植物学技术

As aerobic chemoorganotrophs, most Agrobacterium strains will grow on a wide range of complex and defined media. Methods commonly used for the culture and storage of other chemoorganotrophs will usually work for agrobacteria as well. Problems with culture or strain maintenance will oc ...

A binary vector is a standard tool in the transformation of higher plants mediated by Agrobacterium tumefaciens. It is composed of the borders of T-DNA, multiple cloning sites, replication functions for Escherichia coli and A. tumefaciens, selectable marker genes, reporter genes, and oth ...

The genetic manipulation of Agrobacterium tumefaciens is used to facilitate studies of bacterial gene functions or as a first step in introducing genetic material into transformable plant cells through the use of T-DNA binary vectors. Three methods are commonly used. Transformation ...

Agrobacterium tumefaciens has been widely used to transform numerous plant species. Frequently, investigators want to place more than one gene into Agrobacterium in order to manipulate various bacterial functions during plant genetic transformation. These genes are frequen ...

The ability of Agrobacterium to transform plants and other organisms is under highly regulated genetic control. Two Virulence (Vir) proteins, VirA and VirG, function as a two-component regulatory system to sense particular phenolic compounds synthesized by wounded plant tissues. I ...

Agrobacterium is routinely used as a tool for moving genetic constructs into plant cells. The successful use of Agrobacterium as a tool for the genetic engineering of plant cells often requires the manipulation and analysis of nucleic acids present in recombinant Agrobacterium strains. ...

Medicago truncatula has been developed into a model species for legumes. Large numbers of ESTs have been sequenced, and sequencing of the complete gene-rich space of the genotype Jemalong A17 is in progress. By using cotyledons as explants for Agrobacterium infection and direct shoot forma ...

Agrobacterium-mediated transformation of Nicotiana species, namely, Nicotiana tobaccum and Nicotiana benthamiana, using leaf disks as the target explant has provided the plant community with a valuable tool for rapid evaluation of transgenes in higher plants. This protocol has a ...

Medicago truncatula is a forage crop that has been developed into a model legume. As a model plant, M. truncatula is particularly useful for the study of root endosymbiotic associations, including nodulation and mycorrhizal colonization. The development of different transformation ...

Limitations in transformation capability can be a significant barrier in making advances in our understanding of gene function through the use of transgenics. To this end we have developed both tissue culture and non-tissue culture-based methodologies for the production of transge ...

Arabidopsis floral dip transformation is notable for a number of reasons. First, it is strikingly simple to perform. Agrobacterium is applied to flowering Arabidopsis plants that subsequently set seed, and transgenic plants are then selected among the progeny seedlings. Because no pl ...

Crop improvement is limited by the availability of valuable traits in sexually compatible species. Access to new characters using genetic engineering would be of great value. Barley has been transformed using microprojectile bombardment and by direct gene transfer to protoplasts, ...

Legumes have long been recalcitrant to efficient Agrobacterium tumefaciens-mediated transformation. The choice and use of model legume plants (Medicago truncatula and Lotus japonicus) for molecular studies has triggered extensive studies devoted to the development of eff ...

Agrobacterium tumefaciens-mediated transformation is an effective method for introducing genes into maize. In this chapter, we describe a detailed protocol for genetic transformation of the maize genotype Hi II. Our starting plant material is immature embryos cocultivated with ...

Rice is the world’s most important food crop. Indica-type rice provides the staple food for more than half of the world population. To satisfy the growing demand of the ever-increasing population, more sustained production of indica-type rice is needed. In addition, because of the high per capita ...

Rye (Secale cereale L.) is one of the most recalcitrant plant species for tissue culture and genetic transformation. Embryogenic rye callus loses its ability to regenerate plants quickly in response to high density of Agrobacterium and other stressors. The cocultivation of Agrobacter ...

Agrobacterium-mediated transformation of rice is now used in many laboratories worldwide. Several protocols have been developed and fine-tuned for particular genotypes, including commercial genotypes, making use of either mature seeds or immature embryos as target tissue for ...

This chapter describes a procedure for Agrobacterium-mediated wheat transformation. Freshly isolated immature embryos, precultured immature embryos, or embryogenic calli are inoculated with a disarmed A. tumefaciens strain C58 (ABI) harboring the binary vector pMON18365 ...

High-frequency Agrobacterium tumefaciens-mediated transformation can be obtained in canola by optimizing the preconditioning time of the explant and cocultivation time with A. tumefaciens. A preconditioning time of 72 h and cocultivation of 48 h synergistically increase the t ...

All economically important Brassica species have been successfully transformed using Agrobacterium tumefaciens. Although different tissues have been used as explants, hypocotyls remain the most desirable explants for Brassica tissue culture owing to their amenability to ...