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Plant Germline Development

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Cover of 'Plant Germline Development'

Table of Contents

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    Book Overview
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    Chapter 1 An Introduction to Male Germline Development
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    Chapter 2 Apomixis: Engineering the Ability to Harness Hybrid Vigor in Crop Plants
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    Chapter 3 3D Imaging of Whole-Mount Ovules at Cellular Resolution to Study Female Germline Development in Rice
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    Chapter 4 Live-Cell Imaging of F-Actin Dynamics During Fertilization in Arabidopsis thaliana
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    Chapter 5 Development and Observation of Mature Megagametophyte Cell-Specific Fluorescent Markers
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    Chapter 6 Analysis of Fluorescent Reporter Activity in the Male Germline During Pollen Development by Confocal Microscopy
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    Chapter 7 In Vivo Ploidy Determination of Arabidopsis thaliana Male and Female Gametophytes
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    Chapter 8 Staining and Clearing of Arabidopsis Reproductive Tissue for Imaging of Fluorescent Proteins
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    Chapter 9 Live-Cell Imaging of Auxin and Cytokinin Signaling in Maize Female Gametophytes
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    Chapter 10 Imaging Ca2+ Dynamics in Wild-Type and NADPH Oxidase-Deficient Mutant Pollen Tubes with Yellow Cameleon and Confocal Laser Scanning Microscopy
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    Chapter 11 Immunolocalization of AGPs and Pectins in Quercus suber Gametophytic Structures
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    Chapter 12 Optimization of Cell Spreading and Image Quality for the Study of Chromosomes in Plant Tissues
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    Chapter 13 Whole Mount RNA-FISH on Ovules and Developing Seeds
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    Chapter 14 Analysis of Peroxisome Biogenesis in Pollen by Confocal Microscopy and Transmission Electron Microscopy
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    Chapter 15 Transmission Electron Microscopy (TEM) to Study Histology of Pollen and Pollen Tubes
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    Chapter 16 Isolation of Arabidopsis Pollen, Sperm Cells, and Vegetative Nuclei by Fluorescence-Activated Cell Sorting (FACS)
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    Chapter 17 Isolation of Rice Sperm Cells for Transcriptional Profiling
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    Chapter 18 Manual Isolation of Living Cells from the Arabidopsis thaliana Female Gametophyte by Micromanipulation
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    Chapter 19 Isolation and Detection of Small RNAs from Pollen
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    Chapter 20 Analysis of Proteins Enriched in Rice Gamete
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    Chapter 21 Phosphoprotein Enrichment from Tobacco Mature Pollen Crude Protein Extract
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    Chapter 22 Identification of Cis-Regulatory Modules that Function in the Male Germline of Flowering Plants
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    Chapter 23 RNA-Seq Data Analysis: From Raw Data Quality Control to Differential Expression Analysis
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    Chapter 24 RNA-Seq Data Analysis Protocol: Combining In-House and Publicly Available Data
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    Chapter 25 Impedance Flow Cytometry as a Tool to Analyze Microspore and Pollen Quality
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    Chapter 26 Agrobacterium-Mediated Sorghum Transformation
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    Chapter 27 Use of the Cas9 Orthologs from Streptococcus thermophilus and Staphylococcus aureus for Non-Homologous End-Joining Mediated Site-Specific Mutagenesis in Arabidopsis thaliana
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    Chapter 28 Transgenic Reproductive Cell Ablation
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    Chapter 29 In Vivo Reporters for Protein Half-Life
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    Chapter 30 Conditional Modulation of Biological Processes by Low-Temperature Degrons
Attention for Chapter 13: Whole Mount RNA-FISH on Ovules and Developing Seeds
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Chapter title
Whole Mount RNA-FISH on Ovules and Developing Seeds
Chapter number 13
Book title
Plant Germline Development
Published in
Methods in molecular biology, January 2017
DOI 10.1007/978-1-4939-7286-9_13
Pubmed ID
Book ISBNs
978-1-4939-7285-2, 978-1-4939-7286-9
Authors

Andrea Bleckmann, Thomas Dresselhaus

Abstract

A key element to understand developmental and reproductive processes like germline development, double fertilization, and embryogenesis is the study of cell-specific gene expression patterns which is best analyzed by RNA in situ hybridization. Different visualization techniques have been established to mark either the region of mRNA production (using the classical chromogenic detection system) or the specific localization of mRNAs (using fluorescent labeled probes). In this chapter, we describe and compare whole mount RNA in situ hybridization techniques on ovules and young developing seeds from Arabidopsis thaliana using three different detection systems. The alkaline phosphatase (AP) coupled antibody detecting the antigen labeled probe facilitates the production of a precipitating dye indicating mRNA presence: (1) using BCIP/NBT as substrates, it is converted to a blue staining that can be visualized using differential interference contrast (DIC) microscopy. Alternatively, (2) using Fast-Red as a substrate it is converted to a purple fluorescent staining that can be visualized either by light microscopy or, for a higher cellular resolution, by confocal microscope. To analyze mRNA distribution with subcellular resolution we (3) describe a third, highly sensitive fluorescent detection system, which is based on the enzymatic activity of a peroxidase. In combination with a tyramide signal amplification (TSA) system, it leads to multi-fluorescent labeled antibodies marking the mRNA bound probe locally.

Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 15 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 15 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 4 27%
Researcher 3 20%
Student > Bachelor 2 13%
Other 1 7%
Student > Master 1 7%
Other 1 7%
Unknown 3 20%
Readers by discipline Count As %
Agricultural and Biological Sciences 7 47%
Biochemistry, Genetics and Molecular Biology 4 27%
Unknown 4 27%