植物生理

Plant transformation is the process by which DNA is introduced into plant cells or tissues. The DNA can come from virtually any source. Gene transfer methodology has become part of an essential technology to manipulate plants for both scientific and commercial purposes. Transgenic plants, ...

Agrobacterium rhizogenes-mediated transformation results in the development of hairy roots at the site of infection. The production of hairy roots involves cocultivation of explants with A. rhizogenes and the subsequent selection of hairy roots on hormone-free medium. Hairy roo ...

Particle bombardment, or biolistics, is a commonly used method for genetic transformation of plants and other organisms. Millions of DNA-coated metal particles are shot at target cells or tissues using a biolistic device or gene gun. The DNA elutes off the particles that lodge inside the cells, a ...

An optimized protocol for the electroporation-based transfection of tobacco protoplasts is described that routinely results in transgene expression frequencies approaching 90%. The overall efficiency of the procedure depends collectively on numerous key parameters, in ...

Many researchers use the flowering plant Arabidopsis thaliana to study gene function and basic plant biology. This easy-to-grow, small plant is ideal for genetic studies as it has a relatively simple genome compared to crop plants and its genetic material has been recently sequenced. Anoth ...

The application of floral dipping toward the production of transformed plants has been rather limited. However, this procedure has enabled the successful production of transformed Medicago truncatula plants (a model plant for legume genetics) at efficiencies higher than those ob ...

Production of transgenic plants is gradually becoming routine in cassava biotechnology. Green cotyledons of maturing somatic embryos (somatic cotyledons for short) and friable embryogenic suspensions (FES) are the target tissues for transformation by Agrobacterium or bio ...

Major crop losses occur annually as a result of biotic and abiotic stresses. The ability to hyperexpress foreign proteins, single-step multigene engineering, lack of positive effect and gene silencing, vector sequences and pleiotropic effects have resulted in several hundred-fold ...

Over the last 5 yr, the production of transgenic conifers has been greatly facilitated by the ability to transform somatic embryonal tissues (somatic embryos) via cocultivation with Agrobacterium tumefaciens. This has allowed us to develop protocols for the genetic transformation ...

Most woody fruit species have long juvenile periods that drastically prolong the time required to analyze mature traits. Evaluation of characteristics related to fruits is a requisite to release any new variety into the market. Because of a decline in regenerative and transformation pot ...

Tomato was one of the first crops for which a genetic transformation system was reported involving regeneration by organogenesis from Agrobacterium-transformed explants. Since the initial reports, various factors have been studied that affect the efficiency of tomato transfo ...

Transgenic plant production has been intimately connected to the β-glucuronidase (uidA or GUS) gene used as a reporter marker gene. The enzyme stability and the high sensitivity and amenability of the GUS assay to qualitative (histochemical assay) and to quantitative (fluorometric or s ...

The low efficiencies of most plant transformation methods necessitate the use of selectable marker genes to identify those cells that successfully integrate and express transferred DNA. Genes conferring resistance to various antibiotics or herbicides are commonly used in labo ...

As future biotechnology applications utilize recombinant proteins as commercial products, nondestructive assays will be necessary to determine protein concentrations accurately within plant tissues. Green fluorescent protein (GFP) has been proposed as a potential mar ...

The use of a new mode of selection—positive selection—has been demonstrated to be successful in a large variety of monocot and dicot species. This selection differs from more traditional modes of selection in which compounds such as antibiotics or herbicides are used to kill nontransformed ...

We have developed an efficient system (Multi-Auto-Transformation vectors) for the removal of marker genes and to increase the regeneration frequency of transgenic crops without using antibiotic selection, reducing their possible environmental impact. The MAT vector system is ...

Removal of marker genes improves the design of transgenic plants. Homologous recombination between direct repeats provides a simple method for excising marker genes after transgenic cells and shoots have been isolated. Efficient implementation of the method requires high rates of ...

The development of efficient crop transformation systems has necessitated the development of efficient methods for detailed molecular characterization of putative events. This chapter details the routine use of quantitative real-time polymerase chain reaction to determ ...

Recent years have seen a huge increase and improvement in techniques for analysis of transgene expression in plants. The analysis of RNA frequently provides a vital link between changes in enzyme levels and/or metabolites and the phenotype. This chapter focuses on RNA-based techniques for ...

Matrix attachment regions (MARs) are operationally defined as DNA elements that bind specifically to the nuclear matrix in vitro. When MARs are positioned at the 5′- and 3′-ends of a transgene higher more predictable expression of the transgene results. MARs are increasingly being applied to ...