Microscopy of Physcomitrella patens sperm cells.


BACKGROUND:Archegoniates (bryophytes, ferns and gymnosperms), such as the moss Physcomitrella patens, possess freely motile sperm cells (spermatozoids) which reach the egg cell via surface water. Although these motile flagellated sperm cells are a traditional botanical subject, they have not been thoroughly analysed in the flagship non-seed plant model species P. patens. Protocols are required to determine the behaviour of wild type sperms as a prerequisite for future research such as the characterization of mutants or factors that influence sperm number, morphology, viability and motility. RESULTS:Here, we present protocols for the observation of fixed, as well as live sperms utilizing a standard microscope at intermediate magnifications. Fixed samples can be used for the fast assessment of sperm number and morphology. To determine functionality, the observation of live sperms is required. Protocols for determining both sperm motility and viability are provided, allowing both parameters to be distinguished. CONCLUSIONS:These step-by-step protocols are particularly useful for researchers so far not familiar with the analysis of motile gametes and are meant to aid the establishment and improvement of these analyses in order to stimulate research on spermatogenesis in the moss model species P. patens.


Plant Methods


Plant methods


Horst NA,Reski R




Has Abstract


2017-05-10 00:00:00










  • An efficient transient expression system for gene function analysis in rose.

    abstract:Background:Roses are widely used as garden ornamental plants and cut flowers. Rosa chinensis cv 'Old Blush' has been used as a model genotype in rose studies due to its contribution to recurrent flowering and tea scent traits of modern roses. The deficiency of efficient genetic transformation systems is a handicap limi...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Lu J,Bai M,Ren H,Liu J,Wang C

    更新日期:2017-12-22 00:00:00

  • Use of the non-radioactive SUnSET method to detect decreased protein synthesis in proteasome inhibited Arabidopsis roots.

    abstract:BACKGROUND:In eukaryotic cells, the proteasome maintains homeostasis by selectively degrading regulatory and misfolded proteins, and in doing so contributes to the amino acid pool. Inhibition of the proteasome in yeast and human cells decreases de novo protein synthesis. However, it is not know if proteasome inhibition...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Van Hoewyk D

    更新日期:2016-03-16 00:00:00

  • Barley callus: a model system for bioengineering of starch in cereals.

    abstract:BACKGROUND:Starch is the most important source of calories for human nutrition and the majority of it is produced by cereal farming. Starch is also used as a renewable raw material in a range of industrial sectors. It can be chemically modified to introduce new physicochemical properties. In this way starch is adapted ...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Carciofi M,Blennow A,Nielsen MM,Holm PB,Hebelstrup KH

    更新日期:2012-09-07 00:00:00

  • A new method for functional analysis of plastid EMBRYO-DEFECTIVE PPR genes by efficiently constructing cosuppression lines in Arabidopsis.

    abstract:BACKGROUND:Pentatricopeptide-repeat proteins (PPRs) characterized by tandem arrays of a degenerate 35-amino-acid repeat (PPR motif) can bind a single strand RNA and regulate organelle gene expression at the post-transcriptional level, including RNA cleavage, splicing, editing and stability etc. PPRs are conserved in al...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Chen J,Zhu H,Huang J,Huang W

    更新日期:2020-11-18 00:00:00

  • A rapid, inexpensive, and semi-quantitative method for determining pollen tube extension using fluorescence.

    abstract:BACKGROUND:Pollen tubes extend rapidly when pollen grains are incubated in defined media. Tube extension requires many critical functions of plant cells including molecular signaling, cytoskeleton remodeling, secretion, and cell wall synthesis. Consequently, pollen tube growth has been established as a way to conduct p...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Hartman E,Levy C,Kern DM,Johnson MA,Basu A

    更新日期:2014-01-23 00:00:00

  • Large scale genome skimming from herbarium material for accurate plant identification and phylogenomics.

    abstract:Background:Herbaria are valuable sources of extensive curated plant material that are now accessible to genetic studies because of advances in high-throughput, next-generation sequencing methods. As an applied assessment of large-scale recovery of plastid and ribosomal genome sequences from herbarium material for plant...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Nevill PG,Zhong X,Tonti-Filippini J,Byrne M,Hislop M,Thiele K,van Leeuwen S,Boykin LM,Small I

    更新日期:2020-01-04 00:00:00

  • Analysing complex Triticeae genomes - concepts and strategies.

    abstract::The genomic sequences of many important Triticeae crop species are hard to assemble and analyse due to their large genome sizes, (in part) polyploid genomes and high repeat content. Recently, the draft genomes of barley and bread wheat were reported thanks to cost-efficient and fast NGS technologies. The genome of bar...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Spannagl M,Martis MM,Pfeifer M,Nussbaumer T,Mayer KF

    更新日期:2013-09-06 00:00:00

  • Rapid fluorescent reporter quantification by leaf disc analysis and its application in plant-virus studies.

    abstract:BACKGROUND:Fluorescent proteins are extraordinary tools for biology studies due to their versatility; they are used extensively to improve comprehension of plant-microbe interactions. The viral infection process can easily be tracked and imaged in a plant with fluorescent protein-tagged viruses. In plants, fluorescent ...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Pasin F,Kulasekaran S,Natale P,Simón-Mateo C,García JA

    更新日期:2014-07-05 00:00:00

  • Protocol: genetic transformation of the fern Ceratopteris richardii through microparticle bombardment.

    abstract:BACKGROUND:The inability to genetically transform any fern species has been a major technical barrier to unlocking fern biology. Initial attempts to overcome this limitation were based on transient transformation approaches or achieved very low efficiencies. A highly efficient method of stable transformation was recent...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Plackett AR,Rabbinowitsch EH,Langdale JA

    更新日期:2015-07-03 00:00:00

  • Protocol: An improved high-throughput method for generating tissue samples in 96-well format for plant genotyping (Ice-Cap 2.0).

    abstract:BACKGROUND:We previously developed a high-throughput system called 'Ice-Cap' for growing Arabidopsis seedlings in a 96-well format and rapidly collecting tissue for subsequent DNA extraction and genotyping. While the originally described Ice-Cap method is an effective tool for high-throughput genotyping, one shortcomin...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Clark KA,Krysan PJ

    更新日期:2007-06-12 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

  • Methodology: an optimized, high-yield tomato leaf chloroplast isolation and stroma extraction protocol for proteomics analyses and identification of chloroplast co-localizing proteins.

    abstract:Background:Chloroplasts are critical organelles that perceive and convey metabolic and stress signals to different cellular components, while remaining the seat of photosynthesis and a metabolic factory. The proteomes of intact leaves, chloroplasts, and suborganellar fractions of plastids have been evaluated in the mod...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Bhattacharya O,Ortiz I,Walling LL

    更新日期:2020-09-24 00:00:00

  • Automated phenotyping of plant shoots using imaging methods for analysis of plant stress responses - a review.

    abstract::Current methods of in-house plant phenotyping are providing a powerful new tool for plant biology studies. The self-constructed and commercial platforms established in the last few years, employ non-destructive methods and measurements on a large and high-throughput scale. The platforms offer to certain extent, automa...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Humplík JF,Lazár D,Husičková A,Spíchal L

    更新日期:2015-04-17 00:00:00

  • FRET-SLiM on native autofluorescence: a fast and reliable method to study interactions between fluorescent probes and lignin in plant cell wall.

    abstract:Background:Lignocellulosic biomass is a complex network of polymers making the cell walls of plants. It represents a feedstock of sustainable resources to be converted into fuels, chemicals and materials. Because of its complex architecture, lignocellulose is a recalcitrant material that necessitates some pretreatments...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Terryn C,Paës G,Spriet C

    更新日期:2018-08-27 00:00:00

  • Improved DNase-seq protocol facilitates high resolution mapping of DNase I hypersensitive sites in roots in Arabidopsis thaliana.

    abstract:BACKGROUND:Identifying cis-regulatory elements is critical in understanding the direct and indirect regulatory mechanisms of gene expression. Current approaches include DNase-seq, a technique that combines sensitivity to the nonspecific endonuclease DNase I with high throughput sequencing to identify regions of regulat...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Cumbie JS,Filichkin SA,Megraw M

    更新日期:2015-09-04 00:00:00

  • A novel rind puncture technique to measure rind thickness and diameter in plant stalks.

    abstract:Background:Measurements of rind and culm thickness and stem radius/diameter are important to biomechanical, ecological and physiological plant studies. However, many methods of measuring rind thickness and diameter are labor intensive and induce plant fatality. A novel rind puncture methodology for obtaining measuremen...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Seegmiller WH,Graves J,Robertson DJ

    更新日期:2020-04-01 00:00:00

  • SpikeSegNet-a deep learning approach utilizing encoder-decoder network with hourglass for spike segmentation and counting in wheat plant from visual imaging.

    abstract:Background:High throughput non-destructive phenotyping is emerging as a significant approach for phenotyping germplasm and breeding populations for the identification of superior donors, elite lines, and QTLs. Detection and counting of spikes, the grain bearing organs of wheat, is critical for phenomics of a large set ...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Misra T,Arora A,Marwaha S,Chinnusamy V,Rao AR,Jain R,Sahoo RN,Ray M,Kumar S,Raju D,Jha RR,Nigam A,Goel S

    更新日期:2020-03-18 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 fast, comprehensive and reproducible one-step extraction method for the rapid preparation of polar and semi-polar metabolites, lipids, proteins, starch and cell wall polymers from a single sample.

    abstract:BACKGROUND:The elucidation of complex biological systems requires integration of multiple molecular parameters. Accordingly, high throughput methods like transcriptomics, proteomics, metabolomics and lipidomics have emerged to provide the tools for successful system-wide investigations. Unfortunately, optimized analysi...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Salem MA,Jüppner J,Bajdzienko K,Giavalisco P

    更新日期:2016-11-10 00:00:00

  • Rapid quantification of plant-powdery mildew interactions by qPCR and conidiospore counts.

    abstract:UNLABELLED: BACKGROUND:The powdery mildew disease represents a valuable patho-system to study the interaction between plant hosts and obligate biotrophic fungal pathogens. Numerous discoveries have been made on the basis of the quantitative evaluation of plant-powdery mildew interactions, especially in the context of ...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Weßling R,Panstruga R

    更新日期:2012-08-31 00:00:00

  • Quantifying spatial heterogeneity of chlorophyll fluorescence during plant growth and in response to water stress.

    abstract:BACKGROUND:Effects of abiotic and biotic stresses on plant photosynthetic performance lead to fitness and yield decrease. The maximum quantum efficiency of photosystem II (F v/F m) is a parameter of chlorophyll fluorescence (ChlF) classically used to track changes in photosynthetic performance. Despite recent technical...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Bresson J,Vasseur F,Dauzat M,Koch G,Granier C,Vile D

    更新日期:2015-03-26 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

  • A light in the shadow: the use of Lucifer Yellow technique to demonstrate nectar reabsorption.

    abstract:BACKGROUND:Nectar reabsorption is a widely known phenomenon, related to the strategy of resource-recovery and also to maintain the nectar homeostasis at the nectary. The method currently performed to demonstrate nectar being reabsorbed involves the use of radioactive tracers applied to the nectary. Although this method...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Cardoso-Gustavson P,Robazzi Bignelli Valente Aguiar JM,Ricardo Pansarin E,de Barros F

    更新日期:2013-06-19 00:00:00

  • Isolation of antimicrobial peptides from different plant sources: Does a general extraction method exist?

    abstract::Plants are good sources of biologically active compounds with antimicrobial activity, including polypeptides. Antimicrobial peptides (AMPs) represent one of the main barriers of plant innate immunity to environmental stress factors and are attracting much research interest. There are some extraction methods for isolat...

    journal_title:Plant methods

    pub_type: 杂志文章,评审


    authors: Barashkova AS,Rogozhin EA

    更新日期:2020-10-23 00:00:00

  • An in vitro protocol for rapidly assessing the effects of antimicrobial compounds on the unculturable bacterial plant pathogen, Candidatus Liberibacter asiaticus.

    abstract:Background:Most bacteria are not culturable, but can be identified through molecular methods such as metagenomics studies. Due to specific metabolic requirements and symbiotic relationships, these bacteria cannot survive on typical laboratory media. Many economically and medically important bacteria are unculturable; i...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Krystel J,Shi Q,Shaw J,Gupta G,Hall D,Stover E

    更新日期:2019-07-31 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

  • A new approach to varietal identification in plants by microsatellite high resolution melting analysis: application to the verification of grapevine and olive cultivars.

    abstract:BACKGROUND:Microsatellites are popular molecular markers in many plant species due to their stable and highly polymorphic nature. A number of analysis methods have been described but analyses of these markers are typically performed on cumbersome polyacrylamide gels or more conveniently by capillary electrophoresis on ...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Mackay JF,Wright CD,Bonfiglioli RG

    更新日期:2008-05-19 00:00:00

  • A software tool for the input and management of phenotypic data using personal digital assistants and other mobile devices.

    abstract:BACKGROUND:Plant breeding and genetics demand fast, exact and reproducible phenotyping. Efficient statistical evaluation of phenotyping data requires standardised data storage ensuring long-term data availability while maintaining intellectual property rights. This is state of the art at phenomics centres, which, howev...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Köhl K,Gremmels J

    更新日期:2015-04-07 00:00:00

  • Genome-wide identification of microsatellites in white clover (Trifolium repens L.) using FIASCO and phpSSRMiner.

    abstract:BACKGROUND:Allotetraploid white clover (Trifolium repens L.) is an important forage legume widely cultivated in most temperate regions. Only a small number of microsatellite markers are publicly available and can be utilized in white clover breeding programs. The objectives of this study were to develop an integrated a...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Zhang Y,He J,Zhao PX,Bouton JH,Monteros MJ

    更新日期:2008-07-16 00:00:00

  • Protocol: Streamlined sub-protocols for floral-dip transformation and selection of transformants in Arabidopsis thaliana.

    abstract:: Generating and identifying transformants is essential for many studies of gene function. In Arabidopsis thaliana, a revolutionary protocol termed floral dip is now the most widely used transformation method. Although robust, it involves a number of relatively time-consuming and laborious steps, including manipulating...

    journal_title:Plant methods

    pub_type: 杂志文章


    authors: Davis AM,Hall A,Millar AJ,Darrah C,Davis SJ

    更新日期:2009-02-27 00:00:00