School of Agriculture, Food and Ecosystem Sciences - Theses

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    Effect of elevated carbon dioxide and high temperature on major micronutrients in strawberry
    Balasooriya, Himali ( 2019)
    In this study, four different folate derivatives (tetrahydrofolic acid – THFA, 10-formylfolic acid – 10FFA, 5-formyltetrahydrofolic acid – 5FTHFA, 5-methytetrahydrofolic acid (5MTHFA)) were identified in fresh and freeze-dried strawberry samples. The individual and interaction effects of increased [CO2] and temperature on total folates content were significant (P≤0.05), and the responses were cultivar dependant. Total folate content in strawberries varied from 52.6 ± 5.1 µg to 364.8 ± 16.0 µg/100 g FW in cultivar ‘Albion’ and from 48.6 ± 7.0 µg to 237.4 ± 23.8 µg/100 g FW in cultivar ‘San Andreas’. Although, increased temperature positively affected the total folates content under lower [CO2] levels, the effects turned negative at the highest [CO2] concentration (950 pm). Higher temperature reduced the content of total folates in strawberries by 26% and 13% in cultivar ‘Albion’ and ‘San Andreas’, respectively. Impacts of elevated [CO2], higher temperature and their interactions on total vitamin C content in strawberries were statistically significant (P≤0.05) and the responses were cultivar dependent. Vitamin C contents in cultivar ‘Albion’ and ‘SA’ fresh strawberries were in a range of 59 ± 7 mg to 133 ± 15 mg/100 g FW and 56 ± 9 mg to 132 ± 9 mg/100 g FW, respectively. Increased growth temperature to 30 °C at 650 ppm [CO2] enhanced the amounts of vitamin C significantly (P≤0.05) to a maximum by 123% and 132% in cultivars ‘Albion’ and ‘San Andreas’, respectively. However, that effect wasn’t detected when the CO2 concentration was increased further to 950 ppm, and vitamin C concentrations drastically decreased by 36% and 31% in Albion’ and ‘San Andreas’, respectively. In general, folates and vitamin C contents were significantly (P≤0.05) higher in FD strawberry than fresh fruits. The next step of the study was to study the accessibility of increased polyphenols, vitamin C and folates in the fruits of fresh and frozen strawberries using simulated in vitro gastrointestinal digestion and colonic fermentation. Elevated [CO2] (ambient to 950 ppm) and higher temperature (ambient to 30 °C) enhanced the accessibility of polyphenols, folate and vitamin C in strawberries. Bioaccessibility of Pel-3-Glu increased from 67% to 88% in fresh strawberries when exposed to elevated growth. The exact amounts of individual polyphenols in accessible fraction were significantly (P≤0.05) higher in fresh fruits of strawberries grown under elevated growth conditions. For example, the highest amounts of Pel-3-Glu (19.89±0.4 mg/100 g FW), Pel-3-Rut (2.55±0.5 mg/100 g FW), p-coumaric (0.23±0.02 mg/100 g FW), ferulic (1.33±0.05 mg / 100 g FW), quercetin (1.97±0.2 mg/100 g FW) and p- coumaroyl (0.65±0.05 mg/100 g FW) were detected in fed state simulated gastrointestinal digesta of fresh strawberry grown under elevated growth conditions. Fresh strawberries grown under ambient growth contained 93.09±6.2 µg/100g folates and 18.55±0.5 mg/100g vitamin C as bioaccessible fractions under fed state while, elevated growth enhanced soluble folates and vitamin C up to 188.63±7.5 µg/100g and 30.48±0.3 mg/100g, respectively. Fresh strawberries contained higher amounts of accessible micronutrients than frozen strawberries, while increased bile contents in intestinal fluid (fed state) facilitated the release of bioactive compounds to gastrointestinal fluid. The insoluble fraction of strawberry digests after gastrointestinal digestion was then subjected to in vitro colonic fermentation using human faecal cultures and basal media. The soluble fraction of fermented strawberry digests was extracted to analyse polyphenols, folates and vitamin C. Higher contents of folate (7.90±0.05 µg/100 g FW), vitamin C (33.6±1.0 ng/100 g FW), Pel-3-Glu (2.00±0.14 mg/100 g FW), and p-coumaric (39±5 µg/100 g FW) were observed in soluble fraction of fermented precipitate after simulated gastrointestinal digestion at fasted state in frozen strawberries. These bioactive compounds and their metabolites would play an important role in the human colon by maintaining a healthy environment via scavenging the free radicals. According to the current study, the amount of bioaccessible bioactive compounds in strawberry could vary quantitatively and qualitatively based on growth and storage conditions as well as the status of digestion (fed or fasted state). Increased carbon dioxide and temperature in the growth environment enhanced the bioaccessibility of polyphenols, folates and vitamin C in strawberries. It can be concluded that strawberry fruits grown under elevated [CO2] and temperature may not be visually attractive comparing to normal strawberries. However, considering their nutritional value, those fruits can be promoted as freeze-dried strawberry in value added foods such as dairy products. Additionally, these research outcomes would help the commercial growers to focus on the nutritional aspects of fruits and vegetables grown under such elevated and extreme environmental conditions in the future. However, as a very little information is available concerning the interactive effects of elevated [CO2] and high temperature on fruits and vegetables in the field, more researches are needed to confirm the results from glasshouse studies.
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    Antioxidant defence systems and symptom expression of wheat infected with Barley yellow dwarf virus and grown under elevated CO2
    Vandegeer, Rebecca Kate ( 2016)
    Barley yellow dwarf virus (BYDV) is regarded as the most significant viral pathogen of wheat worldwide. Symptoms produced during viral infection may have an interactive effect with environmental conditions expected under future anthropogenic climate change, including the rising atmospheric CO2 concentration. In particular, antioxidant defence systems – including the key non-enzymatic antioxidants ascorbate and glutathione – play an important role in regulating potentially harmful reactive oxygen species (ROS) produced during plant-virus interactions. However, the role of ascorbate and glutathione during systemic virus infection and growth under elevated CO2 (eCO2) is not well understood. This thesis investigated BYDV infection of three Australian wheat cultivars: the BYDV-susceptible spring wheat ‘Yitpi’, the susceptible winter wheat ‘Revenue’ and the resistant winter wheat ‘Manning’. In addition, the system was investigated under eCO2 to determine any interactions with infection on symptom expression and antioxidant defence capacity. Studies were performed within controlled environment chambers and the field at the Australian Grains Free Air CO2 Enrichment (AGFACE) facility located in the semi-arid grain-growing region of Horsham, Victoria, Australia. The response of plants to virus infection and eCO2 was assessed by measurement of the total concentration and redox state of ascorbate and glutathione. In addition, symptom expression was measured including growth, photosynthesis, stomatal conductance, leaf chlorophyll and nitrogen, and disease incidence and severity. BYDV infection was associated with an imbalance in antioxidative metabolism, which is an indicator of oxidative stress. Greater ROS turnover is the likely cause of the observed decrease in total ascorbate and glutathione and increase in the oxidised fraction of ascorbate after infection. In particular, a decrease in total ascorbate was the most consistent response to infection by all cultivars grown in both chambers and the field. The present research demonstrates that the observed imbalance in non-enzymatic antioxidant metabolism can be used as a marker for oxidative stress during systemic BYDV infection of wheat. The antioxidant response of both the BYDV-susceptible and resistant winter wheat cultivars was similar. Oxidative stress was not influenced by the putatively different virus concentration between these cultivars, but simply by virus infection alone. Infection was also associated with decreased biomass and height in both these cultivars and in both chamber and field studies, which indicates a sensitivity of the resistant cultivar to infection regardless of a putatively lower virus concentration. Despite few interactive effects between virus and eCO2 treatments on symptom expression, eCO2 altered the expression of yellowing disease symptoms in virus-infected plants, although not consistently between cultivars and environmental growing conditions. In addition, although there were significant changes to antioxidants in plants grown under eCO2, results were not consistent between studies. Research into this topic increases our understanding of how plants respond to virus infection and oxidative stress, and how plant-virus interactions may change under future eCO2. With the findings presented in this thesis, I have furthered the knowledge of this area by elucidating the response of ascorbate and glutathione during systemic wheat-BYDV interactions, and reinforced the potential use of these metabolites as markers of oxidative stress.