植物生理

Controlled transcription of biosynthetic genes is one major mechanism regulating alkaloid production in plant cells. This regulation of biosynthetic pathways is achieved by specific transcription factors. Sequence-specific DNA-binding proteins interact with the prom ...

Regulation of gene expression is largely coordinated by a complex network of interactions between transcription factors (TFs), co-factors, and their cognate cis-regulatory elements in the genome. TFs are multidomain proteins that arise evolutionarily through protein domain s ...

The yeast one-hybrid system is widely recognized as a valuable and straightforward technique to study interactions between transcription factors and DNA. By means of one-hybrid screens, transcription factors or other DNA-binding proteins, expressed from cDNA expression libra ...

A powerful method to identify binding sites in target genes is chromatin immunoprecipitation (ChIP), which allows the purification of in vivo formed complexes of a DNA-binding protein and associated DNA. Briefly, the method involves the fixation of plant tissue and the isolation of the tot ...

Metabolomics is a new genomics approach that aims at measuring all or a subset of metabolites in the cell. Several approaches to plant metabolomics are currently used in plant research. These include targeted analysis, metabolite profiling, and metabolic fingerprinting. Metabolic f ...

The rich collection of known genetic information and the recent completion of rice genome sequencing project provided the cereal plant researchers a useful tool to investigate the roles of genes and genomic organization that contribute to numerous agronomic traits. Gramene (http:// ...

Next-generation sequencing has revolutionized biology by exponentially increasing sequencing output while dramatically lowering costs. High-throughput sequence data with shorter reads has opened up new applications such as whole genome resequencing, indel and SNP dete ...

microRNAs (miRNAs) are a new class of small endogenous noncoding regulatory RNAs, which play an important function in plant growth, development, phase change, and response to environmental stress. Identifying miRNAs is the first step for investigating miRNA-mediated gene regulat ...

Quantitative real-time PCR (qRT-PCR), in conjunction with reverse transcriptase, has been used for the systematic measurement of plant physiological changes in gene expression. In the present paper, we describe a qRT-PCR protocol that illustrates the essential technical steps req ...

Gene expression profiling has revolutionized functional genomics research by providing a quick handle on all the transcriptional changes that occur in the cell in response to internal or external perturbations or developmental programs. Microarrays have become the most popular ...

Agroinoculation, first developed as a simple tool to study plant–virus interactions, is a popular method of choice for functional gene analysis of viral genomes. With the explosive growth of genomic information and the development of advanced vectors to dissect plant gene function, this ...

Virus-induced gene silencing (VIGS) is an efficient tool for high throughput reverse genetic screens. VIGS engages the endogenous RNA-silencing machinery of the plant host, and can yield an 85–95% reduction of target transcripts. Gene silencing is rapid, target-specific, and does not re ...

Full-length cDNAs (fl-cDNAs) are important resources for the characterization of gene function, since they contain all the information required for the production of functional RNAs and proteins. Large sets of fl-cDNA clones have been collected from several plant species and have beco ...

Activation tagging is a powerful strategy to find new gene functions, especially from genes that are redundant or show lethal knock-out phenotypes. It has been applied using T-DNA or transposons. En/Spm-I/dSpm engineered transposons are efficient Activation tags in Arabidopsis. An im ...

The process of activation tagging in plants involves the random distribution of plant regulatory sequences throughout the genome. The insertion of a regulatory sequence in the vicinity of an endogenous gene can alter the transcriptional pattern of this gene resulting in a mutant phenot ...

A high throughput drought screen is described for Arabidopsis that is based on a gravimetric method to monitor and control water content of the soil. To screen for plant growth under mild drought conditions, 30% of field capacity can be used, which is equal to 2 g H2O/g dry soil. The screen allows the testing of a l ...

Medicago truncatula has been chosen as one of the two model species for legume molecular genetics and functional genomics studies. With the imminent completion of M. truncatula genome sequencing, availability of large-scale mutant populations becomes a priority. Over the last 5 years, n ...

Insertion mutants offer one of the direct ways to relate a gene to its function by employing forward or reverse genetics approaches. Both T-DNA and transposon insertional mutants are being produced in several crops, including rice, the first cereal with its complete genome sequenced. Trans ...

The central goal of current genomics research in plants, as in other organisms, is to elucidate the functions of every gene. Insertional mutagenesis using known DNA sequences such as T-DNA is a powerful tool in functional genomics. Development of efficient methods for isolating the genomic s ...

With the completion of the rice genome sequencing project, the next major challenge is the large-scale determination of gene function. A systematic phenotypic profiling of mutant collections will provide major insights into gene functions important for crop growth or production. Th ...