Cereal Genomics: Methods and Protocols

  title={Cereal Genomics: Methods and Protocols},
  author={Robert James Henry and Agnelo Furtado},
  journal={Cereal Genomics},
1. Dna Extraction from Vegetative Tissue for Next Generation Sequencing Agnelo Furtado 2. DNA Extraction from Rice Endosperm (Including a Protocol for Extraction of DNA from Ancient Seed Samples) Chiaki Mutou, Katsunori Tanaka, Ryuji Ishikawa 3. RNA Extraction from Cereal Vegetative Tissue Julie A. Pattemore 4. RNA Extraction from Developing or Mature Wheat Seeds Agnelo Furtado 5. cDNA Library Preparation Maarten Kooiker and Gang-Ping Xue 6. Preparation of High Molecular Weight gDNA and… 

Acceleration of the Breeding Program for Winter Wheat

Two approaches that can help plant breeders hasten cultivar development for winter wheat are described, which show the potential to reduce the development of cultivars using six generations of backcrossing with gene introgression by 2 years.

A modular cloning toolkit for genome editing in plants

An expanded cloning toolkit that contains 103 modules encoding a variety of CRISPR/Cas-based nucleases and their corresponding guide RNA backbones is presented that contains a set of modules that enable quick and facile assembly of tRNA-sgRNA polycistronic units without a PCR step involved.

Gene transfer to plants by electroporation: methods and applications

In modern biotechnology, numerical methods have been used in electrotransformation, electroporation-based inactivation, electroextraction, and electroporative biomass drying, where numerical models to iteratively optimize the field homogeneity considering the cell size, shape, number, and electrode positions supplemented by real-time measurements.

The wheat Sr22, Sr33, Sr35 and Sr45 genes confer resistance against stem rust in barley

Assessment of the functionality, in barley, of cloned wheat Sr genes effective against Ug99 found all four genes were found to confer effective stem rust resistance and are therefore a potential source of resistance against wheat stem rust in barley.

Calmodulin HvCaM1 Negatively Regulates Salt Tolerance via Modulation of HvHKT1s and HvCAMTA41[OPEN]

It is proposed that HvCaM1 is involved in regulating Na+ transport, probably via certain class I high-affinity potassium transporter (HvHKT1;5 and HvHK T1;1)-mediated Na+ translocation in roots, which may be a critical factor in the regulation of HKT1s in barley.

Biotechnological advances for improving natural pigment production: a state-of-the-art review

In this review, the innovative methods and strategies for optimization and engineering of both native and heterologous producers of natural pigments are comprehensively summarized.

Transgene expression and plant regeneration in the Brassicaceae Aethionema arabicum

Transformation, der Einbau zusatzlicher DNA Abschnitte, ist eines der wichtigsten Werkzeuge in der modernen Molekularbiologie and Genetik, um Pflanzengenome modifizieren und folglich die Funktion und



DNA extraction from rice endosperm (including a protocol for extraction of DNA from ancient seed samples).

An optimal procedure to process archaeological grain specimens, aged for a couple of thousand years, to isolate DNA from these ancient samples, referred to here as ancient DNA (aDNA), can be successfully amplified by PCR using appropriate primer pairs designed specifically for aDNA amplification.

Transcription Activator-Like Effector Nucleases Enable Efficient Plant Genome Engineering1[W][OA]

The optimized reagents implemented in plant protoplasts should be useful for targeted modification of cells from diverse plant species and using a variety of means for reagent delivery.

Step-by-step protocols for rice gamete isolation

Detailed, step-by-step procedures for collecting intact, unfixed rice egg cells and sperm cells without enzymatic treatments are presented and a general workflow for assessing sample purity by RT-PCR is presented, using primers specific for marker genes preferentially expressed in surrounding cells and tissues.

Preparation of high molecular weight gDNA and bacterial artificial chromosome (BAC) libraries in plants.

This work presents a step-by-step protocol for isolation of HMW DNA and construction of plant BAC libraries, and highlights the importance of knowing the isolation status of intact high molecular weight DNA.

DNA-Free Genetically Edited Grapevine and Apple Protoplast Using CRISPR/Cas9 Ribonucleoproteins

It is demonstrated that direct delivery of purified CRISPR/Cas9 RNPs to the protoplast system enables targeted gene editing and paves the way to the generation of DNA-free genome edited grapevine and apple plants.

Analysis of the bread wheat genome using whole genome shotgun sequencing

It is shown that the hexaploid genome is highly dynamic, with significant loss of gene family members on polyploidization and domestication, and an abundance of gene fragments.

A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes

A simplified and highly efficient transient gene expression system using photosynthetically active rice green tissue protoplasts and its broad applications in protein immunoblot, localization and protein-protein interaction assays are shown.

DNA extraction from vegetative tissue for next-generation sequencing.

This chapter describes a simple two-reagent DNA extraction protocol which yields a high quality and quantity of DNA which can be used for different applications including NGS.

Genetic transformation of wheat: current status and future prospects

The objective of this review is to summarize current successful techniques for wheat transformation and stimulate further research into long-term wheat improvement by genetic engineering approaches.