植物生理

Sequencing of mRNA using next-generation sequencing (NGS) technologies (RNA-seq) has the potential to reveal unprecedented complexity of the transcriptomes. The transcriptome sequencing of an organism provides quick insights into the gene space, opportunity to isolate genes ...

Assessment of gene function oftentimes requires mutant populations that can be screened by forward or reverse genetic analysis. The situation becomes more complicated in polyploidy or paleopolyploid genomes that have two or more copies for most genes. Here we describe a method for engin ...

A growing number of international genome consortia have initiated large-scale sequencing projects for most of the major crop species. This huge amount of information not only boosted genetic and physical mapping research, but it also enabled novel applications on the level of chromoso ...

The primary model legumes to date have been Medicago truncatula and Lotus japonicus. Both species are tractable both genetically and in the greenhouse, and for both, substantial sets of tools and resources for molecular genetic research have been assembled. As sequencing costs have decli ...

Metabolomics is one of the most recent additions to the functional genomics approaches. It involves the use of analytical chemistry techniques to provide high-density data of metabolic profiles. Data is then analyzed using advanced statistics and databases to extract biological in ...

Posttranslational modifications represent a major mechanism by which cells control molecular events. Sample preparation is one of the biggest keys in terms of proteomic analysis, and especially when looking at phosphoproteomics. Here we describe two techniques that are used to ide ...

Application of proteomics is becoming increasingly important to understand the function of genes and their encoding proteins. This is due to not only the poor correlation between the transcript levels and protein accumulation, but also the critical roles of posttranslational modif ...

The recent and massive expansion in plant genomics data has generated a large number of gene sequences for which two seemingly simple questions need to be answered: where do the proteins encoded by these genes localize in cells, and what do they do? One widespread approach to answering the localiza ...

Transient expression is a powerful method for the functional characterization of genes. In this chapter, we outline a protocol for the transient expression of constructs in Medicago truncatula leaves using Agrobacterium tumefaciens infiltration. Using quantitative real-ti ...

Medicago truncatula is a model legume that has all the genomic resources to be suitable as a model for functional genomics. Transformation to produce transgenic plants is part of this toolkit, enabling a spectrum of approaches to study gene function: including knockdown, overexpression, ...

Since the development of molecular markers to tag genes of agronomic traits of interests, molecular markers have played an increasingly significant role in breeding programs. Molecular markers have been implemented for large-scale marker-assisted selection in the breeding pro ...

Medicago truncatula is used widely as a model system for studies of root symbioses, interactions with parasitic nematodes and fungal pathogens, as well as studies of development and secondary metabolism. In Medicago truncatula as well as other legumes, RNA interference (RNAi) coupled w ...

Understanding of developmental processes relies heavily on isolation and functional characterization of relevant genes. The garden pea (Pisum sativum L.) is one of the classic model species in plant genetics and has been used for a wide range of physiological and molecular studies of plant ...

The Lotus Retrotransposon 1 (LORE1) is used for genome-wide mutagenesis of the model legume Lotus japonicus. Characterization of the LORE1 insertion sites in individual mutant lines is critical for development and use of the resource. Here we present guidelines for use of the LORE1 reverse g ...

Gas chromatography–mass spectrometry (GC-MS)-based analysis is an accurate and sensitive method to quantify plant hormones. This method is commonly used for analysis of low-molecular-weight compounds, such as abscisic acid (ABA), gibberellins (GAs), auxins, and brassinoster ...

The plant hormone abscisic acid (ABA) is a major growth regulator mediating several aspects of plant development and stress-induced processes. For the hormone to function within the cell, it must first interact with a proteinaceous receptor to commence transmission of a signal to downstr ...

BRI1, a leucine-rich repeat (CRR)-receptor kinase protein, belongs to the plant hormone brassinosteroid (BR) receptor complex in Arabidopsis thaliana (Arabidopsis). The experiments described in this chapter to carry out BR-binding assays in Arabidopsis plants are largely based ...

The cytokinin class of plant hormones is perceived by transmembrane His-kinases (His-kinases) of the two-component system, otherwise known as the His-Asp phosphorelay system. When cytokinin receptors perceive cytokinins, they are autophosphorylated at a conserved His residu ...

Abscisic acid (ABA) plays a number of key roles in the growth, development, and stress response of plants. For example, it is vital to a plant’s response to drought stress, and is the signalling molecule responsible for closure of the stomata in order to promote water conservation. The hormone is rapidly ...

Analysis of large-scale gene expression data sets is proving to be a powerful tool for gene function prediction, cis-element discovery and hypothesis generation using Arabidopsis thaliana. Public initiatives led by the AtGenExpress Consortium and experiments conducted by indi ...