Development of a multiplex amplicon-sequencing assay to detect low-frequency mutations in poinsettia (Euphorbia pulcherrima) breeding programmes

Poinsettia is an economically important ornamental potted plant in which certain bract colour variants are often obtained by mutation breeding. Previously, in poinsettia, we identified Bract1, a GST gene involved in the sequestration and transport of anthocyanins to the vacuole. This gene carries a short, highly mutable 4-bp repeat in its coding region. Loss of one repeat unit leads to a loss of function for Bract1, and in homozygous mutants, anthocyanin-based coloration is absent, resulting in white or cream-coloured bracts. Although mutation induction through ionizing radiation leads to a high frequency of mutations in Bract1, mutants are difficult to obtain from homozygous red genotypes. In this study, we used Bract1-specific amplicon sequencing as a tool to identify mutations in pools of tissues, which enabled the detection of mutations in dilutions of up to one mutant in 50 nonmutated samples. This approach enabled efficient screening of recalcitrant homozygous genotypes for mutated alleles and the reduction of the mutation load in the application of ionizing radiation in mutation breeding programmes

Extractor for ESI quadrupole TOF tandem MS data enabled for high throughput batch processing

BACKGROUND: Mass spectrometry based proteomics result in huge amounts of data that has to be processed in real time in order to efficiently feed identification algorithms and to easily integrate in automated environments. We present wiff2dta, a tool created to convert MS/MS data obtained using Applied Biosystem's QStar and QTrap 2000 and 4000 series. RESULTS: Comparing the performance of wiff2dta with the standard tools, we find wiff2dta being the fastest solution for extracting spectrum data from ABIs raw file format. wiff2dta is at least 10% faster than the standard tools. It is also capable of batch processing and can be easily integrated in high throughput environments. The program is freely available via , and is also available from Applied Biosystems. CONCLUSIONS: wiff2dta offers the possibility to run as stand-alone application or within a batch process as command-line tool integrated in automation and high-throughput environments. It is more efficient than the state-of-the-art tools provided

Projects of the Center for Energy Research

This inaugural event is dedicated to showcasing the renewable/sustainable energy projects of UNLV faculty, staff, students, and collaborators, as well as other external projects underway statewide and nationally. The development and utilization of new technologies to protect the environment, achieve energy independence, and strengthen the economy will be explored. Speakers and poster-session presenters will provide further insight to many ongoing projects and innovative research ideas. Organized by UNLV’s Office of Strategic Energy Programs, the event offers participants the opportunity to learn about energy projects and will encourage networking and collaboration. This symposium is intended for researchers, educators, students, policy makers, public and private-sector energy and environmental professionals, and citizen

Current projects at the UNLV Center for Energy Research

The 3rd Annual Renewable Energy Symposium took place on the UNLV campus August 11 & 12. The event focused on renewable energy production in Nevada, the US Southwest, and renewable research projects nationwide. The event was a great success with over 200 individuals in attendance

First genome edited poinsettias: targeted mutagenesis of flavonoid 3′-hydroxylase using CRISPR/Cas9 results in a colour shift

The CRISPR/Cas9 system is a remarkably promising tool for targeted gene mutagenesis, and becoming ever more popular for modification of ornamental plants. In this study we performed the knockout of flavonoid 3′-hydroxylase (F3′H) with application of CRISPR/Cas9 in the red flowering poinsettia (Euphorbia pulcherrima) cultivar ‘Christmas Eve’, in order to obtain plants with orange bract colour, which accumulate prevalently pelargonidin. F3′H is an enzyme that is necessary for formation of cyanidin type anthocyanins, which are responsible for the red colour of poinsettia bracts. Even though F3′H was not completely inactivated, the bract colour of transgenic plants changed from vivid red (RHS 45B) to vivid reddish orange (RHS 33A), and cyanidin levels decreased significantly compared with the wild type. In the genetically modified plants, an increased ratio of pelargonidin to cyanidin was observed. By cloning and expression of mutated proteins, the lack of F3′H activity was confirmed. This confirms that a loss of function mutation in the poinsettia F3′H gene is sufficient for obtaining poinsettia with orange bract colour. This is the first report of successful use of CRISPR/Cas9 for genome editing in poinsettia

AN ESTIMATION OF THE PERFORMANCE LIMITS OF DRY COOLING ON TROUGH-TYPE SOLAR THERMAL PLANTS

ABSTRACT The southwestern US is an ideal location for solar power plants due to its abundant solar resource, while there is a difficulty in implementing wet cooling systems due to the shortage of water in this region. Dry cooling could be an excellent solution for this, if it could achieve a high efficiency and low cost as wet cooling. Some dry cooling systems are currently in operation, and investigations of their performance have been reported in the literature. This paper looks into the limits to the power production implicit in dry cooling, assuming that improvements might be made to the system components. Use of higher performance heat transfer surfaces is one such possible improvement. We have developed a model of a fairly typical, but simplified, solar trough plant, and simulated thermodynamic performance of this with the software Gatecycle. We have examined the power generation and cycle efficiency of the plant for the Las Vegas vicinity with conventional wet cooling and conventional dry cooling cases considered separately using this software. TMY2 data are used for this location for this purpose. Similarly, the same studies are carried out for "ideal" cooling systems as a comparison. We assumed that in the ideal dry cooling system, the condensing temperature is the ambient dry bulb temperature, and in the ideal wet cooling system, it is the ambient wet bulb temperature. It turned out that the ideal dry cooling system would significantly outperform the conventional wet cooling system, indicating the possibility of the dry cooling system being able to achieve increased performance levels with component improvements

Hybrid de novo transcriptome assembly of poinsettia (Euphorbia pulcherrima Willd. Ex Klotsch) bracts

Background Poinsettia is a popular and important ornamental crop, mostly during the Christmas season. Its bract coloration ranges from pink/red to creamy/white shades. Despite its ornamental value, there is a lack of knowledge about the genetics and molecular biology of poinsettia, especially on the mechanisms of color formation. We performed an RNA-Seq analysis in order to shed light on the transcriptome of poinsettia bracts. Moreover, we analyzed the transcriptome differences of red- and white-bracted poinsettia varieties during bract development and coloration. For the assembly of a bract transcriptome, two paired-end cDNA libraries from a red and white poinsettia pair were sequenced with the Illumina technology, and one library from a red-bracted variety was used for PacBio sequencing. Both short and long reads were assembled using a hybrid de novo strategy. Samples of red- and white-bracted poinsettias were sequenced and comparatively analyzed in three color developmental stages in order to understand the mechanisms of color formation and accumulation in the species. Results The final transcriptome contains 288,524 contigs, with 33% showing confident protein annotation against the TAIR10 database. The BUSCO pipeline, which is based on near-universal orthologous gene groups, was applied to assess the transcriptome completeness. From a total of 1440 BUSCO groups searched, 77% were categorized as complete (41% as single-copy and 36% as duplicated), 10% as fragmented and 13% as missing BUSCOs. The gene expression comparison between red and white varieties of poinsettia showed a differential regulation of the flavonoid biosynthesis pathway only at particular stages of bract development. An initial impairment of the flavonoid pathway early in the color accumulation process for the white poinsettia variety was observed, but these differences were no longer present in the subsequent stages of bract development. Nonetheless, GSTF11 and UGT79B10 showed a lower expression in the last stage of bract development for the white variety and, therefore, are potential candidates for further studies on poinsettia coloration. Conclusions In summary, this transcriptome analysis provides a valuable foundation for further studies on poinsettia, such as plant breeding and genetics, and highlights crucial information on the molecular mechanism of color formation