The Chloroplast of Chlamydomonas reinhardtii as a Testbed for Engineering Nitrogen Fixation into Plants

Eukaryotic organisms such as plants are unable to utilise nitrogen gas (N₂) directly as a source of this essential element and are dependent either on its biological conversion to ammonium by diazotrophic prokaryotes, or its supply as chemically synthesised nitrate fertiliser. The idea of genetically engineering crops with the capacity to fix N₂ by introduction of the bacterial nitrogenase enzyme has long been discussed. However, the expression of an active nitrogenase must overcome several major challenges: the coordinated expression of multiple genes to assemble an enzyme complex containing several different metal cluster co-factors; the supply of sufficient ATP and reductant to the enzyme; the enzyme’s sensitivity to oxygen; and the intracellular accumulation of ammonium. The chloroplast of plant cells represents an attractive location for nitrogenase expression, but engineering the organelle’s genome is not yet feasible in most crop species. However, the unicellular green alga Chlamydomonas reinhardtii represents a simple model for photosynthetic eukaryotes with a genetically tractable chloroplast. In this review, we discuss the main advantages, and limitations, of this microalga as a testbed for producing such a complex multi-subunit enzyme. Furthermore, we suggest that a minimal set of six transgenes are necessary for chloroplast-localised synthesis of an ‘Fe-only’ nitrogenase, and from this set we demonstrate the stable expression and accumulation of the homocitrate synthase, NifV, under aerobic conditions. Arguably, further studies in C. reinhardtii aimed at testing expression and function of the full gene set would provide the groundwork for a concerted future effort to create nitrogen-fixing crops.     

Effect of high-dose nicotine patch on the characteristics of lapse episodes.

Objective: Nicotine patch improves treatment outcomes, but lapses are still common. To understand the process of relapse on patch, we investigated differences in the antecedents (withdrawal, setting, triggers) of lapse episodes experienced on high-dose (35 mg) nicotine patches versus placebo. Design: Participants were smokers who lapsed during a randomized, double-blind trial of active patches (n = 100) versus placebo (n = 85). Participants used electronic diaries to monitor their smoking, affect, and activities in real time for 5 weeks during their cessation attempt. Results: We analyzed 490 lapse episodes (active: 266; placebo: 224). Lapses on nicotine patch were characterized by significantly lower positive affect and higher negative affect than placebo lapses. Participants treated with high-dose patch were also significantly more likely to lapse in situations involving little or no craving. Situational antecedents of lapses on patch resembled those on placebo. Conclusion: The results suggest that treatment with patch may set a higher threshold for affective stimuli to provoke lapses, but does not change the proximal cues that trigger lapses. This suggests that behavioral relapse-prevention strategies developed for unmedicated smokers should also apply to those treated with nicotine patch. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

N,7,7‐Tricyanoquinomethanimine (TCQMI) Based Organic Magnetic Materials

The synthesis and characterization of a new family of magnetic materials based on the electron accepting cyanocarbon N,7,7‐tricyanoquinomethanimine, TCQMI, its radical anion [TCQMI], and its σ‐dimer, σ‐[TCQMI]₂^2?, are reported. [Fe^IIICp*₂][TCQMI] (where Cp* is pentamethylcyclopentadienide) forms parallel chains of alternating [TCQMI]^?? and [FeCp*₂]^?+ and magnetically orders at 3.4 K as a weak ferromagnet. M[TCQMI]₂?zCH₂Cl₂ (M = V, Fe) are amorphous solids with [TCQMI]^?? coordinated to metal centers through the nitrile groups. The Fe compound magnetically orders as a weak ferromagnet at ≈4 K, whereas the V compound shows no evidence of magnetic ordering. {[Mn^IIITPP]⁺}₂[TCQMI]₂^2? (TPP = tetraphenylporphyrin) results from the reaction of TCQMI with Mn^IITPP(py) due to the formation of the [TCQMI]₂^2? σ‐dimer in situ, and is a weak ferromagnet below 3.7 K. The lack of magnetic ordering in V[TCQMI]₂?zCH₂Cl₂ is not currently understood, and is in strong contrast to V[TCNE]₂?zCH₂Cl₂, which magnetically orders above room temperature.