Non-Chemical Treatments for the Pre- and Post-Harvest Elicitation of Defense Mechanisms in the Fungi–Avocado Pathosystem

The greatest challenge for the avocado (Persea americana Miller) industry is to maintain the quality of the fruit to meet consumer requirements. Anthracnose is considered the most important disease in this industry, and it is caused by different species of the genus Colletotrichum, although other pathogens can be equally important. The defense mechanisms that fruit naturally uses can be triggered in response to the attack of pathogenic microorganisms and also by the application of exogenous elicitors in the form of GRAS compounds. The elicitors are recognized by receptors called PRRs, which are proteins located on the avocado fruit cell surface that have high affinity and specificity for PAMPs, MAMPs, and DAMPs. The activation of defense-signaling pathways depends on ethylene, salicylic, and jasmonic acids, and it occurs hours or days after PTI activation. These defense mechanisms aim to drive the pathogen to death. The application of essential oils, antagonists, volatile compounds, chitosan and silicon has been documented in vitro and on avocado fruit, showing some of them to have elicitor and fungicidal effects that are reflected in the postharvest quality of the fruit and a lower incidence of diseases. The main focus of these studies has been on anthracnose diseases. This review presents the most relevant advances in the use of natural compounds with antifungal and elicitor effects in plant tissues.     

Effect of supplementary manganese on the sporulation of Bacillus endospores analysed by Raman spectroscopy

It is a common practice in microbiology to induce and accelerate sporulation of spore‐forming bacteria by adding small amounts of divalent manganese to the cultivation medium. By micro‐Raman spectroscopy the effect of supplementary divalent manganese during the growth and sporulation of Bacillus spp. bacteria was studied. The spectral alterations in the Raman spectra of single endospores due to this cultivation parameter comprised slight alterations of the bands attributed to intracellular, abundantly present calcium dipicolinate (CaDPA). Those signals suffered a loss of intensity or partial band broadening because of the appearance of new weak signals next to them. Exclusively in Raman spectra of single B. sphaericuss endospores, the band at 1485 cm⁻¹ vanished. The theoretical spectra of CaDPA and manganese dipicolinate (MnDPA) were calculated and compared with the experimental spectra to prove the hypothesis that, while the overall intracellular DPA content decreased, an intracellular assembly of MnDPA in the endospores might also occur. Band shifts of the COO⁻ vibrations in the salt's spectra as well as in the endospore's spectra, and the decrease of the two CaDPA bands, confirmed this proposal. The appearance of the 1030 cm⁻¹ band in all Bacillus spectra as well as the disappearance of the 1485 cm⁻¹ band in the B. sphaericus spectra still needs to be clarified. With the help of two multivariate chemometric methods, these spectral alterations allowed discrimination between single endospores of different Bacillus strains cultivated on normal nutrient agar (NA) and those grown on NA with MnSO₄ · xH₂O addition. With these investigations, a possible strategy is shown to trace back the cultivation environment of matured single endospores. Utilizing the joint concept of micro‐Raman spectroscopy and chemometric analysis, the differentiation between natively grown endospores and those cultivated in a laboratory with the help of manganous salts as a common sporulation accelerator seems accomplishable. Copyright © 2009 John Wiley & Sons, Ltd.     

Disinfection of gram-negative and gram-positive bacteria using DynaJets® hydrodynamic cavitating jets

Cavitating jet technologies (DynaJets®) were investigated as a means of disinfection of gram-negative Escherichia coli, Klebsiellapneumoniae, Pseudomonas syringae, and Pseudomonas aeruginosa, and gram-positive Bacillus subtilis. The hydrodynamic cavitating jets were found to be very effective in reducing the concentrations of all of these species. In general, the observed rates of disinfection of gram-negative species were higher than for gram-positive species. However, different gram-negative species also showed significant differences (P. syringae 6-log₁₀ reduction, P. aeruginosa 2-log₁₀ reduction) under the same conditions. Disinfection of E. coli repeatedly showed five orders of magnitude reduction in concentration within 45-60-min at low nozzle pressure (2.1 bar).Optimization of nozzle design and operating pressures increased disinfection rates per input energy by several orders of magnitude. The power efficiencies of the hydrodynamic cavitating jets were found to be 10-100 times greater than comparable ultrasonic systems.     

Efficient production of poly-γ-glutamic acid by bacillus subtilis ZJU-7

A strain with high poly-γ-glutamic acid (γ-PGA) production was isolated from fermented bean curd, a traditional Chinese food. The strain was named Bacillus subtilis ZJU-7 according to 16s rDNA sequencing and its taxonomic characters. The culture conditions for γ-PGA production were evaluated. The most suitable carbon and nitrogen sources were sucrose and tryptone, respectively. Exogenous l-glutamic acid was necessary for γ-PGA production, and the production of γ-PGA increased on the addition of l-glutamic acid to the medium. In the medium containing 60 g/L of sucrose, 60 g/L of tryptone, 80 g/L of l-glutamic acid, and 10 g/L of NaCl, the yield of γ-PGA reached 54.4 g/L after cultivation at 37°C for 24h, which was the highest γ-PGA production compared with values reported in the literature. The average molecular mass of γ-PGA produced was about 1.24×10⁶ Daltons. B. subtilis ZJU-7 is genetically stable and can synthesize levan instead of γ-PGA without the addition of l-glutamic acid to the medium.     

Cloning, characterization, and expression of a new cry2Ab gene from Bacillus thuringiensis strain 14-1

Bacillus thuringiensis is the major source for transfer of genes to impart insect resistance in transgenic plants. Cry2A proteins of B. thuringiensis are promising candidates for management of resistance development in insects owing to their difference from the currently used Cry1A proteins, in structure and insecticidal mechanism. The cry2Ab gene was found to lack a functional promoter and, hence, is cryptic in nature. The cry2Ab7 gene was cloned from a new indigenous B. thuringiensis strain, 14-1. Nucleotide sequencing of the cry2Ab gene cloned from B. thuringiensis strain 14-1 revealed an open reading frame of 1902 bp. The deduced amino acid sequence of Cry2Ab of B. thuringiensis strain 14-1 showed a variation in three amino acid residues in comparison to the holotype sequence, Cry2Ab1. Expression of the newly cloned cry2Ab gene was studied in an acrystalliferous strain of B. thuringiensis (4Q7) by fusing the cry2Ab gene downstream of cry2Aa promoter and orf1+orf2 sequences. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of a spore-crystal mixture obtained from transformants of B. thuringiensis strain 4Q7 showed production of Cry2Ab protein of about 65 kDa. Alkali solubilized Cry2Ab7 protein showed toxicity against Helicoverpa armigera neonates.     

Temporal and spatial dynamics of blocking and ripening effects on bacterial transport through a porous system: a possible explanation for CFT deviation

The studies on transport of particles across porous systems are based on the Colloid Filtration Theory (CFT). According to CFT, the collision efficiency is constant along the system length [J.N. Ryan, M. Elimelech, Colloids Surf. A: Physicochem. Eng. Aspects 107 (1996) 1–56]. Decreasing values of collision efficiency have been reported, a phenomenon that has been interpreted as a deviation from the CFT [X. Li, T.D. Scheibe, W.P. Johnson, Environ. Sci. Technol. 38 (2004) 5616–5625; N. Tufenkji, J.A. Redman, M. Elimelech, Environ. Sci. Technol. 37 (2003) 616–623; N. Tufenkji, M. Elimelech, Langmuir 20 (2004) 10818–10828; N. Tufenkji, M. Elimelech, Langmuir 21 (2005) 841–852]. This paper presents data on transport of Bacillus megaterium spores through quartz sand columns. The occurrence of consecutive phases of increase and decrease of the values of C/C₀, the effluent spore concentration expressed as a fraction of the influent spore concentration, is reported. These patterns of change in C/C₀ were interpreted as the result of the concomitant occurrence of blocking and ripening, the prevalence of these phenomena in different moments of the experiment, and the spatial distribution of the prevalence of blocking and ripening effects along the porous system. It is argued that this spatial distribution in the predominance of blocking and ripening, what leads to the intensification of ripening at the entrance of the porous system, might be a possible explanation for the reported deviation from the CFT for experimental conditions where ripening and blocking take place.     

Differentiation of strains from the Bacillus cereus group by RFLP‐PFGE genomic fingerprinting

Bacillus mycoides, Bacillus pseudomycoides, Bacillus weihenstephanensis, Bacillus anthracis, Bacillus thuringiensis, and Bacillus cereus belong to the B. cereus group. The last three species are characterized by different phenotype features and pathogenicity spectrum, but it has been shown that these species are genetically closely related. The macrorestriction analysis of the genomic DNA with the NotI enzyme was used to generate polymorphism of restriction profiles for 39 food‐borne isolates (B. cereus, B. mycoides) and seven reference strains (B. mycoides, B. thuringiensis, B. weihenstephanensis, and B. cereus). The PFGE method was applied to differentiate the examined strains of the B. cereus group. On the basis of the unweighted pair group method with the arithmetic mean method and Dice coefficient, the strains were divided into five clusters (types A–E), and the most numerous group was group A (25 strains). A total of 21 distinct pulsotypes were observed. The RFLP‐PFGE analysis was successfully used for the differentiation and characterization of B. cereus and B. mycoides strains isolated from different food products.     

苏云金杆菌4.0718菌株超氧化物歧化酶的纯化和性质研究

经 (NH4 ) 2 SO4 分级沉淀、SephadexG 75凝胶过滤和DEAE 5 2柱层析将苏云金杆菌 4.0 718菌株的超氧化物岐化酶 (SOD)纯化到均一程度 ,酶比活力达 688.0U/mg ,酶得率为 47.9% 该酶经KCN ,H2 O2 ,氯仿 乙醇抑制实验 ,金属元素含量测定和紫外可见吸收光谱测定 ,表明是Fe SOD 经反相高效液相色谱和电喷雾质谱联合分析等方法 ,测得酶分子量为 3 9.4861× 10 6 ,由两个相同亚基组成 ,N 末端氨基酸为丙氨酸     

Biosorption of Heavy Metals by Sol-Gel Immobilized Bacillus sphaericus Cells, Spores and S-Layers

Different types of biocers were prepared by dispersing vegetative cells, spores and surface layer proteins (S-layers) of Bacillus sphaericus JG-A12 in aqueous silica nanosols, gelling or coating on glass, and drying. The enzymatic activity of embedded B. sphaericus cells depends noticeably on the water content of the biocer. The cells are destroyed by drying and shrinkage of the silica network whereas embedded spores retain their ability for germination. The biosorption of uranium and copper of these biocomposites was investigated. Biocers with cells possess the highest metal binding capacity compared to matrices with spores or S-layers. An additional increasement of the metal binding capacity is achieved by using penetration reagents like sorbitol. For renewed use biosorpted uranium and copper can be completely removed from the biocers by using aqueous citric acid. The use of spores as biocomponent offers new interesting possibilities for the preparation of storage-stable bioactive biocers.