Enhancement of in vitro growth and resistance to gray mould of Vitis vinifera co-cultured with plant growth-promoting rhizobacteria

The potential of a plant growth-promoting rhizobacterium, Pseudomonas sp. (strain PsJN), to stimulate the growth and enhancement of the resistance of grapevine (Vitis vinifera L.) transplants to gray mould caused by Botrytis cinerea has been investigated. In vitro inoculation of grapevine plantlets induced a significant plant growth promotion which made them more hardy and vigorous when compared to non-inoculated plantlets. This ability increased upon transplanting. When grown together with B. cinerea, the causal agent of gray mould, significant differences of aggressiveness were observed between the inoculated and non-inoculated plants. The presence of bacteria was accompanied by an induction of plant resistance to the pathogen. The beneficial effect from this plant-microbe association is being postulated.     

Necrotizing activity of five Botrytis cinerea endopolygalacturonases produced in Pichia pastoris

Five Botrytis cinerea endopolygalacturonase enzymes (BcPGs) were individually expressed in Pichia pastoris, purified to homogeneity and biochemically characterized. While the pH optima of the five enzymes were similar (approximately pH 4.5) the maximum activity of individual enzymes differed significantly. For hydrolysis of polygalacturonic acid (PGA), the V(max,app) ranged from 10 to 900 U mg(-1), while the K(m,app) ranged from 0.16 to 0.6 mg ml(-1). Although all BcPGs are true endopolygalacturonases, they apparently have different modes of action. PGA hydrolysis by BcPG1, BcPG2 and BcPG4 leads to the transient accumulation of oligomers with DP < 7, whereas PGA hydrolysis by BcPG3 and BcPG6 leads to the immediate accumulation of monomers and dimers. The necrotizing activity (NA) of all BcPGs was tested separately in tomato, broad bean and Arabidopsis thaliana. They showed different NAs on these plants. BcPG1 and BcPG2 possessed the strongest NA as tissue collapse was observed within 10 min after infiltration of broad bean leaves. The amino acid (aa) D192A substitution in the active site of BcPG2 not only abolished enzyme activity but also the NA, indicating that the NA is dependent on enzyme activity. Furthermore, deletion of the Bcpg2 gene in B. cinerea resulted in a strong reduction in virulence on tomato and broad bean. Primary lesion formation was delayed by approximately 24 h and the lesion expansion rate was reduced by 50-85%. These data indicate that BcPG2 is an important virulence factor for B. cinerea.     

Secondary metabolites influence Arabidopsis/Botrytis interactions: variation in host production and pathogen sensitivity

Numerous studies have suggested that plant/pathogen interactions are partially mediated via plant secondary metabolite production and corresponding pathogen tolerance. However, there are inconsistent reports on the ability of particular compounds to provide resistance to a pathogen. Most of these studies have focused on individual isolates of a given pathogen, suggesting that pathogens vary in their sensitivity to plant-produced toxins. We tested variability in virulence among pathogen isolates, and the impact on this by plant production of, and pathogen tolerance to, secondary metabolites. Botrytis cinerea isolates showed differing sensitivity to purified camalexin, and camalexin-sensitive isolates produced larger lesions on camalexin-deficient Arabidopsis genotypes than on the wild type. In contrast, the camalexin-insensitive isolate produced lesions of similar size on wild-type and camalexin-deficient Arabidopsis. Additional analysis with Arabidopsis secondary metabolite biosynthetic mutants suggests that Botrytis also has variable sensitivity to phenylpropanoids and glucosinolates. Furthermore, Botrytis infection generates a gradient of secondary metabolite responses emanating from the developing lesion, with the Botrytis isolate used determining the accumulation pattern. Collectively, our results indicate that Arabidopsis/Botrytis interactions are influenced at the metabolic level by variations in toxin production in the host and sensitivity in the pathogen.     

中华剑角蝗的营养成分与利用评价

本文分析了中华剑角蝗 Acrida cinerea的营养成分 ,包括蛋白质、氨基酸、总糖、粗脂肪、粗灰分、无机矿物质元素及维生素 B1和 B2 等 ,并在分析其营养价值的基础上进行开发利用评价。     

Immunodetection of Botrytis-specific Invertase in Infected Grapes

Invertase (β-fructofuranosidase, EC 3.2.1.26) activity was shown to be stimulated in grape berries after infection with Botrytis cinerea . By using organism-specific extraction methods, evidence was found proving that both partners contribute to the increase in activity. Qualitative analysis of the extracts by SDS–PAGE showed a new invertase species in the botrytised material, with a molecular weight similar to that of Botrytis invertase (BIT). A method allowing the preparative isolation of homogeneous invertase from liquid cultures of B. cinerea in only three steps (ethanol precipitation, DEAE anion exchange and hydroxyapatite chromatography) with 50% recovery was developed. BIT appeared to be strongly glycosylated; interestingly, the amount of glycan seemed to vary and had a remarkable influence on the chromatographic behaviour of the enzyme. After chemical deglycosylation with TFMS, the BIT peptide was used for the production of polyclonal antibodies in chickens (anti-BIT-IgY). The antibodies recognised glycosylated as well as deglycosylated BIT, a partial denaturation of the protein being necessary for reaction. This indicates that the deglycosylation procedure had been successful and that the antibodies were in fact directed to the peptide moiety. However, after a short incubation at 70°C, native BIT was detectable by anti-BIT-IgY, while still active. Western blotting with extracts of diseased berries confirmed the fungal origin of the new invertase form. The anti-BIT-IgY proved highly specific, although some cross-reaction with a protein in Monilia laxa extracts occurred. The importance of careful immunogen preparation in the production of specific antibodies, and the potential of BIT as a target molecule for the immunological detection of B. cinerea are discussed.     

Developmental failure after experimental activation of insect eggs

Summary

Eggs taken from the genital tracts of Venturia (Nemeritis) canescens (Hym., Ichneum.) and Psychoda cinerea (Dipt., Psychod.) can be activated by brief exposure to distilled water. However, embryogenesis fails soon unless some other conditions are met. These are mechanical deformation of the egg in Venturia, and mating in Psychoda. Development ceases at or soon after meiosis in Psychoda and in most Venturia eggs; the remaining Venturia eggs fail to form an orderly blastoderm although they contain hundreds of nuclei. In Venturia the anomalies are probably due to insufficient activation of some cytoplasmic component(s) while Psychoda eggs fail in the absence of some paternal contribution, possibly the sperm which in this case must enter the oocyte within the follicle.     

Inhibitory effects on microbial growth of Botrytis cinerea and Erwinia carotovora on potato using of a biopolymer chitosan at different molecular weights

The antimicrobial efficiency of chitosan at different molecular weights (5, 37, 57 and 290 kDa) against Botrytis cinerea and Erwinia carotovora on potato (Solanum tuberosum L.) was investigated. In vitro study showed that chitosan of 37 kDa was the most active against E. carotovora (minimum inhibitory concentration (MIC) = 950 mg/L), whereas 5 kDa chitosan was the most active against B.cinerea. Coating of potato tubers with 100, 250 and 500 mg/L significantly decreased the rate of weight loss and chitosan of 37 kDa showed the best effect. The in vivo antibacterial effect indicated that all treatments (500, 1000 and 2000 mg/L) significantly inhibited the growth of E. carotovora compared with the control. The lowest decay incidence was observed with 37 kDa chitosan. However, the antifungal activity against B. cinerea inoculated of leaves showed no decay incidence at 500 and 1000 mg/L with 57 kDa chitosan after 48 h.     

Organic hydroponics induces systemic resistance against the air-borne pathogen,Botrytis cinerea (gray mould)

Here, we propose that organic hydroponics trigger induced systemic resistance (ISR) in lettuce against air-borne Botrytis cinerea, which causes gray mold. We compared effects of organic and chemical hydroponics, assessed presence of ISR elicitors in the hydroponic nutrient solution, and investigated molecular mechanism of ISR. Organic hydroponics significantly reduced gray mold lesions in lettuce (cultivated hydroponically) and cucumber (cultivated in soil and foliar sprayed with nutrient solution). The 1-aminocyclopropane-1-carboxylic acid synthase gene in lettuce and lipoxygenase and ethylene receptor-related gene in cucumber showed heightened expression, suggesting that the jasmonic acid/ethylene (JA/ET)-signaling pathway was involved in ISR for both crops. Low salicylic acid β-glucoside levels confirmed role of the ISR signaling pathway. ISR in both lettuce and cucumbers indicated that elicitors in organic hydroponics were nonhost-specific and that the JA/ET pathway was activated without microbe–root interaction. Thus, organic hydroponics can be an effective method for both soil-borne and air-borne disease control.