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    Aminoacid metabolism in plants : the biosynthesis of [beta]-cyanoalanine and asparagine in lupins : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Biochemistry at Massey University
    (Massey University, 1970) Lever, Michael
    β-Cyanoalanine synthase in etiolated seedlings of Lupinus angustifolius is found mainly in the mitochondrial fraction of the cotyledons and stems. In seedlings developing at 25°C in the dark it reaches a maximal concentration after 5 days, at the same time as asparagine accumulation is most rapid and other physiological changes occur. However, maximal ability to assimilate HCN gas to asparagine develops after 3 to 4 days, before asparagine accumulation begins. A partial purification of lupin β-cyanoalanine synthase is described and its distinction from cysteine synthase confirmed. Carbon-14 labelled substrates were supplied to etiolated seedlings of lupin in order to identify precursors of asparagine. Four carbon acids related to the TCA cycle were readily converted to asparagine i n vivo but not in vitro; the carbon skeleton of aspartate is retained in asparagine. Comparison of the distribution of label in the carbon skeletons of aspartate and asparagine from plants supplied carboxyl labelled fumarate shows a separation of the aspartate pool for asparagine biosynthesis from most cell aspartate. Metobolites that could be expected to give rise to cyanide and β-cyanoalanine are relatively ineffective as asparagine precursors. Thus the β-cyanoalanine pathway is not of major importance in asparagine biosynthesis in lupins.
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    Carbohydrate fractionation and elongation of lupin hypocotyl cell walls : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Botany at Massey University
    (Massey University, 1974) Monro, John Alexander
    The relationship between extensibility, growth rate and carbohydrate composition in different sections of lupin hypocotyl has been investigated. Although significant differences in extensibility were found, the carbohydrate composition of elongating and non-elongating regions were similar when delignified tissue was examined. However, it was subsequently found that the delignification removed all of the wall hydroxyproline, most of the arabinose, and much galactose and that all of these were higher in non-elongating than in elongating hypocotyl. The acid conditions of delignification caused about half of the loss of the sugars but did not cause the loss of hydroxyproline. Extraction of the hypocotyl cell walls with guanidinium thiocyanate and other denaturants, both before and after treatment with dilute acid or sodium methoxide in methanol did not dissolve the hydroxyproline, indicating that compounds containing this amino acids are sprobably covalently linked to insoluble wall constituents other than through acid labile arabinofuranose-hydroxyproline or ester links alone. 10% KOH extracted most of the wall hydroxyproline and hemicellulose largely as non-dialysable material. The hemicellulose thus extracted may be fractionated into hemicelluloses A and B and the latter into linear 1-4 linked polysaccharides and branched polysaccharides. Most of the hydroxyproline containing polymer is co-precipitated with the linear 1-4 linked hemicellulose-B arabinoxylan. When cell walls from elongating and non-elongating hypocotyl sections were compared the hemicellulose-B arabinoxylan fraction from the non-elongating wall had a much higher proportion or arabinose, galactose and hydroxyproline than the same polymer from elongating wall. Extraction of cell walls with 10% KOH at O°C removed about two thirds of the hemicellulose-B but little hydroxyproline. Subsequent treatment with 10% KOH at room temperature removed most of the hydroxyproline and remaining hemicellulose-B. The hemicellulose-B removed at room temperature showed the greatest increases in arabinose and galactose accompanying cessation of elongation. The polysaccharide extracted at 0°C is mainly xylan while that removed at room temperature contains large amounts of galactose and arabinose. The release of galactose at room temperature was accompanied by destruction or serine and appeared to parallel β-elimination of galactosylserine The kinetics of release of arabinose and galactose at room temperature differed. The above and other results are discussed particularly in relation to wall structure and a tentative model for the extensin-polysacchsride complex of lupin hypocotyl cell walls is proposed.
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    An evaluation of lupins (Lupinus spp.) for seed protein production : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy at Massey University, Palmerston North, New Zealand
    (Massey University, 1979) Withers, Neville John
    Since 1972 there has been interest in the greater use of seed protein in grain-based meals for stock. Lupins were one of the crops proposed to fill this requirement. This study was initiated to provide information on the agronomic requirements of Lupinus angustifolius, L. luteus and L. albus for seed production with emphasis on the southern North Island of New Zealand. In addition, some more basic studies on carbon and nitrogen translocation and the response of lupins to water stress were also carried out to provide a better understanding of the lupin plant and its response to its environment. Initially some field experiments were laid down to measure responses to sowing date, plant density, defoliation and cultivar. At wide spacing, L. angustifolius showed an approximately linear decrease in seed yield/plant as sowing date moved from April to October. At normal densities, however, sowing in late July gave the best see yield. Autumn sowings were affected by disease. It was concluded that, in the absence of disease, seed yield was largely determined by the length of the period of favourable environmental conditions between the start of flowering and the finish of reproductive development. This period determined the number of lateral inflorescences produced which, in turn, determined the number of pods producing seed. Pod number was the main component influencing seed yield. Thus, early sowing and reliable summer rainfall or irrigation seem to be the factors determining high lupin seed yields. Responses to density were variable. In one experiment there was no response in seed yield by four cultivars over these sowing times to densities ranging from 50-140 pl/m2. In a further experiment, increases in seed yield were obtained as plant density increased from 25-100 pl/m2. Removal of the main stem growing point early in growth briefly stimulated lateral stem growth but the effect on lateral stem seed yield was insufficient to compensate for the loss of the main stem seeds. There was little difference between the L. angustifolius cultivars Uniharvest, Uniwhite and Unicrop when sown early but, with late spring sowing, Unicrop flowered earlier which was an advantage under dry early summer conditions. In one experiment comparing a range of legume species, L. albus and Pisum sativum produced the highest seed yield but L. albus and L. luteus yielded the most protein per unit area. The peak rate of nitrogen accumulation in all species was similar and the main factor influencing protein yield appeared to be the duration of nitrogen accumulation. Provided each crop utilised similar durations of the growing period, the yield of seed protein/ha from various legume crops is likely to be similar; the main difference being the composition of the seed. It was suggested that, for maximum seed protein yield, indeterminate cultlvars may have some advantage over more determinate cultivars provided appropriate management procedures are adopted. Studies on water stress indicated that it plays an important role by influencing the distribution of assimilate between vegetative and reproductive growth. Mild water stress tended to stop vegetative growth and increase the rate of seed growth. When sufficiently severe, water stress appeared to initiate the senescence of the plant, the timing of which determined the potential seed yield for that situation. Water deficit had its main effect on seed yield by reducing pod number. Other yield components were relatively stable. Day temperatures of 28°C, when imposed early in growth, reduced vegetative and seed yield in L. albus. As the plant developed, however, the adverse effects of high temperature decreased until growth was stimulated during first order lateral flowering. No direct effect of high temperature on pod abscission was apparent and it was suggested that pod loss under high temperatures which have been reported occurred largely because of an associated water stress. A 14C translocation study indicated that most movement of photosynthate in L. albus was into the branch on which the labelled leaf was inserted, or into lower branch orders directly connected to it. Results suggest that, in L. albus cv. Ultra, lower order stems are a more important competitor with the inflorescence for photosynthate than the new, rapidly developing, higher order lateral branches. A possible strategy for growing lupin in a commercially viable situation in the Southern North Island is discussed.
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    Nutritional evaluation of grain legumes for poultry : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Poultry Nutrition at Massey University, Palmerston North, New Zealand
    (Massey University, 2009) Nalle, Catootjie Lusje
    The nutritional value of faba beans (Vicia faba), Australian sweet lupins (Lupinus angustifolius), white lupins (Lupinus albus) and peas (Pisum sativum) grown in New Zealand for broilers were evaluated in terms of their nutritional characteristics, protein quality (protein efficiency ratio), apparent metabolisable energy, apparent ileal digestibility coefficient of amino acids and the effects on bird performance. The effects of dehulling and extrusion cooking on the nutritive value of legumes were also investigated. The first experiment discussed in Chapter 3 evaluated the effect of cultivars on the nutrient profile and protein quality of chickpeas (Cicer arietinum), Australian sweet lupins, peas and soybeans (Glycine max). With the exception of white lupins, cultivars had no effect on the proximate and fibre composition of grain legumes. Starch was the primary carbohydrate component of chickpeas and peas, whilst non-starch polysaccharides were the major carbohydrates in lupins. The legume proteins were deficient in lysine, methionine, cystine and threonine. No differences were found in protein quality between cultivars of the different grain legume species. The lowest weight gain and protein efficiency ratio, in addition to the highest relative pancreatic weight and mortality rate was found in raw soybeans, suggesting that soybeans contained high a concentration of anti-nutritional factors, such as protease inhibitors. Birds fed chickpeas, lupins and peas had a low mortality rate and relative pancreatic weight, confirming that the level of anti-nutrients in these legume seeds was low. The apparent metabolisable energy and apparent ileal digestibility coefficient of amino acids of faba beans, Australian sweet lupins, white lupins and peas were determined in the second experiment (Chapter 4). Cultivar effect on the apparent metabolisable energy values was observed only for faba beans and white lupins. Faba beans, white lupins and peas had comparable apparent metabolisable energy values, but these values were higher than those of Australian sweet lupins, and lower than that of soybean meal. No cultivar differences were found in the apparent ileal digestibility coefficient of amino acids of grain legumes. The apparent ileal digestibility coefficient of amino acids of both lupin species was found to be comparable to that of soybean meal. The effects of feeding diets containing 200 g/kg faba beans, lupins or peas on the performance, digestive tract development and litter quality of broilers were investigated in the third and fourth trials. In the cage trial (Chapter 5), the results showed that the weight gain of birds fed diets containing grain legumes was similar to that of control diet. Feed intake and feed per gain of birds fed diets containing the majority of grain legume cultivars did not differ from those fed the maize-soy diet. Birds fed diets containing faba beans had more dry and friable excreta compared to other treatment diets. The performance of birds fed diets containing 200 g/kg grain legumes during the 35 d grow-out period, in the floor pen trial (Chapter 6), confirmed the results of the cage trial. In this trial, weight gain and feed per gain of birds fed diets without meat meal were superior to those with meat meal. In cage trials, the modification of some segments of digestive tract development was probably due to the dietary NSP. Whilst in floor pen trial, digestive tract development was not influenced by the inclusion of grain legumes. The effect of methodology of determination (direct vs. difference method) on the apparent ileal digestibility coefficient of amino acids of wheat, maize, Australian sweet lupins, peas and soybean meal for broilers was evaluated in the fifth study (Chapter 7). The influence of methodology on apparent ileal digestibility coefficient of amino acids was found to vary amongst the feed ingredients. In general, the apparent ileal digestibility coefficient of amino acids of test ingredients determined by the difference method was higher than those determined by the direct method, suggesting that the use of the direct method may underestimate the apparent ileal digestibility coefficient of amino acids in low and medium protein ingredients. Data reported in Chapter 8 shows that dehulling increased the apparent metabolisable energy values of faba beans and Australian sweet lupins, but it had no beneficial effect on peas. The increase of apparent metabolisable energy values may be attributed to the decrease in non-starch polysaccharides of these legume seeds after dehulling. The removal of hulls increased the amino acid concentrations, but it had no effect on the apparent ileal digestibility coefficient of most amino acids. These results suggest that dehulling of grain legumes would be nutritionally beneficial and, likely to be economical in view of the improved amino acid concentrations and energy values. The final experiment (Chapter 9) demonstrated that extrusion of peas markedly influenced the content of crude protein, non-starch polysaccharides, starch, and trypsin inhibitors. The soluble non-starch polysaccharides and trypsin inhibitor contents of the majority of extruded pea samples were higher than those of raw peas, but insoluble and total non-starch polysaccharides decreased with extrusion. Extrusion had no effect on the apparent ileal protein digestibility and the apparent metabolisable energy of peas, but it increased ileal starch digestibility.