Oil Accumulation Stage Research Articles

Generation and distribution of overpressure in ultra-deep carbonate reservoirs controlled by intra-cratonic strike-slip faults: The Ordovician of Shuntuoguole area in the Tarim Basin

The strike-slip fault-controlled ultra-deep fractured-vuggy carbonate reservoir is an increasingly important exploration target. However, the high heterogeneity of fault-controlled reservoirs and the ineffective methods available for pressure prediction make it difficult to assess the risk of potential overpressure in ultra-deep carbonate reservoirs. Abnormal overpressure is common in the ultra-deep strike-slip fault-controlled carbonate reservoirs in the Shunbei Oilfield of the Tarim Basin. According to measured pressures, petroleum accumulation history, and fault framework, this study investigates the generation mechanisms of overpressure in fault-controlled carbonates and reconstructs the pressure evolution of reservoirs. Finally, the effect of strike-slip faults on current overpressure distribution is discussed. Measured pore pressure data from 90 wells showed that all reservoirs present in the major fault zones are normally pressured, and the reservoirs present far away from the major faults commonly develop overpressures. The distribution patterns of overpressure in the Middle-Lower Ordovician reservoirs coincide with areas with fault reactivation during the Himalayan Period. The results of the comprehensive analysis of petroleum accumulation history, fault activity, and well production suggested that dry gas generation is a primary cause of overpressure in the Middle-Lower Ordovician reservoirs. The relatively low temperature in the reservoir, in addition to the highly compartmentalized nature of overpressure, indicated that these overpressured gas migrate vertically into the Ordovician formation from deep strata along strike-slip faults. The pressure evolution restored by basin modelling and fluid inclusion showed that distinct differences in overpressure magnitude in the different reservoirs were formed at the stage of gas charging but not at the early stages of oil accumulation. The difference in physical properties of the two types of fault-controlled reservoirs resulted in different pressure distributions and oil-bearing levels in the ultra-deep carbonate reservoirs at present. The strike-slip fault zones are the primary pathways by which overpressured fluids migrate from the deeper Cambrian formation into the Ordovician reservoirs and act as barriers to separate the pressure systems in the ultra-deep carbonate reservoirs. These findings are expected to help mitigate the drilling risks with overpressure in deep strike-slip fault-controlled carbonate reservoirs.

Comparative de novo Transcriptome Analysis of Two Cultivars With Contrasting Content of Oil and Fatty Acids During Kernel Development in Torreya grandis.

Vegetable oil is an indispensable nutritional resource for human health and mainly characterized by the composition and content of fatty acids (FAs). As a commercial species of gymnosperm, Torreya grandis produces oil-rich nuts with high unsaturated fatty acids content in the mature kernels. In this study, two cultivars, T. grandis ‘Xifei’ and T. grandis ‘Dielsii,’ with distinct oil content were employed to compare the profiles of FAs accumulation during kernel development. The accumulation rate of oil content was significantly different between ‘Xifei’ and ‘Dielsii.’ Besides, the final oil content of ‘Xifei’ (52.87%) was significantly higher than that of ‘Dielsii’ (41.62%) at maturity. The significant differences in main FAs were observed at almost each kernel development stages between the two cultivars. C16:0, C18:1, and C20:3 FA exhibited different accumulation patterns between cultivars. The content and the initiation of accumulation of C20:3 FA were different between the two cultivars. To explore the molecular mechanism associated with different content of oil and FAs between two cultivars, de novo transcriptome of kernels was compared between ‘Xifei’ (high oil) and ‘Dielsii’ (low oil) at three stages of oil accumulation, respectively. Totally 142,213 unigenes were assembled and 16,379 unigenes with a length of over 1,000 nt were successfully annotated, including 139 unigenes related to FA biosynthesis, elongation, and metabolism. Compared with ‘Dielsii,’ totally 1,476, 2,140, and 1,145 differentially expressed genes (DEGs) were upregulated in ‘Xifei’ at the stage of the initiative, the rapid rise, and the stationary oil accumulation, respectively; the number of downregulated DEGs reached 913, 1,245, and 904, respectively. Relative expressions of 11 DEGs involved in FAs biosynthesis and metabolism were confirmed by RT-qPCR. Abundant differentially expressed transcription factors and pathway DEGs were correlated to oil and FAs according to Pearson’s correlation analysis between transcriptome and metabolites (oil and FAs), suggesting their contributions to the differential oil and FAs between the two cultivars during kernel development of T. grandis. To conclude, our findings can provide novel insights into the developmental differences in metabolites and de novo transcriptome correlated to lipid accumulation and FA synthesis of kernels between cultivars with contrasting oil deposits and demystify the regulatory mechanism of high oil accumulation in T. grandis.

Key oil accumulation periods of ultra-deep fault-controlled oil reservoir in northern Tarim Basin, NW China

A giant fault-controlled oilfield has been found in the ultra-deep (greater than 6000 m) Ordovician carbonate strata in the northern Tarim Basin. It is of great significance for hydrocarbon accumulation study and oil exploitation to determine the key oil accumulation periods. Based on detailed petrographic analysis, fluid inclusion association (FIA) in calcite samples filling in fractures from 12 wells were analyzed, and key accumulation periods of the strike-slip fault-controlled oilfield was studied by combining oil generation periods of the source rocks, formation periods of the fault and traps, and the fluid inclusion data. (1) There are multiple types of FIA, among them, two types of oil inclusions, the type with yellow fluorescence from the depression area and the type with yellow-green fluorescence from the uplift area with different maturities indicate two oil charging stages. (2) The homogenization temperature of the brine inclusions in FIA is mostly affected by temperature rises, and the minimum temperature of brine inclusions symbiotic with oil inclusions is closer to the reservoir temperature during its forming period. (3) FIA with yellow fluorescence all have homogenization temperatures below 50 °C, while the FIA with yellow-green fluorescence have homogenization temperatures of 70–90 °C tested, suggesting two oil accumulation stages in Middle–Late Caledonian and Late Hercynian. (4) The Middle–Late Ordovician is the key formation period of the strike-slip fault, fracture-cave reservoir and trap there. (5) The oil generation peak of the main source rock of the Lower Cambrian is in the Late Ordovician, and the oil accumulation stage is mainly the Late Ordovician in the depression area, but is mainly the Early Permian in the uplift area. The key oil accumulation period of the strike-slip fault-controlled reservoirs is the Late Caledonian, the depression area has preserved the primary oil reservoirs formed in the Caledonian, while the uplift area has secondary oil reservoirs adjusted from the depression area during the Late Hercynian. Oil reservoir preservation conditions are the key factor for oil enrichment in the strike-slip fault zone of northern Tarim, and the Aman transition zone in the depression is richer in oil and gas and has greater potential for exploration and development.

Overexpression of PvFAD3 Gene from Plukenetia volubilis Promotes the Biosynthesis of α-Linolenic Acid in Transgenic Tobacco Seeds.

The ω-3 fatty acid desaturase (FAD3) gene encodes a rate-limiting enzyme in the synthesis of α-linolenic acid. In this study, homologous cloning was used to obtain the full-length sequence of the PvFAD3 gene of Plukenetia volubilis. The full-length DNA sequence was 1871 bp long, with 8 exons and 7 introns. The structural analysis of the amino acid sequence revealed that the PvFAD3 protein contained three histidine-conserved regions and an endoplasmic reticulum retention signal. The real-time reverse transcription-polymerase chain reaction performed for determining the expression patterns of the PvFAD3 gene in different tissues of P. volubilis showed that PvFAD3 expression was highly expressed in the fast oil accumulation stage of seed. The analysis of subcellular localization assay in epidermal cells of tobacco (Nicotiana benthamiana) leaves showed that the PvFAD3 protein was mainly localized in the endoplasmic reticulum. Seed-specific overexpression vectors were constructed, and Agrobacterium-mediated genetic transformation was performed to obtain transgenic tobacco plants overexpressing PvFAD3. The results of fatty acid assays performed using harvested seeds showed a significant increase in α-linolenic acid content, a dramatic decrease in linoleic acid content, and an obvious increase in oil content in transgenic tobacco seeds. Collectively, the PvFAD3 gene of P. volubilis was confirmed as a key enzyme gene for α-linolenic acid synthesis; thus, indicating that the PvFAD3 gene can be used for fatty acid fraction improvement in oilseed plants.

Transcriptomic analysis of \u03b1-linolenic acid content and biosynthesis in Paeonia ostii fruits and seeds

BackgroundPaeonia ostii is a potentially important oilseed crop because its seed yield is high, and the seeds are rich in α-linolenic acid (ALA). However, the molecular mechanisms underlying ALA biosynthesis during seed kernel, seed testa, and fruit pericarp development in this plant are unclear. We used transcriptome data to address this knowledge gap.ResultsGas chromatograph-mass spectrometry indicated that ALA content was highest in the kernel, moderate in the testa, and lowest in the pericarp. Therefore, we used RNA-sequencing to compare ALA synthesis among these three tissues. We identified 227,837 unigenes, with an average length of 755 bp. Of these, 1371 unigenes were associated with lipid metabolism. The fatty acid (FA) biosynthesis and metabolism pathways were significantly enriched during the early stages of oil accumulation in the kernel. ALA biosynthesis was significantly enriched in parallel with increasing ALA content in the testa, but these metabolic pathways were not significantly enriched during pericarp development. By comparing unigene transcription profiles with patterns of ALA accumulation, specific unigenes encoding crucial enzymes and transcription factors (TFs) involved in de novo FA biosynthesis and oil accumulation were identified. Specifically, the bell-shaped expression patterns of genes encoding SAD, FAD2, FAD3, PDCT, PDAT, OLE, CLE, and SLE in the kernel were similar to the patterns of ALA accumulation in this tissue. Genes encoding BCCP, BC, KAS I– III, and FATA were also upregulated during the early stages of oil accumulation in the kernel. In the testa, the upregulation of the genes encoding SAD, FAD2, and FAD3 was followed by a sharp increase in the concentrations of ALA. In contrast, these genes were minimally expressed (and ALA content was low) throughout pericarp development.ConclusionsWe used three tissues with high, moderate, and low ALA concentrations as an exemplar system in which to investigate tissue-specific ALA accumulation mechanisms in P. ostii. The genes and TFs identified herein might be useful targets for future studies of ALA accumulation in the tree peony. This study also provides a framework for future studies of FA biosynthesis in other oilseed plants.

Analysis of fatty acid compositions and differential gene expression in two Iranian olive cultivars during fruit ripening

Olive is one of the important oil-produce fruit species with long history of cultivation. Despite high economic importance of olive, the molecular mechanisms that are involved in the oil production were not investigated completely in this fruit crop. In this study, the trend of oil accumulation, its composition and the transcriptional profiles of two important olive cultivars of Iran (‘Mari’ and ‘Shenge’) was screened during different fruit developmental stages (60, 90, 120, 150 and 180 days after flowering [DAF]). These cultivars showed the same trend for oil accumulation but a quite different fatty acid composition during the growing season. According to results, 60–120 DAF was identified as the most critical stages for oil biosynthesis in these olive cultivars. A cDNA-AFLP procedure was employed to detect the transcripts with differential expression during the critical stages of fatty acid biosynthesis in the fruit mesocarp. Altogether 195 distinguished fragments were identified as the transcripts with differential expression in these cultivars during the stages that were mostly important for oil biosynthesis. Out of which, twenty high quality clear transcript-derived fragments (TDFs) were cloned, sequenced and subjected to homology search analysis in GenBank database. Based on the homology search results, the entire differentially expressed TDFs in current investigation were classified into four main categories including oil production, carbohydrate metabolism, defense responsive and signaling related transcripts. Altogether, the majority of TDFs that were involved in the fatty acid biosynthesis and carbohydrate metabolism had higher expression in ‘Mari’, the cultivar which is superior in oil percentage and its composition. The expression of these TDFs coincided with the critical stages of oil accumulation (60–120 DAF). On the contrary, most of the defense related TDFs had higher expression in ‘Shenge’, an olive cultivar which is highly tolerant to the environmental stresses but inferior in the oil percentage and its composition. In conclusion, our finding provided primary information about the molecular basis of oil production in olive mesocarp and revealed new insight into the mechanisms that may be involved in the stress resistance in this fruit crop and shed some light on the probable candidate transcripts that may contribute to these two processes.

Natural gas accumulation processes of tight sandstone reservoirs in deep formations of Songliao Basin, NE China

The deep formations (underlying strata of the 3rd Member of Quantou Formation, Upper Cretaceous) in Songliao Basin possess a large natural gas potential, with a proven reserve of more than 200 × 109 m3 (706.3bcf). However, due to deep burial depth, multiple tectonic evolutions, and sets of source rocks, complex gas accumulations and different gas reservoir types developed, which may seriously restrict the natural gas exploration and target selection. This study focuses on the hydrocarbon generation, porosity evolution, and hydrocarbon accumulation processes of the tight sandstone gas (TSG) reservoir in the Cretaceous Shahezi Formation. 1) The timing and cumulative amounts of hydrocarbon generation were ascertained, with the Huoshiling being 143–10 Ma and 108.00 mg/g TOC and the Shahezi being 122–44 Ma and 30.00 mg/g TOC, respectively. 2) The mean original porosity of the Shahezi sandstone was 23.11%, and experienced the porosity decreasing of compaction (avg. = 12.57%) and cementation (avg. = 7.07%), and porosity increasing of dissolution (avg. = 3.25%), and the average present porosity is 6.72%. The sandstone densification (porosity <10%) occurred in the late period of Early Cretaceous (~115–105 Ma). 3) Four main stages of hydrocarbon accumulation in the K1sh occurred: I) Early oil accumulation stage (128–114 Ma)，corresponding to the 1st episode of charging; II) Middle oil and gas accumulation stage (111–90 Ma), corresponding to the 2nd episode of charging; III) Late gas accumulation stage (77–67 Ma), corresponding to the 3rd episode of charging; and IV) Adjustment stage (63 Ma–present), corresponding to the uplift stage. The 2nd and 3rd stages were the main gas accumulation stages. 4) Two types of tight sandstone gas reservoirs developed in the K1sh: I) tight conventional gas, and II) tight deep gas. The tight deep gas reservoirs, with favorable potentials, are the main exploration targets in the Shahezi Formation of Songliao Basin.

Transcriptome analysis of metabolic pathways associated with oil accumulation in developing seed kernels of Styrax tonkinensis, a woody biodiesel species

BackgroundStyrax tonkinensis (Pierre) Craib ex Hartwich has great potential as a woody biodiesel species having seed kernels with high oil content, excellent fatty acid composition and good fuel properties. However, no transcriptome information is available on the molecular regulatory mechanism of oil accumulation in developing S. tonkinensis kernels.ResultsThe dynamic patterns of oil content and fatty acid composition at 11 time points from 50 to 150 days after flowering (DAF) were analyzed. The percent oil content showed an up-down-up pattern, with yield and degree of unsaturation peaking on or after 140 DAF. Four time points (50, 70, 100, and 130 DAF) were selected for Illumina transcriptome sequencing. Approximately 73 million high quality clean reads were generated, and then assembled into 168,207 unigenes with a mean length of 854 bp. There were 5916 genes that were differentially expressed between different time points. These differentially expressed genes were grouped into 9 clusters based on their expression patterns. Expression patterns of a subset of 12 unigenes were confirmed by qRT-PCR. Based on their functional annotation through the Basic Local Alignment Search Tool and publicly available protein databases, specific unigenes encoding key enzymes, transmembrane transporters, and transcription factors associated with oil accumulation were determined. Three main patterns of expression were evident. Most unigenes peaked at 70 DAF, coincident with a rapid increase in oil content during kernel development. Unigenes with high expression at 50 DAF were associated with plastid formation and earlier stages of oil synthesis, including pyruvate and acetyl-CoA formation. Unigenes associated with triacylglycerol biosynthesis and oil body development peaked at 100 or 130 DAF.ConclusionsTranscriptome changes during oil accumulation show a distinct temporal trend with few abrupt transitions. Expression profiles suggest that acetyl-CoA formation for oil biosynthesis is both directly from pyruvate and indirectly via acetaldehyde, and indicate that the main carbon source for fatty acid biosynthesis is triosephosphate originating from phosphohexose outside the plastid. Different sn-glycerol-3-phosphate acyltransferases are implicated in diacylglycerol biosynthesis at early versus late stages of oil accumulation. Triacylglycerol biosynthesis may be accomplished by both diacylglycerol and by phospholipid:diacylglycerol acyltransferases.

A genome-wide analysis of the phospholipid: diacylglycerol acyltransferase gene family in Gossypium

BackgroundCotton (Gossypium spp.) is the most important natural fiber crop worldwide, and cottonseed oil is its most important byproduct. Phospholipid: diacylglycerol acyltransferase (PDAT) is important in TAG biosynthesis, as it catalyzes the transfer of a fatty acyl moiety from the sn-2 position of a phospholipid to the sn-3 position of sn-1, 2-diacylglyerol to form triacylglycerol (TAG) and a lysophospholipid. However, little is known about the genes encoding PDATs involved in cottonseed oil biosynthesis.ResultsA comprehensive genome-wide analysis of G. hirsutum, G. barbadense, G. arboreum, and G. raimondii herein identified 12, 11, 6 and 6 PDATs, respectively. These genes were divided into 3 subfamilies, and a PDAT-like subfamily was identified in comparison with dicotyledonous Arabidopsis. All GhPDATs contained one or two LCAT domains at the C-terminus, while most GhPDATs contained a preserved single transmembrane region at the N-terminus. A chromosomal distribution analysis showed that the 12 GhPDAT genes in G. hirsutum were distributed in 10 chromosomes. However, none of the GhPDATs was co-localized with quantitative trait loci (QTL) for cottonseed oil content, suggesting that their sequence variations are not genetically associated with the natural variation in cottonseed oil content. Most GhPDATs were expressed during the cottonseed oil accumulation stage. Ectopic expression of GhPDAT1d increased Arabidopsis seed oil content.ConclusionsOur comprehensive genome-wide analysis of the cotton PDAT gene family provides a foundation for further studies into the use of PDAT genes to increase cottonseed oil content through biotechnology.

Transcriptome analysis reveals crucial genes involved in the biosynthesis of nervonic acid in woody Malania oleifera oilseeds

BackgroundMalania oleifera Chun et Lee (Olacaceae), an evergreen broad-leaved woody tree native to southwest China, is an important oilseed tree. Its seed oil has a high level of nervonic acid (cis-tetracos-15-enoic acid, over 60%), which is essential for human health. M. oleifera seed oil is a promising source of nervonic acid, but little is known about the physiological and molecular mechanisms underlying its biosynthesis.ResultsIn this study, we recorded oil accumulation at four stages of seed development. Using a high-throughput RNA-sequencing technique, we obtained 55,843 unigenes, of which 29,176 unigenes were functionally annotated. By comparison, 22,833 unigenes had a two-fold or greater expression at the fast oil accumulation stage than at the initial stage. Of these, 198 unigenes were identified as being functionally involved in diverse lipid metabolism processes (including de novo fatty acid synthesis, carbon chain elongation and modification, and triacylglycerol assembly). Key genes (encoding KCS, KCR, HCD and ECR), putatively responsible for nervonic acid biosynthesis, were isolated and their expression profiles during seed development were confirmed by quantitative real-time PCR analysis. Also, we isolated regulatory factors (such as WRI1, ABI3 and FUS3) that are putatively involved in the regulation of oil biosynthesis and seed development.ConclusionOur results provide novel data on the physiological and molecular mechanisms of nervonic acid biosynthesis and oil accumulation in M. oleifera seeds, and will also serve as a starting point for biotechnological genetic engineering for the production of nervonic acid resources.

Developing a set of strong intronic promoters for robust metabolic engineering in oleaginous Rhodotorula (Rhodosporidium) yeast species.

BackgroundRed yeast species in the Rhodotorula/Rhodosporidium genus are outstanding producers of triacylglyceride and cell biomass. Metabolic engineering is expected to further enhance the productivity and versatility of these hosts for the production of biobased chemicals and fuels. Promoters with strong activity during oil-accumulation stage are critical tools for metabolic engineering of these oleaginous yeasts.ResultsThe upstream DNA sequences of 6 genes involved in lipid biosynthesis or accumulation in Rhodotorula toruloides were studied by luciferase reporter assay. The promoter of perilipin/lipid droplet protein 1 gene (LDP1) displayed much stronger activity (4–11 folds) than that of glyceraldehyde-3-phosphate dehydrogenase gene (GPD1), one of the strongest promoters known in yeasts. Depending on the stage of cultivation, promoter of acetyl-CoA carboxylase gene (ACC1) and fatty acid synthase β subunit gene (FAS1) exhibited intermediate strength, displaying 50–160 and 20–90% levels of GPD1 promoter, respectively. Interestingly, introns significantly modulated promoter strength at high frequency. The incorporation of intron 1 and 2 of LDP1 (LDP1in promoter) enhanced its promoter activity by 1.6–3.0 folds. Similarly, the strength of ACC1 promoter was enhanced by 1.5–3.2 folds if containing intron 1. The intron 1 sequences of ACL1 and FAS1 also played significant regulatory roles. When driven by the intronic promoters of ACC1 and LDP1 (ACC1in and LDP1in promoter, respectively), the reporter gene expression were up-regulated by nitrogen starvation, independent of de novo oil biosynthesis and accumulation. As a proof of principle, overexpression of the endogenous acyl-CoA-dependent diacylglycerol acyltransferase 1 gene (DGA1) by LDP1in promoter was significantly more efficient than GPD1 promoter in enhancing lipid accumulation.ConclusionIntronic sequences play an important role in regulating gene expression in R. toruloides. Three intronic promoters, LDP1in, ACC1in and FAS1in, are excellent promoters for metabolic engineering in the oleaginous and carotenogenic yeast, R. toruloides. Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-016-0600-x) contains supplementary material, which is available to authorized users.

Fatty acid composition of developing tree peony (Paeonia section Moutan DC.) seeds and transcriptome analysis during seed development.

BackgroundTree peony (Paeonia section Moutan DC.) is known for its excellent ornamental and medicinal values. In 2011, seeds from P. ostii have been identified as novel resource of α-linolenic acid (ALA) for seed oil production and development in China. However, the molecular mechanism on biosynthesis of unsaturated fatty acids in tree peony seeds remains unknown. Therefore, transcriptome data is needed to better understand the underlying mechanisms.ResultsIn this study, lipid accumulation contents were measured using GC-MS methods across developing tree peony seeds, which exhibited an extraordinary ALA content (49.3%) in P. ostii mature seeds. Transcriptome analysis was performed using Illumina sequencing platform. A total of 144 million 100-bp paired-end reads were generated from six libraries, which identified 175,874 contigs. In the KEGG Orthology enrichment of differentially expressed genes, lipid metabolism pathways were highly represented categories. Using this data we identified 388 unigenes that may be involved in de novo fatty acid and triacylglycerol biosynthesis. In particular, three unigenes (SAD, FAD2 and FAD8) encoding fatty acid desaturase with high expression levels in the fast oil accumulation stage compared with the initial stage of seed development were identified.ConclusionsThis study provides the first comprehensive genomic resources characterizing tree peony seeds gene expression at the transcriptional level. These data lay the foundation for further understanding of molecular mechanism responsible for lipid biosynthesis and the high unsaturated fatty acids (especially ALA) accumulation. Meanwhile, it provides theoretical base for potential oilseed application in the respect of n-6 to n-3 ratio for human diets and future regulation of target healthy components of oils.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1429-0) contains supplementary material, which is available to authorized users.

The effect of water stress on super-high- density 'Koroneiki' olive oil quality.

Over the last two decades, the area of cultivated super-high-density olive orchards has increased rapidly. Water stress is an important tool in super-high-density orchards to reduce tree growth and promote suitability for overhead mechanical harvesters. Little is known regarding the effect of water stress in super-high-density orchards on oil quality parameters. In this study the effect of irrigation rate on oil quality parameters was evaluated in a six-year-old super-high-density 'Koreneiki' olive orchard for five consecutive seasons. Five water status levels, determined by irrigating in order to maintain various midday stem water potential threshold values (-1.5, -2, -2.5, -3 and -4 MPa), were applied during the oil accumulation stage. The MUFA/PUFA ratio and free fatty acid content generally decreased as a function of increasing tree water stress. In most seasons a reduction in polyphenols was found with decreasing irrigation level. Peroxide value was not affected by the water stress level. The present study demonstrates that limiting irrigation and exposure of olive trees to water stress in a super-high-density orchard lowers free fatty acid content and therefore benefits oil quality. However, the decreased MUFA/PUFA ratio and the reduction in polyphenol content that were also found under increased water stress negatively influence oil quality.

Transcriptome-Wide Identification and Characterization of MicroRNAs from Castor Bean (Ricinus communis L.)

BackgroundMicroRNAs (miRNAs) are endogenously encoded small RNAs that post-transcriptionally regulate gene expression and play essential roles in numerous developmental and physiological processes. Currently, little information on the transcriptome and tissue-specific expression of miRNAs is available in the model non-edible oilseed crop castor bean (Ricinus communis L.), one of the most important non-edible oilseed crops cultivated worldwide. Recent advances in sequencing technologies have allowed the identification of conserved and novel miRNAs in many plant species. Here, we used high-throughput sequencing technologies to identify and characterize the miRNAs in castor bean.ResultsFive small RNA libraries were constructed for deep sequencing from root tips, leaves, developing seeds (at the initial stage, seed1; and at the fast oil accumulation stage, seed2) and endosperms in castor bean. High-throughput sequencing generated a large number of sequence reads of small RNAs in this study. In total, 86 conserved miRNAs were identified, including 63 known and 23 newly identified. Sixteen miRNA isoform variants in length were found from the conserved miRNAs of castor bean. MiRNAs displayed diverse organ-specific expression levels among five libraries. Combined with criteria for miRNA annotation and a RT-PCR approach, 72 novel miRNAs and their potential precursors were annotated and 20 miRNAs newly identified were validated. In addition, new target candidates for miRNAs newly identified in this study were proposed.ConclusionsThe current study presents the first high-throughput small RNA sequencing study performed in castor bean to identify its miRNA population. It characterizes and increases the number of miRNAs and their isoforms identified in castor bean. The miRNA expression analysis provides a foundation for understanding castor bean miRNA organ-specific expression patterns. The present study offers an expanded picture of miRNAs for castor bean and other members in the family Euphorbiaceae.

Expression analysis of VfDGAT2 in various tissues of the tung tree and in transgenic yeast

The tung tree (Vernicia fordii Hemsl.; Vf) has great potential as an industrial crop owning to its seed oil that has multiple uses. Diacylglycerol acyltransferases (DGATs) catalyze the last and most committed step of triacylglycerol (TAG) biosynthesis. In order to examine the physiological role of the VfDGAT2 gene in the tung tree, we characterized its expression profiles in different tung tissues/organs and seeds at different developmental stages. Oil content and α-eleostearic acid production during seed development were also examined. Expression studies showed that VfDGAT2 was expressed in all tissues tested, with the highest expression in developing seeds where the expression was about 19-fold more than that in leaves. VfDGAT2 showed temporal-specific expression during seed development and maturation. Notably, the expression of VfDGAT2 in developing seeds was found to be consistent with tung oil accumulation and α-eleostearic acid production. The expression level of VfDGAT2 was lower in the early stages of oil accumulation and α-eleostearic acid biosynthesis, rapidly increased during the peak periods of fatty acid synthesis in August, and then decreased during completion of the accumulation period at the end of September. When the VfDGAT2 gene was transferred to the oleaginous yeast Rhodotorula glutinis, its expression was detected along with fatty acid products. The results showed that VfDGAT2 was highly expressed in transgenic yeast clones, and the total fatty acid content in one of these clones, VfDGAT2-3, was 7.8-fold more than that in the control, indicating that VfDGAT2 contributed to fatty acid accumulation into TAG and might be a target gene for improving tung oil composition through genetic engineering.

Transcriptome analysis of Sacha Inchi (Plukenetia volubilis L.) seeds at two developmental stages

BackgroundSacha Inchi (Plukenetia volubilis L., Euphorbiaceae) is a potential oilseed crop because the seeds of this plant are rich in unsaturated fatty acids (FAs). In particular, the fatty acid composition of its seed oil differs markedly in containing large quantities of α-linolenic acid (18C:3, a kind of ω-3 FAs). However, little is known about the molecular mechanisms responsible for biosynthesis of unsaturated fatty acids in the developing seeds of this species. Transcriptome data are needed to better understand these mechanisms.ResultsIn this study, de novo transcriptome assembly and gene expression analysis were performed using Illumina sequencing technology. A total of 52.6 million 90-bp paired-end reads were generated from two libraries constructed at the initial stage and fast oil accumulation stage of seed development. These reads were assembled into 70,392 unigenes; 22,179 unigenes showed a 2-fold or greater expression difference between the two libraries. Using this data we identified unigenes that may be involved in de novo FA and triacylglycerol biosynthesis. In particular, a number of unigenes encoding desaturase for formation of unsaturated fatty acids with high expression levels in the fast oil accumulation stage compared with the initial stage of seed development were identified.ConclusionsThis study provides the first comprehensive dataset characterizing Sacha Inchi gene expression at the transcriptional level. These data provide the foundation for further studies on molecular mechanisms underlying oil accumulation and PUFA biosynthesis in Sacha Inchi seeds. Our analyses facilitate understanding of the molecular mechanisms responsible for the high unsaturated fatty acids (especially α-linolenic acid) accumulation in Sacha Inchi seeds.

The effects of crop load and irrigation rate in the oil accumulation stage on oil yield and water relations of ‘Koroneiki’ olives

The interactions between irrigation rates applied during the oil accumulation stage and crop load were studied in a six-year-old very-high-density Koroneiki (Olea europaea L.) orchard. Five irrigation rates, determined as thresholds of midday stem water potential, were applied from July 1st until harvest in 2008 and 2009 and from July 1st to the end of September in 2010. Oil yield increased with increasing crop load in all the irrigation treatments. Oil yield did not respond to increasing irrigation at very low crop load and the higher the crop load the higher the response to irrigation. There was no response to irrigation at the lowest crop loads, but the higher the irrigation rate the higher the oil yield at high crop loads. The predicted commercial oil yield at common fruit counts increased from 1.99 t/ha at the lowest irrigation rate to 3.06 t/ha at the highest irrigation rate. Stomatal conductance decreased with decreasing stem water potential but leveled off at 30–60 mmol m−2 s−1 at stem water potential values lower than −4.0 MPa. High crop load increased stomatal conductance and decreased stem water potential relative to low crop load at low and medium irrigation rates. The effect of crop load on water relations became evident by the end of August and was well pronounced at the beginning of October. Physiological and irrigation water management implications related to the interactions between tree water status and crop load are discussed.

Timing of nitrogen application to macadamias. 1. Tree nitrogen status and vegetative growth

Nitrogen (690 g/tree.annum) was applied to 12-year-old macadamia (Macadamia integrifoloia Maiden and Betche) trees in subtropical Queensland (lat. 26�S.) at either pre-floral initiation, inflorescence development, premature nut drop or at the oil accumulation stage. Control trees received a total of 230 g of nitrogen (N) split into equal monthly applications. Leaf N concentrations in May (vegetative and reproductive dormancy) was lower in control trees (approx. 1.2%), which were visually N-deficient during winter and spring. Leaf N concentrations of all other treatments were approx. 1.3-1.4% N and leaves remained a healthy green colour throughout the year. Low N status in control trees suppressed vegetative growth (indicated by changes in trunk girth, tree height and canopy volume), effectively manipulating the vegetative:reproductive balance in favour-of the latter. The desirability of small, but frequent applications of N fertiliser to macadamias is indicated, although the optimum rate is likely to be greater than that applied to control trees in this study. It is suggested that determining tree N requirements using standard leaf nutrient levels should not be done in isolation. Tree health and vegetative growth in relation to yield, and time of sampling, particularly in relation to time of application of N and flushing peaks, should also be considered.

Constraining Diagenesis Using Fluid Inclusions in Cements from Petroleum Basins: Examples from North Sea and Middle East: ABSTRACT

Cements in petroleum reservoirs commonly contain inclusions filled with brine, hydrocarbon, or mixtures of both. The physicochemical conditions of cementation as well as the quality of the inclusion oil can be derived from microthermometric and fluorescence microspectrophotometric analysis of fluid inclusions. These data, when integrated with petrographic, geochemical, engineering, and burial history data, can constrain the timing and nature of both diagenetic events and oil migration. Brine and oil-bearing inclusions occur in quartz overgrowths from the Brent Sandstone, Northwest Hutton field, North Sea. Fluorescence analysis indicates that the inclusion oil is similar to the reservoired oil. Inclusion data define the reservoir conditions that existed during the initial stages of oil accumulation and concurrent quartz cementation. The abnormally high pressure of inclusion entrapment suggests that the reservoir was overpressured. Paleo-overpressuring probably helped preserve reservoir porosity by slowing compaction-related loss of porosity in coarse-grained reservoir sands. Brine, oil, and three-phase (brine, oil, and vapor) inclusions occur in burial-related calcite and dolomite cements from a Lower Cretaceous (Thamama) carbonate reservoir in the Middle East. Inclusion data indicate that these cements were precipitated from and simultaneously trapped an invading basinal brine that contained an immiscible component of hydrocarbon. The intersection of brine and oilmore » isochores in pore-throat space allows a unique determination of the time and depth of initial reservoir fill-up. The fill-up appears to coincide with the onset of peak oil generation from the source rocks believed to have supplied these reservoirs.« less

Changes in lipid class and fatty acid composition during maturation of mesocarp of oil palm (Elaeis guineensis) variety dura

AbstractThe deposition of oil, major lipid classes, and the constituent fatty acids of the oil palm fruit mesocarp were investigated to gain a better insight into the accumulation of these components. The oils were extracted with chloroform‐methanol 1:2 (v/v) and were fractionated by silicic acid column chromatography into classes. Fatty acids were analysed by gas‐liquid chromatography. Shortly after anthesis, lipids, mainly phospho‐ and glyco‐lipids, were a small proportion of the mesocarp. With time the lipid content increased, until at maturity it accounted for about 60% of fresh mesocarp weight. Approximately 97% of the lipid at maturity was neutral lipid, essentially triacylglycerol. Maximum oil quantity was obtained at 22 weeks after anthesis. The active lipid accumulation period was from weeks 18 to 22 after pollination. Contrary to previously published data, palmitic, oleic and linoleic acids were characteristically present at all stages of oil accumulation. Linolenic acid was present in the neutral lipids from immature mesocarp of the oil palm fruit, but was probably diluted out in mature palms. Appreciable amounts of linolenic acid were present in the polar lipid fractions.