last step

5. Culture of plant cells into trees
The plant cell is cultured by tissue culture methods and is converted into a tree. All the cells of this tree contain the Nif gene and they can fix free nitrogen.

This plasmid is isolated and is cleaved open at a site in the T-DNA segment with the help of a specific restriction endonuclease. Now the isolated Nif gene is spliced into the opening with the help of the enzyme ligase. Thus the Nif gene is incorporated into the Ti plasmid. This Nif gene, integrated with the plasmid is called recombinant DNA.

3. Cloning the Nif gene

Cloning is a process by which hundreds of copies of Nif genes are produced. For this, the recombinant plasmid is introduced into the Agrobacterium. As these cells divide the plasmid and the Nif genes are also replicated. Thus thousands of copies of Nif genes are produced.

4. Introduction into the plant genome

Now the bacterium containing the recombinant plasmid is introduced into the plant cell. In the cell, the T-DNA containing the Nif gene of the plasmid gets separated and is transferred to the plant-cell genome. It gets incorporated with the plant genome and replicates along with the plant genome. Thus the plant genome gets the property of nitrogen fixation.

1. Isolation of Nif gene

Nif genes are present in the nitrogen fixing bacteria Rhizobium. The bacterial cell is opened by the enzyme Lysozyme. The DNA is treated with a specific restriction endonuclease.
It produces DNA fragments with sticky ends. These fragments carry Nif genes.

2. Integration of Nif gene in the vector

steps

1. Isolation of Nif gene

Nif genes are present in the nitrogen fixing bacteria Rhizobium. The bacterial cell is opened by the enzyme Lysozyme. The DNA is treated with a specific restriction endonuclease.
It produces DNA fragments with sticky ends. These fragments carry Nif genes.

2. Integration of Nif gene in the vector

Mechanism of genetic engineering

Mechanism of genetic engineering
The mechanism of genetic engineering can be studied by transferring a Nif gene (nitrogen fixing gene) from a bacterium to a dicotyledonous plant. The Nif genes help the bacteria to convert free N2 into nitrates. But dicotyledonous plants do not contain Nif genes; so they cannot use free N2. When Nif genes are transferred from bacteria to plants, the plants get the ability to use free N2. So the plant need not be given artificial fertilizers. This saves money labour and increases the yield.

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ABSTRACT

The number of canola varieties released, since the enactment of Plant Breeders' Rights (PBR) legislation in Canada in 1990, has increased substantively. The effect of genetic enhancements, including PBR, on canola yield and its variance is estimated for canola production regions of Manitoba using the Just-Pope production function. Diagnostic tests reveal the presence of heteroscedasticity in the data. The percentage of varieties seeded with PBR has a negligible effect on canola yield. Hybrid and herbicide tolerant varieties (HY/HT) seeded on over 90% of canola production area increases average yield by about 6.8%. Increased spatial diversity (more varieties planted on fewer acres) has reduced canola yield by up to 1.7%. Temporal diversity (turnover of varieties) has no significant impact on yield. Nitrogen and sulphur fertilizers are yield increasing. The variance of yield is not impacted by PBR, HY/HT, other technology measures, or by fertilizer other than potassium. Canola breeding programs have not compromised canola yield stability.

Depuis l'adoption de la Loi sur la protection des obtentions végétales (LPOV) au Canada en 1990, le nombre de variétés de canola mises au point a augmenté considérablement. L'effet des améliorations génétiques, y compris l'effet de la LPOV, sur le rendement du canola et sa variance a été estimé dans les régions du Manitoba productrices de canola à l'aide de la fonction de production Just-Pope. Des tests de diagnostic ont révélé la présence d'hétéroscédasticité dans les données. Le pourcentage de variétés ensemencées depuis la LPOV a un effet négligeable sur le rendement de canola. Les variétés hybrides et les variétés tolérantes aux herbicides (HY-HT) ensemencées sur plus de 90% des superficies cultivées en canola ont augmenté le rendement moyen d'environ 6.8%. L'accroissement de la diversité spatiale (plus de variétés sur moins de superficies) a diminué le rendement du canola jusqu'à 1.7%. La diversité temporelle (rotation des variétés) n'a aucun impact significatif sur le rendement. Les engrais azotés et soufrés augmentent les rendements. La variance du rendement n'est pas influencée par la LPOV, ni les HY-HT, ni les autres mesures technologiques ni les engrais autres que le potassium. Les programmes de sélection du canola n'ont pas compromis la stabilité du rendement du canola.

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