Isolation of a gene preferentially expressed in mature anthers of rice (Oryza sativa L.)

Using monoclonal antibodies raised against pollen-specific proteins, we have isolated a cDNA clone, designatedOry-Cl from a rice anther cDNA expression library. A transcript corresponding to theOry-Cl gene showed preferential expression in anthers. This transcript was not detected in any vegetative tissues analysed. RNA gel blot analysis of different developmental stages of anthers showed that theOry-Cl gene is expressed at later stages of pollen development. In situ hybridisation showed that theOry-Cl transcript is only present in mature pollen.

The effect of type of cereal on the glycaemic response of two traditional Sri Lankan food items

Pittu and roti are two traditional food items consumed by Sri Lankan people mostly for breakfast or dinner. Rice (Oryza sativa L.) and kurakkan (Eleucine coracana L.) are two types of cereal that can be used to prepare them. The determination of blood glucose elevating effect (glycaemic response) of pittu and roti prepared from rice flour and kurakkan flour was the objective of this study. Proximate composition of Bg 403 rice flour and kurakkan flour was determined and the available carbohydrate content of the two types of cereal was calculated. Pittu and roti were prepared from each flour, following traditional methods and given to eight young healthy adult volunteers. Each subject was given a weighed portion of pittu or roti equivalent to 50 g available carbohydrate as the test food. As the standard food 50 g glucose was given orally. After a 12 hrs overnight fast on the assigned day each subject was given either the standard food or the test food and blood glucose was measured in capillary blood at fasting (0), 15, 30, 45, 60, 90 and 120 min after the consumption of food. The incremental area under the glycaemic response curve (IAUC) for each test food was expressed as a percentage of IAUC of the standard food taken by the same subject and the average value of subjects was taken as the glycemic index (GI) for the test food. Proximate analysis revealed that percentage moisture, crude fat, crude fibre, crude protein and minerals of rice flour and kurakkan flour were 13.0, 1.7, 0.42, 10.3, 0.88 and 13.2, 1.9, 4.4, 8.7 and 2.8, respectively. Accordingly the available carbohydrate percentage of rice flour and kurakkan flour were 73.7 and 69.0, respectively. The GI of pittu and roti, prepared using Bg 403 rice flour were 52 and 64 and that of kurakkan flour were 71 and 80 respectively. Based on the GI, it can be suggested that pittu is better for health than roti, while rice flour is better than kurakkan flour to prepare these. The basis of recommending kurkkan flour based products for diabetic people has to be re-examined in the light of these findings.

Bioprocessing of hemp hurd (Cannabis sativa) for biofuel production

 This project is based on production of biofuel from lignocellulosic waste. Hemp hurds were used for the production of ethanol. This was achieved by treating hemp at high temperature followed by enzymatic reaction and fermentation. Different techniques were used to detect the changes occurred on hemp structure and product quantification.

Understanding physicochemical changes in pretreated and enzyme hydrolysed hemp (Cannabis sativa) biomass for biorefinery development

The physicochemical properties of hemp biomass structure to pretreatment and enzymatic hydrolysis were investigated to improve upon reducing sugar production for biofuel development. Sodium hydroxide pretreated biomass (SHPB) yielded maximum conversion of holocellulose into reducing sugar (72 %). Scanning electron microscopy (SEM) revealed that enzymatic hydrolysis generated regular micropores in the fragmented biomass structure. The thermogravimetric analysis (TGA) curve suggested the degradation of hemicellulose and cellulose, which conformed well to the subsequent nuclear magnetic resonance (NMR) studies indicating the presence of α- and β-glucose (28.4 %) and α- and β-xylose (10.7 %), the major carbohydrate components commonly found in hydrolysis products of hemicellulose and cellulose. Attenuated total reflection Fourier transform infrared (ATR-FTIR) spectra showed stretching modes of the lignin acetyl group, suggesting the loosening of the polymer matrix and thus the exposure of the cellulose polymorphs. X-ray diffraction pattern indicated that enzymatic hydrolysis caused a higher crystallinity index (36.71), due to the fragmentation of amorphous cellulose leading to the reducing sugar production suitable for biofuel development.

Enrichment of cellulosic waste hemp (Cannabis sativa) hurd into non-toxic microfibres

In this study a largely available lignocellulose feedstock hemp (Cannabis sativa), obtained as an industrial waste, was used for cellulose extraction. The extraction of cellulose microfibres from hemp biomass was conducted by alkaline treatment and an acidification process. The extracted cellulose microfibres were characterised using Fourier-transformed infrared spectroscopy (FTIR), Scanning electron microscopy (SEM), thermogravimetric analysis (TGA) and X-ray diffraction (XRD).The viability of the study was determined by growing human fibroblasts on the preparation which resulted in being non-toxic; indicating its potential in preparing biological scaffolds. Upon enzymatic hydrolysis of the cellulose microfibre using cellulase from Trichoderma reesei, a maximum of 909 mg/g of reducing sugars were obtained, which endorses its suitability for biofuel production.

Genome sequence of Ensifer arboris strain LMG 14919T, a microsymbiont of the legume Prosopis chilensis growing in Kosti, Sudan

Ensifer arboris LMG 14919T is an aerobic, motile, Gram-negative, non-spore-forming rod that can exist as a soil saprophyte or as a legume microsymbiont of several species of legume trees. LMG 14919T was isolated in 1987 from a nodule recovered from the roots of the tree Prosopis chilensis growing in Kosti, Sudan. LMG 14919T is highly effective at fixing nitrogen with P. chilensis (Chilean mesquite) and Acacia senegal (gum Arabic tree or gum acacia). LMG 14919T does not nodulate the tree Leucena leucocephala, nor the herbaceous species Macroptilium atropurpureum, Trifolium pratense, Medicago sativa, Lotus corniculatus and Galega orientalis. Here we describe the features of E. arboris LMG 14919T, together with genome sequence information and its annotation. The 6,850,303 bp high-quality-draft genome is arranged into 7 scaffolds of 12 contigs containing 6,461 protein-coding genes and 84 RNA-only encoding genes, and is one of 100 rhizobial genomes sequenced as part of the DOE Joint Genome Institute 2010 Genomic Encyclopedia for Bacteria and Archaea-Root Nodule Bacteria (GEBA-RNB) project.

Genome sequence of Ensifer meliloti strain WSM1022, a highly effective microsymbiont of the model legume Medicago truncatula A17

Ensifer meliloti WSM1022 is an aerobic, motile, Gram-negative, non-spore-forming rod that can exist as a soil saprophyte or as a legume microsymbiont of Medicago. WSM1022 was isolated in 1987 from a nodule recovered from the roots of the annual Medicago orbicularis growing on the Cyclades Island of Naxos in Greece. WSM1022 is highly effective at fixing nitrogen with M. truncatula and other annual species such as M. tornata and M. littoralis and is also highly effective with the perennial M. sativa (alfalfa or lucerne). In common with other characterized E. meliloti strains, WSM1022 will nodulate but fixes poorly with M. polymorpha and M. sphaerocarpos and does not nodulate M. murex. Here we describe the features of E. meliloti WSM1022, together with genome sequence information and its annotation. The 6,649,661 bp high-quality-draft genome is arranged into 121 scaffolds of 125 contigs containing 6,323 protein-coding genes and 75 RNA-only encoding genes, and is one of 100 rhizobial genomes sequenced as part of the DOE Joint Genome Institute 2010 Genomic Encyclopedia for Bacteria and Archaea-Root Nodule Bacteria (GEBA-RNB) project.

Was monochloramine responsible for widespread lettuce crop failures at a major recycled water irrigation scheme?

In 2008, vegetable growers observed stunted lettuce plants showing signs of chlorosis and wilting. It was suspected that monochloramine in the recycled water used for irrigation, in combination with extreme environmental conditions (high irrigation water salinity and extreme heat), was responsible for these crop failures. A series of glasshouse studies was conducted to evaluate the impact of monochloramine concentration alone on iceberg lettuce seedlings, as well as in combination with high salinity and hot ambient temperatures. Monochloramine concentrations up to 9 and 15 mg L−1 Cl2 for continuous and initial irrigation only, respectively, did not affect the weight of iceberg lettuce heads (p > 0.05), while the combination of monochloramine (4–5 mg L−1 Cl2) and salinity (3,500 μS cm−1) did not significantly affect harvest measurements (p > 0.05). We therefore conclude that it is unlikely that monochloramine was responsible for the observed crop failures.

Suitability of magnetic nanoparticle immobilised cellulases in enhancing enzymatic saccharification of pretreated hemp biomass

Previous research focused on pretreatment of biomass, production of fermentable sugars and their consumption to produce ethanol. The main goal of the work was to economise the production process cost of fermentable sugars. Therefore, the objective of the present work was to investigate enzyme hydrolysis of microcrystalline cellulose and hemp hurds (natural cellulosic substrate) using free and immobilised enzymes. Cellulase from Trichoderma reesei was immobilised on an activated magnetic support by covalent binding and its activity was compared with that of the free enzyme to hydrolyse microcrystalline cellulose and hemp hurds on the basis of thermostability and reusability.

Omega-3 fatty acid production from enzyme saccharified hemp hydrolysate using a novel marine thraustochytrid strain

In this work, a newly isolated marine thraustochytrid strain, Schizochytrium sp. DT3, was used for omega-3 fatty acid production by growing on lignocellulose biomass obtained from local hemp hurd (Cannabis sativa) biomass. Prior to enzymatic hydrolysis, hemp was pretreated with sodium hydroxide to open the biomass structure for the production of sugar hydrolysate. The thraustochytrid strain was able to grow on the sugar hydrolysate and accumulated polyunsaturated fatty acids (PUFAs). At the lowest carbon concentration of 2%, the PUFAs productivity was 71% in glucose and 59% in the sugars hydrolysate, as a percentage of total fatty acids. Saturated fatty acids (SFAs) levels were highest at about 49% of TFA using 6% glucose as the carbon source. SFAs of 41% were produced using 2% of SH. This study demonstrates that SH produced from lignocellulose biomass is a potentially useful carbon source for the production of omega-3 fatty acids in thraustochytrids, as demonstrated using the new strain, Schizochytrium sp. DT3.

Genome sequence of Ensifer medicae Di28; an effective N2-fixing microsymbiont of Medicago murex and M. polymorpha

Ensifer medicae Di28 is an aerobic, motile, Gram-negative, non-spore-forming rod that can exist as a soil saprophyte or as a legume microsymbiont of Medicago spp. Di28 was isolated in 1998 from a nodule recovered from the roots of M. polymorpha growing in the south east of Sardinia (Italy). Di28 is an effective microsymbiont of the annual forage legumes M. polymorpha and M. murex and is capable of establishing a partially effective symbiotic association with the perennial M. sativa. Here we describe the features of E. medicae Di28, together with genome sequence information and its annotation. The 6,553,624 bp standard draft genome is arranged into 104 scaffolds of 104 contigs containing 6,394 protein-coding genes and 75 RNA-only encoding genes. This rhizobial genome is one of 100 sequenced as part of the DOE Joint Genome Institute 2010 Genomic Encyclopedia for Bacteria and Archaea-Root Nodule Bacteria (GEBA-RNB) project.

Screening of cannabinoids in industrial-grade hemp using two-dimensional liquid chromatography coupled with acidic potassium permanganate chemiluminescence detection

Widely known for its recreational use, the cannabis plant also has the potential to act as an antibacterial agent in the medicinal field. The analysis of cannabis plants/products in both pharmacological and forensic studies often requires the separation of compounds of interest and/or accurate identification of the whole cannabinoid profile. In order to provide a complete separation and detection of cannabinoids, a new two-dimensional liquid chromatography method has been developed using acidic potassium permanganate chemiluminescence detection, which has been shown to be selective for cannabinoids. This was carried out using a Luna 100 Å CN column and a Poroshell 120 EC-C18 column in the first and second dimensions, respectively. The method has utilized a large amount of the available separation space with a spreading angle of 48.4° and a correlation of 0.66 allowing the determination of more than 120 constituents and mass spectral identification of ten cannabinoids in a single analytical run. The method has the potential to improve research involved in the characterization of sensitive, complex matrices.