Influence of Genome-Specific Granule-Bound Starch Synthase I (GBSSI/Waxy) on Starch Composition, Structure and In Vitro Enzymatic Hydrolysis in Wheat (Triticum aestivum L.)

Wheat grain quality and consumption is influenced by its constituents structure and concentrations. In the first part of the dissertation, six Canadian bread wheat cultivars; four (CDC Teal, AC Superb, AC Barrie, AC Splendor) belonging to the Canada Western Red Spring (CWRS), and two (AC Foremost, and AC Crystal) to the Canada Prairie Spring Red (CPSR) market classes were characterized for the relationship between their starch constituents and starch in vitro enzymatic hydrolysis. CPSR cultivars with relatively longer amylopectin chains of DP 37-45, reduced chain lengths of DP 15-18, and a low volume percent of small C-type starch granules, had reduced starch in vitro enzymatic hydrolysis rates. In the second part of the dissertation, near-isogenic wheat lines differing at the Waxy locus were analyzed for the influence of genome-specific granule-bound starch synthase I (GBSSI/Waxy; Wx-A, Wx-B, Wx-D) on starch composition, structure and starch in vitro enzymatic hydrolysis. Amylose concentration was more severely affected in genotypes with GBSSI missing from two genomes (double nulls) than from one genome (single nulls) of wheat, indicating dosage dependent amylose synthesis. Subtle differences in amylopectin chain length distribution were observed among non-waxy, partial and completely waxy starches, suggesting a non-limiting role of genome-specific GBSSI for amylopectin synthesis. A suppressive role of Wx-D on the short chain phenotype of wheat amylopectin was observed. In addition, Wx-D increased the volume percentage of large A-type starch granules and reduced starch hydrolysis index. Thus, among the waxy isoproteins, Wx-D might be the major contributor for reducing the rate of in vitro starch enzymatic hydrolysis in wheat. In the third part of the dissertation, endosperm starch’s physicochemical properties and structure during grain development in wheat waxy-null genotypes were analyzed. The study was conducted with pure starch isolated from wheat grains at 3-30 days post anthesis (DPA), at three day intervals. Changes in amylopectin structure were observed until 12 DPA, suggesting the formation of a basic amylopectin skeleton by this stage. A differential influence of waxy isoproteins on amylopectin structure formation has been suggested, with Wx-B and Wx-D affecting short glucan chains of DP 6-8 at 3 and 6 DPA, Wx-A being effective at 9 and 12 DPA, and Wx-D affecting DP 18-25 chains from 18-30 DPA

Enzymatic clarification of carrot juice by using response surface methodology

 Carrot juice was treated with pectinase at various enzyme concentrations (0.01% to 0.1%), process temperatures (350C to 550C) and incubation time (40 to 120 min).  The effect of these enzyme treatments on filterability, clarity, turbidity and viscosity of the juice were studied by employing a second order central composite design.  The coefficient of determination (R2) values for filterability, clarity, turbidity and viscosity were greater than 0.85.  Statistical analysis showed that filterability, clarity, viscosity and turbidity were significantly correlated to enzyme concentration, incubation temperature and incubation time.  Enzyme concentration was the most important factor affecting the characteristics of the carrot juice as it exerted a highly significant influence on all the dependent variables.  An increase in process time and/or concentration of enzyme treatment was associated with an increase in filterability and clarity, and decrease in turbidity and viscosity.  Based on response surface and contour plots, the optimum conditions for clarifying carrot juice were 0.092% enzyme concentration, incubation temperature of 54.20C and incubation time of 119 min. Keywords: carrot juice, enzyme clarification, response surface methodology  

Development of Barley (<i>Hordeum vulgare</i> L.) Lines with Altered Starch Granule Size Distribution

Microscope analysis of starches prepared from 139 barley genotypes identified a Japanese genotype, Kinai Kyoshinkai-2 (KK-2), with altered starch granule size distribution. Compared to normal barley starch, KK-2 produced consistently higher volumes of starch granules with 5–15 μm diameter and reduced volumes of starch granules with >15 μm diameter when grown in different environments. A cross between KK-2 and normal starch cultivar CDC Kendall was made and led to the production of 154 F₅ lines with alterations to the normal 7:3:1 distribution for A-:B-:C-type starch granule volumes. Three F₅ lines showed unimodal starch granule size distribution due to apparent lack of very small (<5.0 μm diameter) C-type starch granules, but the phenotype was accompanied by reduced grain weight and total starch concentration. Five F₅ lines produced a significantly larger population of large (>15 μm diameter) A-type starch granules as compared to normal starch and showed on average a 10:4:1 distribution for A-:B-:C-type starch granule volumes. The unusual starch phenotypes displayed by the F₅ lines confirm starch granule size distribution in barley can be genetically altered

Braving the attitude of altitude: Caragana jubata at work in cold desert of Himalaya

The present work was conducted to understand the basis of adaptation in Caragana jubata in its niche environment at high altitude cold desert of Himalaya. Molecular data showed predominance of genes encoding chaperones and those involved in growth and development at low temperature (LT), a major cue operative at high altitude. Importantly, these genes expressed in C. jubata in its natural habitat. Their homologues in Arabidopsis thaliana, Oryza sativa, and Glycine max did not exhibit similar trend of gene expression at LT. Constitutive expression and a quick up-regulation of the above genes suggested the ability of C. jubata to adjust its cellular machinery to maintain growth and development in its niche. This was reflected in LT50 (the temperature at which 50% injury occurred) and LT mediated photosynthetic acclimatory response. Such molecular and physiological plasticity enables C. jubata to thrive in the high altitude cold desert of Himalayas