Category Archives: 2017 Vol 29 June

COMPARING POLLINATION BAG TYPES FOR MICRO-ENVIRONMENTAL PARAMETERS INFLUENCING SEED PRODUCTION IN OIL PALM

For oil palm seed production, the pollination bag must prevent contamination by unintended pollen to ensure the genetic integrity. An investigation was undertaken by the seed production unit of Dami Oil Palm Research Station (DAMI OPRS) in Papua New Guinea to compare the impact of the type of pollination bags on both the genetic integrity and seed yield in the commercial seed production environment. Three pollination bag types [canvas, high density polyethylene (HDPE) and polyester] were compared during pollination and blank pollination exercises. The production of seeds and results for intactness, presence of insects, impact of water and economic parameters such as average time taken for isolating, demonstrated that the polyester bags outperformed the HDPE and canvas bags. Bunches isolated with polyester bags produced 13% more seeds per bag than the HDPE bags and 6% more seeds per bag than those using canvas. The canvas bag took longer to apply than the polyester or HDPE bags, and was not averting presence of insects. Both HDPE and canvas bags showed significant collapse when wet. The resulting economics on seed production profitability and risk of contaminations which would alter commercial reputation indicate that the polyester bags are a better choice than the canvas and HDPE bags.

CONSTRUCTION OF A VECTOR CONTAINING HYGROMYCIN (HPT) GENE DRIVEN BY DOUBLE 35S (2XCAMV35S) PROMOTER FOR OIL PALM TRANSFORMATION

Transformation vector construction is one of the important disciplines for plant genetic transformation
studies. A series of vector consisting of hygromycin phosphotransferase (hpt) gene as the selective marker and green fluorescent protein (GFP) as the visual reporter gene, under the control of double cauliflower mosaic virus 35S (2XCaMV35S) promoter has been engineered for transformation into oil palm cells. These genes were cloned into different types of cloning and expression vectors. The cloning was carried out by using restriction enzyme digestion and ligation method. Five intermediate vectors have been created for insertion of 2XCaMV35S-HPT-35ST and 2XCaMV35S-GFP-35ST into modified pBINPLUS backbone vector for particle bombardment and Agrobacterium-based transformation protocols. All vectors were sequenced to confirm the integrity of DNA region. The vectors were later transformed into oil palm embryogenic calli using biolistic device. The viability of the vectors was initially evaluated by transient GFP fluorescence expression observed under fluorescence microscope. It was demonstrated that the 2XCaMV35S promoter was able to drive the expression of gfp as gene in oil palm calli.

THE USE OF Arabidopsis thaliana MODEL SYSTEM FOR TESTING OIL PALM PROMOTER: CASE STUDY ON OIL PALM MT3-A PROMOTER

A previous study on transient expression of oil palm tissues has shown that the oil palm metallothioneinlike type 3 (MT3-A) gene promoter is specifically expressed in the mesocarp and not in other tissues. This study was conducted to determine whether or not Arabidopsis can be used as a model system to study oil palm promoter. Functional characterisation of the oil palm MT3-A promoter was performed using promoter::GUS fusion analysis in transgenic Arabidopsis. The localisation of β-glucuronidase (GUS) expression in several different tissues of transgenic Arabidopsis homozygous lines driven by the oil palm MT3-A promoter was determined. Histochemical GUS analysis in transgenic Arabidopsis revealed the highest expression in the cotyledon and hypocotyls as well as at the early stage of plant development and gradually decreased as the plant grew. Lower expression was detected in organs of mature plant and no expression was observed in reproductive tissues. This observation may suggest that MT3-A promoter might be involved during early stage of plant development. Since we use Arabidopsis which is neither a fruit nor mesocarp plant to check mesocarp-specific promoter it might not give a faithful pattern of expression but it may be suited to study oil palm specific promoter derived from other tissues.

ANALYSIS OF GENE EXPRESSION BY EST FROM SSH LIBRARY IN DWARF OIL PALM

Six subtracted cDNA libraries were constructed from PS1 and AG1 oil palm breeding lines using Suppression Subtractive Hybridisation (SSH) method. Six putative clones identified to be associated
in dwarfism encodes for brassinosteroid biosynthesis-like protein (DWF1), brassinosteroid insensitive
1-associated receptor kinase 1 precursor (BRI1), late elongated hypocotyl protein (LHY), gibberellin receptor (GID1), sterol 24-methyltransferase 1 (SMT1) and E3 ubiquitin-protein ligase (E3Ub) were analysed. These candidate dwarfing genes are involved in various stages of brassinosteroids (BR), gibberellins (GA) biosynthesis and signaling pathways for plants growth and development. The expression levels of candidate genes were validated by quantitative Real-time PCR (qRT-PCR) by two reference genes, manganese superoxide dismutase (PD569) and predicted protein IFH-1 like (EA1332). Higher expression level of BRI1, LHY and SMT1 genes were observed in dwarf palms compared to standard palms with normalised fold-difference of 2.3285, 1.5620 and 4.9044, respectively. Statistical analysis of all potential transcripts showed further evidence of SMT1 as a potential molecular marker for the screening of dwarf palm planting materials with highly significant expression levels in dwarf palms compared to standard palms; however, the expression levels of BRI and LHY genes were statistically insignificant.

THIAMINE BIOSYNTHESIS GENE EXPRESSION ANALYSIS IN Elaeis guineensis DURING INTERACTIONS WITH Hendersonia toruloidea

Thiamine plays an indispensable role as a cofactor in many metabolic reactions in all living organisms. The active form, thiamine pyrophosphate (TPP) functions as a cofactor in important metabolic reactions including glycolysis, pentose phosphate pathway and the tricarboxylic acid cycle. Besides that, thiamine is also associated with the induction of systemic acquired resistance (SAR) in plants. Hendersonia toruloidea is an endophytic fungus originally isolated from oil palm roots which have been shown to have biocontrol activity. Seven months old oil palm seedlings were inoculated with H. toruloidea and the response on the expression of thiamine biosynthesis genes were investigated. Microscopy analysis was performed to visualise the colonisation of the fungus. Ribonucleic acid (RNA) was extracted from oil palm leaves at Day 1, 7, 15 and 30 post-treatment. Quantitative Real-time PCR (qRT-PCR) was performed to measure the level of expression of four key thiamine biosynthesis genes, namely THI4, THIC, TH1 and TPK. The results showed up to 12-fold increase in the expression of all gene transcripts at Day 1 and 7 post-treatment. This work provides first evidence of enhancement of thiamine biosynthesis by endophytic colonisation in oil palm seedlings.

METABOLOMICS UNRAVEL DIFFERENCES BETWEEN CAMEROON Dura AND DELI Dura OIL PALM (Elaeis guineensis Jacq.) GENETIC BACKGROUNDS AGAINST BASAL STEM ROT

Metabolomics is emerging as a powerful tool for screening of metabolites in both phenotyping and diagnostic analyses in plants. Metabolomics coupled with multivariate statistical analysis has provided a fast and unbiased comparative investigation of the metabolite composition in oil palm root. Basal stem rot (BSR) caused by the white rot fungus, Ganoderma boninense is a common oil palm disease in south-east Asia. Little is known about the disease mechanism and there is yet no satisfactory treatment for it. Besides agronomic practice of reducing the disease inoculum in the field, breeding for resistance to the disease is probably the most practical control. This study investigated metabolomic differences in oil palm planting materials with varying susceptibility to G. boninense from Cameroon and Deli genetic backgrounds. Liquid chromatography-quadrupole/ time-of-flight-mass spectrometry (LC-Q/TOF-MS) data was subjected to multivariate statistical analysis and discovered that the palms could be categorised into two clusters linked to Cameroon and Deli oil palms. Changes in metabolites involved in phenylpropanoid pathway and primary metabolism including shikimic acid, glucose and malic acid were observed. The comprehensive and unbiased strategy for metabolite data analysis presented here can be adopted to screen wider oil palm germplasm for those potentially useful for breeding BSR-resistant palms.

VARIATIONS OF δ13C AND δ15N IN OIL PALM TREE ORGANS: AN INSIGHT INTO C AND N DISTRIBUTION

Many studies have shown that seasonal variation in fruit bunch production occurs in relation to climatic factors and internal trophic conditions. As the annual fruit bunch production is important for planters, there is a need to get some insights into the carbon (C) and nitrogen (N) distribution in oil palm tree which are key factors in delineating plant C fluxes and nutrient cycling dynamics. Hence, this study utilises stable C and N isotope analyses to assess possible variations in the δ13C and δ15N signature in oil palm organs. Our study found that the average δ13C of each oil palm tree organs are; root: –27.1‰, frond: –27.9‰, leaflets: –29.1‰, mesocarp: –28.6‰ and kernel: –28.7‰. The δ13C results showed an enrichment of 13C in heterotrophic organs compared to leaflets. Whilst the average δ15N values of oil palm tree organs are; root: 4.4‰, frond: 8.9‰, leaflets: 1.6‰, mesocarp: 10.3‰ and kernel: 6.8‰. The δ15N results also showed 15N enrichment in heterotrophic organs compared to leaflets. In conclusion, there were clear isotopic differences between autotrophic leaflets and heterotrophic organs. This study demonstrates the applicability of stable isotope analysis (SIA) in providing novel insights into C and N distribution in oil palm tree whereby both factors are key in the understanding of oil palm physiology which in turn is crucial in maintaining a productive fruit bunch yield annually.

APOPTOSIS GENE NETWORK REGULATED BY DELTA-TOCOTRIENOL IN K562 CHRONIC MYELOID LEUKAEMIA CELLS

Palm oil is rich in various health-beneficial phytonutrients. The vitamin E and the carotenoids make up a large portion of these phytonutrients. The tocotrienols are a relatively newer class of vitamin E isoforms that are actively being researched for its various health promoting properties. In this study, the anti-cancer properties of the delta-tocotrienol (δ-T3) isoform were investigated on a leukemic cell line. A preliminary cytotoxicity assay was carried out to identify δ-T3’s potency in inducing cell death. Following this a real time-based gene array experiment was carried out to distinguish the apoptosis genes regulated by δ-T3 in the K562 leukemic cell line. Results show that treatment with δ-T3 significant regulated the expression of several genes that promote apoptosis such as TP53, TP73, HRK, MCL1, CASP7, CASP8, DAPK1, PYCARD, FASLG and TNFRSF9. These findings suggest that palm δ-T3 can exert anti-cancer activities in chronic myeloid leukemic cells

OIL PALM ROOTS COLONISATION BY Ganoderma boninense: AN INSIGHT STUDY USING SCANNING ELECTRON MICROSCOPY

Oil palm (Elaeis guineensis Jacq.) is an oil crop widely cultivated throughout South-east Asia countries. Unfortunately, the industry is badly affected by Basal Stem Rot (BSR) disease, caused by soil-borne pathogen, Ganoderma boninense, which is the most devastating and widespread disease in oil palm. Although some investigations have been conducted, complete understanding on BSR infection is yet to be achieved. In this article, observation on roots of inoculated oil palm seedlings under scanning electron microscopy showed G. boninense colonisation predominantly formed on root surface, proving endophytic colonisation primarily through the hair base, crossing epidermal cells, and followed by spreading into cortical cells which a possible pathway leading to vascular system. It also showed the formation of highly dense hypal network within the colonised area, mainly in the epidermal and cortical cells. By six months of inoculation, cell compartment of infected roots was lost and noticeable changes in the root cells structure were observed. The present study provides additional insights in the infection process and pathogen route inside the host tissue and may be useful for development of disease control strategies.

A GREEN METHOD TO PRODUCE BIODIESEL FROM PALM OLEIN OIL

The current transesterification method to produce biodiesel from palm oil uses NaOH as the homogeneous catalyst. NaOH has several disadvantages. It is corrosive, can not be reused, and separation from biodiesel is time-consuming. The objective of the present work was to replace NaOH with a greener and efficient heterogeneous catalyst, calcium diglyceroxide. The potential contamination of biodiesel by Ca2+ was minimised by incorporating Na2CO3 into the transesterification mixture. The work aimed to identify the optimum reaction conditions which would minimise the production cost of biodiesel complying with the quality prescribed by ASTM standards. Optimum conditions were oil: methanol ratio of 1:6 (molar), catalyst 0.2% (w/w oil), Na2CO3 2.5% (w/w oil), and 3 hr reaction period at 65°C. The yield was 92.2%. All the studied properties of biodiesel except kinematic viscosity conformed with recommended levels by ASTM. Reusing the catalyst which was used twice previously, increased the biodiesel yield from 92.2% to 94.3%. The production of good quality biodiesel from palm olein by this novel method is greener. It is more efficient because the catalyst is not corrosive and easy to separate. Also, the catalyst can be reused at least twice, thus minimising the amounts of chemicals used.

ACTIVATED CARBON FROM OIL PALM BIOMASS AS POTENTIAL ADSORBENT FOR PALM OIL MILL EFFLUENT TREATMENT

The study aims to produce renewable and green activated carbon (AC) from palm biomass through pyrolysis and the activation process. The study also aims to evaluate the application of AC as adsorbent in palm oil mill effluent treatment (POME). This is in order to reduce the pollutant levels in its final discharge. The AC was prepared from empty fruit bunch (EFB), by pyrolysis (to produce bio-char), followed by physical activation. The bio-chars with higher carbon content were selected for the preparation of the AC. The physical activation process was carried out by purging with carbon dioxide (100 cm3 min-1) at 10°C min-1 heating rate for 30 min until the AC reaches the desired activation temperature. The activation temperatures studied ranged from 500°C to 900°C in quartz tube furnace. The optimum activation temperature i.e. 800°C gave the maximum specific surface area, (SBET) of 937 m2 g-1. Batch adsorption experiments applying the prepared adsorbent to synthetic dye yielded adsorption data well fitted to the Langmuir isotherm model. The AC produced from EFB performed better in dye removal achieving a maximum adsorption capacity of 333.3 mg g-1. The EFB-based AC was able to reduce the biochemical oxygen demand of POME. These studies suggested that the EFB AC could be useful as a low cost alternative adsorbent in wastewater treatment.

CULTIVATION OF MICROALGAE IN MEDIUM CONTAINING PALM OIL MILL EFFLUENT AND ITS CONVERSION INTO BIOFUEL

The production of renewable energy has become an important element in worldwide energy policy aimed to reduce greenhouse gases caused by fossil fuels. Biodiesel derived from transesterification of vegetable oil or animal fats, with alcohol in the presence of catalyst has been considered as one of the alternative resources to replace fossil fuels. However, the biodiesel produced from these materials may not be enough for total replacement; hence microalgae are used as another potential alternative due to their high photosynthetic efficiency and biomass productivity. In this study, a microalgae species i.e. Chlorella vulgaris UMACC 001 having high oil to biomass ratio was cultured, harvested and characterised. It showed very high growth rate, μ (0.29 per day) and biomass productivity (0.14 mg litre per day). In addition, the resulting algal oil showed almost equal amount of saturated fatty acid (48.9%) and unsaturated fatty acid (51.1%). When converted to biodiesel, the ester content was 68.9% and comparable with that produced in previous study (71.0%). Thus, Chlorella vulgaris can be considered as a potential feedstock for biofuel production in the future.

REVIEW OF BIOFUEL PRODUCTION – FEEDSTOCK, PROCESSES AND MARKETS – Review Article

Transportation fuels derived from biomass are an alternative to fossil-based fuels. Due to the fact that globally the available biomass resources are limited and considering the fact that they are also used as food materials there is an increasingly stronger need to use such organic matter as efficiently as possible (i.e. biorefinery approach). Against this background the overall goal of this article is to discuss the overall available biomass on a global scale. The article will also touch on the technologies to efficiently convert this biomass into transportation biofuels useable within the existing markets for such fuels and are presented in detail according to the current state of technology. Based on this, the global market for biofuels is discussed against the background of the available biomass resources. Finally the most important findings are summarised and presented.