Transformation In Petunia

Transformation in Petunia Introduction

Plant transformation technology has become an adaptable platform for cultivar improvement as well as for studying gene function in plants and the discovery of novel techniques to transfer genes to plant cells provided the prerequisite for the practical use of genetic engineering in crop modification and improvement aiming to satisfy specific needs such as yield improvement, resistance to diseases and climate change. ( Abdul et Allah 2009)

Petunia has played a central role in transformation research since the earliest reports of plant transformation. It was a key model system when the first definitive accounts of Agrobacterium-mediated transformation and

When seedlings of a white-flowered plant were treated with DNA (M.W. ≈2–88×105) from a red-flowered Petunia, the recovery of up to 2730 “transformed plants” was reported (Hess 1969a, b).

Petunia and its specialty

Petunia is genus of 35 species of flowering plants of southern America origin, closely related to tobacco, tomatoes, chili peppers in the family of Solanaceae. The popular flower of the same name derived its epithet from the French, which took the word petun, meaning "tobacco, Most of the varieties seen in gardens are P. Hybrida( P.* atkinsiana also known as P.Hybrida).

Petunias can tolerate relatively harsh conditions and hot climates. They need at least five hours of sunlight every day. They grow well in low humidity, moist soil. Young plants can be grown from seeds. In drier regions, the plants should be watered daily. Maximum growth occurs in late spring.

Petunia has been studied since around 1830, and there are some early papers that mention specific flower developmental

Transformation via microinjection consists of introducing DNA into the nucleus or cytoplasm through a glass microcapillary-injection pipette.

The pipettes help in holding and immobilizing the cells while exogenous DNAs are being introduced into the nucleus. ( MONIKA RAKOCZY-TROJANOWSKA,2002).

Step 1. Isolation of Evacuolated protoplast

Evacuolated protoplasts were isolated from a cell line of P. hybrida Vilm. The evacuolated protoplasts at a concentration of 5 X 10S/ml were resuspended in regeneration medium which contains Murashige and Skoog's salts and vitamins, 2 g/1 bacto-peptone, I mg/1 napthyleneacetic acid (NAA), 2 mg/1 benzylaminopurine (BA), 68 g/1 glucose, 250 rag/1 sucrose, 20 ml/1 coconut milk and 6.5 g/l agarose at pH 6.0.

The evacuolated protoplasts were put on feeder plates which serve to allow cells to grow at a lower density.

Step 2. Injection of suspended chromosomes into Evacuolated