Browsing by Author "Hapuarachchi, N.S."

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    Agricultural, Economical and Ecological Importance of Phoenix pusilla (Ceylon Date Palm) and Its Future Research Perspectives
    (Uva Wellassa University of Sri Lanka, 2018) Hapuarachchi, N.S.; Warnakula, L.; Cooray, R.; Dayarathne, D.A.R.K.; Safeena, M.I.S.
    Phoenix pusilla, commonly known as Ceylon date palm (Indi) is a native palm species to Sri Lanka, belonging to the Family Arecaceae and Genus Phoenix. It is closely related but distinct from Phoenix dactylifera (date palm) by many morphological features. P.pusilla grows naturally in many parts of Northern and Eastern regions in Sri Lanka, and occasionally raised as an ornamental crop. It is considered as an underutilized fruit crop, and an untouched genetic resource, facing a severe genetic loss due to the negligence of the public and lack of scientific research. The aim of the review was to study the existing knowledge on agricultural, economical and ecological importance of P.pusilla. An extensive literature search was carried out in "Google scholar", "NCBI" and "AGRIS" databases using search terms "Ceylon date palm" and "P.pusilla" which resulted total of seven research articles. One study depicted that it is a multipurpose plant with many valuable traits, but with an inconsiderable attention within the country and globally. Three research papers showed the importance of its edible fruit in treating fever, hyperdipsia, consumption, cardiac and general debility, burning sensation, seminal weakness, and gasteropathy. Furthermore, leaflets have been used to weave mats, brooms and baskets. It is ecologically important as a nursing plant in improving degraded tropical dry evergreen forest landscapes, by conserving soil and maintaining favorable conditions for the growth of woody plants. It is highly drought tolerant and has been used to pollinate P.datylifera which has produced commercially valuable dwarf hybrids bearing seedless, edible fruits. In conclusion, further studies are essential to understand the physiology of P.pusilla to biotic and abiotic stresses, its ecological adaptations, nutritional and medicinal properties and phylogenetic relationships for the development as a cash crop and to improve other Phoenix species through interspecies hybridization.
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    Comparison of DNA Isolation and Dominant and Co-dominant Molecular Markers to Reveal the Genetic Sex of Gallus domesticus (Domestic Chickens)
    (Uva Wellassa University of Sri Lanka, 2018) Nirmali, W.K.R.; Warnakula, L.; Cooray, R.; Hapuarachchi, N.S.; Magamage, M.
    The whole blood of chickens contains nucleated erythrocytes and thrombocytes which yield high amount of DNA thus cause many troubles during DNA extractions. Optimization of DNA extraction from avian blood is important to yield high quality DNA and is vital for the success of all downstream applications. Determination of genetic sex of chickens is an important aspect in the avian research as well as in layer industry. Among the various methods of sex determination, Polymerase Chain Reaction (PCR) based methods are considered as most accurate and inexpensive. PCR methods have been developed based on the amplification of sex chromosome linked dominant and co-dominant loci in the chicken genome to distinguish between sexes. Success of this PCR based genetic sexing depends on the optimization of the PCR conditions and the reliability and reproducibility of molecular markers. Therefore this study was aimed at investigating the optimum conditions of DNA isolation from chicken blood and to compare the reproducibility of one co-dominant and two dominant sex markers to be validated as a tool for sex determination in avian research. Six different extraction procedures including manual and solution based commercial purification kit were evaluated. Efficacy of procedures was assessed with different combinations of initial blood, lysis buffer, and protein denaturant in related to the DNA yield and purity. Three primer sets namely CHD1, HUR 0423 and HUR 0424 were evaluated for the genetic sexing of chicken by polymerase chain reaction. The study results showed that an initial volume of 10 µL blood yields a significantly high DNA with high purity. Dominant marker HUR0424 showed to be a reliable marker system for the genetic sexing of domestic chickens over co-dominant markers.
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    Identification of Vectors for In-vivo Protein Overexpression in Introducing Drought Resistance: A Review
    (Uva Wellassa University of Sri Lanka, 2019) Cooray, R.; Dissanayake, Y.; Warnakula, L.; Hapuarachchi, N.S.
    The prolonged lack of rainfall is a major abiotic stress affecting crop productivity in Sri Lanka. In vivo protein overexpression to induce drought resistance is frequently studied globally however; the selection of a most suitable vector for gene transfection has been a challenge for high protein yield. This study aims to review such possible vectors, through a thorough published literature survey at ‘Google scholar’ and ‘PubMed’ with search terms ‘drought resist*’, ‘drought tolerant*’, ‘vector’, ‘overexpression’. A late embryogenesis abundant protein gene OsLEA3-1 overexpressed with three binary expression constructs (OsLEA3-S/A/H), by inserting the full length cDNA into backbone vectors pCAMBIA1301-S/A/H respectively, with double CaMV 35S, rice Actin1 and HVA1-like promoter, followed by Agrobacterium transformation. All other constructs OsLEA3-S/-H had higher grain yield than wild type under stress except OsLEA3-A. Over expression levels are 63% 56% and 46% for OsLEA3-S/A/H, respectively. Protein coding region of OsbZIP72 in vector pCAMBIA1300S to construct pCAMBIA1300S-OsbZIP72; and a promoter of drought tolerance rice cultivar IRAT109 in a pCAMBIA1381xb-GFP vector to control GFP expression and Arabidopsis BnLEA4-1 in pGEMT-BnLEA4-1 vector transfected via Agrobacterium are few examples at global scale. However, no published studies in Sri Lanka were found during the literature search. Therefore, cloning of drought tolerant gene to a crop plant followed by in vivo overexpression to stimulate the drought tolerance is a promising biotechnological advancement in agriculture. Literature showed vectors can be modified to fit to an individual crop product for in vivo overexpression of drought tolerant genes. It is recommended to extend the review to analyse characteristics for a suitable vector include compatibility, linker region, and screening genes to develop a complete guide for selection of vector in in vivo protein overexpression for drought resistance.
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    Importance of Genetic Diversity and Phytochemical Assessment of Madhuca spp. in Sri Lanka: A Review
    (Uva Wellassa University of Sri Lanka, 2019) Withana, W.V.E.; Hapuarachchi, N.S.; Cooray, R.; Dissanayake, Y.; Warnakula, L. W
    Madhuca is a large evergreen tree belonging to the family Sapotaceae distributed in many Asian countries including Sri Lanka. The aim of this review was to identify potential applications of Madhuca spp.in Sri Lanka to be developed into a research prospective in genetic and phytochemistry analysis. Literature survey was done using “Google Scholar” search engine and “PubMed” database using search terms “Madhuca”,” Genetics”,” Phytochemistry”. Altogether twenty-two research studies were retrieved on phytochemical analysis, pharmacological profiles but fewer in its other utilizations and genetic analysis. Five research articles were found on phytochemistry studies and three articles were about modified DNA isolation methods and genetic diversity analysis using Madhuca spp. in India. Studies showed that all parts of the Madhuca tree carry a number of medicinal properties and is rich in secondary metabolites. Literature evidence depicted it as a multipurpose forest tree, source of food and nutrition, pharmaceutical ingredient, bio-fertilizer and bio-fuel. This search identified seven Madhuca species currently found in Sri Lanka of which four of them are endemic. Being a plant species with a wide scope of potential applications, to date there are only one research available in published literature on phytochemical analysis of four Madhuca species found in Sri Lanka. None of supporting materials were found on genetic studies in genus Madhuca in Sri Lanka. Hence this raises the demand for such phytochemical analysis of different Madhuca spp. in Sri Lanka. Developing methods to extract genetic data, molecular authentication studies via molecular barcoding to fill the gaps in phenotypic and genotypic characterisation, storing the genetic data in databases for public access for future research purposes and efficient sustainable germplasm management will widen the research scope and potential application development of this invaluable plant in Sri Lanka.
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    Protein overexpression in Different E. call Strains for Industrial Scale Drug Development in Sri Lanka
    (Uva Wellassa University of Sri Lanka, 2018) Cooray, R.; Hapuarachchi, N.S.; Warnakula, L.
    Research and industrial scale protein overexpression for pharmaceutical products uses Escherichia con (E. coli) strains due to their well-characterised genome and easy manipulation. This review addresses the current scope of E. coli strains for protein overexpression, at global scale and in Sri Lanka, and the feasibility of development of pharmaceutical protein manufacturing in Sri Lanka, through a published literature at PubMed and Google Scholar. We identified that globally E. coli BL21 is the most common host strain due to properties such as mutation of Lon Protease coding gene and hsdSB gene that degrade foreign and extracellular proteins, respectively. It is developed into BL21(DE3) for T7 expression system, C41(DE3) for toxic and C43(DE3) for highly toxic and membrane proteins, hence, useful for large Glade of enzymes in system biology. In K-12 lineage, the AD494 and GrigamiTM strains were developed with thioredoxin reductase mutation enhancing disulphide bond formation in cytoplasm. HMS 174 stain with recA mutation carries higher plasmid stability. In Sri Lanka, however, only few published data were available, mainly focused on pathogenicity of E. coli with small published research at local institutes for overexpression of growth hormone like protein of Setaria digitata, multiepitope IgG/IgM proteins, in E. coli BL21(DE3). Although, this evidenced the availability of required technology and assets in Sri Lanka for necessary protocols, protein products such as pharmaceutical drugs being imported at high cost than developing research to optimise protein overexpression in the country. Through this review, we suggest the optimum utilisation of available laboratory infrastructure, personnel asserts and protocols by developing research not only with BL21(DE3), but other strains for specific protein expressions, where the research can be developed for industrialising to meet the demands of the pharmaceutical industry and many other biomedical researches in Sri Lanka.