Genetic transformation of Bacopa monnieri by wild type strains of Agrobacterium rhizogenes stimulates production of bacopa saponins in transformed calli and plants.

Genetic transformation of Bacopa monnieri by wild type strains of Agrobacterium rhizogenes stimulates production of bacopa saponins in transformed calli and plants.

Majumdar S, Garai S, Jha S.2011plant cell rep30:941-954

The major advantage of this approach is the rapidity and technical simplicity of A. rhizogenes transformation. Instead of the 4-6 months-time required to regenerate transgenic plants following A. tumefaciens-based transformation (see chapter “Agrobacterium tumefaciens-mediated transformation”), production of Ri T-DNA transgenic roots takes only a few weeks. Thus, this technique is particularly well adapted to RNAi-based approaches to study gene function (Limpens et al. 2004), performing functional promoter analyses (Boisson-Dernier et al. 2005) and as a means for rapidly evaluating the complementation of plant mutants withcandidate genes (e.g. Endre et al. 2002; Levy et al. 2004), when the genetic determinant of the corresponding mutation is root-determined.

Production of Hairy Root Cultures and Transgenic Plants by Agrobacterium rhizogenes-Mediated Transformation

Production of Hairy Root Cultures and Transgenic Plants

by Agrobacterium rhizogenes-Mediated Transformation

Mary C. Christey and Robert H. Braun

Summary

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 hormonefree medium. 

Hairy roots have many applications for research including secondary product production and for the study of biochemical pathways. In addition, transgenic plants regenerated from hairy roots often show an altered phenotype due to the presence of the rol genes. In this chapter we describe how to produce and grow hairy root cultures, how to regenerate shoots from these hairy roots, and how to conduct molecular analysis of these cultures.

Key Words: Agrobacterium rhizogenes; hairy roots; Ri phenotype; rol genes; transformation.

Plant transformation 1

1. What is a transgene/transgenic plant?

2. What are some differences between physical and biological methods for DNA introduction into plant cells?

3. What are some ways that the biological method for DNA introduction (Agrobacterium) has been improved over the years?

4. How is gene introduction performed with the model plant, Arabidopsis? Is this technique widely applied to other plants?

5. How do the DNA integration patterns differ in plant cells, transformed via Agrobacterium and particle bombardment?

6. Can you think of additional methods for DNA introduction into plant cells?

Formula

Tissue culture + DNA delivery and integration = transgenic plants

Methods of delivering DNA into plant cells

Biological

Agrobacterium

Other bacteria

Viruses

Physical

Particle bombardment

Electroporation 

Silicon carbide whiskers

Carbon nanofibers

Purification of Agrobacterium rhizogenes protein (GALLS) required for gene transfer to plants

Agrobacterium damages at least 1.4 million dollars worth of agriculture per year in California and Oregon alone.

Agrobacterium is the only known prokaryote to transfer genes to a eukaryote.

Agrobacterium is used to transfer DNA into plant cells

Agrobacterium tumefaciens and Agrobacterium rhizogenes infect wounded plants and transfer plasmid DNA (T-DNA) and virulence (Vir) proteins into plant cells. 

Acne

Acne is a skin condition that causes pimples or "zits." This includes whiteheads, blackheads, and red, inflammed patches of skin (such as cysts).

Acne vulgaris (cystic acne or simply acne) is a common human skin disease, characterized by areas of skin with seborrhea (scaly red skin), comedones (blackheads and whiteheads), papules (pinheads), pustules (pimples), nodules (large papules) and possibly scarring.[1] Acne affects mostly skin with the densest population of sebaceous follicles; these areas include the face, the upper part of the chest, and the back. Severe acne is inflammatory, but acne can also manifest in noninflammatory forms.[2] The lesions are caused by changes in pilosebaceous units, skin structures consisting of a hair follicle and its associated sebaceous gland, changes that require androgen stimulation.