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The effects of dark chilling on CO2 assimilation, chlorophyll a fluorescence kinetics and nitrogen fixation were compared in two Glycine max (L.) Merr. genotypes. The aim was to elucidate the mechanisms by which photosynthesis was inhibited as well as identification of selection criteria for dark chilling tolerance. Seedlings were dark chilled (8 degrees C) for 9 consecutive nights but kept at normal day temperatures (28 degrees C). CO2 gas exchange analysis indicated that photosynthesis in Maple Arrow was inhibited largely as a result of stomatal limitation, while in Fiskeby V, it indicated inhibition of the mesophyll reactions. Increased intercellular CO2 concentration and decreased carboxylation efficiency suggested loss of Rubisco activity in Fiskeby V, although no effect on the KM (CO2) of Rubisco was observed. Quantification and deconvolution of the Chl a fluorescence transients into several phenomenological and biophysical parameters (JIP-test) revealed large genotypic differences in the response of PSII to dark chilling. These parameters differentially changed in the two genotypes during the progression of the chilling treatment. Among them, the performance index, reflecting several responses of the photochemical apparatus, provided the best preliminary overall assessment of the genotypes. In contrast, the quantum yield of primary photochemistry varphiPo (FV/FM) was quite insensitive. The recovery of most of the JIP-test parameters in Maple Arrow after 6 and 9 nights of dark chilling was a major genotypic difference. Genotypic differences were also observed with regard to the ureide response and N2 fixation appeared to be more sensitive to dark chilling than CO2 assimilation. The JIP-test provided information consistent with results derived from CO2 assimilation and N2 fixation studies suggesting that it can substitute the much more time-consuming methods for the detection of chilling stress and can well satisfy the requirements of a rapid and accurate screening method.

4,4-Diphenyl-2,5-cyclohexadienone (1) crystallized as four conformational polymorphs and a record number of 19 crystallographically independent molecules have been characterized by low-temperature X-ray diffraction: form A (P2(1), Z'=1), form B (P1, Z'=4), form C (P1, Z'=12), and form D (Pbca, Z'=2). We have now confirmed by variable-temperature powder X-ray diffraction that form A is the thermodynamic polymorph and B is the kinetic form of the enantiotropic system A-D. Differences in the packing of the molecules in these polymorphs result from different acidic C-H donors approaching the C=O acceptor in C-H...O chains and in synthons I-III, depending on the molecular conformation. The strength of the C-HO interaction in a particular structure correlates with the number of symmetry-independent conformations (Z') in that polymorph, that is, a short C-HO interaction leads to a high Z' value. Molecular conformation (Econf) and lattice energy (Ulatt) contributions compensate each other in crystal structures A, B, and D resulting in very similar total energies: Etotal of the stable form A=1.22 kcal mol(-1), the metastable form B=1.49 kcal mol(-1), and form D=1.98 kcal mol(-1). Disappeared polymorph C is postulated as a high-Z', high-energy precursor of kinetic form B. Thermodynamic form A matches with the third lowest energy frame based on the value of Ulatt determined in the crystal structure prediction (Cerius2, COMPASS) by full-body minimization. Re-ranking the calculated frames on consideration of both Econf (Spartan 04) and Ulatt energies gives a perfect match of frame #1 with stable structure A. Diphenylquinone 1 is an experimental benchmark used to validate accurate crystal structure energies of the kinetic and thermodynamic polymorphs separated by <0.3 kcal mol(-1) (approximately 1.3 kJ mol(-1)).

a b s t r a c tEdited by J van Staden Keywords: A. hybridus A. hypochondriacus Antioxidative enzymes A. tricolor Cell membrane stability Leaf area Leaf water status Metabolic response Osmoregulation Proline Relative water content Water stressTraditional crops are extremely important for food production in low income, food-deficit countries (LIFDCs) where they continue to be maintained by socio-cultural preferences and traditional uses. Significant potential exists to improve these crops, one of which is to select for improved productivity during moisture stress conditions. Germplasm of Amaranthus tricolor, Amaranthus hypochondriacus and Amaranthus hybridus were subjected to various screening methods to measure metabolic and physiological changes during water stress. The activities of enzymes involved in the oxygen-scavenging system during abiotic stress conditions (superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione reductase (GR)), free proline production, leaf area (LA), cell membrane stability (CMS), leaf water potential (LWP) and relative water content (RWC) were measured in these three amaranth species during induced water stress. This study showed significant differences in metabolic responses during water deficit of the three species tested. Moisture stress and a decrease in RWC and LWP were first experienced in A. hybridus and A. hypochondriacus, followed by A. tricolor. There was an indirect correlation between leaf water status (RWC and LWP), enzyme activity, proline production and leaf area. The combined effect of GR, APX and SOD could ensure higher levels of regulation of the toxic effect of H 2 O 2 which could be associated with drought tolerance in Amaranthus. Distinct differences in onset of proline accumulation and the amount of accumulated in leaves upon induced water stress was noticed for the three amaranth species tested. Proline accumulation during water stress conditions in amaranth seems to be indirect and could possibly have a protective role apart from osmoregulation during stress conditions. This contention is supported by the decrease in leaf area and high cell membrane stability for two of the species tested. This study forms part of a project aimed at the development of improved traditional crops to contribute to food production and quality for subsistence farmers in areas with low precipitation or variable rainfall patterns.