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Questions and Answers
What are some challenges faced by Agrobacterium-mediated transformation in plants?
somaclonal variations, oxidative burst, infection time, and DNA methylation
What is the purpose of the in planta Argo-infiltration transformation method?
to avoid tissue culture-based obstacles and enable efficient, fast, and economic transformation of plants
Which plasmids were isolated from the parent bacterium E. coli (DH5α) and introduced into Agrobacterium tumefaciens strains LBA4404?
pB121, pBCSK_His3_II, and pSIM24
How was the seedling establishment of Basmati-370 carried out?
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How was the confirmation of transformants done?
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What is the main advantage of in planta Argo-infiltration transformation method over tissue culture-based transformation?
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What concentration of ascorbic acid showed maximum transformation efficiency for pB121 plasmid in rice plants?
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Which plasmid showed maximum transformation efficiency at 6mM and 10mM concentrations of ascorbic acid in rice plants?
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What was the purpose of isolating plasmids from E. coli (DH5α) and introducing them into Agrobacterium tumefaciens strains LBA4404?
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What was used to confirm the presence of transformants in rice plants?
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Study Notes
Challenges in Agrobacterium-mediated Transformation
- Agrobacterium-mediated transformation in plants faces several challenges.
In Planta Agro-infiltration Transformation Method
- The purpose of the in planta Agro-infiltration transformation method is to introduce genes into plants.
Plasmid Isolation and Introduction
- Plasmids pBI121 and pCAMBIA1301 were isolated from the parent bacterium E. coli (DH5α) and introduced into Agrobacterium tumefaciens strains LBA4404.
Seedling Establishment of Basmati-370
- Seedling establishment of Basmati-370 was carried out by germinating seeds in a controlled environment.
Confirmation of Transformants
- The presence of transformants in rice plants was confirmed using PCR (Polymerase Chain Reaction) analysis.
Advantages of In Planta Agro-infiltration Transformation Method
- The main advantage of the in planta Agro-infiltration transformation method over tissue culture-based transformation is its higher efficiency and reduced labor requirements.
Optimal Ascorbic Acid Concentration for Transformation Efficiency
- A concentration of 10mM ascorbic acid showed maximum transformation efficiency for pBI121 plasmid in rice plants.
Plasmid Transformation Efficiency
- The pBI121 plasmid showed maximum transformation efficiency at 6mM and 10mM concentrations of ascorbic acid in rice plants.
Purpose of Plasmid Isolation and Introduction
- The purpose of isolating plasmids from E. coli (DH5α) and introducing them into Agrobacterium tumefaciens strains LBA4404 was to generate a competent Agrobacterium strain for plant transformation.
Confirmation of Transformants in Rice Plants
- PCR (Polymerase Chain Reaction) analysis was used to confirm the presence of transformants in rice plants.
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Description
Test your knowledge on plant transformation methods, including Agrobacterium-mediated transformation and in planta Argo-infiltration transformation. Learn about the challenges and benefits of each method.
