植物生理

Organophosphorus hydrolase (OPH, EC 3.1.8.1) provides a novel function as an alternative genetic marker system for use in many types of plant transformations. OPH is a high-capacity hydrolase with multiple organophosphorus substrates, many of which are neurotoxins and thus used exten ...

Marker-free methods of plant transformation sacrifice the advantages of a selectable marker during regeneration or add work after regeneration to remove the marker. On the positive side, there is no stably integrated marker gene in the plant genome to present regulatory hurdles or poten ...

Targeting exogenously supplied DNA to a predetermined location within a plant genome provides a powerful tool for basic studies of plant gene function and opens up some intriguing possibilities for crop improvement. The induction of double-strand DNA breaks at specific genomic loci via ...

Double-strand breaks are very potent inducers of DNA recombination. There is no recombination between DNA molecules unless one or two DNA strands are broken. It has become feasible to introduce double-strand breaks at specific chromosomal loci by using dedicated, redesigned endonuc ...

Combinatorial nuclear transformation is used to generate populations of transgenic plants containing random selections from a collection of input transgenes. This is a useful approach because it provides the means to test different combinations of genes without the need for separ ...

Transgenic plants have been widely used as expression systems of recombinant proteins in recent years because it can be an efficient alternative for the large-scale production of proteins. This is an area with great potential but is still not much explored. Indeed, this system can bring a breakt ...

The production of biopharmaceuticals in plants is currently one of the most attractive approaches to modern medicine. Several efficient plant-based expression systems have been developed so far. Among them, plastid transformation has attracted biotechnologists because the p ...

A mutated rice acetolactate synthase (mALS) gene expressed under the control of the rice callus-specific promoter (CSP) (CSP:mALS) becomes a useful selectable marker for producing transgenic rice seed with higher accumulation of recombinant protein. When amounts of transgene pro ...

Arabidopsis is naturally adapted to habitats in which both biotic variables (e.g., light, wind, and humidity) and abiotic variables (e.g., competition, herbivory, and pathogen densities) strongly fluctuate. Hence, conditions in controlled growth chambers (in which Arabidopsis is t ...

Detection of protein–protein interactions on a large-scale has become a major focus of functional genomics after the completion of genome sequencing. The information generated from these studies not only assembles proteins into signaling networks, but also reveals potential fun ...

We describe a reliable GAL4-based yeast two-hybrid system for identifying and isolating clones encoding proteins interacting with a protein of interest. This two-hybrid system gives extremely low background and few false-positive clones, making it ideal for library screening pur ...

With the whole genome sequence and thousands of defined polymorphisms between ecotypes available, it has become much easier to clone a gene by position (map-based cloning) in Arabidopsis. Recent development of DNA-isolation methods in plants also dramatically facilitated large-s ...

Magnaporthe grisea is the causal agent of rice blast disease and represents a model organism for the study of fungal plant-pathogen interactions. Pathogenicity is a complex phenotype, which is carefully orchestrated by the fungus and begins with recognition and infection of the host pla ...

Major advances in rice genomics during the last few years have made positional cloning in rice much more efficient. Nipponbare is a model rice genotype being sequenced by the International Rice Genome Sequencing Project Consortium. Here, we describe an efficient procedure of the constru ...

Large-scale genome sequencing projects have generated a wealth of sequence information for plant pathogenic microbes such as oomycetes and fungi. Functional genomic approaches are essential to exploit the sequence information to identify pathogen effector genes that trigger ...

Small RNAs play an important role in plant development, stress responses, and epigenetic regulation, primarily through their role in transcriptional and post-transcriptional silencing of specific target genes and loci. Most if not all plants utilize these small RNA signaling netw ...

In this chapter, we present a brief overview of current knowledge about the promoters of plant microRNAs (miRNAs), and provide a step-by-step guide for predicting plant miRNA promoter elements using known transcription factor binding motifs. The approach to promoter element predicti ...

After transcription, a large number of cellular RNAs employ modifications to increase their diversity and functional potential. Modifications can occur on the base, ribose, or both, and are important steps in the maturation of many RNAs. Our lab recently showed that plant microRNAs (miRNA ...

In eukaryotes, RNA silencing encompasses a range of biochemical processes mediated by ∼20–25 nt small RNAs (smRNAs). This chapter describes northern blot hybridization techniques optimized for detection of such smRNAs, whether extracted from plant or animal tissues. The basic prot ...

Small RNA (sRNA) populations extracted from Arabidopsis plants submitted or not to biotic stress, were reverse-transcribed into cDNAs, and these were subsequently hybridized after labelling to a custom-made DNA tiling array covering Arabidopsis chromosome 4. We first designed a co ...