Crystal Structure of an Indole-3-Acetic Acid Amido Synthetase from Grapevine Involved in Auxin Homeostasis

Auxins are important for plant growth and development, including the control of fruit ripening. Conjugation to amino acids by indole-3-acetic acid (IAA)-amido synthetases is an important part of auxin homeostasis. The structure of the auxin-conjugating Gretchen Hagen3-1 (GH3-1) enzyme from grapevine (Vitis vinifera), in complex with an inhibitor (adenosine-5′-[2-(1H-indol-3-yl)ethyl]phosphate), is presented. Comparison with a previously published benzoate-conjugating enzyme from Arabidopsis thaliana indicates that grapevine GH3-1 has a highly similar domain structure and also undergoes a large conformational change during catalysis. Mutational analyses and structural comparisons with other proteins have identified residues likely to be involved in acyl group, amino acid, and ATP substrate binding. Vv GH3-1 is a monomer in solution and requires magnesium ions solely for the adenlyation reaction. Modeling of IAA and two synthetic auxins, benzothiazole-2-oxyacetic acid (BTOA) and 1-naphthaleneacetic acid (NAA), into the active site indicates that NAA and BTOA are likely to be poor substrates for this enzyme, confirming previous enzyme kinetic studies. This suggests a reason for the increased effectiveness of NAA and BTOA as auxins in planta and provides a tool for designing new and effective auxins.     

Where's the ecology in molecular ecology?

Molecular techniques have had a profound impact in biology. Major disciplines, including evolutionary biology, now consistently utilize molecular tools. In contrast, molecular techniques have had a more limited impact in ecology. This discrepancy is surprising. Here, we describe the unexpected paucity of ecological research in the field colloquially referred to as 'molecular ecology.' Publications over the past 15 years from the journals Ecology , Evolution and Molecular Ecology reveal that much of the research published under the molecular ecology banner is in fact evolutionary in nature, and that comparatively little ecological research incorporates molecular tools. This failure to more broadly utilize molecular techniques in ecology is alarming because several promising lines of ecological inquiry could benefit from molecular approaches. Here we summarize the use of molecular tools in ecology and evolution, and suggest several ways to renew the ecological focus in 'molecular ecology'.     

The evolution of antennal courtship in diplazontine parasitoid wasps (Hymenoptera,Ichneumonidae, Diplazontinae)

Background

As predicted by theory, traits associated with reproduction often evolve at a comparatively high speed. This is especially the case for courtship behaviour which plays a central role in reproductive isolation. On the other hand, courtship behavioural traits often involve morphological and behavioural adaptations in both sexes; this suggests that their evolution might be under severe constraints, for instance irreversibility of character loss. Here, we use a recently proposed method to retrieve data on a peculiar courtship behavioural trait, i.e. antennal coiling, for 56 species of diplazontine parasitoid wasps. On the basis of a well-resolved phylogeny, we reconstruct the evolutionary history of antennal coiling and associated morphological modifications to study the mode of evolution of this complex character system.

Results

Our study reveals a large variation in shape, location and ultra-structure of male-specific modifications on the antennae. As for antennal coiling, we find either single-coiling, double-coiling or the absence of coiling; each state is present in multiple genera. Using a model comparison approach, we show that the possession of antennal modifications is highly correlated with antennal coiling behaviour. Ancestral state reconstruction shows that both antennal modifications and antennal coiling are highly congruent with the molecular phylogeny, implying low levels of homoplasy and a comparatively low speed of evolution. Antennal coiling is lost on two independent occasions, and never reacquired. A zero rate of regaining antennal coiling is supported by maximum parsimony, maximum likelihood and Bayesian approaches.

Conclusions

Our study provides the first comparative evidence for a tight correlation between male-specific antennal modifications and the use of the antennae during courtship. Antennal coiling in Diplazontinae evolved at a comparatively low rate, and was never reacquired in any of the studied taxa. This suggests that the loss of antennal coiling is irreversible on the timescale examined here, and therefore that evolutionary constraints have greatly influenced the evolution of antennal courtship in this group of parasitoid wasps. Further studies are needed to ascertain whether the loss of antennal coiling is irreversible on larger timescales, and whether evolutionary constraints have influenced courtship behavioural traits in a similar way in other groups.

     

Identification and characterization of two families of F420H2‐dependent reductases from Mycobacteria that catalyse aflatoxin degradation

Aflatoxins are polyaromatic mycotoxins that contaminate a range of food crops as a result of fungal growth and contribute to serious health problems in the developing world because of their toxicity and mutagenicity. Although relatively resistant to biotic degradation, aflatoxins can be metabolized by certain species of Actinomycetales. However, the enzymatic basis for their breakdown has not been reported until now. We have identified nine Mycobacterium smegmatis enzymes that utilize the deazaflavin cofactor F₄₂₀H₂ to catalyse the reduction of the α,β‐unsaturated ester moiety of aflatoxins, activating the molecules for spontaneous hydrolysis and detoxification. These enzymes belong to two previously uncharacterized F₄₂₀H₂ dependent reductase (FDR‐A and ‐B) families that are distantly related to the flavin mononucleotide (FMN) dependent pyridoxamine 5′‐phosphate oxidases (PNPOxs). We have solved crystal structures of an enzyme from each FDR family and show that they, like the PNPOxs, adopt a split barrel protein fold, although the FDRs also possess an extended and highly charged F₄₂₀H₂ binding groove. A general role for these enzymes in xenobiotic metabolism is discussed, including the observation that the nitro‐reductase Rv3547 from Mycobacterium tuberculosis that is responsible for the activation of bicyclic nitroimidazole prodrugs belongs to the FDR‐A family.     

钙离子对PSII颗粒放氧活性的影响

外加5 mmol/L Ca~(2 )可以使菠菜PSⅡ颗粒的放氧活性增高。PSⅡ颗粒经EGTA透析、低pH值、光照、2 mol/L NaCl等处理后,放氧活性下降,同时,这些颗粒的钙含量也相应降低。但当外加 5 mmol/L Ca~(2 )时,可使这些颗粒全部或部分地恢复放氧活性。PSⅡ颗粒中存在的钙对放氧起着重要作用;钙在PSⅡ颗粒中的结合位点不止一个,其结合状态有紧密和松散之别。     

The ultrastructure of immobilised desiccated cells of the cyanobacterium Nostoc commune UTEX 584

Cells of the cyanobacterium Nostoc commune UTEX 584 were immobilised, subjected to acute matric water stress (ψ_m = −128 MPa) and then desiccated. Their ultrastructure was investigated by the use of an anhydrous fixation procedure. Although shrunken and bleached, the integrity of the vegatative cells at the ultrastructural level was apparently preserved. The ease with which certain cyanobacterial cells can recover from desiccation may be consequent upon the maintenance of cellular organisation at the ultrastructural level.     

Heterocyst structure in Chlorogloea fritschii

Summary Heterocysts of Chlorogloea fritschii were studied with the aid of light and electron microscopy. Two main types of heterocyst were recognized. One of these, termed here the H.1 cell, is the only type in filaments, but occurs also among endospore groups. The other, the H.3 cell, is restricted to endospore groups. The transition from one type of heterocyst to the other, therefore, does not coincide exactly with the transition in arrangement of vegetative cells from filaments to endospores. Both types of heterocyst differ considerably from other published accounts of heterocysts.The differences between H.1 and H.3 cells reside mainly in the arrangement, and relative development or disappearance of the various cell organelles.A summary is given of other types of cell seen with the electron microscope which share some features with typical heterocysts. Some of these would probably be recognized as heterocysts by light microscopy, whereas others would not.     

Soil arthropods as indicators of water stress in Antarctic terrestrial habitats?

Abiotic features of Antarctic terrestrial habitats, particularly low temperatures and limited availability of liquid water, strongly influence the ecophysiology and life histories of resident biota. However, while temperature regimes of a range of land microhabitats are reasonably well characterized, much less is known of patterns of soil water stress, as current technology does not allow measurement at the required scale. An alternative approach is to use the water status of individual organisms as a proxy for habitat water status and to sample over several years from a population to identify seasonal or long‐term patterns. This broad generalization for terrestrial invertebrates was tested on arthropods in the maritime Antarctic. We present analyses of a long‐term data set of body water content generated by monthly sampling for 8–11 years of seven species of soil arthropods (four species of Acari, two Collembola and one Diptera) on maritime Antarctic Signy Island, South Orkney Islands. In all species, there was considerable within‐ and between‐sample variability. Despite this, clear seasonal patterns were present in five species, particularly the two collembolans and a prostigmatid mite. Analyses of monthly water content trends across the entire study period identified several statistically significant trends of either increase or decrease in body water content, which we interpret in the context of regional climate change. The data further support the separation of the species into two groups as follows: firstly, the soft‐bodied Collembola and Prostigmata, with limited cuticular sclerotization, which are sensitive to changes in soil moisture and are potentially rapid sensors of microhabitat water status, secondly, more heavily sclerotized forms such as Cryptostigmata (=Oribatida) and Mesostigmata mites, which are much less sensitive and responsive to short‐term fluctuations in soil water availability. The significance of these findings is discussed and it is concluded that annual cycles of water content were driven by temperature, mediated via radiation and precipitation, and constituted reliable indicators of habitat moisture regimes. However, detailed ecophysiological studies are required on particular species before such information can be used to predict over long timescales.