Identification of the bona fide DHDPS from a common plant pathogen

Agrobacterium tumefaciens is a Gram‐negative soil‐borne bacterium that causes Crown Gall disease in many economically important crops. The absence of a suitable chemical treatment means there is a need to discover new anti‐Crown Gall agents and also characterize bona fide drug targets. One such target is dihydrodipicolinate synthase (DHDPS), a homo‐tetrameric enzyme that catalyzes the committed step in the metabolic pathway yielding meso‐diaminopimelate and lysine. Interestingly, there are 10 putative DHDPS genes annotated in the A. tumefaciens genome, including three whose structures have recently been determined (PDB IDs: 3B4U, 2HMC, and 2R8W). However, we show using quantitative enzyme kinetic assays that nine of the 10 dapA gene products, including 3B4U, 2HMC, and 2R8W, lack DHDPS function in vitro. A sequence alignment showed that the product of the dapA7 gene contains all of the conserved residues known to be important for DHDPS catalysis and allostery. This gene was cloned and the recombinant product expressed and purified. Our studies show that the purified enzyme (i) possesses DHDPS enzyme activity, (ii) is allosterically inhibited by lysine, and (iii) adopts the canonical homo‐tetrameric structure in both solution and the crystal state. This study describes for the first time the structure, function and allostery of the bona fide DHDPS from A. tumefaciens, which offers insight into the rational design of pesticide agents for combating Crown Gall disease. Proteins 2014; 82:1869–1883. © 2014 Wiley Periodicals, Inc.     

Diseases of the gladiolus: I. Control of hard rot, due to Septoria gladioli Passer., by fungicidal treatment of the corms

Losses from hard rot, measured by an arbitrary disease index, were reduced by treating the dehusked corms before planting with mercuric chloride (with or without the addition of 10% hydrochloric acid), mercurous chloride (calomel), three proprietary mercury compounds (Aretan, Uspulun and Ceresan) and one proprietary non-mercury compound (Folosan). Calomel was the least effective. All the treatments were relatively less effective when corms with definite lesions were treated.
The weight of clean corms produced per old corm planted (weight index) was usually increased by all the fungicides tried, but calomel and Ceresan were less satisfactory than the others.
Mercuric chloride (3 hr. steep in a 0.1% solution) was not rendered more effective by the addition of hydrochloric acid nor by a preliminary dip in methylated spirits to facilitate wetting, while the addition of a proprietary wetting compound (Agral) was definitely harmful to the corms and usually less effective than mercuric chloride alone. Increase in time of steeping or concentration of mercuric chloride was not beneficial and was sometimes harmful. Reduction in time of steeping to 1 hr. gave promising results.
Treatment in November had some advantages over treatment in March.
All the mercury compounds tended to delay flowering, this being most marked in the presence of the wetting compound. Stunted foliage and poor quality flowers resulted from the use of Ceresan.     

Modulation of peroxisomal and microsomal fatty acid oxidation by acetone. A comparative study between liver and kidney

The effect of acetone consumption on some microsomal and peroxisomal activities was studied in rat kidney and these results were compared with data from former investigations in liver. Acetone increased the microsomal lauric acid hydroxylation, the aminopyrine N-demethylation catalyzed by cytochrome P450 and the microsomal UDP-glucuronyltransferase activity. Also, acetone increased the peroxisomal β-oxidation of palmitoyl CoA and catalase activities in kidney. These studies suggest that acetone is a common inducer of the microsomal and peroxisomal fatty acid oxidation, as previously shown in both starved and ethanol treated rats. Our results support the hypothesis that microsomal fatty acid ω-hydroxylation results in the generation of substrates being supplied for peroxisomal β-oxidation. We propose that the final purpose of these linked fatty acid oxidations could be the catabolism of fatty acids or the generation of a substrate for the synthesis of glucose from fatty acids. This pathway would be triggered by acetone treatment in a similar way in liver and kidney.     

Notes on basal rot of narcissus

Control of basal rot of narcissus following hot-water treatment against eelworm was equally good when 0.5% formalin was included in the bath or was used as a cold or warm steep immediately afterwards. When the formalin steep was delayed control was less good. All methods of applying formalin in connexion with hot-water treatments used in these experiments were equally harmless to growth and flowering of the bulbs.