Biological control of the root-knot nematode Meloidogyne javanica by Trichoderma harzianum

The filamentous fungi Trichoderma spp. is currently developed as biocontrol agents against many plant pathogens. Recent studies have shown that these fungi are able to infect nematode eggs and juveniles. In this research, biological control of root-knot nematode (Meloidogyne javanica) by Trichoderma harzianum BI was investigated in greenhouse and laboratory experiments. Results showed that different concentrations (10²–10⁸ spores/ml) of T. harzianum BI decreased nematode infection and other parameters significantly, compared to control. T. harzianum BI was able to penetrate nematode egg mass matrix and significantly decreased nematode egg hatching level. Specific activities of resistance-related enzymes, namely peroxidase (POX), polyphenol oxidase (PPO) and phenylalanine ammonia lyase (PAL) increased significantly in T. harzianum BI inoculated plants. Maximum activities of POX, PPO and PAL were observed at the 5, 5 and 6 days after inoculation, respectively. Chitinase activity was also increased in culture filtrates of T. harzianum BI grown on wheat bran moistened with salt solution supplemented with colloidal chitin or nematode eggs. Maximum activity of chitinase was recorded at the 4 days after inoculation, in media supplemented with colloidal chitin (1.15 U/min per ml) and nematode eggs (0.85 U/min per ml). Results suggested that direct parasitism of eggs through the increase in extracellular chitinase activity, which would be indicator of eggs infection capability, and inducing plant defense mechanisms leading to systemic resistance are two main suppression mechanisms used by T. harzianum BI against nematode.     

Interactions between the arbuscular mycorrhizal fungus Glomus mosseae and plant growth-promoting fungi and their significance for enhancing plant growth and suppressing damping-off of cucumber (Cucumis sativus L.)

This study investigated the interactions between the arbuscular mycorrhizal fungus (AMF) Glomus mosseae and the plant growth-promoting fungi (PGPF) Penicillium simplicissimum GP17-2 and Trichoderma harzianum GT3-2 in relation to their colonization of roots and the rhizosphere of cucumber (Cucumis sativus L.), and their effect on plant growth and suppression of the damping-off pathogen Rhizoctonia solani. Combined inoculation of T. harzianum with G. mosseae increased the percentage of AMF root colonization, but the presence of P. simplicissimum had no significant effect on mycorrhizal formation. The existence of G. mosseae decreased the population development of T. harzianum in and/or around the roots, however, growth of P. simplicissimum was not affected. Both PGPF isolates were effective in increasing the plant shoot and root dry weight, G. mosseae, however, was not. Nevertheless, co-inoculation of G. mosseae with T. harzianum synergistically enhanced plant growth. A comparable additive effect on plant growth was not observed when P. simplicissimum was combined with G. mosseae. Treating plants with the PGPF either simultaneously with the pathogen or prior to pathogen infection suppressed damping-off disease. G. mosseae was significantly effective in reducing disease severity when inoculated prior to the pathogen but not when inoculated simultaneously with the pathogen. Interestingly, the levels of disease protection achieved by the single inoculation of P. simplicissimum, T. harzianum or G. mosseae were significantly increased by combined inoculation of each PGPF with G. mosseae.     

Interactions between Onychiurus armatus and Trichoderma harzianum in take-all disease suppression in a simple experimental system

In a simple experimental system with wheat plants, interactions between the collembolan Onychiurus armatus and Trichoderma harzianum, a soil-borne fungus parasite of several plant pathogenic fungi, were studied in the presence of Gaeumannomyces graminis var. tritici, one of the most important foot and root fungal pathogens of cereals world-wide. Trichoderma harzianum was used according to two different modalities: fungal inoculum applied to seed or mixed with substrate. The isolate of T. harzianum proved to be ineffective against the pathogenic fungus, independent of the application modality, as shown in the experiments where this fungus was used alone, whereas Collembola used alone significantly reduced disease severity. However, the mode by which T. harzianum was applied significantly influenced the disease control ability of Collembola. In fact, only when T. harzianum was coated to seed were springtails able to reduce the disease caused by G. graminis var. tritici.     

The Relationship Study of Biomass,Situation, and Artificial Control: the Degradation of NP Using Estuary-Derived Fungi

Cultural situation generally played a crucial role on biodegradation mechanism establishment of pollutant. Extensive studies had focused on the optimization of cultivation environment based on in situ conditions. However, there were still few reports on the effects of artificial control on microbial growth and degradation. In this work, the relationship of biomass, situation, and artificial control was explored through the biodegradation of nonylphenols as standard containments by four trains named A. niger, A. terreus SHPP01, A. terreus NIH2624, and T. aff. harzianum from the estuary sediment of Jiaozhou Bay. Various culture conditions covering mineral salt medium, glucose synergistic medium, and carbon rich complex medium had been used to quest the relationship. As a conclusion, different strains usually showed different mechanisms within the same media. The correlation ship between biomass and degradation and removal rate was positive, and the natural medium was usually the best choice for microbial study in situ simulation. Hence, our study provided a valuable reference for the realization of more efficient and rapid biodegradation of pollutants.     

Interactive effects of Nemarioc-AL phytonematicide,Steinernema feltiae and Trichoderma harzianum on the reproduction of Meloidogyne incognita race 2 under greenhouse conditions

Interactive effects of alternatives to methyl bromide on root-knot (Meloidogyne species) nematodes could provide information on whether to use the products combined or singularly. The objective of this study was to investigate the interactive effects of Nemarioc-AL phytonematicide, Steinernema feltiae and Trichoderma harzianum on eggs and reproductive factors of Meloidogyne incognita race 2 under greenhouse conditions. The interactive effects of Nemarioc-AL phytonematicide (N), S. feltiae (S) and T. harzianum (T) on population densities of M. incognita were examined in a 2?×?2?×?2 factorial experiment in pots filled with steam-pasteurised soil. Each tomato seedling was inoculated with 2000 eggs and second-stage juveniles (J2), with phytonematicide and biocontrol agents applied at commercial rates. At 56 days after the treatments, the second-order (N₁S₁T₁) interaction reduced eggs and the reproductive factor (RF) in roots by 100% and 93%, respectively. Similarly, the first-order (N₁S₀T₁) interaction reduced the two variables by 92% and 72%, respectively, whereas N₁S₁T₀ interaction had 63% and 55% reduction effects, respectively. Nemarioc-AL phytonematicide was compatible with S. feltiae and T. harzianum and could, therefore, be used collectively to manage population densities of M. incognita.     

Fermentation of sugar beet waste by Aspergillus niger facilitates growth and P uptake of external mycelium of mixed populations of arbuscular mycorrhizal fungi

Sugar beet waste has potential value as a soil amendment and this work studied whether fermentation of the waste by Aspergillus niger would influence the growth and P uptake of arbuscular mycorrhizal (AM) fungi. Plants were grown in compartmentalised growth units, each with a root compartment (RC) and two lateral root-free compartments (RFC). One RFC contained untreated soil while the other RFC contained soil, which was uniformly mixed with sugar beet waste, either untreated (SB) or degraded by A. niger (ASB) in a rock phosphate (RP)-supplied medium. The soil in each pair of RFC was labelled with ³³P and ³²P in order to measure P uptake by the AM fungal mycelium, of which length density was also measured. Whole cell fatty acid (WCFA) signatures were used as biomarkers of the AM fungal mycelium and other soil microorganisms. The amount of biomarkers of saprotrophic fungi and both Gram-positive and Gram-negative bacteria was higher in SB than in ASB treatments. Whilst ASB increased growth and activity of AM mycelium, SB had the opposite effect. Moreover, shoot P content was increased by the addition of ASB, and by inoculation with AM fungi. Modification of soil microbial structure and production of exudates by A. niger, as a consequence of fermentation process of sugar beet waste, could possibly explain the increase of AM growth in ASB treatments. On the other hand, the highest P uptake was a result of the solubilisation of rock phosphate by A. niger during the fermentation.     

Effets de quelques pesticides sur la colonisation d'un substrat par le Trichoderma harzianum rifai en presence des autres champignons du sol

The effects of some pesticides upon substrate colonization by Trichoderma harzianum Rifai in the presence of other soil fungi.A herbicide (propyzamide) and 5 fungicides (benomyl, quintozene, vinclozolin. thiram, prothiocarb) were added at various concentrations to two Trichoderma harziunum-enriched garden soils. After thorough mixing, followed by solidifying with water agar, discs of these mixtures were cut with a cork-borer and transferred to a non specific nutrient medium (PDA). The development of T. harzianum on the agar, around each implant, was estimated after incubation for 1 week at 28°C.In the presence of other soil fungi, T. harzianum development was scarcely or not at all modified by prothiocarb, propyzamide and quintozene. It was highly depressed by benomyl. Vinclozolin was only slightly inhibitory. In our experimental conditions, T. harzianum development was strongly enhanced by thiram, even at a high dosage (300 μg ml^?1).The limits and the advantages of the method are estimated and the results are discussed in view of practical applications.     

Evaluation of bioremediation and detoxification potentiality of Aspergillus niger for removal of hexavalent chromium in soil microcosm

Aspergillus niger isolated from soil of leather tanning effluent had higher activity to remove chromium then the other fungal isolates. The potency of A. niger was evaluated in shake flask culture by absorption of chromium at pH 6, temperature 30 °C. The toxicity of chromium evaluated in petriplates and soil microcosm seed bioassay test had indicated increase in toxicity with the higher concentration of chromate. A. niger introduced in soil microcosm (40% moisture content) with different concentration of chromate (250, 500, 1000, 1500 and 2000 ppm) removed more than 70% chromium in soil contaminated by 250 and 500 ppm of chromate. However, chromium-contaminated soil (2000 ppm of potassium chromate) mixed with compost (5% and 10%) significantly removed chromium in presence of fungus, A. niger. The results of chromate toxicity in the wheat plants revealed that the peroxidases was induced due to increase of metal stress which was reversed in soil microcosm amended with compost.     

甘薯长喙壳侵染对甘薯块根抗氧化酶活性的影响

以易感黑斑病甘薯品种“徐薯18”为试验材料, 分别接种分离自石榴、甘薯和芋头的甘薯长喙壳(Ceratocystis fimbriata), 研究其保护酶活性及丙二醛(MDA)含量变化。结果显示, 非亲和性菌株[分离自石榴的甘薯长喙壳(CQS2)和分离自芋头的甘薯长喙壳(YP1)]和亲和性菌株[分离自甘薯的甘薯长喙壳(SP1)]接种的甘薯块根苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性, 与对照处理相比均有升高, 且接种CQS2 和YP1 的处理始终高于接种SP1 的处理; 非亲和性菌株和亲和性菌株接种的甘薯块根MDA 含量均呈先上升后下降趋势, 且接种CQS2 和YP1 的处理在接种25 h 时MDA 含量明显高于接种SP1 的处理, 接种SP1 的处理在接种后期(50 h)的MDA 含量降至与对照相当水平。由此表明: 不同寄主的甘薯长喙壳侵染甘薯块根后, 非亲和性接种菌株(CQS2 和YP1)比亲和性接种菌株(SP1)能增强POD、SOD和CAT 酶活性及诱导较多的MDA, 从而增强病原菌侵染的抗性。     

Effects of Single and Combined Inoculations with Azospirillum brasilense and Trichoderma harzianum on Seedling Growth or Yield Parameters of Wheat (Triticum vulgaris L., Giza 168) and Corn (Zea mays L., Hybrid 310)

Inoculation of wheat and corn grains with formulations of Azospirillum brasilense significantly increased seedling growth parameters of wheat and corn compared to untreated controls. Inoculation with Azospirillum and supplemental Trichoderma harzianum free or coimmobilized in calcium alginate resulted in significant increase in all plant growth parameters in addition to improving plant nutrient-content [phosphorus (P), potassium (K), and calcium (Ca)]. Grain treatments with T. harzianum alone or in a combination with A. brasilense were protected from invasion by Fusarium in a pot experiment. Nitrogen (N) fixation was investigated by A. brasilense free or double inoculated with T. harzianum in soil amended with different C-sources; also, phosphate solubilization was tested by these two organisms. Single and double inoculation with A. brasilense and/or T. harzianum improved wheat yield growth parameters in addition to seed protein; therefore, immobilized and coimmobilized formulations could be used as biofertilizer and biopesticide, and might be recommended to avoid the extensive use of the agrochemicals.     

APPLICATION OF TRICHODERMA HARZIANUM T22 AS A BIOFERTILIZER POTENTIAL IN MAIZE GROWTH

This study was conducted to investigate the effect of Trichoderma harzianum (T22) on the growth and development of maize (Zea mays) plants. T. harzianum was applied to the grains in two different treatments, either by inoculating the soil with air-dried mycelia of T. harzianum or by treating the seeds with different concentrations of the metabolic solution (as the culture supernatant of T. harzianum) before sowing. Comparisons were made between the two treatments to determine if any beneficial effects for improving the growth of maize plants occurred. Results revealed that application of the highest concentration of air-dried mycelia and all concentrations of the metabolic solution of T. harzianum caused increases in all measured parameters which include growth parameters, chlorophyll content, starch content, nucleic acids content, total protein content and phytohormone content of maize plants but the magnitude of these increases was much more pronounced in case of treating the seeds with the metabolic solution of T. harzianum.     

Production of chlamydospores and conidia by Trichoderma spp in liquid and solid growth media

Isolates of Trichoderma spp grew and produced chlamydospores as well as conidia in molasses-corn steep liquor (M-CSL), sucrose nitrate (SN), and glucose tartrate (GT) media. In M-SCL, isolates of T. hamatum, T. viride, and T. harzianum formed 10.4, 5.9 and 1.1 × 10⁸ chlamydospores g^?1 dry weight of mycelium. Fewer chlamydospores formed in SN and GT. Although T. harzianum formed the least number of chlamydospores, it produced the highest number of conidia in all three media. Molasses-corn steep liquor was superior to SN or GT in supporting development of both spore types. Spore production was not influenced by initial pH of the media or by continuous maintenance of the media at pH 4 or 7. Equal numbers of chlamydospores were formed in liquid media incubated in shake or static culture. Conidia formation, however, was stimulated in static culture. Chlamydospores and conidia of several naturally occurring isolates and induced biotypes of Trichoderma spp were abundantly produced on a variety of solid substrates moistened with liquid nutrients or water, preferably at pH 4. Bran, cornmeal, and peanut hull meal were better than eight other solid substrates for production of chlamydospores and conidia. A u.v.-induced, benomyl-resistant biotype of T. viride (T-1-R9) formed 22 × 10⁷ and 18 × 10⁸ chlamydospores and conidia, respectively, g^?1 of bran-SN. The ratio of conidia to chlamydospores was always greater in solid than liquid media. In solid media, 10 times more conidia than chlamydospores were formed, whereas in liquid only two or three times more were formed. Chlamydospores from liquid and solid fermentations were viable (ca 80%) and fresh chlamydospores germinated well (ca 75%) on nutrient agar. Although dried chlamydospores were viable, as determined with tetrazolium bromide, their germination on agar was poor (ca 8%).     

Astigmatid mite growth and fungi preference (Acari: Acaridida): Comparisons in laboratory experiments

The relationship between palatability and suitability of fungi for sustaining mite population growth was tested in the laboratory. The effect of mite fungal preference on spore dispersion was also studied. Eight species of microscopic fungi, Alternaria alternata, Aspergillus niger, Aspergillus versicolor, Cladosporium cladosporioides, Eurotium amstelodami var. amstelodami, E. amstelodami var. montevidensis, Mycocladus corymbifer, and Penicillium aurantiogriseum differed in their attractiveness as food and in their suitability to sustain population growth of three astigmatid mites, Acarus siro, Lepidoglyphus destructor and Tyrophagus putrescentiae. The preference for a particular species of fungus and its suitability for sustaining population growth were not correlated. Observed differences among mite species in their population growth were more likely associated with their ability to digest fungi. Fungi were divided into four groups based on their palatability and suitability to sustain population growth: (i) preferred food and suitable for mite growth (e.g. A. alternata, C. cladosporioides); (ii) preferred, but unsuitable (e.g. E. amstelodami var. amstelodami); (iii) avoided, but suitable (e.g. A. versicolor, M. corymbifer and E. amstelodami var. montevidensis); and (iv) avoided and unsuitable (e.g. A. niger). With respect to fungal spore dispersion, mites had the least effect on “avoided and unsuitable” fungal species while preferred fungi were most influenced by mite grazing.     

生活污泥对白菜供磷和土壤磷状况的影响

采用好气培养和盆栽试验以探明污泥磷的肥效,降低污泥施用导致的土壤磷累积引起的环境风险。结果表明,单施污泥土壤有效磷含量和白菜吸磷量均显著低于施用磷酸一铵和鸡粪处理;施用污泥后有利于增加白菜生长后期土壤磷酸酶的活性和土壤Olsen-P含量。在白菜等产量条件下,单施污泥处理土壤中Olsen-P残留量显著高于污泥与化肥混施处理;在P2O5施用量为902~70 kg/hm2时,污泥堆肥磷的肥效为磷酸一铵的25%左右。     

溶磷真菌液体培养条件下对磷矿粉的溶解效果

Rock phosphate （RP） is a low efficiency P fertilizer that is directly applied to the soil and can be solubilized by phosphate-solubilizing microorganisms （PSMs） in fermentation or soil conditions. This study investigated dynamic solubilization of 2 concentrations of rock phosphate in a liquid culture with different dosages of glucose by two fungal isolates,Aspergillus niger P39 and Penicillium oxalicum P66, from soybean and wheat rhizosphere soil. Although during the 20 day culture period A. niger P39 had a stronger ability to acidify the culture media than P. ozalicum P66, soluble P concentrations at glucose dosages of 30 and 50 g L^-1 with RP of 15 g L^-1 in the culture solution were much higher by P. oxalicum P66. The greater effectiveness of P. oxalicum P66 compared to A. niger P39 in the solubilization of RP was strongly associated with the production of organic acids. This study suggested that for RP solubilization the type rather than the concentration of PSM-produced organic acids was more important.     

Effect of microbial culture filtrates on the growth of sal (Shorea robusta gaertn.) leaf litter fungi

Experiments were made on the nature of fungal competition on sal leaf-litter with emphasis on the antibiotic action of culture filtrates. The culture filtrates of Mortierella subtilissima, Aspergillus candidus, A. flavus, A. niger, Penicillium rubrum, Papulaspora sp., Staphylococcus aureus and Bacillus subtilis were found most effective in suppressing the growth of various leaf-litter fungi. Dominant fungi which displayed best competitive tolerance to the staling products of culture filtrates were Aspergillus flavus, A. niger, A. sclerotiorum, A. terreus and Trichoderma harzianum.     

Evaluation of Compatibility of Rice Starch and Pectins by Glass Transition and Sub‐Tg Endotherms and the Effect of Compatibility on Gel Viscosity and Water Loss

Differences in molecular structure and hydrophilicity may affect the compatibility of food components in a highly concentrated solution. Mixtures of TNuS19 rice starch (RS) and pectins with three different degrees of esterification (22, 64, and 92%) were used as a model system to evaluate the components' compatibility in a low‐moisture system. When analyzed individually by differential scanning calorimetry (DSC), RS, low methoxyl pectin (LMP), intermediate methoxyl pectin (IMP), and high methoxyl pectin (HMP) showed the presence of a glass transition temperature (T_g) at 75.2, 96.2, 96.4, and 93.5°C, respectively. Among mixtures, the compatible RS‐HMP exhibited only a single T_g between the T_g values of the two components, whereas the incompatible RS‐LMP showed two T_g values that were close to those of the individual components. The sub‐T_g endotherms of all three mixtures (1:1) were lower than the means of the corresponding components. The degree of decrease was more pronounced in the RS‐HMP mixture than in the others. The above results imply that the interaction, which led to close contact between side chains of the two components, was more intense in the compatible RS‐HMP mixture than in the RS‐IMP and RS‐LMP mixtures. The decrease of the sub‐T_g endotherm can be used as an index to evaluate the degree of compatibility as well as the interaction occurring between the two molecules. The above findings were further verified by dynamic mechanical analyses. Both viscosity and water retention of the compatible RS‐HMP mixed gel were significantly higher than those of the RS‐IMP and RS‐LMP mixed gels. This evidence further suggests that RS and HMP are compatible and exhibit a strong intermolecular interaction that increases gel viscosity and decreases water loss during high‐temperature heating.     

Medically used radionuclides in sewage sludge

The concentration of medically used radionuclides has been studied in sludge from the sewage treatment plant serving the borough of Malmo. In this area all nuclear medicine procedures are carried out in one hospital and almost all patients live in the borough. Therefore, the input of medically used radionuclides into the sewage system can be estimated with good accuracy. Samples of digested sludge have been taken once or twice a week during half a year. Iodine-131 (physical half life (T) = R.05 d) was detected in all samples. The ¹³¹I-activity concentration due to medical use varied between (0.03±0.01) and (0.12±0.02) nCi kg^?1. The ratio between the total output of ¹³¹I via the sludge and an adherent input of the radionuclide into the sewage system was determined to (2.6 ± 0.6) × 10^?3, which is equivalent to a ratio of (2 ± 1) × 10^?2 for stable I. Occasionally measurable activities of ¹⁹⁸Au (T=2.7 d) and ²⁰¹T1(T=3.l d) have been found. The radioactivity concentration of medically used radionuclides in the sludge is low and constitutes no health problems for the persons involved. The sludge however has proved to be a very sensitive and suitable integrator of radioactive material released from a large urban area.     

Parasitism of sclerotia of Sclerotium rolfsii by trichoderma harzianum

The ability of Trichoderma harzianum isolate 203 to attack the soil-borne plant pathogen Sclerotium rolfsii is apparently connected with the production by the isolates of chitinase and β-(1,3)-glucanase inside the attacked sclerotia during parasitism.SEM and TEM micrographs show that the mycoparasite degraded walls of sclerotial cells and the attacked cells lost their cytoplasmic contents. It is assumed that T. harzianum utilizes sclerotial cell contents thus enabling it to sporulate intensively on the sclerotial surface and inside the digested cells.     

Effect of some rare earth elements on the growth and lanthanide accumulation in different Trichoderma strains

Accumulation of rare earth elements (REE) in the soil may be due to the use of REE enriched fertilizers and to contamination by REE containing wastes. Although widely used in China for soil and foliar dressing of crops, little is known about the effect of REE applications on the soil microbial community. The effect of REE on the growth of biological control strains of Trichoderma atroviride and Trichoderma harzianum was investigated in vitro using either a mix of different REE containing different amounts of lanthanum, cerium, praseodymium, neodymium, gadolinium nitrate and lanthanum nitrate alone in comparison to treatments with potassium nitrate and water. In plate tests applied concentrations ranged from 0.1 mM to 300 mM for lanthanum and REE mix and from 0.1 mM to 900 mM for the potassium solution. In liquid culture tests applied concentrations ranged from 0.001 mM to 100 mM for lanthanum and REE mix and from 0.003 mM to 900 mM for the potassium solution. ICP-MS, TEM and TEM X-ray microanalysis were used to study the accumulation of REE in fungal biomass. All the Trichoderma strains showed a good tolerance to the presence of REE in the culture media. Some growth enhancing effects were observed in liquid cultures of T. harzianum strains but not in T. atroviride. Accumulation of REE in fungal biomass, both at intracellular level and in the extracellular matrix, was observed.