Genetic improvement of Trichoderma ability to induce systemic resistance

The beneficial applications of Trichoderma spp. in agriculture include not only the control of plant pathogens, but also the improvement of plant growth, micronutrient availability, and plant tolerance to abiotic stress. In addition, it has been suggested that these fungi are able to increase plant disease resistance by activating induced systemic resistance (ISR) . The mode of action of these beneficial fungi in the Trichoderma -plant-pathogen interaction are many, complex and not completely understood. Numerous lytic enzymes have been characterized, the encoding genes (ech42 gluc78, nag1 from T. atroviride strain P1) cloned, and their role in biocontrol demonstrated. The corresponding biocontrol-related inducible promoters have been used in a reporter system based on the Aspergillus niger glucose oxidase gene (goxA) to monitor biocontrol activity. Glucose oxidase catalyzes the oxygen-dependent oxidation of D-glucose to D-glucono-1,5-lactone and hydrogen peroxide; this latter compound is known to have an antifungal effect and activate the plant defence cascade, thus increasing resistance to pathogen attack. T. atroviride P1 transformants with various promoters gox were tested as seed coating treatments on bean seeds planted in soil infested with a soilborne fungal pathogen. Successively, the emergent leaves were inoculated with a foliar pathogen to determine the effect of the GOX transformants on biocontrol and resistance to pathogen attack. Inoculations with the P1-GOX transformants not only reduced disease symptoms caused by a soil pathogen, but also the lesions of various foliar pathogens applied far from the Trichoderma colonization, thus activating ISR. A similar approach is being use to genetically improve T. harzianum T22, a rhizosphere competent and commercially marketed strain not transformed yet, by using four different gox gene constructs under the control of constitutive and inducible promoters. Plasmids have been introduced in Trichoderma by protoplasts co-transformation. hygromicin resistant progeny selected, and mitotically stable transformants analysed to confirm the presence of the novel enzyme activity. Progenies are being tested for biocontrol ISR inducing activity.

Induced Resistance of Tomato against Gray Mold (Botrytis cinerea) by Salicylic Acid (SA)

[ Objective ] The paper was to explore the induced resistance of tomato against gray mold （Botrytis cinerea） by salicylic acid. [ Method ] SA was used as an inducer to treat tomato seedlings, the effects of SA on mycelial diameter and spore germination of B. cinerea were studied, and the changes of 4 defense enzyme activities containing catalase （CAT）, peroxidase （POD）, polyphenol oxidase （PPO） and phenylalanine ammonia lyase （PAL）, as well as malondialdehyde （MDA） content during the production process of induced resistance were also measured. [ Result] SA had no inhibitory effect against spore germination and myce- lial growth of B. cinerea within the concentration range, and the relative induced effect had different degrees of improvement after treatment. The induced effect was the best as B. cinerea was challenged to inoculate at the third day after using 150 mg/L SA in tomato plants, and the duration of resistance was 10 -15 d. After treated by SA, CAT, POD, PPO and PAL first increased and then decreased in systemic induced resistance against B. cinerea, which were significantly higher than control. Meanwhile, MDA content showed ascendant trend in wavy line form. [ Conclusion ] The use of SA within a certain concentration range is safe; CAT, POD, PPO and PAL activities have positive correlation with induced resistance against B. cinerea, the increase of MDA content also has close relationship with the imvrovement of disease resistance.

Bacillus cereus AR156 primes induced systemic resistance by suppressing miR825/825 and activating defense-related genes in Arabidopsis

Small RNAs play an important role in plant immune responses. However, their regulatory function in induced systemic resistance（ISR） is nascent. Bacillus cereus AR156 is a plant growth-promoting rhizobacterium that induces ISR in Arabidopsis against bacterial infection. Here,by comparing small RNA profiles of Pseudomonas syringae pv. tomato（Pst） DC3000-infected Arabidopsis with and without AR156 pretreatment, we identified a group of Arabidopsis micro RNAs（mi RNAs） that are differentially regulated by AR156 pretreatment. mi R825 and mi R825 are two mi RNA generated from a single mi RNA gene.Northern blot analysis indicated that they were significantly downregulated in Pst DC3000-infected plants pretreated with AR156, in contrast to the plants without AR156 pretreatment. mi R825 targets two ubiquitin-protein ligases,while mi R825 targets toll-interleukin-like receptor（TIR）-nucleotide binding site（NBS） and leucine-rich repeat（LRR）type resistance（R） genes. The expression of these target genes negatively correlated with the expression of mi R825 and mi R825. Moreover, transgenic plants showing reduced expression of mi R825 and mi R825 displayed enhanced resistance to Pst DC3000 infection, whereas transgenic plants overexpressing mi R825 and mi R825 were more susceptible. Taken together, our data indicates that Bacillus cereus AR156 pretreatment primes ISR to Pst infection by suppressing mi R825 and mi R825 and activating the defense related genes they targeted.

Effect of Chaetomium globosum ND35 on Plant Growth and Preliminary Study of Its Biocontrol Efficacy

[Objective] The aim was to study effect of Chaetomium globosum ND35 on plant growth and preliminary study of its biocontrol efficacy, and provide basis for popularization and application of this strain.[ Method] With endophytic fungus C. globosum ND35 as a tested strain, effect of C. globosumND35 on plant growth and its biocontrol on five plant diseases were investigated in the greenhouse and field,[Result]The results showed that ND35 promoted growth of lateral root and diameter of breast height of poplar. ND35 can induce poplar to resist Poplar Valsa Canker caused by Valsa sordida and Poplar Rust caused by Melampsora puplicola. ND35 was also able to induce tomato and bean to resist Botrytis cinera. Biocontrol of Bean Stem Rot Rhizoctonia by ND35 was effective as well. [Conclusion] Induced systemic resistance by endophytic C. globosum ND35 plays an important role in biocontrol of plant diseases.

Effects of arbuscular mycorrhizal fungi on carbon assimilation and ecological stoichiometry of maize under combined abiotic stresses

Arbuscular mycorrhizal fungi(AMF)enhance plant tolerance to abiotic stresses like salinity and improve crop yield.However,their effects are variable,and the underlying cause of such variation remains largely unknown.This study aimed to assess how drought modifed the effect of AMF on plant resistance to high calcium-saline stress.A pot experiment was performed to examine how AMF inoculation affects the growth,photosynthetic activity,nutrient uptake and carbon(C),nitrogen(N)and phosphorus(P)stoichiometric ratio(C:N:P)of maize under high calcium stress and contrasting water conditions.The results showed that high calcium stress signifcantly reduced mycorrhizal colonization,biomass accumulation,C assimilation rate and C:N stoichiometric ratio in plant tissues.Besides,the adverse effects of calcium stress on photosynthesis were exacerbated under drought.AMF inoculation profoundly alleviated such reductions under drought and saline stress.However,it barely affected maize performance when subjected to calcium stress under well-watered conditions.Moreover,watering changed AMF impact on nutrient allocation in plant tissues.Under well-watered conditions,AMF stimulated P accumulation in roots and plant growth,but did not induce leaf P accumulation proportional to C and N,resulting in increased leaf C:P and N:P ratios under high calcium stress.In contrast,AMF decreased N content and the N:P ratio in leaves under drought.Overall,AMF inoculation improved maize resistance to calcium-salt stress through enhanced photosynthesis and modulation of nutrient stoichiometry,particularly under water defcit conditions.These results highlighted the regulatory role of AMF in carbon assimilation and nutrient homeostasis under compound stresses,and provide signifcant guidance on the improvement of crop yield in saline and arid regions.

Does drought stress modify the effects of plant-growth promoting rhizobacteria on an aboveground chewing herbivore？

Soil microbes have important effects on the interactions of plants with their environment, by promoting plant growth, inducing resistance to pests or by conferring tolerance to abiotic stress. However, their effects are variable and the factors responsible for this variation are mainly unknown. Our aim was to assess how drought stress modifies the effect of the nonpathogenic rhizobacterium Pseudomonas simiae WCS417r on plant growth and resistance against the generalist leaf-chewing caterpillar Mamestra brassicae. We studied Arabidopsis tha/iana Col-0 plants, as well as mutants altered in the biosyn- thesis of the phytohormones jasmonic acid （JA） and abscisic acid （ABA）. Caterpillars did not prefer rhizobacteria-treated plants, independently of drought stress. Rhizobacteria colonization had a variable effect on caterpillar performance, which ranged from positive in one experiment to neutral in a second one. Drought had a consistent negative effect on herbivore performance; however, it did not modify the effect of rhizobacteria on herbivore performance. The effect of drought on herbivore performance was JA-mediated （confirmed with the use of the dde2-2 mutant）, but it was still present in the ABA-deficient mutant aba2-1. Plant biomass was reduced by both drought and herbivory but it was enhanced by rhizobacterial colonization. Pseudomonas simiae WCS417r is able to promote plant growth even when plants are suffering herbivory. Nevertheless, the microbial effect on the herbivore is variable, independently of drought stress. To get the best possible outcome from the rhizobacteria-plant mutualism it is important to understand which other factors may be responsible for its context-dependency.