Diverse molecular markers have been used to analyze the genetic diversity of plant pathogens. Compared with traditional fingerprinting methods, multiple loci variable number of tandem repeat analyses (MLVAs) have gained importance recently due to their reproducibility, high discriminatory power, ease of performance, low cost, and throughput potential. These characteristics are desirable for continuous pathogen monitoring, especially for pathogens with relatively low genetic diversity, and for disease epidemiology studies. Genetic diversity studies of Xanthomonas phaseoli pv. manihotis, which is the causal agent of cassava bacterial blight, have shown variability and changes in the bacterial population over time. Thus, an easy and fast method needs to be developed to type populations of this pathogen in different countries of the world, especially on small scales. In this study, we developed an MLVA scheme to analyze X. phaseoli pv. manihotis variability on a local scale. The MLVA-15 scheme comprises 15 variable number of tandem repeat loci grouped into four multiplex polymerase chain reaction pools. We showed that the MLVA-15 scheme had slightly higher discriminatory ability at the locality level when compared with amplified fragment length polymorphisms. The MLVA-15 scheme allowed for an accurate determination of the number of genotypes in the sample and showed reproducibility and portability. Additionally, this scheme could be used to analyze numerous strains in a reasonable timeframe. The MLVA-15 scheme was highly specific to X. phaseoli but up to eight tandem repeat loci could be amplified from other Xanthomonas spp. Finally, we assessed the utility of the scheme for analyses of X. phaseoli pv. manihotis genetic variability in the Colombian Caribbean region. MLVA-15 distinguished 88.9% of the haplotypes in our sample. Strains originating from the same field and isolated at the same time could be discriminated. In this study, the advantages of the MLVA-15 scheme targeting 6- or 7-bp repeats were demonstrated. Moreover, this scheme was a fast method that was appropriate for routine monitoring of X. phaseoli pv. manihotis populations on a local scale and, thus, was useful for addressing epidemiological questions.
Cassava (Manihot esculenta), an important multipurpose crop, represents a food source for millions of people and a source of industrial raw material in tropical regions. A limitation in its production is cassava bacterial blight (CBB), a disease caused by Xanthomonas axonopodis pv. manihotis (Xam). CBB is spread mainly by propagation of infected cuttings, therefore early pathogen detection and disease diagnosis are crucial to prevent its dispersal and the consequent negative impact on the crop. A previously reported detection method was inefficient at detecting isolates of Xam more recently collected in cassava fields in Colombia. Consequently, a new method for pathogen detection was developed using multiplex nested PCR. A set of primers was selected amongst a group designed for five housekeeping gene sequences (rpoB, gltA, ftsZ, gyrB and groEL), and for the region of the gene encoding for the C-terminal portion of TALE1 Xam , a protein involved in virulence of Xam. In this work, a multiplex nested PCR based on the set of primers RpoB and Cterm was optimized to achieve an effective detection of Xam. The developed method successfully detects the presence of Xam in cassava plants collected in infected fields.
Energy and the environment will always play key roles in society. The climate emergency cannot be ruled out to enable the transition for a clean energy future. Currently, non-renewable energy resources are declining, therefore is important to continuously explore renewable resources. Biomass is a renewable resource that can be applied to reduce climate changes and to accomplhish emission policies. Cellulose is the most abundant type of biomass worldwide, which can be transformed into biofuels and potential building block platform molecules (e.g furfural) throughout biological or chemical methods. Furfural can be synthetized from cellulose using hydrolysis and dehydration reactions. Furfural has a furan ring and carbonyl functional group which makes it an important intermediary to produce higher value-added molecules at industrial level. These molecules include gasoline, diesel and jet fuel. However, furfural can also be transformed by hydrogenation, oxidation, decarboxylation and condensation reactions. The selective hydrogenation of furfural produces furfuryl alcohol, an important industrial compound, which is widely employed in the production of resins, fibers, and is considered an essential product for pharmaceutical applications. On the other hand, the oxidation of furfural produces furoic acid which is appliedin the agrochemical industry, where it is commonly transformed to furoyl chloride which is finally used in the production of drugs and insecticides. The oxidation and reduction of furfural can carry out through heterogeneous and homogeneous catalysis, and biocatalysis. Selectivity is an important issue in furfural hydrogenation and oxidation reactions since different products can be obtained by using monometallic or bimetallic catalysts and/or different catalyst supports. In biocatalysis approach, different enzymes, complete cells, tools of modern biotechnology, DNA sequencing, regulation of metabolic networks, overexpression of genes that encode enzymes of interest and optimization of the cellular properties of the microorganism are used. Herein, a review on the current status of furfuryl alcohol and furoic acid production from furfural by heterogeneous catalysis and biocatalysis has been studied. The stability, selectivity and activity of catalystsalong with the different furfural oxidation and reduction conditions have been pointed out. Additionally, the main enzymes, microorganisms and mechanism involved in the furfural degradation process have also been discussed.
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