植物学技术

Laser microdissection is a powerful tool to obtain cell-specific isolates from complex tissue samples. This chapter outlines how to prepare plant material for microdissection and methods to extract and measure high-quality RNA suitable for a variety of different downstream appli ...

Microarrays provide the ability to quantitatively measure the abundance of specific RNA transcripts through sample hybridization to a solid-state grid of oligonucleotides or amplicons. The prospect of measuring the entire transcriptome is extremely alluring, but as with any ex ...

Protocols for the proteome analysis of Arabidopsis tissues based on 2 D gel electrophoresis are given for leaves and roots as well as for seeds. The protocols contain a detailed description of the sample preparation step and of the solubilization of proteins. We then describe our protocols for the ...

Proteome analysis is becoming a powerful tool of discovery-driven research, with investigations ranging from whole organisms to specific subcellular compartments. Especially for the latter, efficient and robust methods for protein purification are the prerequisite for obt ...

Proteomics is a very powerful approach to link the information containe3|d in sequenced genomes, such as Arabidopsis, to the functional knowledge provided by studies of plant cell compartments. However, membrane proteomics remains a challenge. One way to bring into view the complex mixt ...

Large-scale and high-throughput approaches play an essential role in the study of the highly complex biological systems. In plant biology, obtaining the complete genome sequence of the model plant Arabidopsis thaliana has definitively encouraged the incorporation of a completely ...

The identification of protein phosphorylation sites has always been a challenging task, traditionally involving large amounts of radioactive phosphorus and high-performance liquid chromatography separation and Edman sequencing of phosphopeptides. The rapid develo ...

Metabolite profiling is the multiparallel relative quantification of a mixture of compounds or compound classes using chromatography and universal detection technologies (GC-MS, LC-MS). In this respect it is an extension of classical single-target methods from which it can be dist ...

A highly sensitive and accurate multiplex gas chromatography-mass spectrometry (GC-MS/MS)-technique is reported for the quantitative analysis of acidic phytohormones in Arabidopsis thaliana and other plant species. The optimized setup allows the routine processing and an ...

The size of the chloroplast DNA molecule of Arabidopsis thaliana has been determined to be about 153 kb (1). In this chapter, the goal is to provide a step—by—step laboratory procedure for isolating chloroplast DNA from Arabidopsis thaliana with minimal nuclear or mitochondrial DNA contami ...

Plant mttochondrtal genomes vary m size and complextty. Both can be rapidly estimated by analysis of the restriction pattern of purified mitochondrial DNA (mtDNA). A prerequisite to purify mtDNA with little contamination of nuclear and also of plastid DNA is the isolatton of mitochondrta ...

The preparation of high quality (i.e., intact) total RNA from biological samples 1s the primary step in the study of gene expression. It is the procedure that bridges the interface between the experimental manipulation of the hvmg system, and the subsequent analysis of effects through molecul ...

Only structurally intact chloroplasts with a functional envelope exhibit metabolic activities comparable to those of the original tissues. Therefore, the most important objectives that should guide the isolation of chloroplasts from any plant species are then morphological a ...

Plant cells contam different subcellular compartments, which serve distinct phystologrcal functions. One of these organelles, the mitochondrion, provides most of the nonphotosynthetrc energy required m the cell. Mitochondria contain their own genome(s), but encode only a small ...

The generation of mutations is the most basic element of genetic analysis. Mutagenesis is the process by which heritable alterattons in the genome of an organism, mutations, are produced. In order to conduct genetic analysis, at least two alleles for a given locus must exist and the facile product ...

Successful extraction of DNA is an essential and time consuming step in many plant molecular biology procedures. The classical approaches of plant DNA isolation are often designed to produce large amounts of DNA of high molecular weight with sufficient purity. Since the polymerase chain r ...

The cytogenetrcal analysis of a species enables the correlation of the structure, number and morphology of its chromosomes with heredity and variation. Most accessions of Arabidopszs thaliana L. have a diploid (2n) chromosome number of 10 (Fig. 1). The small genome size, convenient for many as ...

The genetic behavior of Arabidupsis is not different from that of other diploid organisms, and therefore one can find the topics of this chapter in almost any handbook of genetics. It is only owing to the specific (reproductive) biology of a species that sometimes a certain analytical procedure is ...

With large-scale sequencing projects (1,2), thousands of unique sequences ESTs; expressed sequence tags) have been generated. In addition, hundreds of sequences have been identified based on then homology with a gene of known interest or from subtractive hybridization procedures. F ...

To locate (map) genes a populatton segregating for such genes is required. In many cases, a specrfic population 1s constructed to map a particular trait or locus, e.g., identified as a mutant. However, in other sttuattons the gene(s) to be mapped do not show a specific phenotype and only polymorphtsms at t ...