The advent of biotechnology-derived, herbicide-resistant crops has revolutionized farming practices in many countries. Facile, highly effective, environmentally sound, and profitable weed control methods have been rapidly adopted by crop producers who value the benefits associated with biotechnology-derived weed management traits. But a rapid rise in the populations of several troublesome weeds that are tolerant or resistant to herbicides currently used in conjunction with herbicide-resistant crops may signify that the useful lifetime of these economically important weed management traits will be cut short. We describe the development of soybean and other broadleaf plant species resistant to dicamba, a widely used, inexpensive, and environmentally safe herbicide. The dicamba resistance technology will augment current herbicide resistance technologies and extend their effective lifetime. Attributes of both nuclear- and chloroplast-encoded dicamba resistance genes that affect the potency and expected durability of the herbicide resistance trait are examined.

Dicamba O-demethylase is a multicomponent enzyme from Pseudomonas maltophilia, strain DI-6, that catalyzes the conversion of the widely used herbicide dicamba (2-methoxy-3,6-dichlorobenzoic acid) to DCSA (3,6-dichlorosalicylic acid). We recently described the biochemical characteristics of the three components of this enzyme (i.e. reductase DIC , ferredoxin DIC , and oxygenase DIC ) and classified the oxygenase component of dicamba O-demethylase as a member of the Rieske nonheme iron family of oxygenases. In the current study, we used N-terminal and internal amino acid sequence information from the purified proteins to clone the genes that encode dicamba O-demethylase. Two reductase genes (ddmA1 and ddmA2) with predicted amino acid sequences of 408 and 409 residues were identified. The open reading frames encode 43.7-and 43.9-kDa proteins that are 99.3% identical to each other and homologous to members of the FAD-dependent pyridine nucleotide reductase family. The ferredoxin coding sequence (ddmB) specifies an 11.4-kDa protein composed of 105 residues with similarity to the adrenodoxin family of [2Fe-2S] bacterial ferredoxins. The oxygenase gene (ddmC) encodes a 37.3-kDa protein composed of 339 amino acids that is homologous to members of the Phthalate family of Rieske non-heme iron oxygenases that function as monooxygenases. Southern analysis localized the oxygenase gene to a megaplasmid in cells of P. maltophilia. Mixtures of the three highly purified recombinant dicamba O-demethylase components overexpressed in Escherichia coli converted dicamba to DCSA with an efficiency similar to that of the native enzyme, suggesting that all of the components required for optimal enzymatic activity have been identified. Computer modeling suggests that oxygenase DIC has strong similarities with the core ␣ subunits of naphthalene 1,2-dioxygenase. Nonetheless, the present studies point to dicamba O-demethylase as an enzyme system with its own unique combination of characteristics.The herbicide dicamba (2-methoxy-3,6-dichlorobenzoic acid) has been used to effectively control broadleaf weeds in crops such as corn and wheat for almost 40 years. Like a number of other chlorinated organic compounds, dicamba does not persist in the soil because it is efficiently metabolized by a consortium of soil bacteria under both aerobic and anaerobic conditions (1-4). Studies with different soil types treated with dicamba have demonstrated that 3,6-dichlorosalicylic acid (DCSA), 1 a compound without herbicidal activity, is a major product of the microbial degradation process (2, 3, 5). Soil samples taken from a single site exposed to dicamba for several years yielded a number of bacterial species capable of utilizing dicamba as a sole carbon source (6). These soil microorganisms could completely mineralize dicamba to carbon dioxide, water, and chloride ion (7). Studies on the metabolism of dicamba in the cells of one of these bacteria, the DI-6 strain of Pseudomonas maltophilia, showed that DCSA is a major degradation product (7,8).We have be...

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Herbicide-tolerant crops have been widely and rapidly adopted by farmers in several countries due to enhanced weed control, lower labor and production costs, increased environmental benefits, and gains in profitability. Soon to be introduced transgenic soybean and cotton varieties tolerant to treatments with the herbicide dicamba offer prospects for excellent broadleaf weed control in these broadleaf crops. Because monocots such as maize (Zea mays) can be treated with dicamba only during a limited window of crop development and because crop injury is sometimes observed when conditions are unfavorable, transgenic maize plants have been produced and tested for higher levels of tolerance to treatment with dicamba. Maize plants expressing the gene encoding dicamba monooxygenase (DMO) linked with an upstream chloroplast transit peptide (CTP) display greatly enhanced tolerance to dicamba applied either pre-emergence or postemergence. Comparisons of DMO coupled to CTPs derived from the Rubisco small subunit from either Arabidopsis thaliana or Z. mays showed that both allowed production of transgenic maize plants tolerant to treatment with levels of dicamba (i.e., 27 kg/ha) greatly exceeding the highest recommended rate of 0.56 kg/ha.

Fatty acid transport protein 2 (FATP2) is highly expressed in the liver, small intestine, and kidney, where it functions in both the transport of exogenous long-chain fatty acids and the activation of very-long-chain fatty acids. Here, using a murine model, we investigated the phenotypic impacts of deleting FATP2, followed by a transcriptomic analysis using unbiased RNA-Seq to identify concomitant changes in the liver transcriptome. WT and FATP2-null (Fatp2−/−) mice (5 weeks) were maintained on a standard chow diet for 6 weeks. The Fatp2−/− mice had reduced weight gain, lowered serum triglyceride, and increased serum cholesterol levels and attenuated dietary fatty acid absorption. Transcriptomic analysis of the liver revealed 258 differentially expressed genes in male Fatp2−/− mice and a total of 91 in female Fatp2−/− mice. These genes mapped to the following gene ontology categories: fatty acid degradation, peroxisome biogenesis, fatty acid synthesis, and retinol and arachidonic acid metabolism. Targeted RT-quantitative PCR verified the altered expression of selected genes. Of note, most of the genes with increased expression were known to be regulated by peroxisome proliferator–activated receptor α (PPARα), suggesting that FATP2 activity is linked to a PPARα-specific proximal ligand. Targeted metabolomic experiments in the Fatp2−/− liver revealed increases of total C16:0, C16:1, and C18:1 fatty acids; increases in lipoxin A4 and prostaglandin J2; and a decrease in 20-hydroxyeicosatetraenoic acid. We conclude that the expression of FATP2 in the liver broadly affects the metabolic landscape through PPARα, indicating that FATP2 provides an important role in liver lipid metabolism through its transport or activation activities.