Biological, chemical and mathematical sciences

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Browsing Biological, chemical and mathematical sciences by Subject "Agricultural biotechnology"

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    Agrobacterium tumefaciens - mediated transformation of nicotiana benthamiana with dehalogenase gene resistant to monochloroacetic acid
    (Universiti Teknologi Malaysia, 2017) Mohamed, Elizah
    Weeds give adverse effects to crops because of the competition to get nutrients, light and moisture. Many farmers used broad-spectrum herbicide such as monocloroacetic acid (MCA) which is effective at killing a wide range of weeds. Unfortunately, broad-spectrum of herbicide can also kill valuable crops and cause significant losses in agricultural productivity. One of the solutions to this problem is by developing herbicide resistant plant using dehalogenase D (dehD) gene isolated from Rhizobium sp. RC1. A dehD gene encoding dehalogenase enzyme that has the capability to degrade monochloroacetic acid (MCA) was isolated and characterized from Rhizobium sp. RC1. dehD gene was used as herbicide resistance gene and selectable marker gene in Nicotiana benthamiana plant transformation. The 798 bp dehD gene was inserted into pCAMBIA 1305.2 under the control of the Cauliflower Mosaic Virus 35S (CaMV35S) promoter and designated as pCAMdehD, with a total size of 10,592 bp. A few parameters of Agrobacterium tumefaciens-mediated transformation were optimized including hygromycin concentration (40 μg/mL of hygromycin), and the MCA toxicity level to N. benthamiana at tissue culture (60 μg/L of MCA) and whole plant stage (2.0 g/L of MCA). pCAMdehD was introduced into N. benthamiana via Agrobacterium mediated transformation method. Based on the screening of the transformants on MS media containing 60 μg/L MCA, the results showed that N. benthamiana was successfully transformed with dehalogenase D gene with 50 % of transformation efficiency. The integration and expression of dehD gene in N. benthamiana were confirmed by PCR, Southern Blotting and reverse transcription PCR. Analysis of leaf-painting assay revealed that transgenic N. benthamiana (T1) was resistant to 4.0 g/L MCA compared to 2.0 g/L for non-transformed plants control. The Chi Square analyses of five transgenic plants (T1), suggested that the dehD gene was segregated according to Mendelian 3:1 ratio. These findings showed that transgenic N. benthamiana plant resistant to MCA herbicide was successfully produced
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    Optimization of total phenolic contents in pyroligneous acid from oil palm kernel shell and its bioactivities
    (Universiti Teknologi Malaysia, 2017) Mahmud, Khoirun Nisa
    Processing of oil palm fruit into valuable commodities resulted in the generation of large volumes of solid biomass which contributes to various environmental problems. Current practice of leaving the biomass at the plantation area to putrefy, does not contribute to the economy of the nation even though huge potential of high-value biocompounds are present in these biomass. From the available processes for biomass management, pyrolysis offers excellent solution due to quick process and production of useful by-products which can be utilized further such as pyroligneous acid (PA). Nevertheless, very few studies were reported regarding the optimization of total phenolic contents (TPC) in PA using statistical approach. In this study, optimization of pyrolysis process was carried out to maximize the production of PA with highest TPC from palm kernel shell (PKS) and its bioactivities using experimental design including two-level fractional factorial design and subsequently Response Surface Methodology (RSM) using central composite design (CCD). From the results, the following condition were obtained; final temperature of 429 °C, heating rate of 1.34 °C/min, holding time of 39 min, gas flow rate of 0.42 L/min and condensing temperature of 7.2 °C. PA produced in optimized (OPPA-PKS) and unoptimized (IPA-PKS) conditions were extracted using ethyl acetate (EA) and fractions containing highest TPC (F15-17 EA-OPPA-PKS was 673.49 ± 6.05 µg GAE/mg; F13-15 EA-IPA-PKS was 866.84 ± 54.28 µg GAE/mg) were subsequently characterized for its bioactivities. Results of antioxidant assay for F15-17 EA-OPPA-PKS as follows; DPPH was 61.63 ± 1.19 %, TEAC was 1101.83 ± 4.59 µg Trolox/mg, FRAP was 11.80 ± 0.41 mmol Fe(II)/mg, hydroxyl radical scavenging was IC50 283.85 ± 0.90 µg/mL, and superoxide radical scavenging was IC50 491.49 ± 5.99 µg/mL. For F13-17 EA-IPA-PKS, the results of antioxidant assay as follows; DPPH was 75.34 ± 3.40 %, TEAC was 1346.48 ± 5.29 µg Trolox/mg, FRAP was 11.80 ± 0.41 mmol Fe(II)/mg, hydroxyl radical scavenging was IC50 270.34 ± 4.88 µg/mL and superoxide radical scavenging was IC50 472.32 ± 1.87 µg/mL. The viability of cell was more than 94.29 % for F15-17 EA-OPPA-PKS and more than 93.08 % for F13-17 EA-IPA-PKS. The nitric oxide production for F15-17 EA-OPPA-PKS and F13-17 EA-IPA-PKS were 6.65% and 6.55% respectively. The results of bioactivities suggested potential application of PA as alternative source for synthetic antioxidants and anti-inflammatory drugs. The suitability of the model employed and the success of RSM in optimizing the pyrolysis conditions to obtain the high TPC in the PA from PKS, PA fractions as natural-based source for antioxidant and anti-inflammatory compound(s), and the feasibility of agricultural biomass as new source of bioactive compounds are the important findings in this study