植物生理

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 ...

AFLP− (KeyGene, Wagenmgen, The Netherlands), is a DNA fingerprmtmg technique that visualizes DNA restriction fragments by polymerase chain reaction (PCR) amphfication (1,2). Routinely, 50-100 restriction fragments are amphfied simultaneously and detected on denaturing po ...

The AFLP− (Keygene, Wageningen, The Netherlands) technology allows the simultaneous amplification of many restriction fragments in a single polymerase chain reaction (PCR). Thus, the technology can be used to very efficiently detect restriction fragment polymorphisms. As an exa ...

Plant genetic maps can be constructed either wrth phenotyprc or molecular markers. Historically, genetic mapping utrlrzed vtsrble markers, but it is difficult to examme many such markers m a single cross. The recognmon that distantly related individuals differ m DNA sequence througho ...

Except the fact that part of T-DNA- or transposon-induced mutations can be directly identified via the T-DNA or transposon tag (see Chapters 32–34) non-tagged mutations and mutations induced by chemicals or radiation need to be mapped to chromosomal regions as a first step towards their furth ...

In vrvo footprintmg is a method of observmg DNA/protein mteractions of a gene within the context of the livmg cell. In other words, in vivo footprintmg provides an opportunity to observe conformational changes in chromatm associated with the transcriptional machinery of a cell as it responds ...

The development of technologies that allow the introduction and functional expression of foreign genes in plant cells has extended in less than two decades to the production of transgenic plants with improved insect and disease resistance, seeds and fruits with enhanced nutritional q ...