Graduation date: 1998

No description available.

Haloxyfop-methyl 2-[4-[ [3-chloro-5-(trifluoromethyl)-2-pyridinyl] oxy] phenoxy] propanoate, fluazifop-butyl (±)-2-[4-[ [5-(trifluoromethyl)-2-pyridinyl] oxy] phenoxy] propanoate, and sethoxydim 2- [1-(ethoxyimino)butyl]-5- [2-(ethylthio)propyl] -3-hydroxy-2-cyclohexen-1-one modified chloroplast pigment, growth, anatomy, and ultrastructure of pre- and postemergent-treated wheat (Triticum aestivum 'Pavon 76') seedlings. Seedlings were grown under 16 h photoperiod (105 μE/M²/S) at 32/24 C day/night temperature. Pigment, growth and anatomical studies were made on pre- and postemergent-treated seedlings. Postemergent treatments were applied to 6-day-old seedlings. Carotenoid levels were reduced by 90 to 94% in preemergent-treated seedlings at 5 x 10⁻⁵ M for each herbicide. Postemergent application of 10⁻³ M haloxyfop-methyl and sethoxydim caused a reduction of 11 and 15% in carotenoid concentration than the control, respectively. Also, they produced a significant reduction in chlorophyll a and total chlorophyll concentration. Preemergent treatment with 5 x 10⁻⁶ M of each herbicide caused a reduction of 78 to 91% in leaf fresh weight and 72 to 84% reduction in root dry weight as compared to the control. Height of the seedlings was reduced by 79 to 86% of the control. Postemergent treatments with 10⁻³ M of each herbicide reduced the length of the second leaf by 98%, whereas length of the first leaf was reduced by 36% following haloxyfop-methyl treatment and 45% following fluazifop-butyl or sethoxydim treatments. Fresh weight of leaf tissue treated with 10⁻³ M of each herbicide was reduced by 40%. Preemergent treatment of the seedlings with 3 x 10⁻⁶ and 5 x 10⁻⁶ M haloxyfop-methyl or with 5 x 10⁻⁶ M fluazifop-butyl or sethoxydim caused 35 to 50% enlargement of mesophyll cells as compared to the control. These cells appeared to contain less cytoplasm. Post- emergent application of 10⁻³ M of each herbicide caused swelling of mesophyll cells. Ultrastructural studies showed that plastids of seedlings germinated in 5 x 10⁻⁶ M of each herbicide were disrupted, swollen, and lacked internal thylakoids but contained masses of plastoglobuli. The chloroplasts of postemergent-treated seedlings had fewer granal and intergranal thylakoids, but the injury to plastids was less severe than in preemergent treatments.

Considerable research has been concentrated on the foliage application of Cycoce, whereas little attention has focused on its use in seed application. Imbibition of caryopses of wheat (Triticum aestivum L.) in (2-chloroethyl) trimethylammonium chloride (Cycocel/CCC) resulted in a reduction in germination rate and a stunting of seedling growth. This was consistent with the appearance of a thicker stem, with shorter leaves than the control. Chlorophyll content per unit weight of leaf and numbers of tillers initiated both increased in comparison to controls. However, Cycocel treatment was accompanied by an enhanced survival capacity of seedlings when they were subjected to water limitation under growth room conditions. Seedlings grown from caryopses imbibed in 0.4 % Cycocel and exposed to a 7-day cycle of watering remained green and turgid beyond the time when the controls had wilted and died. Seedlings, at the fourth leaf stage, showed no wilting 23 days after watering had been withheld totally. Enhanced survival was also noted in seedlings when caryopses had been imbibed in Cycocel for 24 hours followed by a period of dry storage for up to 2 weeks prior to sowing. Enhanced survival was also detected as a carryover into the tillers, and the subsequent GEN2 seedlings, derived from GEN1plants. Cycocel pre-treatment did not alter Stomatal Index to an extent which could lead to enhanced water use efficiency, nor to the accumulation of proline to bring compatible solutes into equilibrium. However, the pre-treatment with Cycocel was found to enhance growth as a counter to water stress, and maintained fresh and dry weight of shoots when examined under the polyethylene glycol (PEG) induced stress. At the anatomical level the microscopic investigation of the GEN2 leaf tissues, derived from GENl plant, revealed a fully turgid cell structure, cells did not collapse and tissues were not distorted after the extraction of chlorophyll when compared with the control of the continuous watering as well as the moderate watering regime. Additionally, the cortex of GENl plants derived from pre-treated caryopses, showed a bright fluorescing unidentified deposition under Nutrient Solution (NS). This was more much greater when sampled from the NS + PEG treatments. From the results achieved Cycocel pre-treatment of caryopses appeared to have potential in manipulating wheat plants against water limitation. However, the mechanism (s) by which this can be achieved was not fully resolved in this study. Further investigations are required at the biochemistry and molecular level, particularly in relation to potential carryover effects between generations.

Phenylketonuria and hyperphenylalanemia are autosomal recessive inborn errors of phenylalanine metabolism that are caused by mutations in the phenylalanine hydroxylase gene. Due to the stringency of the present dietary therapy, alternative treatments are being studied. Phenylalanine ammonia-lyase (PAL) is one of the potential dietary supplements for these patients. PAL is a well-studied plant enzyme which breaks down phenylalanine into trans-cinnamic acid and ammonia (Camm and Towers, 1973). It is found in the cytoplasm of the plant cells and is naturally encapsulated by plant cell walls which may protect it against the acidic pH environment in the gastrointestinal tract. It presumably degrades ingested Phe that circulates in the intestinal lumen. In this study, red spring wheat seedlings (Triticum aestivum L.) found to contain high PAL activity naturally were investigated as a potential alternative oral therapy. Specifically, the objectives were (1) to evaluate different drying methods on generating concentrated and dried preparation of wheat seedlings containing high levels of PAL activity; (2) to examine the retention of PAL activity over three months of storage under various storage conditions; (3) to determine the stability of PAL activity in simulated human digestion condition to establish if further study of using plant source enzyme in vivo is warranted. Freeze-drying (FD) was found to have retained the most activity (>90 % recovery dry wt basis) compared to air-drying (AD) and vacuum-microwave drying (VMD) for both leaf and residual seed/root samples. Pre-freezing of leaf tissues at -18 °C before FD significantly retained the highest PAL activity compared to pre-freezing at -25 °C, -35 °C, and -80 °C (P<0.05). Over three months of storage, 60-80 % of PAL activity was recovered in leaf and —100 % was recovered in residual seed/root tissues after storage at -20 °C. After in vitro protein digestion, 36% and 42 % of PAL activity was recovered in fresh leaf and root tissues respectively; however, FD tissues were found to be susceptible to proteases and acidic environment and no activity was recovered after three hours of in vitro protein digestion. High performance liquid chromatography (HPLC) analysis of the residual Phe after in vitro protein digestion confirmed that fresh tissues had significantly higher conversion of Phe than that of FD tissues. Together, these results suggest that red spring wheat seedlings may have potential as a dietary supplement for phenylketonuric patients while further study to enhance PAL activity in plant preparations is required.

Phenylketonuria and hyperphenylalanemia are autosomal recessive inborn errors of phenylalanine metabolism that are caused by mutations in the phenylalanine hydroxylase gene. Due to the stringency of the present dietary therapy, alternative treatments are being studied. Phenylalanine ammonia-lyase (PAL) is one of the potential dietary supplements for these patients. PAL is a well-studied plant enzyme which breaks down phenylalanine into trans-cinnamic acid and ammonia (Camm and Towers, 1973). It is found in the cytoplasm of the plant cells and is naturally encapsulated by plant cell walls which may protect it against the acidic pH environment in the gastrointestinal tract. It presumably degrades ingested Phe that circulates in the intestinal lumen. In this study, red spring wheat seedlings (Triticum aestivum L.) found to contain high PAL activity naturally were investigated as a potential alternative oral therapy. Specifically, the objectives were (1) to evaluate different drying methods on generating concentrated and dried preparation of wheat seedlings containing high levels of PAL activity; (2) to examine the retention of PAL activity over three months of storage under various storage conditions; (3) to determine the stability of PAL activity in simulated human digestion condition to establish if further study of using plant source enzyme in vivo is warranted. Freeze-drying (FD) was found to have retained the most activity (>90 % recovery dry wt basis) compared to air-drying (AD) and vacuum-microwave drying (VMD) for both leaf and residual seed/root samples. Pre-freezing of leaf tissues at -18 °C before FD significantly retained the highest PAL activity compared to pre-freezing at -25 °C, -35 °C, and -80 °C (P<0.05). Over three months of storage, 60-80 % of PAL activity was recovered in leaf and —100 % was recovered in residual seed/root tissues after storage at -20 °C. After in vitro protein digestion, 36% and 42 % of PAL activity was recovered in fresh leaf and root tissues respectively; however, FD tissues were found to be susceptible to proteases and acidic environment and no activity was recovered after three hours of in vitro protein digestion. High performance liquid chromatography (HPLC) analysis of the residual Phe after in vitro protein digestion confirmed that fresh tissues had significantly higher conversion of Phe than that of FD tissues. Together, these results suggest that red spring wheat seedlings may have potential as a dietary supplement for phenylketonuric patients while further study to enhance PAL activity in plant preparations is required.

Phenylketonuria and hyperphenylalanemia are autosomal recessive inborn errors of phenylalanine metabolism that are caused by mutations in the phenylalanine hydroxylase gene. Due to the stringency of the present dietary therapy, alternative treatments are being studied. Phenylalanine ammonia-lyase (PAL) is one of the potential dietary supplements for these patients. PAL is a well-studied plant enzyme which breaks down phenylalanine into trans-cinnamic acid and ammonia (Camm and Towers, 1973). It is found in the cytoplasm of the plant cells and is naturally encapsulated by plant cell walls which may protect it against the acidic pH environment in the gastrointestinal tract. It presumably degrades ingested Phe that circulates in the intestinal lumen. In this study, red spring wheat seedlings (Triticum aestivum L.) found to contain high PAL activity naturally were investigated as a potential alternative oral therapy. Specifically, the objectives were (1) to evaluate different drying methods on generating concentrated and dried preparation of wheat seedlings containing high levels of PAL activity; (2) to examine the retention of PAL activity over three months of storage under various storage conditions; (3) to determine the stability of PAL activity in simulated human digestion condition to establish if further study of using plant source enzyme in vivo is warranted. Freeze-drying (FD) was found to have retained the most activity (>90 % recovery dry wt basis) compared to air-drying (AD) and vacuum-microwave drying (VMD) for both leaf and residual seed/root samples. Pre-freezing of leaf tissues at -18 °C before FD significantly retained the highest PAL activity compared to pre-freezing at -25 °C, -35 °C, and -80 °C (P<0.05). Over three months of storage, 60-80 % of PAL activity was recovered in leaf and —100 % was recovered in residual seed/root tissues after storage at -20 °C. After in vitro protein digestion, 36% and 42 % of PAL activity was recovered in fresh leaf and root tissues respectively; however, FD tissues were found to be susceptible to proteases and acidic environment and no activity was recovered after three hours of in vitro protein digestion. High performance liquid chromatography (HPLC) analysis of the residual Phe after in vitro protein digestion confirmed that fresh tissues had significantly higher conversion of Phe than that of FD tissues. Together, these results suggest that red spring wheat seedlings may have potential as a dietary supplement for phenylketonuric patients while further study to enhance PAL activity in plant preparations is required. / Land and Food Systems, Faculty of / Graduate