L-rhamnose-1-dehydrogenase gene and L-rhamnose catabolism in the yeast Pichia stipitis

The purpose of this work was to identify some of the genes of the catabolic route of L-rhamnose in the yeast Pichia stipitis. There are at least two distinctly different pathways for L-rhamnose catabolism. The one described in bacteria has phosphorylated intermediates and the enzymes and the genes of this route have been described. The pathway described in yeast does not have phosphorylated intermediates. The intermediates and the enzymes of this pathway are known but none of the genes have been identified. The work was started by purifying the L-rhamnose dehydrogenase, which oxidates L-rhamnose to rhamnonic acid-gamma-lactone. NAD is used as a cofactor in this reaction. A DEAE ion exchange column was used for purification. The active fraction was further purified using a non-denaturing PAGE and the active protein identified by zymogram staining. In the last step the protein was separated in a SDS-PAGE, the protein band trypsinated and analysed by MALDI-TOF MS. This resulted in the identification of the corresponding gene, RHA1, which was then, after a codon change, expressed in Saccharomyces cerevisiae. Also C- or N-terminal histidine tags were added but as the activity of the enzyme was lost or strongly reduced these were not used. The kinetic properties of the protein were analysed in the cell extract. Substrate specifity was tested with different sugars; L-rhamnose, L-lyxose and L-mannose were oxidated by the enzyme. Vmax values were 180 nkat/mg, 160 nkat/mg and 72 nkat/mg, respectively. The highest affinity was towards L-rhamnose, the Km value being 0.9 mM. Lower affinities were obtained with L-lyxose, Km 4.3 mM, and L-mannose Km 25 mM. Northern analysis was done to study the transcription of RHA1 with different carbon sources. Transcription was observed only on L-rhamnose suggesting that RHA1 expression is L-rhamnose induced. A RHA1 deletion cassette for P. stipitis was constructed but the cassette had integrated randomly and not targeted to delete the RHA1 gene. Enzyme assays for L-lactaldehyde dehydrogenase were done similarly to L-rhamnose dehydrogenase assays. NAD is used as a cofactor also in this reaction where L-lactaldehyde is oxidised to L-lactate. The observed enzyme activities were very low and the activity was lost during the purification procedures.

Sitting l-r Rabbi Dudley Weinberg, Leo Baeck, Joseph H. Cohen, Dr. Joshua Liebman; Standing l-r Max Feldberg, Louis Salvage, Rabbi Beryl D. Cohon, Harry Penn, Abraham Schiller; Boston Portraits; Groups; Men

Page 58 of the "American Jewish Cavalcade" scrapbook of Leo Baeck in New York found in ROS 10 Folder 3

Leo Baeck with New York Mayor William O'Dwyer and Union of American Hebrew Congregations President Maurice N. Eisendrath, Oscar M. Lazrus and Rabbi Daniel L. Davis Portraits; Groups; Men

Page 12 of the "American Jewish Cavalcade" scrapbook of Leo Baeck in New York found in ROS 10 Folder 3

Phosphine resistance in Sitophilus oryzae (L.) from eastern Australia: Inheritance, fitness and prevalence

The inheritance and fitness of phosphine resistance was investigated in an Australian strain of the rice weevil, Sitophilus oryzae (L.), as well as its prevalence in eastern Australia. This type of knowledge may provide insights in to the development of phosphine resistance in this species with the potential for better management. This strain was 12.2 × resistant at the LC50 level based on results for adults exposed for 20 h. Data from the testing of F1 adults from the reciprocal crosses (R♀ × S♂ and S♀ × R♂) showed that resistance was autosomal and inherited as an incompletely recessive trait with a degree of dominance of -0.88. The dose-response data for the F1 × S and F1 × R test crosses, and the F2 progeny were compared with predicted dose-response assuming monogenic recessive inheritance, and the results were consistent with resistance being conferred by one major gene. There was no evidence of fitness cost based on the frequency of susceptible phenotypes in hybridized populations that were reared for seven generations without exposure to phosphine. Lack of fitness cost suggests that resistant alleles will tend to persist in field populations that have undergone selection even if selection pressure is removed. Discriminating dose tests on 107 population samples collected from farms from 2006 to 2010 show that populations containing insects with the weak resistant phenotype are common in eastern Australia, although the frequency of resistant phenotypes within samples was typically low. The prevalence of resistance is a warning that this species has been subject to considerable selection pressure and that effective resistance management practices are needed to address this problem. Crown Copyright © 2014.

Field evaluation of transgenic pineapple (Ananas comosus (L.) Merr.) cv. ‘Smooth Cayenne’ for resistance to blackheart under subtropical conditions

A comparative analysis of transgenic pineapple lines transformed with a polyphenol oxidase (PPO) gene (ppo) and the untransformed cultivar ‘Smooth Cayenne’ was made from plants grown in a series of field trials under cool subtropical conditions in southeast Queensland. In the four field trials where blackheart was recorded, all of the control lines expressed blackheart on each occasion and exhibited the greatest incidence (50%) and severity (34%) of symptoms. Irrespective of the gene transfer method or the gene construct used, 38% of the lines produced were regarded as blackheart resistant, having no blackheart symptoms in two or more trials. Five blackheart resistant transgenic lines consistently performed as well as or better than control plants in terms of fruit characteristics and quality.

Dr. J. L. Magnes

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X-ray studies on crystalline complexes involving amino acids. IV. The structure of L-arginine L-ascorbate

L-Arginine ascorbate, C6HIsN40+.C6H706, a 1"1 crystalline complex between the amino acid arginineand the vitamin ascorbic acid, crystallizes in the monoclinic space group P21 with two formula units in a cell of dimensions a = 5.060 (8), b = 9.977 (9), c = 15.330 (13) A, fl = 97.5 (2) °. The structure was solved by the symbolic addition procedure and refined to an R of 0.067 for 1501 photographically observed reflec- tions. The conformation of the arginine molecule in the structure is different from any observed so far. The present structure provides the first description of the ascorbate anion unaffected by the geometrical constraints and disturbances imposed by the requirements of metal coordination. The lactone group and the deprotonated enediol group in the anion are planar and the side chain assumes a conformation which appears to be sterically the most favourable. In the crystals, the arginine molecules and the ascorbate anions aggregate separately into alternating layers. The molecules in the arginine layer are held together by interactions involving a-amino and ~t-carboxylate groups, a situation analogous to that found in proteins. The two layers of unlike molecules are interconnected primarily through the interactions of the side-chain guanidyl group of arginine with the ascorbate ion. These involve a specific ion-pair interaction accompanied by two convergent hydrogen bonds and another pair of nearly parallel hydrogen bonds.

Studies on crystalline complexes involving amino acids. V. The structure of L-serine-L-ascorbic acid

L-Serine-L-ascorbic acid, C3HTNOa. C6HsO6, a 1:1 complex between the amino acid serine and the vitamin ascorbic acid, crystallizes in the orthorhombic space group P2~2~2~ with four formula units in a cell of dimensions a = 5.335(3), b = 8.769(2), c = 25.782 (5) A. The structure was solved by direct methods and refined by full-matrix least squares to an R of 0.036 for 951 observed reflections. Both molecules are neutral in the structure. The conformation of the serine molecule is different from that observed in the crystal structures of L-serine, DL-serine and L-serine monohydrate. The enediol group in the ascorbic acid molecule is planar, whereas significant departures from planarity are observed in the lactone group. The conformation of this molecule is similar to that observed in arginine ascorbate. The unlike molecules aggregate into separate columns in the crystal structure. The columns are held together by hydrogen bonds. Among these, a pair of hydrogen bonds between the enediol group of ascorbic acid and the carboxylate group of serine provides a possible model for a specific interaction between ascorbic acid and a carboxylate ion.

Damage assessment of structures with uncertainty by using mode-shape curvatures and fuzzy logic

Uncertainties associated with the structural model and measured vibration data may lead to unreliable damage detection. In this paper, we show that geometric and measurement uncertainty cause considerable problem in damage assessment which can be alleviated by using a fuzzy logic-based approach for damage detection. Curvature damage factor (CDF) of a tapered cantilever beam are used as damage indicators. Monte Carlo simulation (MCS) is used to study the changes in the damage indicator due to uncertainty in the geometric properties of the beam. Variation in these CDF measures due to randomness in structural parameter, further contaminated with measurement noise, are used for developing and testing a fuzzy logic system (FLS). Results show that the method correctly identifies both single and multiple damages in the structure. For example, the FLS detects damage with an average accuracy of about 95 percent in a beam having geometric uncertainty of 1 percent COV and measurement noise of 10 percent in single damage scenario. For multiple damage case, the FLS identifies damages in the beam with an average accuracy of about 94 percent in the presence of above mentioned uncertainties. The paper brings together the disparate areas of probabilistic analysis and fuzzy logic to address uncertainty in structural damage detection.

Responses of Super Rice (Oryza sativa L.) to Different Planting Methods for Grain Yield and Nitrogen-Use Efficiency in the Single Cropping Season

To break the yield ceiling of rice production, a super rice project was developed in 1996 to breed rice varieties with super high yield. A two-year experiment was conducted to evaluate yield and nitrogen (N)-use response of super rice to different planting methods in the single cropping season. A total of 17 rice varieties, including 13 super rice and four non-super checks (CK), were grown under three N levels [0 (N0), 150 (N150), and 225 (N225) kg ha−1] and two planting methods [transplanting (TP) and direct-seeding in wet conditions (WDS)]. Grain yield under WDS (7.69 t ha−1) was generally lower than TP (8.58 t ha−1). However, grain yield under different planting methods was affected by N rates as well as variety groups. In both years, there was no difference in grain yield between super and CK varieties at N150, irrespective of planting methods. However, grain yield difference was dramatic in japonica groups at N225, that is, there was an 11.3% and 14.1% average increase in super rice than in CK varieties in WDS and TP, respectively. This suggests that high N input contributes to narrowing the yield gap in super rice varieties, which also indicates that super rice was bred for high fertility conditions. In the japonica group, more N was accumulated in super rice than in CK at N225, but no difference was found between super and CK varieties at N0 and N150. Similar results were also found for N agronomic efficiency. The results suggest that super rice varieties have an advantage for N-use efficiency when high N is applied. The response of super rice was greater under TP than under WDS. The results suggest that the need to further improve agronomic and other management practices to achieve high yield and N-use efficiency for super rice varieties in WDS.

Refining low-volume, high-concentration herbicide applications to control Chromolaena odorata (L.) King & Robinson (Siam weed) in remote areas

Chromolaena odorata (L.) King and Robinson (Siam weed) is a highly invasive plant and a high priority for control in north Queensland. It can be effectively treated using high-volume, groundbased herbicide spray equipment, but operational information shows that this control method becomes increasingly difficult in areas where vehicle access is prevented by rougher terrain. Low-volume, high-concentration herbicide applications have proven capable of causing high mortality in these remote situations. Two trials were undertaken between May 2010 and May 2012 to refine effective rates of aminopyralid/fluroxypyr, fluroxypyr and metsulfuron-methyl, only using low-volume, high-concentration applications on Siam weed. Fluroxypyr on its own was as effective as aminopyralid/fluroxypyr as both herbicides caused 95-100% mortality at overlapping rates containing 5 to 18.85 g a.i. L-1 of fluroxypyr. Metsulfuron-methyl caused 100% mortality when applied at 3 and 6 g a.i. L-1. Effective control was achieved with approximately 16 to 22 mL of the solutions per plant, so a 5 L mixture in a backpack could treat 170 to 310 adult plants. There are several options for treating Siam weed on the ground and the choice of methods reflects the area, plant density and accessibility of the infestation. Control information from Siam weed field crews shows that low volume, high concentration herbicide applications applied using a splatter gun are a more efficient method for controlling larger, denser remote infestations than physical removal. By identifying effective herbicides that are applied through low-volume equipment, these trials provide an additional and more efficient tool for controlling Siam weed in remote areas.

Polarized Ethylenes: Structures of (l,3-Dimethyl-2-imidazolidinylidene)malononitrile and (l,3-Dimethyl-2-perhydropyrimidinylidene)malononitrile

Crystal structures of the title compounds, (I) and (II), have been determined by three-dimensional diffraction methods. Crystals of CsHIoN 4 (I) are monoclinic, space group P21/a with Z = 4, Mr= 162, a = 7.965 (1), b = 16.232 (2), c = 7.343 (1) A, fl = 113.54 (1) °, V = 890.7 A 3, D,n = 1.218, D x = 1.208 gcm -3, g(Cu Ka, 2 = 1.5418/~) = 6.47 em -1, F(000) = 344. The crystals of C9H12N4 (II) are orthorhombic, space group P21en, with Z = 4, Mr = 176, a = 7.983 (3), b = 8.075 (2), c = 14.652 (3) ./k, V = 44.43/~3, Dm= 1.219, D x = 1.237 g cm -3, #(Mo Ka, ). = 0.7107 ,/k) = 0.868 cm -1, F(000) = 376. Both structures were solved by direct methods and refined to R = 5.8% for (I) and 5.3 % for (II). The C-C double-bond distances are 1.407 (3) in (I) and 1.429 (6)/~ in (II), appreciably longer than normal. The steric and push-pull effects result in rotation about the C=C bond, the rotation angles being 20.2 (3) in (I) and 31.5 (6) o in (II).

Effects of light conditions and plant density on growth and reproductive biology of Cascabela thevetia (L.) Lippold

Cascabela thevetia (L.) Lippold (Apocynaceae) is an invasive woody weed that has formed large infestations at several locations in northern Australia. Understanding the reproductive biology of C. thevetia is vital to its management. This paper reports results of a shade house experiment that determined the effects of light conditions (100% or 30% of natural light) and plant densities (one, two, four or eight plants per plot) on the growth, time to flowering and seed formation, and monthly pod production of two C. thevetia biotypes (peach and yellow). Shaded plants were significantly larger when they reached reproductive maturity than plants grown under natural light. However, plants grown under natural light flowered earlier (268 days compared with 369 days) and produced 488 more pods per pot (a 5-fold increase) over 3 years. The yellow biotype was slightly taller at reproductive maturity but significantly taller and with significantly greater aboveground biomass at the end of the study. Both biotypes flowered at a similar time under natural light and low plant densities but the yellow biotype was quicker to seed (478 versus 498 days), produced significantly more pods (364 versus 203 pods) and more shoot growth (577 g versus 550 g) than the peach biotype over 3 years. Higher densities of C. thevetia tended to significantly reduce the shoot and root growth by 981 g and 714 g per plant across all light conditions and biotypes over 3 years and increase the time taken to flower by 140 days and produce seeds by 184 days. For land managers trying to prevent establishment of C. thevetia or to control seedling regrowth once initial infestations have been treated, this study indicates that young plants have the potential to flower and produce seeds within 268 and 353 days, respectively. However, with plant growth and reproduction most likely to be slower under field conditions, annual surveillance and control activities should be sufficient to find and treat plants before they produce seeds and replenish soil seed banks. The most at-risk part of the landscape may be open areas that receive maximum sunlight, particularly within riparian habitats where plants would consistently have more favourable soil moisture conditions.