) of P. fluorescens did not influence colonization success. However, in complex microcosms, an inoculum of 10³ cells was insufficient to ensure colonization by P. fluorescens, while 10⁶ cells resulted in colonization of liquid and beads. Simple microcosm studies, utilizing only a few species, were poor models for complex natural systems. In complex enrichment systems, colonization of surfaces resulted in higher numbers of organisms but did not noticeably promote persistence. Adhesiveness of a particular organism may be a relatively minor factor influencing its ability to colonize solid surfaces in complex natural environments.     

High-Fat Diet: Bacteria Interactions Promote Intestinal Inflammation Which Precedes and Correlates with Obesity and Insulin Resistance in Mouse

Background

Obesity induced by high fat (HF) diet is associated with inflammation which contributes to development of insulin resistance. Most prior studies have focused on adipose tissue as the source of obesity-associated inflammation. Increasing evidence links intestinal bacteria to development of diet-induced obesity (DIO). This study tested the hypothesis that HF western diet and gut bacteria interact to promote intestinal inflammation, which contributes to the progression of obesity and insulin resistance.

Methodology/Principal Findings

Conventionally raised specific-pathogen free (CONV) and germ-free (GF) mice were given HF or low fat (LF) diet for 2–16 weeks. Body weight and adiposity were measured. Intestinal inflammation was assessed by evaluation of TNF-α mRNA and activation of a NF-κB^EGFP reporter gene. In CONV but not GF mice, HF diet induced increases in body weight and adiposity. HF diet induced ileal TNF-α mRNA in CONV but not GF mice and this increase preceded obesity and strongly and significantly correlated with diet induced weight gain, adiposity, plasma insulin and glucose. In CONV mice HF diet also resulted in activation of NF-κB^EGFP in epithelial cells, immune cells and endothelial cells of small intestine. Further experiments demonstrated that fecal slurries from CONV mice fed HF diet are sufficient to activate NF-κB^EGFP in GF NF-κB^EGFP mice.

Conclusions/Significance

Bacteria and HF diet interact to promote proinflammatory changes in the small intestine, which precede weight gain and obesity and show strong and significant associations with progression of obesity and development of insulin resistance. To our knowledge, this is the first evidence that intestinal inflammation is an early consequence of HF diet which may contribute to obesity and associated insulin resistance. Interventions which limit intestinal inflammation induced by HF diet and bacteria may protect against obesity and insulin resistance.     

Generation of novel cationic antimicrobial peptides from natural non-antimicrobial sequences by acid-amide substitution

Background

The accessory gene regulator (agr) is a quorum sensing cluster of genes which control colonization and virulence in Staphylococcus aureus. We evaluated agr function in community- (CA) and healthcare-associated (HA) MRSA, to compare the pharmacodynamics and bactericidal activity of vancomycin against agr functional and dysfunctional HA-MRSA and CA-MRSA.

Methods

40 clinical isolates of MRSA from the Canadian Nosocomial Infection Surveillance Program were evaluated for delta-haemolysin production, as a surrogate marker of agr function. Time kill experiments were performed for vancomycin at 0 to 64 times the MIC against an initial inoculum of 10⁶ and 10⁸ cfu/ml of agr functional and dysfunctional CA-MRSA and HA-MRSA and these data were fit to a hill-type pharmacodynamic model.

Results

15% isolates were agr dysfunctional, which was higher among HA-MRSA (26.3%) versus CA-MRSA (4.76%). Against a low initial inoculum of 10⁶ cfu/ml of CA-MRSA, vancomycin pharmacodynamics were similar among agr functional and dysfunctional strains. However, against a high initial inoculum of 10⁸ cfu/ml, killing activity was notably attenuated against agr dysfunctional CA-MRSA (USA400) and HA-MRSA (USA100). CA-MRSA displayed a 20.0 fold decrease in the maximal reduction in bacterial counts (Emax) which was 3.71 log₁₀ CFU/ml for agr functional vs. 2.41 log₁₀ CFU/ml for agr dysfunctional MRSA (p = 0.0007).

Conclusions

Dysfunction in agr was less common among CA-MRSA vs. HA-MRSA. agr dysfunction demonstrated an impact on vancomycin bactericidal activity and pharmacodynamics against a high initial inoculum of CA-MRSA and HA-MRSA, which may have implications for optimal antimicrobial therapy against persistent, difficult to treat MRSA infections.     

Competition amongst rhizobial strains for the colonization and nodulation of two tropical legumes

Summary Paired rhizobial strains supplied in several proportions were used to study inter-strain competition in association with Macroptilium atropurpureum (DC) URB (siratro) and Stylosanthes guianensis (Aubl.) Swartz (Stylo, line I R I 1022). Plants raised from surface sterilized seed, were grown on agar in large cotton-wool plugged tubes, and populations and inter-strain ratios determined in the inoculum and on the root at several times after inoculation. The nodules were mapped in order of appearance and the strains they contained identified at harvest.Related substrains and strains of similar growth habit competed more with each other in the colonization of the root surface than did a fast-growing strain in association with a typical slow grower. Capacity amongst slow-growing strains to dominate a paired competitor in the colonization of the root was a strain characteristic and was not affected by host. It was unrelated to effectiveness in the rhizobium-host association.In 5 of the 7 cases nodulation success could be related quantitatively to root-surface representation and a competitive index calculated; in the remainder one of each pair overwhelmed the other over a wide range of inoculum ratios. It was not possible to relate competitive nodulating success to any single feature of the host: rhizobium symbiosis. In the two most striking cases relationship between competitiveness and N₂-fixing effectiveness was reversed; in others competitiveness difference was as great between equally effective as between strains of differing effectiveness. In the case of Stylo there was a marked dominance of an ineffective over an effective competitor, which might be attributed to greater compatibility, as indicated by faster nodulaton by the ineffective strain. This last result argues against use of mixed inocula including any strain ineffective on any of the hosts for which the inoculum is recommended.Work completed as part requirement for the degree of M. Sc., University of New South Wales;     

Consequences of inoculation with native arbuscular mycorrhizal fungi for root colonization and survival of <Emphasis Type="Italic">Artemisia tridentata</Emphasis> ssp. <Emphasis Type="Italic">wyomingensis</Emphasis> seedlings after transplanting

In arid environments, the propagule density of arbuscular mycorrhizal fungi (AMF) may limit the extent of the plant–AMF symbiosis. Inoculation of seedlings with AMF could alleviate this problem, but the success of this practice largely depends on the ability of the inoculum to multiply and colonize the growing root system after transplanting. These phenomena were investigated in Artemisia tridentata ssp. wyomingensis (Wyoming big sagebrush) seedlings inoculated with native AMF. Seedlings were first grown in a greenhouse in soil without AMF (non-inoculated seedlings) or with AMF (inoculated seedlings). In spring and fall, 3-month-old seedlings were transplanted outdoors to 24-L pots containing soil from a sagebrush habitat (spring and fall mesocosm experiments) or to a recently burned sagebrush habitat (spring and fall field experiments). Five or 8 months after transplanting, colonization was about twofold higher in inoculated than non-inoculated seedlings, except for the spring field experiment. In the mesocosm experiments, inoculation increased survival during the summer by 24 % (p?=?0.011). In the field experiments, increased AMF colonization was associated with increases in survival during cold and dry periods; 1 year after transplanting, survival of inoculated seedlings was 27 % higher than that of non-inoculated ones (p?<?0.001). To investigate possible mechanisms by which AMF increased survival, we analyzed water use efficiency (WUE) based on foliar ¹³C/¹²C isotope ratios (δ ¹³C). A positive correlation between AMF colonization and δ ¹³C values was observed in the spring mesocosm experiment. In contrast, inoculation did not affect the δ ¹³C values of fall transplanted seedlings that were collected the subsequent spring. The effectiveness of AMF inoculation on enhancing colonization and reducing seedling mortality varied among the different experiments, but average effects were estimated by meta-analyses. Several months after transplanting, average AMF colonization was in proportion 84 % higher in inoculated than non-inoculated seedlings (p?=?0.0042), while the average risk of seedling mortality was 42 % lower in inoculated than non-inoculated seedlings (p?=?0.047). These results indicate that inoculation can increase AMF colonization over the background levels occurring in the soil, leading to higher rates of survival.     

Studies on the Physiology of Nodule Formation: VIII. The Influence of the Size of the Rhizosphere Population of Nodule Bacteria on Root Hair Infection in Clover

With an adequate inoculum the number of infected root hairsin three species of clover (Trifolium parviflorum, T. patensand T. glomeratum) increased exponentially with time in twophases; the increase was rapid during the first 8-10 days beforenodulation begins, but slower afterwards. T. parviflorum hadmost infections and T. glomeratum the fewest. Experiments on varying inoculum size, using an avirulent mutanstrain of Rhizobhtm trifolii as diluent, showed that root-hairinfection was differentially limited by inoculum size duringthe two phases. Infection in all three species was about doubledby doubling the density of the virulent bacteria in the rhizospherebefore nodulation begins. After nodulation bacterial densityhad to be increased much more than twice to double the numberof infections. This increase in the infecting population wasinversely related to the numbers of infections formed on thethree host species. Early infection and nodulation were promoted by high bacterialdensity in the rhizosphere.     

Obesity-induced hypertension develops in young rats independently of the renin-angiotensin-aldosterone system

A correlation exists between obesity and hypertension. In the currently available models of diet-induced obesity, the treatment of rats with a high fat (HF) diet does not begin until adulthood. Our aim was to develop and characterize a model of pre-pubescent obesity-induced hypertension. Male Sprague-Dawley rats were fed a HF diet (35% fat) for 10 weeks, beginning at age 3 weeks. Blood pressure was measured by tail-cuff, and a terminal blood sample was obtained to measure fasting blood glucose, insulin, plasma renin, aldosterone, thiobarbitutic acid reactive substances (TBARS), and free 8-isoprostanes levels. The vascular reactivity in the aorta was assessed using a myograph. Blood pressure was increased in rats fed the HF diet (HF, 161 +/- 2 mm Hg vs. control, 137 +/- 2 mm Hg, P < 0.05). Blood glucose (HF, 155 +/- 4 mg/dL vs. control, 123 +/- 5 mg/dL, P < 0.05), insulin (HF, 232 +/- 63 pM vs. control, 60 +/- 11 pM, P < 0.05), TBARS (expressed as nM of malondialdehyde [MDA]/ml [HF, 1.8 +/- 0.37 nM MDA/ml vs. control 1.05 +/- 0.09 nM MDA/ml, P < 0.05]), and free 8-isoprostanes (HF, 229 +/- 68 pg/ml vs. control, 112 +/- 9 pg/ml, P < 0.05) levels were elevated in the HF diet group. Interestingly, plasma renin and aldosterone levels were not different between the groups. The maximum vasoconstriction to phenylephrine (10(-4) M) was increased in the HF diet group (HF, 26.1 +/- 1.5 mN vs. control 22.3 +/- 1.2 mN, P < 0.05). In conclusion, pre-pubescent rats become hypertensive and have increased oxidative stress and enhanced vasoconstriction when fed a HF diet. Surprisingly, this occurs without the increase in renin or aldosterone levels seen in the adult models of diet-induced obesity.     

Induced terpene accumulation in Norway spruce inhibits bark beetle colonization in a dose-dependent manner

Background

Tree-killing bark beetles (Coleoptera, Scolytinae) are among the most economically and ecologically important forest pests in the northern hemisphere. Induction of terpenoid-based oleoresin has long been considered important in conifer defense against bark beetles, but it has been difficult to demonstrate a direct correlation between terpene levels and resistance to bark beetle colonization.

Methods

To test for inhibitory effects of induced terpenes on colonization by the spruce bark beetle Ips typographus (L.) we inoculated 20 mature Norway spruce Picea abies (L.) Karsten trees with a virulent fungus associated with the beetle, Ceratocystis polonica (Siem.) C. Moreau, and investigated induced terpene levels and beetle colonization in the bark.

Results

Fungal inoculation induced very strong and highly variable terpene accumulation 35 days after inoculation. Trees with high induced terpene levels (n = 7) had only 4.9% as many beetle attacks (5.1 vs. 103.5 attacks m⁻²) and 2.6% as much gallery length (0.029 m m⁻² vs. 1.11 m m⁻²) as trees with low terpene levels (n = 6). There was a highly significant rank correlation between terpene levels at day 35 and beetle colonization in individual trees. The relationship between induced terpene levels and beetle colonization was not linear but thresholded: above a low threshold concentration of ∼100 mg terpene g⁻¹ dry phloem trees suffered only moderate beetle colonization, and above a high threshold of ∼200 mg terpene g⁻¹ dry phloem trees were virtually unattacked.

Conclusion/Significance

This is the first study demonstrating a dose-dependent relationship between induced terpenes and tree resistance to bark beetle colonization under field conditions, indicating that terpene induction may be instrumental in tree resistance. This knowledge could be useful for developing management strategies that decrease the impact of tree-killing bark beetles.     

Effect of peat-bran inoculum ofTrichoderma species on biological control ofRhizoctonia solani in lettuce

A range of known biocontrol or plant growth-stimulating species ofTrichoderma orGliocladium were grown on peat-bran substrate to yield between 5×10⁷–3×10¹⁰ colony forming units (cfu's)g^–1 substrate after 14 days growth. Inocula were incorporated into peat:sand potting compost infested withRhizoctonia solani to give 7–8 × 10⁴ cfu's of antagonist g^–1 compost and assessed for biological control activity using lettuce seedlings. Six of the eight antagonists decreased daming-off and three of these consistently increased yield in comparison withR. solani treatment alone.Subsequently, peat-bran inoculum ofT. harzianum isolate TH1 was incorporated at 0.5% w/v intoR. solani infested potting compost. Both autoclaved and nonautoclaved inoculum ofT. harzianum TH1 decreased disease and increased yield. Incorporation of ethyl acetate-extracted autoclaved inoculum ofT. harzianum TH1 resulted in similar levels of biocontrol and improved plant growth as did incorporation of nonautoclaved and autoclavedT. harzianum TH1 inoculum. The need to standardize inocula and controls is emphasized.     

Competition studies withRhizobium trifolii in laboratory experiments

Summary Competition studies were carried out in soil cores by comparing the nodule forming ability of antibiotic resistance marked strains, inoculated at three inoculum levels onTrifolium repens versus an effective indigenous Rhizobium population of 1.5×10⁵/gm. It was seen that the indigenous marked isolate G1067 formed a high proportion of nodules sampled (>90%) at all three inoculum levels (10⁵, 10⁷ and 10⁹ cells/seed) wherein the introduced foreign strain G1032 formed >50% of the nodules at the highest inoculum level.In a test tube experiment, competition for nodule sites was examined by inoculating mixtures of twoR. trifolii strains at different input levels on two cultivars ofTrifolium repens var. Huia and Titan. It was seen that G1032 was less competitive than G1006 and G1067 on cv. Huia but was more competitive on cv. Titan than the other two strains.