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ItemEffect of elevated carbon dioxide and high temperature on major micronutrients in strawberryBalasooriya, 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.