1. KAUTIR (Kerala Agricultural University Theses Information and Retrieval)

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    Antioxidant and anticarcinogenic potential of jackfruit based ready-to-cook (RTC) curry mixes
    (Department of Community Science, College of Agriculture,Vellayani, 2019) Gayathri Mohan; Suma Divakar
    The research work entitled, “Antioxidant and anti-carcinogenic potential of jackfruit based Ready-To-Cook (RTC) curry mixes” was conducted at College of Agriculture, Vellayani during 2017-2019, with the objective to ascertain the antioxidant and anti-carcinogenic properties of the jackfruit based curry mixes (‘Avial’ mix, ‘Koottu’ mix and ‘Ularth’ mix) which were developed earlier at the Department of Community Science, using raw jackfruit (Koozha type). Methanol, ethanol, petroleum ether and acetone were used to extract the bioactive compounds of the powdered mixes for phytochemical screening, antioxidant analysis and verification of anti-carcinogenic property. The results of the phytochemical analysis indicated the presence of tannins, flavonoids, saponins, steroids, phenolic compounds, cardiac glycosides and anthraquinones in the three mixes; whereas the analyses revealed the absence of alkaloids and phlotobatinins. Quantitative analysis of antioxidants revealed that the beta carotene content of the RTC mixes was in the range of 0.07-1.16 mg/100g. Beta carotene content was found to be more in ‘Avial’ mix (1.16mg/100g). Ascorbic acid content was also found to be higher for ‘Avial’ mix (38.45 mg) and the lowest content was found for ‘Ularth’ mix (15.34 mg). The saponin content was found to be highest in ‘Avial’ mix (6.65 per cent) and lowest in ‘Ularth’ mix (5.00 per cent). The tannin content of ‘Ularth’ mix (11.57 mg) was observed to be on par with ‘Avial’ mix (11.52 mg). Total phenol content was found to be more in ‘Koottu’ mix (21.53 mg/100g) and the total flavonoid content of RTC mixes was seen to be in the range between 0.86-3.25 mg/100g. There was significant difference in the lectin content of ‘Avial’ mix (0.35%), ‘Koottu’ mix (0.56%) and ‘Ularth’ mix (0.75%). Alkaloids and selenium were found to be absent in the RTC mixes. In case of mineral analyses, Copper content was found to be higher in ‘Ularth’ mix (2.57 µg); zinc content was in the range of 4.65-6.55 µg and was found to be higher for ‘Koottu’ mix (6.55 µg) and lower for ‘Ularth’ mix (4.65 µg). Manganese content was found to be higher in ‘Koottu’ mix (5.30 µg) while, it was observed to be on par in ‘Avial’ mix (2.55 µg) and ‘Ularth’ mix (2.51 µg). The highest iron content was found in ‘Ularth’ mix (0.92 µg). Antioxidant activity in the present study revealed that ‘Avial’ mix had the highest DPPH activity with an IC50 value of 33.81 µg/ml. Total antioxidant activity was found to be more for ‘Avial’ mix (41.44 per µg/ml), followed by ‘Koottu’ mix (42.41 µg/ml) and ‘Ularth’ mix (43.45 µg/ml). ‘Avial’ mix showed more ferric reducing capacity while ‘Ularth’ mix had the least capacity in this regard. ABTS radical scavenging activities of RTC mixes ranged between 34.84-46.69 µg/ml. ABTS radical scavenging activity was observed to be higher for ‘Avial’ mix (34.84 µg/ml) and lower values were noted for ‘Ularth’ mix (40.52 µg/ml). Hydroxyl radical scavenging activity of RTC mixes was found to range between 50.55-52.55 µg/ml. ‘Avial’ mix showed higher superoxide radical scavenging activity with an IC50 value of 48.54 µg/ml and the lowest superoxide radical scavenging activity was observed in ‘Ularth’ mix (60.73 µg/ml). However, the highest nitric oxide scavenging activity was observed for ‘Ularth’ mix (14.11 µg/ml) followed by ‘Koottu’ mix (30.92 µg/ml) and ‘Avial’ mix (36.58 µg/ml). Among the three jackfruit based Ready-To-Cook (RTC) curry mixes ‘Avial’ mix was found to have higher antioxidant property and was therefore taken up for studying the anti-cancer property. Cytotoxicity of ‘Avial’ mix was estimated by MTT assay on Human breast adenocarcinoma cells (MCF-7) and 17 per cent of loss in cell viability was observed. Findings of the present study revealed that the jackfruit based RTC mixes comprised of various health promoting components such as beta carotene, ascorbic acid, saponins, tannins, total phenols, total flavonoids and lectins they also had potent antioxidant activities. Thus the curry mixes, in particular the ‘Avial’ mix can be promoted as functional food which will help to enhance its commercialization.
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    Isolation and characterisation of cDNA encoding dihydroflavonol 4- reductase gene from Orchid Dendrobium variety Sonia17
    (Department of Plant Biotechnology, College of Agriculture, Vellayani, 2009) Saritha, V S; Soni, K B
    The thesis entitled “Isolation and characterization of cDNA encoding dihydroflavonol 4-reductase gene from Dendrobium orchid variety Sonia 17” was conducted at the Department of Plant Biotechnology, College of Agriculture, Vellayani, Thiruvananthapuram during 2006-2008 with an objective of characterizing one of the key genes involved in anthocyanin biosynthesis, viz., dihydroflavonol 4-reductase from flower buds of Dendrobium Sonia 17 orchid. Reverse Transcription-PCR technique was used in the present study for the amplification of cDNA encoding dfr gene. Heterologous forward and reverse primers were designed based on the gene sequences of Cymbidium orchid and Oryza sativa using Primer 3 software. Total RNA isolated from the flower buds using TRIZOL reagent yielded good quality intact RNA, with no genomic DNA contamination and showed an A260/A280 ratio of 1.6-2.0. The cDNA synthesized from purified mRNA was amplified using gene specific primers designed. All the primers included in the study yielded reproducible amplification. The primer sets A (Af and Ar) and C (Cf and Cr) which were designed based on the dfr gene sequence of Cymbidium orchid yielded amplified fragments of size approximately 400-500 bp and 100 bp respectively. Primer set B (Bf and Br) synthesized from Oryza sativa dfr gene sequences yielded an amplified product of size nearly 300 bp. The cDNA amplified with primer set A was cloned in pGEM-T Easy Vector and multiplied in competent E. Coli strain DH5α for sequencing. The transformed E. Coli colonies were grown on L.B agar plates and selected by blue – white screening. Three white colonies were randomly selected and plasmid DNA was isolated from transformed clones. The presence of insert was checked by PCR analysis of the plasmid DNA with gene specific primer (primer set A). The agarose gel electrophoresis showed the presence of amplified fragment. Sequencing of two clones at Chromous Biotech Pvt. Ltd. using T7 and SP6 primer failed to amplify the plasmid DNA. This work was an initial step towards isolation and characterization of dihydroflavonol 4-reductase gene in Dendrobium variety Sonia 17. The primers designed in this study could successfully amplify the cDNA encoding dfr. PCR reactions gave good results but the sequencing was not satisfactory. For the better understanding of the dfr sequences, the sequencing reaction needs to be repeated with other clones.