Isolation and characterization of a 2,4-dichlorophenoxyacetic acid-degrading soil bacterium

1989 ◽  
Vol 31 (1) ◽  
Author(s):  
G�nther Ditzelm�ller ◽  
Michael Loidl ◽  
Franz Streichsbier
Keyword(s):  
Soil Bacterium ◽  
Dichlorophenoxyacetic Acid ◽  
Isolation And Characterization ◽  
2,4 Dichlorophenoxyacetic Acid

scholarly journals Isolation and Characterization of Halotolerant 2,4- Dichlorophenoxyacetic Acid Degrading Bacteria From Sulfidic, Low Salinity Salt Springs

Fine Focus ◽  
2014 ◽  
Vol 1 (1) ◽  
pp. 39-50
Author(s):  
Michael G. Willis ◽  
David S. Treves
Keyword(s):  
Bacterial Communities ◽  
Dichlorophenoxyacetic Acid ◽  
Growth Strategies ◽  
Low Salinity ◽  
Isolation And Characterization ◽  
Degrading Bacteria ◽  
History Of ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Tfda Gene

The bacterial communities at two sulfidic, low salinity springs with no history of herbicide contamination were screened for their ability to grow on 2,4-dichlorophenoxyacetic acid (2,4-D). Nineteen isolates, closely matching the genera Bacillus, Halobacillus, Halomonas, Georgenia and Kocuria, showed diverse growth strategies on NaCl-supplemented and NaCl-free 2,4-D medium. The majority of isolates were halotolerant, growing best on nutrient rich broth with 0% or 5% NaCl; none of the isolates thrived in medium with 20% NaCl. The tfdA gene, which codes for an a – ketoglutarate dioxygenase and catalyzes the first step in 2,4-D degradation, was detected in nine of the salt spring isolates. The tfdAa gene, which shows ~60% identity to tfdA, was present in all nineteen isolates. Many of the bacteria described here were not previously associated with 2,4-D degradation suggesting these salt springs may contain microbial communities of interest for bioremediation.

Characterization of the Reproductive Mode in Guayule In Vitro

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 483a-483
Author(s):  
Roy N. Keys ◽  
Dennis T. Ray ◽  
David A. Dierig
Keyword(s):  
Field Experiments ◽  
Reproductive Mode ◽  
Dichlorophenoxyacetic Acid ◽  
Initial Field ◽  
Breeding Lines ◽  
Reproductive Modes ◽  
Desert Southwest ◽  
2,4 Dichlorophenoxyacetic Acid

Guayule (Parthenium argentatum Gray, Asteraceae) is a latex-producing perennial desert shrub that is potentially of economic importance as an industrial crop for the desert Southwest. It is known to possess complex reproductive modes. Diploids are predominantly sexual and self-incompatible, while polyploids show a range of apomictic potential and self-compatibility. This paper describes the development of a relatively rapid and simple technique for characterizing reproductive modes of breeding lines of P. argentatum. Initial field experiments were based on an auxin test used successfully to characterize reproductive mode in the Poaceae. The application of 2,4-dichlorophenoxyacetic acid inhibited embryo formation in P. argentatum, but this was not the case with other auxins tested. Results of field experiments were ambiguous because: 1) the floral structure of P. argentatum is such that auxins might not have penetrated to the ovules, and 2) there was potential self-fertilization by pollen released within isolation bags. Therefore, in vitro culture of flower heads was tested because it provided much better control of environmental conditions, growth regulator application, and pollen release. Auxin alone, or in combination with gibberellic acid or kinetin, inhibited parthenogenesis in vitro. Embryo production did not vary using two substantially different nutrient media. In vitro flower head culture using a (Nitsch and Nitsch) liquid nutrient medium without growth regulators, enabled characterization of the reproductive mode of seven breeding lines, ranging from predominantly sexual to predominantly apomictic. The results of this technique were substantiated using RAPD analyzes of progeny arrays from controlled crosses.

scholarly journals Characterization of Hapten-Protein Conjugates: Antibody Generation and Immunoassay Development for Chlorophenoxyacetic Acid Pesticides

2009 ◽  
Vol 92 (6) ◽  
pp. 1773-1779 ◽  
Author(s):  
Robin C Boro ◽  
K Vikas Singh ◽  
C Raman Suri
Keyword(s):  
Inhibitory Concentration ◽  
Dichlorophenoxyacetic Acid ◽  
Protein Conjugation ◽  
Protein Conjugates ◽  
Highly Sensitive ◽  
Ic50 Values ◽  
Chlorophenoxyacetic Acid ◽  
2,4 Dichlorophenoxyacetic Acid

Abstract The generation of specific and sensitive antibodies against small molecules is greatly dependent upon the characteristics of the hapten-protein conjugates. In this study, we report a new fluorescence-based method for the characterization of hapten-protein conjugates. The method is based on an effect promoted by hapten-protein conjugation density upon the fluorescence intensity of the intrinsic tryptophan chromophore molecules of the protein. The proposed methodology is applied to quantify the hapten-protein conjugation density for two different chlorophenoxyacetic acid pesticides, 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4-dichlorophenoxybutyric acid (2,4-DB), coupled to carrier protein. Highly sensitive anti-2,4-D and anti-2,4-DB antibodies were obtained using these well-characterized hapten-protein conjugates. The generated antibodies were used in an immunoassay format demonstrating inhibitory concentration (IC50) values equal to 30 and 7 ng/mL for 2,4-D and 2,4-DB, respectively. Linearity was observed in the concentration range between 0.1500 ng/mL with LODs around 4 and 3 ng/mL for 2,4-D and 2,4-DB, respectively, in standard water samples. The proposed method was successfully applied for the determination of the extent of hapten-protein conjugation to produce specific antibodies for immunoassay development against pesticides.

scholarly journals Purification and Characterization of Two Enantioselective α-Ketoglutarate-Dependent Dioxygenases, RdpA and SdpA, from Sphingomonas herbicidovorans MH

2006 ◽  
Vol 72 (7) ◽  
pp. 4853-4861 ◽  
Author(s):  
Tina A. Müller ◽  
Thomas Fleischmann ◽  
Jan Roelof van der Meer ◽  
Hans-Peter E. Kohler
Keyword(s):  
Amino Acids ◽  
Divalent Cations ◽  
Propanoic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Purification And Characterization ◽  
Subgroup Ii ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Acid Herbicides ◽  
Growth Experiments

ABSTRACT α-Ketoglutarate-dependent (R)-dichlorprop dioxygenase (RdpA) and α-ketoglutarate-dependent (S)-dichlorprop dioxygenase (SdpA), which are involved in the degradation of phenoxyalkanoic acid herbicides in Sphingomonas herbicidovorans MH, were expressed and purified as His6-tagged fusion proteins from Escherichia coli BL21(DE3)(pLysS). RdpA and SdpA belong to subgroup II of the α-ketoglutarate-dependent dioxygenases and share the specific motif HXDX24TX131HX10R. Amino acids His-111, Asp-113, and His-270 and amino acids His-102, Asp-104, and His 257 comprise the 2-His-1-carboxylate facial triads and were predicted to be involved in iron binding in RdpA and SdpA, respectively. RdpA exclusively transformed the (R) enantiomers of mecoprop [2-(4-chloro-2-methylphenoxy)propanoic acid] and dichlorprop [2-(2,4-dichlorophenoxy)propanoic acid], whereas SdpA was specific for the (S) enantiomers. The apparent Km values were 99 μM for (R)-mecoprop, 164 μM for (R)-dichlorprop, and 3 μM for α-ketoglutarate for RdpA and 132 μM for (S)-mecoprop, 495 μM for (S)-dichlorprop, and 20 μM for α-ketoglutarate for SdpA. Both enzymes had high apparent Km values for oxygen; these values were 159 μM for SdpA and >230 μM for RdpA, whose activity was linearly dependent on oxygen at the concentration range measured. Both enzymes had narrow cosubstrate specificity; only 2-oxoadipate was able to replace α-ketoglutarate, and the rates were substantially diminished. Ferrous iron was necessary for activity of the enzymes, and other divalent cations could not replace it. Although the results of growth experiments suggest that strain MH harbors a specific 2,4-dichlorophenoxyacetic acid-converting enzyme, tfdA-, tfdAα-, or cadAB-like genes were not discovered in a screening analysis in which heterologous hybridization and PCR were used.

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