Expanding the bioluminescent reporter toolkit for plant science with NanoLUC.


Background:Protein data over circadian time scale is scarce for clock transcription factors. Further work in this direction is required for refining quantitative clock models. However, gathering highly resolved dynamics of low-abundance transcription factors has been a major challenge in the field. In this work we provide a new tool that could help this major issue. Bioluminescence is an important tool for gathering data on circadian gene expression. It allows data collection over extended time periods for low signal levels, thanks to a large signal-to-noise ratio. However, the main reporter so far used, firefly luciferase (FLUC), presents some disadvantages for reporting total protein levels. For example, the rapid, post-translational inactivation of this luciferase will result in underestimation of protein numbers. A more stable reporter protein could in principle tackle this issue. We noticed that NanoLUC might fill this gap, given its reported brightness and the stability of both enzyme and substrate. However, no data in plant systems on the circadian time scale had been reported. Results:We tested NanoLUC activity under different scenarios that will be important for generating highly quantitative data. These include enzyme purification for calibration curves, expression in transient plant systems, stable transgenic plants and in planta time series over circadian time scales. Furthermore, we show that the difference in substrate use between firefly luciferase and NanoLUC allows tracking of two different reporters from the same samples. We show this by exploring the impact of a BOAp:BOA-NanoLUC construct transformed into a Col-0 CCA1p:FLUC background. Conclusions:We concluded that NanoLUC reporters are compatible with established instrumentation and protocols for firefly luciferase. Overall, our results provide guidelines for researchers gathering dynamic protein data over different time scales and experimental setups.


Plant Methods


Plant methods


Urquiza-García U,Millar AJ




Has Abstract


2019-07-08 00:00:00










  • De novo assembly of potential linear artificial chromosome constructs capped with expansive telomeric repeats.

    abstract:BACKGROUND:Artificial chromosomes (ACs) are a promising next-generation vector for genetic engineering. The most common methods for developing AC constructs are to clone and combine centromeric DNA and telomeric DNA fragments into a single large DNA construct. The AC constructs developed from such methods will contain ...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Lin L,Koo DH,Zhang W,St Peter J,Jiang J

    更新日期:2011-04-15 00:00:00

  • Integrating field-based heat tents and cyber-physical system technology to phenotype high night-time temperature impact on winter wheat.

    abstract:Background:Many agronomic traits have been bred into modern wheat varieties, but wheat (Triticum aestivum L.) continues to be vulnerable to heat stress, with high night-time temperature (HNT) stress shown to have large negative impact on yield and quality. Global mean temperature during the day is consistently warming ...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Hein NT,Wagner D,Bheemanahalli R,Šebela D,Bustamante C,Chiluwal A,Neilsen ML,Jagadish SVK

    更新日期:2019-04-24 00:00:00

  • Rapid diagnosis of nitrogen status in rice based on Fourier transform infrared photoacoustic spectroscopy (FTIR-PAS).

    abstract:Background:An effective and expeditious approach to assess plant nitrogen status is urgently needed in rice production and management as the conventional chemical methods are laborious and time-consuming. Results:Fourier transform infrared photoacoustic spectroscopy (FTIR-PAS) was used to record the spectra of rice le...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Wu K,Du C,Ma F,Shen Y,Liang D,Zhou J

    更新日期:2019-08-19 00:00:00

  • Chlorothalonil: an effective bacteriostatic agent for bud induction of Acacia auriculiformis under open condition (non-axenic).

    abstract:Background:Open tissue culture technique could be simplified by using different bacteriostatic agents. There is a great difference in the bacteriostatic effects of different antimicrobial agents on various explants. However, there is no report about the effective bacteriostatic agent for open tissue culture of Acacia a...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Huang L,Wang H,Shahid MQ,Zeng B

    更新日期:2019-01-25 00:00:00

  • Striga parasitizes transgenic hairy roots of Zea mays and provides a tool for studying plant-plant interactions.

    abstract:BACKGROUND:Striga species are noxious root hemi-parasitic weeds that debilitate cereal production in sub-Saharan Africa (SSA). Control options for Striga are limited and developing Striga resistant crop germplasm is regarded as the best and most sustainable control measure. Efforts to improve germplasm for Striga resis...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Runo S,Macharia S,Alakonya A,Machuka J,Sinha N,Scholes J

    更新日期:2012-06-21 00:00:00

  • Inoculation insensitive promoters for cell type enriched gene expression in legume roots and nodules.

    abstract:BACKGROUND:Establishment and maintenance of mutualistic plant-microbial interactions in the rhizosphere and within plant roots involve several root cell types. The processes of host-microbe recognition and infection require complex signal exchange and activation of downstream responses. These molecular events coordinat...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Gavrilovic S,Yan Z,Jurkiewicz AM,Stougaard J,Markmann K

    更新日期:2016-01-22 00:00:00

  • Real-time, non-destructive and in-field foliage yield and growth rate measurement in perennial ryegrass (Lolium perenne L.).

    abstract:Background:In-field measurement of yield and growth rate in pasture species is imprecise and costly, limiting scientific and commercial application. Our study proposed a LiDAR-based mobile platform for non-invasive vegetative biomass and growth rate estimation in perennial ryegrass (Lolium perenne L.). This included de...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Ghamkhar K,Irie K,Hagedorn M,Hsiao J,Fourie J,Gebbie S,Hoyos-Villegas V,George R,Stewart A,Inch C,Werner A,Barrett B

    更新日期:2019-07-10 00:00:00

  • DiNAR: revealing hidden patterns of plant signalling dynamics using Differential Network Analysis in R.

    abstract:Background:Progress in high-throughput molecular methods accompanied by more complex experimental designs demands novel data visualisation solutions. To specifically answer the question which parts of the specifical biological system are responding in particular perturbation, integrative approach in which experimental ...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Zagorščak M,Blejec A,Ramšak Ž,Petek M,Stare T,Gruden K

    更新日期:2018-08-30 00:00:00

  • The advantages and limitations of trait analysis with GWAS: a review.

    abstract::Over the last 10 years, high-density SNP arrays and DNA re-sequencing have illuminated the majority of the genotypic space for a number of organisms, including humans, maize, rice and Arabidopsis. For any researcher willing to define and score a phenotype across many individuals, Genome Wide Association Studies (GWAS)...

    journal_title:Plant methods

    pub_type: 杂志文章,评审


    authors: Korte A,Farlow A

    更新日期:2013-07-22 00:00:00

  • SHiNeMaS: a web tool dedicated to seed lots history, phenotyping and cultural practices.

    abstract:Motivation:In 2005, researchers from the French National Research Institute for Agriculture, Food and Environment (Institut national de recherche pour l'agriculture, l'alimentation et l'environnement, INRAE) started a collaboration with the French farmers' seed network Réseau Semences Paysannes (RSP) on bread wheat par...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: De Oliveira Y,Burlot L,Dawson JC,Goldringer I,Madi D,Rivière P,Steinbach D,van Frank G,Thomas M

    更新日期:2020-07-23 00:00:00

  • Methods for transient assay of gene function in floral tissues.

    abstract:BACKGROUND:There is considerable interest in rapid assays or screening systems for assigning gene function. However, analysis of gene function in the flowers of some species is restricted due to the difficulty of producing stably transformed transgenic plants. As a result, experimental approaches based on transient gen...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Shang Y,Schwinn KE,Bennett MJ,Hunter DA,Waugh TL,Pathirana NN,Brummell DA,Jameson PE,Davies KM

    更新日期:2007-01-08 00:00:00

  • Improving the sorting efficiency of maize haploid kernels using an NMR-based method with oil content double thresholds.

    abstract:BACKGROUND:Maize haploid breeding technology can be used to rapidly develop homozygous lines, significantly shorten the breeding cycle and improve breeding efficiency. Rapid and accurate sorting haploid kernels is a prerequisite for the large-scale application of this technology. At present, the automatic haploid sorti...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Qu Y,Liu Z,Zhang Y,Yang J,Li H

    更新日期:2021-01-06 00:00:00

  • An UPLC-MS/MS method for highly sensitive high-throughput analysis of phytohormones in plant tissues.

    abstract:UNLABELLED: BACKGROUND:Phytohormones are the key metabolites participating in the regulation of multiple functions of plant organism. Among them, jasmonates, as well as abscisic and salicylic acids are responsible for triggering and modulating plant reactions targeted against pathogens and herbivores, as well as resis...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Balcke GU,Handrick V,Bergau N,Fichtner M,Henning A,Stellmach H,Tissier A,Hause B,Frolov A

    更新日期:2012-11-22 00:00:00

  • Convergence of developmental mutants into a single tomato model system: 'Micro-Tom' as an effective toolkit for plant development research.

    abstract:BACKGROUND:The tomato (Solanum lycopersicum L.) plant is both an economically important food crop and an ideal dicot model to investigate various physiological phenomena not possible in Arabidopsis thaliana. Due to the great diversity of tomato cultivars used by the research community, it is often difficult to reliably...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Carvalho RF,Campos ML,Pino LE,Crestana SL,Zsögön A,Lima JE,Benedito VA,Peres LE

    更新日期:2011-06-29 00:00:00

  • Protocol: a rapid and economical procedure for purification of plasmid or plant DNA with diverse applications in plant biology.

    abstract:: Research in plant molecular biology involves DNA purification on a daily basis. Although different commercial kits enable convenient extraction of high-quality DNA from E. coli cells, PCR and agarose gel samples as well as plant tissues, each kit is designed for a particular type of DNA extraction work, and the cost ...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Li JF,Li L,Sheen J

    更新日期:2010-01-14 00:00:00

  • High-throughput quantification of more than 100 primary- and secondary-metabolites, and phytohormones by a single solid-phase extraction based sample preparation with analysis by UHPLC-HESI-MS/MS.

    abstract:BACKGROUND:Plant metabolites are commonly functionally classified, as defense- or growth-related phytohormones, primary and specialized metabolites, and so forth. Analytical procedures for the quantifications of these metabolites are challenging because the metabolites can vary over several orders of magnitude in conce...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Schäfer M,Brütting C,Baldwin IT,Kallenbach M

    更新日期:2016-05-26 00:00:00

  • Simple extraction methods that prevent the artifactual conversion of chlorophyll to chlorophyllide during pigment isolation from leaf samples.

    abstract:BACKGROUND:When conducting plant research, the measurement of photosynthetic pigments can provide basic information on the physiological status of a plant. High-pressure liquid chromatography (HPLC) is becoming widely used for this purpose because it provides an accurate determination of a variety of photosynthetic pig...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Hu X,Tanaka A,Tanaka R

    更新日期:2013-06-19 00:00:00

  • Methanol fixation of plant tissue for Scanning Electron Microscopy improves preservation of tissue morphology and dimensions.

    abstract:BACKGROUND:It is well known that preparation of biological (plant and animal) tissues for Scanning Electron Microscopy (SEM) by chemical fixation and critical point drying results in shrinkage of tissues, often by up to 20-30%, depending on the tissue type and fixation protocol used. We sought to identify a protocol th...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Talbot MJ,White RG

    更新日期:2013-10-02 00:00:00

  • Kinome profiling of Arabidopsis using arrays of kinase consensus substrates.

    abstract:BACKGROUND:Kinome profiling aims at the parallel analysis of kinase activities in a cell. Novel developed arrays containing consensus substrates for kinases are used to assess those kinase activities. The arrays described in this paper were already used to determine kinase activities in mammalian systems, but since sub...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Ritsema T,Joore J,van Workum W,Pieterse CM

    更新日期:2007-02-12 00:00:00

  • Correction to: Soybean iron deficiency chlorosis high throughput phenotyping using an unmanned aircraft system.

    abstract::[This corrects the article DOI: 10.1186/s13007-019-0478-9.]. ...

    journal_title:Plant methods

    pub_type: 已发布勘误


    authors: Dobbels AA,Lorenz AJ

    更新日期:2019-10-10 00:00:00

  • Automatic segmentation and measurement methods of living stomata of plants based on the CV model.

    abstract:Background:The stomata of plants mainly regulate gas exchange and water dispersion between the interior and external environments of plants and play a major role in the plants' health. The existing methods of stomata segmentation and measurement are mostly for specialized plants. The purpose of this research is to deve...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Li K,Huang J,Song W,Wang J,Lv S,Wang X

    更新日期:2019-07-03 00:00:00

  • Suitable transfection methods for single particle tracing in plant suspension cells.

    abstract:BACKGROUND:A multitude of different imaging systems are already available to image genetically altered RNA species; however, only a few of these techniques are actually suitable to visualize endogenous RNA. One possibility is to use fluorescently-labelled and hybridization-sensitive probes. In order to yield more infor...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Göhring J,Fulcher N,Schilcher K,Barta A,Jacak J

    更新日期:2014-05-31 00:00:00

  • Testing deep placement of an 15N tracer as a method for in situ deep root phenotyping of wheat, barley and ryegrass.

    abstract:Background:Deep rooting is one of the most promising plant traits for improving crop yield under water-limited conditions. Most root phenotyping methods are designed for laboratory-grown plants, typically measuring very young plants not grown in soil and not allowing full development of the root system. Results:This s...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Chen S,Svane SF,Thorup-Kristensen K

    更新日期:2019-12-09 00:00:00

  • The Rg1 allele as a valuable tool for genetic transformation of the tomato 'Micro-Tom' model system.

    abstract:BACKGROUND:The cultivar Micro-Tom (MT) is regarded as a model system for tomato genetics due to its short life cycle and miniature size. However, efforts to improve tomato genetic transformation have led to protocols dependent on the costly hormone zeatin, combined with an excessive number of steps. RESULTS:Here we re...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Pino LE,Lombardi-Crestana S,Azevedo MS,Scotton DC,Borgo L,Quecini V,Figueira A,Peres LE

    更新日期:2010-10-07 00:00:00

  • Rapid, high efficiency virus-mediated mutant complementation and gene silencing in Antirrhinum.

    abstract:Background:Antirrhinum (snapdragon) species are models for genetic and evolutionary research but recalcitrant to genetic transformation, limiting use of transgenic methods for functional genomics. Transient gene expression from viral vectors and virus-induced gene silencing (VIGS) offer transformation-free alternatives...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Tan Y,Bukys A,Molnár A,Hudson A

    更新日期:2020-10-27 00:00:00

  • Development of an innovative and sustainable one-step method for rapid plant DNA isolation for targeted PCR using magnetic ionic liquids.

    abstract:Background:Nowadays, there is an increasing demand for fast and reliable plant biomolecular analyses. Conventional methods for the isolation of nucleic acids are time-consuming and require multiple and often non-automatable steps to remove cellular interferences, with consequence that sample preparation is the major bo...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Marengo A,Cagliero C,Sgorbini B,Anderson JL,Emaus MN,Bicchi C,Bertea CM,Rubiolo P

    更新日期:2019-03-09 00:00:00

  • Non-destructive, high-content analysis of wheat grain traits using X-ray micro computed tomography.

    abstract:Background:Wheat is one of the most widely grown crop in temperate climates for food and animal feed. In order to meet the demands of the predicted population increase in an ever-changing climate, wheat production needs to dramatically increase. Spike and grain traits are critical determinants of final yield and grain ...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Hughes N,Askew K,Scotson CP,Williams K,Sauze C,Corke F,Doonan JH,Nibau C

    更新日期:2017-11-01 00:00:00

  • An efficient system composed of maize protoplast transfection and HPLC-MS for studying the biosynthesis and regulation of maize benzoxazinoids.

    abstract:Background:Insect herbivory poses a major threat to maize. Benzoxazinoids are important anti-insect secondary metabolites in maize, whose biosynthetic pathway has been extensively studied. However, yet little is known about how benzoxazinoids are regulated in maize, partly due to lack of mutant resources and recalcitra...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Gao L,Shen G,Zhang L,Qi J,Zhang C,Ma C,Li J,Wang L,Malook SU,Wu J

    更新日期:2019-11-28 00:00:00

  • Computer aided data acquisition tool for high-throughput phenotyping of plant populations.

    abstract:BACKGROUND:The data generated during a course of a biological experiment/study can be sometimes be massive and its management becomes quite critical for the success of the investigation undertaken. The accumulation and analysis of such large datasets often becomes tedious for biologists and lab technicians. Most of the...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Vankadavath RN,Hussain AJ,Bodanapu R,Kharshiing E,Basha PO,Gupta S,Sreelakshmi Y,Sharma R

    更新日期:2009-12-10 00:00:00

  • Genotype-independent Agrobacterium rhizogenes-mediated root transformation of chickpea: a rapid and efficient method for reverse genetics studies.

    abstract:Background:Chickpea (Cicer arietinum L.), an important legume crop is one of the major source of dietary protein. Developing an efficient and reproducible transformation method is imperative to expedite functional genomics studies in this crop. Here, we present an optimized and detailed procedure for Agrobacterium rhiz...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Aggarwal PR,Nag P,Choudhary P,Chakraborty N,Chakraborty S

    更新日期:2018-07-06 00:00:00